Enhanced human periodontal ligament stem cell viability and osteogenic differentiation on two implant materials: An experimental in vitro study

Enhanced human periodontal ligament stem cell... | F1000Research "use strict";function _typeof(t){return(_typeof="function"==typeof Symbol&&"symbol"==typeof Symbol.iterator?function(t){return typeof t}:function(t){return t&&"function"==typeof Symbol&&t.constructor===Symbol&&t!==Symbol.prototype?"symbol":typeof t})(t)}!function(){var t=function(){var t,e,o=[],n=window,r=n;for(;r;){try{if(r.frames.__tcfapiLocator){t=r;break}}catch(t){}if(r===n.top)break;r=r.parent}t||(!function t(){var e=n.document,o=!!n.frames.__tcfapiLocator;if(!o)if(e.body){var r=e.createElement("iframe");r.style.cssText="display:none",r.name="__tcfapiLocator",e.body.appendChild(r)}else setTimeout(t,5);return!o}(),n.__tcfapi=function(){for(var t=arguments.length,n=new Array(t),r=0;r 3&&2===parseInt(n[1],10)&&"boolean"==typeof n[3]&&(e=n[3],"function"==typeof n[2]&&n[2]("set",!0)):"ping"===n[0]?"function"==typeof n[2]&&n[2]({gdprApplies:e,cmpLoaded:!1,cmpStatus:"stub"}):o.push(n)},n.addEventListener("message",(function(t){var e="string"==typeof t.data,o={};if(e)try{o=JSON.parse(t.data)}catch(t){}else o=t.data;var n="object"===_typeof(o)&&null!==o?o.__tcfapiCall:null;n&&window.__tcfapi(n.command,n.version,(function(o,r){var a={__tcfapiReturn:{returnValue:o,success:r,callId:n.callId}};t&&t.source&&t.source.postMessage&&t.source.postMessage(e?JSON.stringify(a):a,"*")}),n.parameter)}),!1))};"undefined"!=typeof module?module.exports=t:t()}(); dataLayer = dataLayer || []; // Standard GTM initialization - Google Consent Mode handles consent automatically (function(w,d,s,l,i){w[l]=w[l]||[];w[l].push({'gtm.start': new Date().getTime(),event:'gtm.js'});var f=d.getElementsByTagName(s)[0], j=d.createElement(s),dl=l!='dataLayer'?'&l='+l:'';j.async=true;j.src= 'https://www.googletagmanager.com/gtm.js?id='+i+dl+ '>m_auth=hzk0Vc3qFsQYhCrIoHz68A>m_preview=env-1>m_cookies_win=x';f.parentNode.insertBefore(j,f); })(window,document,'script','dataLayer','GTM-MWFK8L5J'); ;window.NREUM||(NREUM={});NREUM.init={distributed_tracing:{enabled:true},privacy:{cookies_enabled:true},ajax:{deny_list:["bam.nr-data.net"]}}; ;NREUM.loader_config={accountID:"438030",trustKey:"438030",agentID:"772317073",licenseKey:"97f8f67f26",applicationID:"772317073"} ;NREUM.info={beacon:"bam.nr-data.net",errorBeacon:"bam.nr-data.net",licenseKey:"97f8f67f26",applicationID:"772317073",sa:1} ;/*! For license information please see nr-loader-spa-1.236.0.min.js.LICENSE.txt */ (()=>{"use strict";var e,t,r={5763:(e,t,r)=>{r.d(t,{P_:()=>l,Mt:()=>g,C5:()=>s,DL:()=>v,OP:()=>T,lF:()=>D,Yu:()=>y,Dg:()=>h,CX:()=>c,GE:()=>b,sU:()=>_});var n=r(8632),i=r(9567);const o={beacon:n.ce.beacon,errorBeacon:n.ce.errorBeacon,licenseKey:void 0,applicationID:void 0,sa:void 0,queueTime:void 0,applicationTime:void 0,ttGuid:void 0,user:void 0,account:void 0,product:void 0,extra:void 0,jsAttributes:{},userAttributes:void 0,atts:void 0,transactionName:void 0,tNamePlain:void 0},a={};function s(e){if(!e)throw new Error("All info objects require an agent identifier!");if(!a[e])throw new Error("Info for ".concat(e," was never set"));return a[e]}function c(e,t){if(!e)throw new Error("All info objects require an agent identifier!");a[e]=(0,i.D)(t,o),(0,n.Qy)(e,a[e],"info")}var u=r(7056);const d=()=>{const e={blockSelector:"[data-nr-block]",maskInputOptions:{password:!0}};return{allow_bfcache:!0,privacy:{cookies_enabled:!0},ajax:{deny_list:void 0,enabled:!0,harvestTimeSeconds:10},distributed_tracing:{enabled:void 0,exclude_newrelic_header:void 0,cors_use_newrelic_header:void 0,cors_use_tracecontext_headers:void 0,allowed_origins:void 0},session:{domain:void 0,expiresMs:u.oD,inactiveMs:u.Hb},ssl:void 0,obfuscate:void 0,jserrors:{enabled:!0,harvestTimeSeconds:10},metrics:{enabled:!0},page_action:{enabled:!0,harvestTimeSeconds:30},page_view_event:{enabled:!0},page_view_timing:{enabled:!0,harvestTimeSeconds:30,long_task:!1},session_trace:{enabled:!0,harvestTimeSeconds:10},harvest:{tooManyRequestsDelay:60},session_replay:{enabled:!1,harvestTimeSeconds:60,sampleRate:.1,errorSampleRate:.1,maskTextSelector:"*",maskAllInputs:!0,get blockClass(){return"nr-block"},get ignoreClass(){return"nr-ignore"},get maskTextClass(){return"nr-mask"},get blockSelector(){return e.blockSelector},set blockSelector(t){e.blockSelector+=",".concat(t)},get maskInputOptions(){return e.maskInputOptions},set maskInputOptions(t){e.maskInputOptions={...t,password:!0}}},spa:{enabled:!0,harvestTimeSeconds:10}}},f={};function l(e){if(!e)throw new Error("All configuration objects require an agent identifier!");if(!f[e])throw new Error("Configuration for ".concat(e," was never set"));return f[e]}function h(e,t){if(!e)throw new Error("All configuration objects require an agent identifier!");f[e]=(0,i.D)(t,d()),(0,n.Qy)(e,f[e],"config")}function g(e,t){if(!e)throw new Error("All configuration objects require an agent identifier!");var r=l(e);if(r){for(var n=t.split("."),i=0;i {r.d(t,{D:()=>i});var n=r(50);function i(e,t){try{if(!e||"object"!=typeof e)return(0,n.Z)("Setting a Configurable requires an object as input");if(!t||"object"!=typeof t)return(0,n.Z)("Setting a Configurable requires a model to set its initial properties");const r=Object.create(Object.getPrototypeOf(t),Object.getOwnPropertyDescriptors(t)),o=0===Object.keys(r).length?e:r;for(let a in o)if(void 0!==e[a])try{"object"==typeof e[a]&&"object"==typeof t[a]?r[a]=i(e[a],t[a]):r[a]=e[a]}catch(e){(0,n.Z)("An error occurred while setting a property of a Configurable",e)}return r}catch(e){(0,n.Z)("An error occured while setting a Configurable",e)}}},6818:(e,t,r)=>{r.d(t,{Re:()=>i,gF:()=>o,q4:()=>n});const n="1.236.0",i="PROD",o="CDN"},385:(e,t,r)=>{r.d(t,{FN:()=>a,IF:()=>u,Nk:()=>f,Tt:()=>s,_A:()=>o,il:()=>n,pL:()=>c,v6:()=>i,w1:()=>d});const n="undefined"!=typeof window&&!!window.document,i="undefined"!=typeof WorkerGlobalScope&&("undefined"!=typeof self&&self instanceof WorkerGlobalScope&&self.navigator instanceof WorkerNavigator||"undefined"!=typeof globalThis&&globalThis instanceof WorkerGlobalScope&&globalThis.navigator instanceof WorkerNavigator),o=n?window:"undefined"!=typeof WorkerGlobalScope&&("undefined"!=typeof self&&self instanceof WorkerGlobalScope&&self||"undefined"!=typeof globalThis&&globalThis instanceof WorkerGlobalScope&&globalThis),a=""+o?.location,s=/iPad|iPhone|iPod/.test(navigator.userAgent),c=s&&"undefined"==typeof SharedWorker,u=(()=>{const e=navigator.userAgent.match(/Firefox[/\s](\d+\.\d+)/);return Array.isArray(e)&&e.length>=2?+e[1]:0})(),d=Boolean(n&&window.document.documentMode),f=!!navigator.sendBeacon},1117:(e,t,r)=>{r.d(t,{w:()=>o});var n=r(50);const i={agentIdentifier:"",ee:void 0};class o{constructor(e){try{if("object"!=typeof e)return(0,n.Z)("shared context requires an object as input");this.sharedContext={},Object.assign(this.sharedContext,i),Object.entries(e).forEach((e=>{let[t,r]=e;Object.keys(i).includes(t)&&(this.sharedContext[t]=r)}))}catch(e){(0,n.Z)("An error occured while setting SharedContext",e)}}}},8e3:(e,t,r)=>{r.d(t,{L:()=>d,R:()=>c});var n=r(2177),i=r(1284),o=r(4322),a=r(3325);const s={};function c(e,t){const r={staged:!1,priority:a.p[t]||0};u(e),s[e].get(t)||s[e].set(t,r)}function u(e){e&&(s[e]||(s[e]=new Map))}function d(){let e=arguments.length>0&&void 0!==arguments[0]?arguments[0]:"",t=arguments.length>1&&void 0!==arguments[1]?arguments[1]:"feature";if(u(e),!e||!s[e].get(t))return a(t);s[e].get(t).staged=!0;const r=[...s[e]];function a(t){const r=e?n.ee.get(e):n.ee,a=o.X.handlers;if(r.backlog&&a){var s=r.backlog[t],c=a[t];if(c){for(var u=0;s&&u {let[t,r]=e;return r.staged}))&&(r.sort(((e,t)=>e[1].priority-t[1].priority)),r.forEach((e=>{let[t]=e;a(t)})))}function f(e,t){var r=e[1];(0,i.D)(t[r],(function(t,r){var n=e[0];if(r[0]===n){var i=r[1],o=e[3],a=e[2];i.apply(o,a)}}))}},2177:(e,t,r)=>{r.d(t,{c:()=>f,ee:()=>u});var n=r(8632),i=r(2210),o=r(1284),a=r(5763),s="nr@context";let c=(0,n.fP)();var u;function d(){}function f(e){return(0,i.X)(e,s,l)}function l(){return new d}function h(){u.aborted=!0,u.backlog={}}c.ee?u=c.ee:(u=function e(t,r){var n={},c={},f={},g=!1;try{g=16===r.length&&(0,a.OP)(r).isolatedBacklog}catch(e){}var p={on:b,addEventListener:b,removeEventListener:y,emit:v,get:x,listeners:w,context:m,buffer:A,abort:h,aborted:!1,isBuffering:E,debugId:r,backlog:g?{}:t&&"object"==typeof t.backlog?t.backlog:{}};return p;function m(e){return e&&e instanceof d?e:e?(0,i.X)(e,s,l):l()}function v(e,r,n,i,o){if(!1!==o&&(o=!0),!u.aborted||i){t&&o&&t.emit(e,r,n);for(var a=m(n),s=w(e),d=s.length,f=0;fn,p:()=>i});var n=r(2177).ee.get("handle");function i(e,t,r,i,o){o?(o.buffer([e],i),o.emit(e,t,r)):(n.buffer([e],i),n.emit(e,t,r))}},4322:(e,t,r)=>{r.d(t,{X:()=>o});var n=r(5546);o.on=a;var i=o.handlers={};function o(e,t,r,o){a(o||n.E,i,e,t,r)}function a(e,t,r,i,o){o||(o="feature"),e||(e=n.E);var a=t[o]=t[o]||{};(a[r]=a[r]||[]).push([e,i])}},3239:(e,t,r)=>{r.d(t,{bP:()=>s,iz:()=>c,m$:()=>a});var n=r(385);let i=!1,o=!1;try{const e={get passive(){return i=!0,!1},get signal(){return o=!0,!1}};n._A.addEventListener("test",null,e),n._A.removeEventListener("test",null,e)}catch(e){}function a(e,t){return i||o?{capture:!!e,passive:i,signal:t}:!!e}function s(e,t){let r=arguments.length>2&&void 0!==arguments[2]&&arguments[2],n=arguments.length>3?arguments[3]:void 0;window.addEventListener(e,t,a(r,n))}function c(e,t){let r=arguments.length>2&&void 0!==arguments[2]&&arguments[2],n=arguments.length>3?arguments[3]:void 0;document.addEventListener(e,t,a(r,n))}},4402:(e,t,r)=>{r.d(t,{Ht:()=>u,M:()=>c,Rl:()=>a,ky:()=>s});var n=r(385);const i="xxxxxxxx-xxxx-4xxx-yxxx-xxxxxxxxxxxx";function o(e,t){return e?15&e[t]:16*Math.random()|0}function a(){const e=n._A?.crypto||n._A?.msCrypto;let t,r=0;return e&&e.getRandomValues&&(t=e.getRandomValues(new Uint8Array(31))),i.split("").map((e=>"x"===e?o(t,++r).toString(16):"y"===e?(3&o()|8).toString(16):e)).join("")}function s(e){const t=n._A?.crypto||n._A?.msCrypto;let r,i=0;t&&t.getRandomValues&&(r=t.getRandomValues(new Uint8Array(31)));const a=[];for(var s=0;s {r.d(t,{Bq:()=>n,Hb:()=>o,oD:()=>i});const n="NRBA",i=144e5,o=18e5},7894:(e,t,r)=>{function n(){return Math.round(performance.now())}r.d(t,{z:()=>n})},7243:(e,t,r)=>{r.d(t,{e:()=>o});var n=r(385),i={};function o(e){if(e in i)return i[e];if(0===(e||"").indexOf("data:"))return{protocol:"data"};let t;var r=n._A?.location,o={};if(n.il)t=document.createElement("a"),t.href=e;else try{t=new URL(e,r.href)}catch(e){return o}o.port=t.port;var a=t.href.split("://");!o.port&&a[1]&&(o.port=a[1].split("/")[0].split("@").pop().split(":")[1]),o.port&&"0"!==o.port||(o.port="https"===a[0]?"443":"80"),o.hostname=t.hostname||r.hostname,o.pathname=t.pathname,o.protocol=a[0],"/"!==o.pathname.charAt(0)&&(o.pathname="/"+o.pathname);var s=!t.protocol||":"===t.protocol||t.protocol===r.protocol,c=t.hostname===r.hostname&&t.port===r.port;return o.sameOrigin=s&&(!t.hostname||c),"/"===o.pathname&&(i[e]=o),o}},50:(e,t,r)=>{function n(e,t){"function"==typeof console.warn&&(console.warn("New Relic: ".concat(e)),t&&console.warn(t))}r.d(t,{Z:()=>n})},2587:(e,t,r)=>{r.d(t,{N:()=>c,T:()=>u});var n=r(2177),i=r(5546),o=r(8e3),a=r(3325);const s={stn:[a.D.sessionTrace],err:[a.D.jserrors,a.D.metrics],ins:[a.D.pageAction],spa:[a.D.spa],sr:[a.D.sessionReplay,a.D.sessionTrace]};function c(e,t){const r=n.ee.get(t);e&&"object"==typeof e&&(Object.entries(e).forEach((e=>{let[t,n]=e;void 0===u[t]&&(s[t]?s[t].forEach((e=>{n?(0,i.p)("feat-"+t,[],void 0,e,r):(0,i.p)("block-"+t,[],void 0,e,r),(0,i.p)("rumresp-"+t,[Boolean(n)],void 0,e,r)})):n&&(0,i.p)("feat-"+t,[],void 0,void 0,r),u[t]=Boolean(n))})),Object.keys(s).forEach((e=>{void 0===u[e]&&(s[e]?.forEach((t=>(0,i.p)("rumresp-"+e,[!1],void 0,t,r))),u[e]=!1)})),(0,o.L)(t,a.D.pageViewEvent))}const u={}},2210:(e,t,r)=>{r.d(t,{X:()=>i});var n=Object.prototype.hasOwnProperty;function i(e,t,r){if(n.call(e,t))return e[t];var i=r();if(Object.defineProperty&&Object.keys)try{return Object.defineProperty(e,t,{value:i,writable:!0,enumerable:!1}),i}catch(e){}return e[t]=i,i}},1284:(e,t,r)=>{r.d(t,{D:()=>n});const n=(e,t)=>Object.entries(e||{}).map((e=>{let[r,n]=e;return t(r,n)}))},4351:(e,t,r)=>{r.d(t,{P:()=>o});var n=r(2177);const i=()=>{const e=new WeakSet;return(t,r)=>{if("object"==typeof r&&null!==r){if(e.has(r))return;e.add(r)}return r}};function o(e){try{return JSON.stringify(e,i())}catch(e){try{n.ee.emit("internal-error",[e])}catch(e){}}}},3960:(e,t,r)=>{r.d(t,{K:()=>a,b:()=>o});var n=r(3239);function i(){return"undefined"==typeof document||"complete"===document.readyState}function o(e,t){if(i())return e();(0,n.bP)("load",e,t)}function a(e){if(i())return e();(0,n.iz)("DOMContentLoaded",e)}},8632:(e,t,r)=>{r.d(t,{EZ:()=>u,Qy:()=>c,ce:()=>o,fP:()=>a,gG:()=>d,mF:()=>s});var n=r(7894),i=r(385);const o={beacon:"bam.nr-data.net",errorBeacon:"bam.nr-data.net"};function a(){return i._A.NREUM||(i._A.NREUM={}),void 0===i._A.newrelic&&(i._A.newrelic=i._A.NREUM),i._A.NREUM}function s(){let e=a();return e.o||(e.o={ST:i._A.setTimeout,SI:i._A.setImmediate,CT:i._A.clearTimeout,XHR:i._A.XMLHttpRequest,REQ:i._A.Request,EV:i._A.Event,PR:i._A.Promise,MO:i._A.MutationObserver,FETCH:i._A.fetch}),e}function c(e,t,r){let i=a();const o=i.initializedAgents||{},s=o[e]||{};return Object.keys(s).length||(s.initializedAt={ms:(0,n.z)(),date:new Date}),i.initializedAgents={...o,[e]:{...s,[r]:t}},i}function u(e,t){a()[e]=t}function d(){return function(){let e=a();const t=e.info||{};e.info={beacon:o.beacon,errorBeacon:o.errorBeacon,...t}}(),function(){let e=a();const t=e.init||{};e.init={...t}}(),s(),function(){let e=a();const t=e.loader_config||{};e.loader_config={...t}}(),a()}},7956:(e,t,r)=>{r.d(t,{N:()=>i});var n=r(3239);function i(e){let t=arguments.length>1&&void 0!==arguments[1]&&arguments[1],r=arguments.length>2?arguments[2]:void 0,i=arguments.length>3?arguments[3]:void 0;return void(0,n.iz)("visibilitychange",(function(){if(t)return void("hidden"==document.visibilityState&&e());e(document.visibilityState)}),r,i)}},1214:(e,t,r)=>{r.d(t,{em:()=>v,u5:()=>N,QU:()=>S,_L:()=>I,Gm:()=>L,Lg:()=>M,gy:()=>U,BV:()=>Q,Kf:()=>ee});var n=r(2177);const i="nr@original";var o=Object.prototype.hasOwnProperty,a=!1;function s(e,t){return e||(e=n.ee),r.inPlace=function(e,t,n,i,o){n||(n="");var a,s,c,u="-"===n.charAt(0);for(c=0;c 2?n-2:0),o=2;o {r(A[T],e,w),r(E[T],e,w)})),r(l._A,"fetch",y),t.on(y+"end",(function(e,r){var n=this;if(r){var i=r.headers.get("content-length");null!==i&&(n.rxSize=i),t.emit(y+"done",[null,r],n)}else t.emit(y+"done",[e],n)})),t}const O={},j=["pushState","replaceState"];function S(e){const t=function(e){return(e||n.ee).get("history")}(e);return!l.il||O[t.debugId]++||(O[t.debugId]=1,s(t).inPlace(window.history,j,"-")),t}var P=r(3239);const C={},R=["appendChild","insertBefore","replaceChild"];function I(e){const t=function(e){return(e||n.ee).get("jsonp")}(e);if(!l.il||C[t.debugId])return t;C[t.debugId]=!0;var r=s(t),i=/[?&](?:callback|cb)=([^&#]+)/,o=/(.*)\.([^.]+)/,a=/^(\w+)(\.|$)(.*)$/;function c(e,t){var r=e.match(a),n=r[1],i=r[3];return i?c(i,t[n]):t[n]}return r.inPlace(Node.prototype,R,"dom-"),t.on("dom-start",(function(e){!function(e){if(!e||"string"!=typeof e.nodeName||"script"!==e.nodeName.toLowerCase())return;if("function"!=typeof e.addEventListener)return;var n=(a=e.src,s=a.match(i),s?s[1]:null);var a,s;if(!n)return;var u=function(e){var t=e.match(o);if(t&&t.length>=3)return{key:t[2],parent:c(t[1],window)};return{key:e,parent:window}}(n);if("function"!=typeof u.parent[u.key])return;var d={};function f(){t.emit("jsonp-end",[],d),e.removeEventListener("load",f,(0,P.m$)(!1)),e.removeEventListener("error",l,(0,P.m$)(!1))}function l(){t.emit("jsonp-error",[],d),t.emit("jsonp-end",[],d),e.removeEventListener("load",f,(0,P.m$)(!1)),e.removeEventListener("error",l,(0,P.m$)(!1))}r.inPlace(u.parent,[u.key],"cb-",d),e.addEventListener("load",f,(0,P.m$)(!1)),e.addEventListener("error",l,(0,P.m$)(!1)),t.emit("new-jsonp",[e.src],d)}(e[0])})),t}var k=r(5763);const H={};function L(e){const t=function(e){return(e||n.ee).get("mutation")}(e);if(!l.il||H[t.debugId])return t;H[t.debugId]=!0;var r=s(t),i=k.Yu.MO;return i&&(window.MutationObserver=function(e){return this instanceof i?new i(r(e,"fn-")):i.apply(this,arguments)},MutationObserver.prototype=i.prototype),t}const z={};function M(e){const t=function(e){return(e||n.ee).get("promise")}(e);if(z[t.debugId])return t;z[t.debugId]=!0;var r=n.c,o=s(t),a=k.Yu.PR;return a&&function(){function e(r){var n=t.context(),i=o(r,"executor-",n,null,!1);const s=Reflect.construct(a,[i],e);return t.context(s).getCtx=function(){return n},s}l._A.Promise=e,Object.defineProperty(e,"name",{value:"Promise"}),e.toString=function(){return a.toString()},Object.setPrototypeOf(e,a),["all","race"].forEach((function(r){const n=a[r];e[r]=function(e){let i=!1;[...e||[]].forEach((e=>{this.resolve(e).then(a("all"===r),a(!1))}));const o=n.apply(this,arguments);return o;function a(e){return function(){t.emit("propagate",[null,!i],o,!1,!1),i=i||!e}}}})),["resolve","reject"].forEach((function(r){const n=a[r];e[r]=function(e){const r=n.apply(this,arguments);return e!==r&&t.emit("propagate",[e,!0],r,!1,!1),r}})),e.prototype=a.prototype;const n=a.prototype.then;a.prototype.then=function(){var e=this,i=r(e);i.promise=e;for(var a=arguments.length,s=new Array(a),c=0;c e())),t};function m(e,t){i.inPlace(t,["onreadystatechange"],"fn-",E)}function b(){var e=this,t=r.context(e);e.readyState>3&&!t.resolved&&(t.resolved=!0,r.emit("xhr-resolved",[],e)),i.inPlace(e,f,"fn-",E)}if(function(e,t){for(var r in e)t[r]=e[r]}(o,p),p.prototype=o.prototype,i.inPlace(p.prototype,J,"-xhr-",E),r.on("send-xhr-start",(function(e,t){m(e,t),function(e){h.push(e),a&&(y?y.then(A):u?u(A):(w=-w,x.data=w))}(t)})),r.on("open-xhr-start",m),a){var y=c&&c.resolve();if(!u&&!c){var w=1,x=document.createTextNode(w);new a(A).observe(x,{characterData:!0})}}else t.on("fn-end",(function(e){e[0]&&e[0].type===d||A()}));function A(){for(var e=0;e {r.d(t,{t:()=>n});const n=r(3325).D.ajax},6660:(e,t,r)=>{r.d(t,{A:()=>i,t:()=>n});const n=r(3325).D.jserrors,i="nr@seenError"},3081:(e,t,r)=>{r.d(t,{gF:()=>o,mY:()=>i,t9:()=>n,vz:()=>s,xS:()=>a});const n=r(3325).D.metrics,i="sm",o="cm",a="storeSupportabilityMetrics",s="storeEventMetrics"},4649:(e,t,r)=>{r.d(t,{t:()=>n});const n=r(3325).D.pageAction},7633:(e,t,r)=>{r.d(t,{Dz:()=>i,OJ:()=>a,qw:()=>o,t9:()=>n});const n=r(3325).D.pageViewEvent,i="firstbyte",o="domcontent",a="windowload"},9251:(e,t,r)=>{r.d(t,{t:()=>n});const n=r(3325).D.pageViewTiming},3614:(e,t,r)=>{r.d(t,{BST_RESOURCE:()=>i,END:()=>s,FEATURE_NAME:()=>n,FN_END:()=>u,FN_START:()=>c,PUSH_STATE:()=>d,RESOURCE:()=>o,START:()=>a});const n=r(3325).D.sessionTrace,i="bstResource",o="resource",a="-start",s="-end",c="fn"+a,u="fn"+s,d="pushState"},7836:(e,t,r)=>{r.d(t,{BODY:()=>A,CB_END:()=>E,CB_START:()=>u,END:()=>x,FEATURE_NAME:()=>i,FETCH:()=>_,FETCH_BODY:()=>v,FETCH_DONE:()=>m,FETCH_START:()=>p,FN_END:()=>c,FN_START:()=>s,INTERACTION:()=>l,INTERACTION_API:()=>d,INTERACTION_EVENTS:()=>o,JSONP_END:()=>b,JSONP_NODE:()=>g,JS_TIME:()=>T,MAX_TIMER_BUDGET:()=>a,REMAINING:()=>f,SPA_NODE:()=>h,START:()=>w,originalSetTimeout:()=>y});var n=r(5763);const i=r(3325).D.spa,o=["click","submit","keypress","keydown","keyup","change"],a=999,s="fn-start",c="fn-end",u="cb-start",d="api-ixn-",f="remaining",l="interaction",h="spaNode",g="jsonpNode",p="fetch-start",m="fetch-done",v="fetch-body-",b="jsonp-end",y=n.Yu.ST,w="-start",x="-end",A="-body",E="cb"+x,T="jsTime",_="fetch"},5938:(e,t,r)=>{r.d(t,{W:()=>o});var n=r(5763),i=r(2177);class o{constructor(e,t,r){this.agentIdentifier=e,this.aggregator=t,this.ee=i.ee.get(e,(0,n.OP)(this.agentIdentifier).isolatedBacklog),this.featureName=r,this.blocked=!1}}},9144:(e,t,r)=>{r.d(t,{j:()=>m});var n=r(3325),i=r(5763),o=r(5546),a=r(2177),s=r(7894),c=r(8e3),u=r(3960),d=r(385),f=r(50),l=r(3081),h=r(8632);function g(){const e=(0,h.gG)();["setErrorHandler","finished","addToTrace","inlineHit","addRelease","addPageAction","setCurrentRouteName","setPageViewName","setCustomAttribute","interaction","noticeError","setUserId"].forEach((t=>{e[t]=function(){for(var r=arguments.length,n=new Array(r),i=0;i 1?r-1:0),i=1;i {e.exposed&&e.api[t]&&o.push(e.api[t](...n))})),o.length>1?o:o[0]}(t,...n)}}))}var p=r(2587);function m(e){let t=arguments.length>1&&void 0!==arguments[1]?arguments[1]:{},m=arguments.length>2?arguments[2]:void 0,v=arguments.length>3?arguments[3]:void 0,{init:b,info:y,loader_config:w,runtime:x={loaderType:m},exposed:A=!0}=t;const E=(0,h.gG)();y||(b=E.init,y=E.info,w=E.loader_config),(0,i.Dg)(e,b||{}),(0,i.GE)(e,w||{}),(0,i.sU)(e,x),y.jsAttributes??={},d.v6&&(y.jsAttributes.isWorker=!0),(0,i.CX)(e,y),g();const T=function(e,t){t||(0,c.R)(e,"api");const h={};var g=a.ee.get(e),p=g.get("tracer"),m="api-",v=m+"ixn-";function b(t,r,n,o){const a=(0,i.C5)(e);return null===r?delete a.jsAttributes[t]:(0,i.CX)(e,{...a,jsAttributes:{...a.jsAttributes,[t]:r}}),x(m,n,!0,o||null===r?"session":void 0)(t,r)}function y(){}["setErrorHandler","finished","addToTrace","inlineHit","addRelease"].forEach((e=>h[e]=x(m,e,!0,"api"))),h.addPageAction=x(m,"addPageAction",!0,n.D.pageAction),h.setCurrentRouteName=x(m,"routeName",!0,n.D.spa),h.setPageViewName=function(t,r){if("string"==typeof t)return"/"!==t.charAt(0)&&(t="/"+t),(0,i.OP)(e).customTransaction=(r||"http://custom.transaction")+t,x(m,"setPageViewName",!0)()},h.setCustomAttribute=function(e,t){let r=arguments.length>2&&void 0!==arguments[2]&&arguments[2];if("string"==typeof e){if(["string","number"].includes(typeof t)||null===t)return b(e,t,"setCustomAttribute",r);(0,f.Z)("Failed to execute setCustomAttribute.\nNon-null value must be a string or number type, but a type of was provided."))}else(0,f.Z)("Failed to execute setCustomAttribute.\nName must be a string type, but a type of was provided."))},h.setUserId=function(e){if("string"==typeof e||null===e)return b("enduser.id",e,"setUserId",!0);(0,f.Z)("Failed to execute setUserId.\nNon-null value must be a string type, but a type of was provided."))},h.interaction=function(){return(new y).get()};var w=y.prototype={createTracer:function(e,t){var r={},i=this,a="function"==typeof t;return(0,o.p)(v+"tracer",[(0,s.z)(),e,r],i,n.D.spa,g),function(){if(p.emit((a?"":"no-")+"fn-start",[(0,s.z)(),i,a],r),a)try{return t.apply(this,arguments)}catch(e){throw p.emit("fn-err",[arguments,this,"string"==typeof e?new Error(e):e],r),e}finally{p.emit("fn-end",[(0,s.z)()],r)}}}};function x(e,t,r,i){return function(){return(0,o.p)(l.xS,["API/"+t+"/called"],void 0,n.D.metrics,g),i&&(0,o.p)(e+t,[(0,s.z)(),...arguments],r?null:this,i,g),r?void 0:this}}function A(){r.e(439).then(r.bind(r,7438)).then((t=>{let{setAPI:r}=t;r(e),(0,c.L)(e,"api")})).catch((()=>(0,f.Z)("Downloading runtime APIs failed...")))}return["actionText","setName","setAttribute","save","ignore","onEnd","getContext","end","get"].forEach((e=>{w[e]=x(v,e,void 0,n.D.spa)})),h.noticeError=function(e,t){"string"==typeof e&&(e=new Error(e)),(0,o.p)(l.xS,["API/noticeError/called"],void 0,n.D.metrics,g),(0,o.p)("err",[e,(0,s.z)(),!1,t],void 0,n.D.jserrors,g)},d.il?(0,u.b)((()=>A()),!0):A(),h}(e,v);return(0,h.Qy)(e,T,"api"),(0,h.Qy)(e,A,"exposed"),(0,h.EZ)("activatedFeatures",p.T),T}},3325:(e,t,r)=>{r.d(t,{D:()=>n,p:()=>i});const n={ajax:"ajax",jserrors:"jserrors",metrics:"metrics",pageAction:"page_action",pageViewEvent:"page_view_event",pageViewTiming:"page_view_timing",sessionReplay:"session_replay",sessionTrace:"session_trace",spa:"spa"},i={[n.pageViewEvent]:1,[n.pageViewTiming]:2,[n.metrics]:3,[n.jserrors]:4,[n.ajax]:5,[n.sessionTrace]:6,[n.pageAction]:7,[n.spa]:8,[n.sessionReplay]:9}}},n={};function i(e){var t=n[e];if(void 0!==t)return t.exports;var o=n[e]={exports:{}};return r[e](o,o.exports,i),o.exports}i.m=r,i.d=(e,t)=>{for(var r in t)i.o(t,r)&&!i.o(e,r)&&Object.defineProperty(e,r,{enumerable:!0,get:t[r]})},i.f={},i.e=e=>Promise.all(Object.keys(i.f).reduce(((t,r)=>(i.f[r](e,t),t)),[])),i.u=e=>(({78:"page_action-aggregate",147:"metrics-aggregate",242:"session-manager",317:"jserrors-aggregate",348:"page_view_timing-aggregate",412:"lazy-feature-loader",439:"async-api",538:"recorder",590:"session_replay-aggregate",675:"compressor",733:"session_trace-aggregate",786:"page_view_event-aggregate",873:"spa-aggregate",898:"ajax-aggregate"}[e]||e)+"."+{78:"ac76d497",147:"3dc53903",148:"1a20d5fe",242:"2a64278a",317:"49e41428",348:"bd6de33a",412:"2f55ce66",439:"30bd804e",538:"1b18459f",590:"cf0efb30",675:"ae9f91a8",733:"83105561",786:"06482edd",860:"03a8b7a5",873:"e6b09d52",898:"998ef92b"}[e]+"-1.236.0.min.js"),i.o=(e,t)=>Object.prototype.hasOwnProperty.call(e,t),e={},t="NRBA:",i.l=(r,n,o,a)=>{if(e[r])e[r].push(n);else{var s,c;if(void 0!==o)for(var u=document.getElementsByTagName("script"),d=0;d {s.onerror=s.onload=null,clearTimeout(h);var i=e[r];if(delete e[r],s.parentNode&&s.parentNode.removeChild(s),i&&i.forEach((e=>e(n))),t)return t(n)},h=setTimeout(l.bind(null,void 0,{type:"timeout",target:s}),12e4);s.onerror=l.bind(null,s.onerror),s.onload=l.bind(null,s.onload),c&&document.head.appendChild(s)}},i.r=e=>{"undefined"!=typeof Symbol&&Symbol.toStringTag&&Object.defineProperty(e,Symbol.toStringTag,{value:"Module"}),Object.defineProperty(e,"__esModule",{value:!0})},i.j=364,i.p="https://js-agent.newrelic.com/",(()=>{var e={364:0,953:0};i.f.j=(t,r)=>{var n=i.o(e,t)?e[t]:void 0;if(0!==n)if(n)r.push(n[2]);else{var o=new Promise(((r,i)=>n=e[t]=[r,i]));r.push(n[2]=o);var a=i.p+i.u(t),s=new Error;i.l(a,(r=>{if(i.o(e,t)&&(0!==(n=e[t])&&(e[t]=void 0),n)){var o=r&&("load"===r.type?"missing":r.type),a=r&&r.target&&r.target.src;s.message="Loading chunk "+t+" failed.\n("+o+": "+a+")",s.name="ChunkLoadError",s.type=o,s.request=a,n[1](s)}}),"chunk-"+t,t)}};var t=(t,r)=>{var n,o,[a,s,c]=r,u=0;if(a.some((t=>0!==e[t]))){for(n in s)i.o(s,n)&&(i.m[n]=s[n]);if(c)c(i)}for(t&&t(r);u {i.r(o);var e=i(3325),t=i(5763);const r=Object.values(e.D);function n(e){const n={};return r.forEach((r=>{n[r]=function(e,r){return!1!==(0,t.Mt)(r,"".concat(e,".enabled"))}(r,e)})),n}var a=i(9144);var s=i(5546),c=i(385),u=i(8e3),d=i(5938),f=i(3960),l=i(50);class h extends d.W{constructor(e,t,r){let n=!(arguments.length>3&&void 0!==arguments[3])||arguments[3];super(e,t,r),this.auto=n,this.abortHandler,this.featAggregate,this.onAggregateImported,n&&(0,u.R)(e,r)}importAggregator(){let e=arguments.length>0&&void 0!==arguments[0]?arguments[0]:{};if(this.featAggregate||!this.auto)return;const r=c.il&&!0===(0,t.Mt)(this.agentIdentifier,"privacy.cookies_enabled");let n;this.onAggregateImported=new Promise((e=>{n=e}));const o=async()=>{let t;try{if(r){const{setupAgentSession:e}=await Promise.all([i.e(860),i.e(242)]).then(i.bind(i,3228));t=e(this.agentIdentifier)}}catch(e){(0,l.Z)("A problem occurred when starting up session manager. This page will not start or extend any session.",e)}try{if(!this.shouldImportAgg(this.featureName,t))return void(0,u.L)(this.agentIdentifier,this.featureName);const{lazyFeatureLoader:r}=await i.e(412).then(i.bind(i,8582)),{Aggregate:o}=await r(this.featureName,"aggregate");this.featAggregate=new o(this.agentIdentifier,this.aggregator,e),n(!0)}catch(e){(0,l.Z)("Downloading and initializing ".concat(this.featureName," failed..."),e),this.abortHandler?.(),n(!1)}};c.il?(0,f.b)((()=>o()),!0):o()}shouldImportAgg(r,n){return r!==e.D.sessionReplay||!1!==(0,t.Mt)(this.agentIdentifier,"session_trace.enabled")&&(!!n?.isNew||!!n?.state.sessionReplay)}}var g=i(7633),p=i(7894);class m extends h{static featureName=g.t9;constructor(r,n){let i=!(arguments.length>2&&void 0!==arguments[2])||arguments[2];if(super(r,n,g.t9,i),("undefined"==typeof PerformanceNavigationTiming||c.Tt)&&"undefined"!=typeof PerformanceTiming){const n=(0,t.OP)(r);n[g.Dz]=Math.max(Date.now()-n.offset,0),(0,f.K)((()=>n[g.qw]=Math.max((0,p.z)()-n[g.Dz],0))),(0,f.b)((()=>{const t=(0,p.z)();n[g.OJ]=Math.max(t-n[g.Dz],0),(0,s.p)("timing",["load",t],void 0,e.D.pageViewTiming,this.ee)}))}this.importAggregator()}}var v=i(1117),b=i(1284);class y extends v.w{constructor(e){super(e),this.aggregatedData={}}store(e,t,r,n,i){var o=this.getBucket(e,t,r,i);return o.metrics=function(e,t){t||(t={count:0});return t.count+=1,(0,b.D)(e,(function(e,r){t[e]=w(r,t[e])})),t}(n,o.metrics),o}merge(e,t,r,n,i){var o=this.getBucket(e,t,n,i);if(o.metrics){var a=o.metrics;a.count+=r.count,(0,b.D)(r,(function(e,t){if("count"!==e){var n=a[e],i=r[e];i&&!i.c?a[e]=w(i.t,n):a[e]=function(e,t){if(!t)return e;t.c||(t=x(t.t));return t.min=Math.min(e.min,t.min),t.max=Math.max(e.max,t.max),t.t+=e.t,t.sos+=e.sos,t.c+=e.c,t}(i,a[e])}}))}else o.metrics=r}storeMetric(e,t,r,n){var i=this.getBucket(e,t,r);return i.stats=w(n,i.stats),i}getBucket(e,t,r,n){this.aggregatedData[e]||(this.aggregatedData[e]={});var i=this.aggregatedData[e][t];return i||(i=this.aggregatedData[e][t]={params:r||{}},n&&(i.custom=n)),i}get(e,t){return t?this.aggregatedData[e]&&this.aggregatedData[e][t]:this.aggregatedData[e]}take(e){for(var t={},r="",n=!1,i=0;i t.max&&(t.max=e),e 2&&void 0!==arguments[2])||arguments[2];super(e,r,j.t,n),c.il&&((0,t.OP)(e).initHidden=Boolean("hidden"===document.visibilityState),(0,N.N)((()=>(0,s.p)("docHidden",[(0,p.z)()],void 0,j.t,this.ee)),!0),(0,O.bP)("pagehide",(()=>(0,s.p)("winPagehide",[(0,p.z)()],void 0,j.t,this.ee))),this.importAggregator())}}var P=i(3081);class C extends h{static featureName=P.t9;constructor(e,t){let r=!(arguments.length>2&&void 0!==arguments[2])||arguments[2];super(e,t,P.t9,r),this.importAggregator()}}var R,I=i(2210),k=i(1214),H=i(2177),L={};try{R=localStorage.getItem("__nr_flags").split(","),console&&"function"==typeof console.log&&(L.console=!0,-1!==R.indexOf("dev")&&(L.dev=!0),-1!==R.indexOf("nr_dev")&&(L.nrDev=!0))}catch(e){}function z(e){try{L.console&&z(e)}catch(e){}}L.nrDev&&H.ee.on("internal-error",(function(e){z(e.stack)})),L.dev&&H.ee.on("fn-err",(function(e,t,r){z(r.stack)})),L.dev&&(z("NR AGENT IN DEVELOPMENT MODE"),z("flags: "+(0,b.D)(L,(function(e,t){return e})).join(", ")));var M=i(6660);class B extends h{static featureName=M.t;constructor(r,n){let i=!(arguments.length>2&&void 0!==arguments[2])||arguments[2];super(r,n,M.t,i),this.skipNext=0;try{this.removeOnAbort=new AbortController}catch(e){}const o=this;o.ee.on("fn-start",(function(e,t,r){o.abortHandler&&(o.skipNext+=1)})),o.ee.on("fn-err",(function(t,r,n){o.abortHandler&&!n[M.A]&&((0,I.X)(n,M.A,(function(){return!0})),this.thrown=!0,(0,s.p)("err",[n,(0,p.z)()],void 0,e.D.jserrors,o.ee))})),o.ee.on("fn-end",(function(){o.abortHandler&&!this.thrown&&o.skipNext>0&&(o.skipNext-=1)})),o.ee.on("internal-error",(function(t){(0,s.p)("ierr",[t,(0,p.z)(),!0],void 0,e.D.jserrors,o.ee)})),this.origOnerror=c._A.onerror,c._A.onerror=this.onerrorHandler.bind(this),c._A.addEventListener("unhandledrejection",(t=>{const r=function(e){let t="Unhandled Promise Rejection: ";if(e instanceof Error)try{return e.message=t+e.message,e}catch(t){return e}if(void 0===e)return new Error(t);try{return new Error(t+(0,D.P)(e))}catch(e){return new Error(t)}}(t.reason);(0,s.p)("err",[r,(0,p.z)(),!1,{unhandledPromiseRejection:1}],void 0,e.D.jserrors,this.ee)}),(0,O.m$)(!1,this.removeOnAbort?.signal)),(0,k.gy)(this.ee),(0,k.BV)(this.ee),(0,k.em)(this.ee),(0,t.OP)(r).xhrWrappable&&(0,k.Kf)(this.ee),this.abortHandler=this.#e,this.importAggregator()}#e(){this.removeOnAbort?.abort(),this.abortHandler=void 0}onerrorHandler(t,r,n,i,o){"function"==typeof this.origOnerror&&this.origOnerror(...arguments);try{this.skipNext?this.skipNext-=1:(0,s.p)("err",[o||new F(t,r,n),(0,p.z)()],void 0,e.D.jserrors,this.ee)}catch(t){try{(0,s.p)("ierr",[t,(0,p.z)(),!0],void 0,e.D.jserrors,this.ee)}catch(e){}}return!1}}function F(e,t,r){this.message=e||"Uncaught error with no additional information",this.sourceURL=t,this.line=r}let U=1;const q="nr@id";function G(e){const t=typeof e;return!e||"object"!==t&&"function"!==t?-1:e===c._A?0:(0,I.X)(e,q,(function(){return U++}))}function V(e){if("string"==typeof e&&e.length)return e.length;if("object"==typeof e){if("undefined"!=typeof ArrayBuffer&&e instanceof ArrayBuffer&&e.byteLength)return e.byteLength;if("undefined"!=typeof Blob&&e instanceof Blob&&e.size)return e.size;if(!("undefined"!=typeof FormData&&e instanceof FormData))try{return(0,D.P)(e).length}catch(e){return}}}var X=i(7243);class W{constructor(e){this.agentIdentifier=e,this.generateTracePayload=this.generateTracePayload.bind(this),this.shouldGenerateTrace=this.shouldGenerateTrace.bind(this)}generateTracePayload(e){if(!this.shouldGenerateTrace(e))return null;var r=(0,t.DL)(this.agentIdentifier);if(!r)return null;var n=(r.accountID||"").toString()||null,i=(r.agentID||"").toString()||null,o=(r.trustKey||"").toString()||null;if(!n||!i)return null;var a=(0,_.M)(),s=(0,_.Ht)(),c=Date.now(),u={spanId:a,traceId:s,timestamp:c};return(e.sameOrigin||this.isAllowedOrigin(e)&&this.useTraceContextHeadersForCors())&&(u.traceContextParentHeader=this.generateTraceContextParentHeader(a,s),u.traceContextStateHeader=this.generateTraceContextStateHeader(a,c,n,i,o)),(e.sameOrigin&&!this.excludeNewrelicHeader()||!e.sameOrigin&&this.isAllowedOrigin(e)&&this.useNewrelicHeaderForCors())&&(u.newrelicHeader=this.generateTraceHeader(a,s,c,n,i,o)),u}generateTraceContextParentHeader(e,t){return"00-"+t+"-"+e+"-01"}generateTraceContextStateHeader(e,t,r,n,i){return i+"@nr=0-1-"+r+"-"+n+"-"+e+"----"+t}generateTraceHeader(e,t,r,n,i,o){if(!("function"==typeof c._A?.btoa))return null;var a={v:[0,1],d:{ty:"Browser",ac:n,ap:i,id:e,tr:t,ti:r}};return o&&n!==o&&(a.d.tk=o),btoa((0,D.P)(a))}shouldGenerateTrace(e){return this.isDtEnabled()&&this.isAllowedOrigin(e)}isAllowedOrigin(e){var r=!1,n={};if((0,t.Mt)(this.agentIdentifier,"distributed_tracing")&&(n=(0,t.P_)(this.agentIdentifier).distributed_tracing),e.sameOrigin)r=!0;else if(n.allowed_origins instanceof Array)for(var i=0;i 2&&void 0!==arguments[2])||arguments[2];super(r,n,Z.t,i),(0,t.OP)(r).xhrWrappable&&(this.dt=new W(r),this.handler=(e,t,r,n)=>(0,s.p)(e,t,r,n,this.ee),(0,k.u5)(this.ee),(0,k.Kf)(this.ee),function(r,n,i,o){function a(e){var t=this;t.totalCbs=0,t.called=0,t.cbTime=0,t.end=E,t.ended=!1,t.xhrGuids={},t.lastSize=null,t.loadCaptureCalled=!1,t.params=this.params||{},t.metrics=this.metrics||{},e.addEventListener("load",(function(r){_(t,e)}),(0,O.m$)(!1)),c.IF||e.addEventListener("progress",(function(e){t.lastSize=e.loaded}),(0,O.m$)(!1))}function s(e){this.params={method:e[0]},T(this,e[1]),this.metrics={}}function u(e,n){var i=(0,t.DL)(r);i.xpid&&this.sameOrigin&&n.setRequestHeader("X-NewRelic-ID",i.xpid);var a=o.generateTracePayload(this.parsedOrigin);if(a){var s=!1;a.newrelicHeader&&(n.setRequestHeader("newrelic",a.newrelicHeader),s=!0),a.traceContextParentHeader&&(n.setRequestHeader("traceparent",a.traceContextParentHeader),a.traceContextStateHeader&&n.setRequestHeader("tracestate",a.traceContextStateHeader),s=!0),s&&(this.dt=a)}}function d(e,t){var r=this.metrics,i=e[0],o=this;if(r&&i){var a=V(i);a&&(r.txSize=a)}this.startTime=(0,p.z)(),this.listener=function(e){try{"abort"!==e.type||o.loadCaptureCalled||(o.params.aborted=!0),("load"!==e.type||o.called===o.totalCbs&&(o.onloadCalled||"function"!=typeof t.onload)&&"function"==typeof o.end)&&o.end(t)}catch(e){try{n.emit("internal-error",[e])}catch(e){}}};for(var s=0;s 1?e[1]=i:e.push(i)}else e[0]&&e[0].headers&&s(e[0].headers,n)&&(this.dt=n);function s(e,t){var r=!1;return t.newrelicHeader&&(e.set("newrelic",t.newrelicHeader),r=!0),t.traceContextParentHeader&&(e.set("traceparent",t.traceContextParentHeader),t.traceContextStateHeader&&e.set("tracestate",t.traceContextStateHeader),r=!0),r}}function x(e,t){this.params={},this.metrics={},this.startTime=(0,p.z)(),this.dt=t,e.length>=1&&(this.target=e[0]),e.length>=2&&(this.opts=e[1]);var r,n=this.opts||{},i=this.target;"string"==typeof i?r=i:"object"==typeof i&&i instanceof Y?r=i.url:c._A?.URL&&"object"==typeof i&&i instanceof URL&&(r=i.href),T(this,r);var o=(""+(i&&i instanceof Y&&i.method||n.method||"GET")).toUpperCase();this.params.method=o,this.txSize=V(n.body)||0}function A(t,r){var n;this.endTime=(0,p.z)(),this.params||(this.params={}),this.params.status=r?r.status:0,"string"==typeof this.rxSize&&this.rxSize.length>0&&(n=+this.rxSize);var o={txSize:this.txSize,rxSize:n,duration:(0,p.z)()-this.startTime};i("xhr",[this.params,o,this.startTime,this.endTime,"fetch"],this,e.D.ajax)}function E(t){var r=this.params,n=this.metrics;if(!this.ended){this.ended=!0;for(var o=0;o 2&&void 0!==arguments[2])||arguments[2];super(e,t,we.t,r),this.importAggregator()}}new class{constructor(e){let t=arguments.length>1&&void 0!==arguments[1]?arguments[1]:(0,_.ky)(16);c._A?(this.agentIdentifier=t,this.sharedAggregator=new y({agentIdentifier:this.agentIdentifier}),this.features={},this.desiredFeatures=new Set(e.features||[]),this.desiredFeatures.add(m),Object.assign(this,(0,a.j)(this.agentIdentifier,e,e.loaderType||"agent")),this.start()):(0,l.Z)("Failed to initial the agent. Could not determine the runtime environment.")}get config(){return{info:(0,t.C5)(this.agentIdentifier),init:(0,t.P_)(this.agentIdentifier),loader_config:(0,t.DL)(this.agentIdentifier),runtime:(0,t.OP)(this.agentIdentifier)}}start(){const t="features";try{const r=n(this.agentIdentifier),i=[...this.desiredFeatures];i.sort(((t,r)=>e.p[t.featureName]-e.p[r.featureName])),i.forEach((t=>{if(r[t.featureName]||t.featureName===e.D.pageViewEvent){const n=function(t){switch(t){case e.D.ajax:return[e.D.jserrors];case e.D.sessionTrace:return[e.D.ajax,e.D.pageViewEvent];case e.D.sessionReplay:return[e.D.sessionTrace];case e.D.pageViewTiming:return[e.D.pageViewEvent];default:return[]}}(t.featureName);n.every((e=>r[e]))||(0,l.Z)("".concat(t.featureName," is enabled but one or more dependent features has been disabled (").concat((0,D.P)(n),"). This may cause unintended consequences or missing data...")),this.features[t.featureName]=new t(this.agentIdentifier,this.sharedAggregator)}})),(0,T.Qy)(this.agentIdentifier,this.features,t)}catch(e){(0,l.Z)("Failed to initialize all enabled instrument classes (agent aborted) -",e);for(const e in this.features)this.features[e].abortHandler?.();const r=(0,T.fP)();return delete r.initializedAgents[this.agentIdentifier]?.api,delete r.initializedAgents[this.agentIdentifier]?.[t],delete this.sharedAggregator,r.ee?.abort(),delete r.ee?.get(this.agentIdentifier),!1}}}({features:[J,m,S,class extends h{static featureName=oe;constructor(t,r){if(super(t,r,oe,!(arguments.length>2&&void 0!==arguments[2])||arguments[2]),!c.il)return;const n=this.ee;let i;(0,k.QU)(n),this.eventsEE=(0,k.em)(n),this.eventsEE.on(se,(function(e,t){this.bstStart=(0,p.z)()})),this.eventsEE.on(ae,(function(t,r){(0,s.p)("bst",[t[0],r,this.bstStart,(0,p.z)()],void 0,e.D.sessionTrace,n)})),n.on(ce+ne,(function(e){this.time=(0,p.z)(),this.startPath=location.pathname+location.hash})),n.on(ce+ie,(function(t){(0,s.p)("bstHist",[location.pathname+location.hash,this.startPath,this.time],void 0,e.D.sessionTrace,n)}));try{i=new PerformanceObserver((t=>{const r=t.getEntries();(0,s.p)(te,[r],void 0,e.D.sessionTrace,n)})),i.observe({type:re,buffered:!0})}catch(e){}this.importAggregator({resourceObserver:i})}},C,xe,B,class extends h{static featureName=de;constructor(e,r){if(super(e,r,de,!(arguments.length>2&&void 0!==arguments[2])||arguments[2]),!c.il)return;if(!(0,t.OP)(e).xhrWrappable)return;try{this.removeOnAbort=new AbortController}catch(e){}let n,i=0;const o=this.ee.get("tracer"),a=(0,k._L)(this.ee),s=(0,k.Lg)(this.ee),u=(0,k.BV)(this.ee),d=(0,k.Kf)(this.ee),f=this.ee.get("events"),l=(0,k.u5)(this.ee),h=(0,k.QU)(this.ee),g=(0,k.Gm)(this.ee);function m(e,t){h.emit("newURL",[""+window.location,t])}function v(){i++,n=window.location.hash,this[ve]=(0,p.z)()}function b(){i--,window.location.hash!==n&&m(0,!0);var e=(0,p.z)();this[pe]=~~this[pe]+e-this[ve],this[ye]=e}function y(e,t){e.on(t,(function(){this[t]=(0,p.z)()}))}this.ee.on(ve,v),s.on(be,v),a.on(be,v),this.ee.on(ye,b),s.on(ge,b),a.on(ge,b),this.ee.buffer([ve,ye,"xhr-resolved"],this.featureName),f.buffer([ve],this.featureName),u.buffer(["setTimeout"+le,"clearTimeout"+fe,ve],this.featureName),d.buffer([ve,"new-xhr","send-xhr"+fe],this.featureName),l.buffer([me+fe,me+"-done",me+he+fe,me+he+le],this.featureName),h.buffer(["newURL"],this.featureName),g.buffer([ve],this.featureName),s.buffer(["propagate",be,ge,"executor-err","resolve"+fe],this.featureName),o.buffer([ve,"no-"+ve],this.featureName),a.buffer(["new-jsonp","cb-start","jsonp-error","jsonp-end"],this.featureName),y(l,me+fe),y(l,me+"-done"),y(a,"new-jsonp"),y(a,"jsonp-end"),y(a,"cb-start"),h.on("pushState-end",m),h.on("replaceState-end",m),window.addEventListener("hashchange",m,(0,O.m$)(!0,this.removeOnAbort?.signal)),window.addEventListener("load",m,(0,O.m$)(!0,this.removeOnAbort?.signal)),window.addEventListener("popstate",(function(){m(0,i>1)}),(0,O.m$)(!0,this.removeOnAbort?.signal)),this.abortHandler=this.#e,this.importAggregator()}#e(){this.removeOnAbort?.abort(),this.abortHandler=void 0}}],loaderType:"spa"})})(),window.NRBA=o})(); window.jQuery || document.write(' ') CKEDITOR_BASEPATH='https://f1000research.com/js/vendor/ckeditor/' window.reactTheme = 'research'; window.MathJax = { CommonHTML: { linebreaks: { automatic: true } }, 'HTML-CSS': { linebreaks: { automatic: true } }, SVG: { linebreaks: { automatic: true } }, AuthorInit: function() { MathJax.Hub.Register.MessageHook('End Process', function () { let timeout = false; // holder for timeout id const delay = 250; // delay after event is "complete" to run callback const reflowMath = function() { const dispFormulas = document.querySelectorAll('.disp-formula.panel'); if (!dispFormulas) { return; } for (const dispFormula of dispFormulas) { const child = dispFormula.querySelector('.MathJax_Preview').nextSibling.firstChild; const isMultiline = MathJax.Hub.getAllJax(dispFormula)[0].root.isMultiline; if (dispFormula.offsetWidth < child.offsetWidth || isMultiline) { MathJax.Hub.Queue(['Rerender', MathJax.Hub, dispFormula]); } } }; window.addEventListener('resize', function() { clearTimeout(timeout); // clear the timeout timeout = setTimeout(reflowMath, delay); // start timing for event "completion" }); }); }, }; if (window.location.hash == '#_=_'){ window.location = window.location.href.split('#')[0] } !function(f,b,e,v,n,t,s){if(f.fbq)return;n=f.fbq=function() {n.callMethod? n.callMethod.apply(n,arguments):n.queue.push(arguments)} ;if(!f._fbq)f._fbq=n; n.push=n;n.loaded=!0;n.version='2.0';n.queue=[];t=b.createElement(e);t.async=!0; t.src=v;s=b.getElementsByTagName(e)[0];s.parentNode.insertBefore(t,s)}(window, document,'script','https://connect.facebook.net/en_US/fbevents.js'); fbq('init', '1641728616063202'); fbq('track', "PixelInitialized", {}); (function(h,o,t,j,a,r){ h.hj=h.hj||function(){(h.hj.q=h.hj.q||[]).push(arguments)}; h._hjSettings={hjid:2318163,hjsv:6}; a=o.getElementsByTagName('head')[0]; r=o.createElement('script');r.async=1; r.src=t+h._hjSettings.hjid+j+h._hjSettings.hjsv; a.appendChild(r); })(window,document,'https://static.hotjar.com/c/hotjar-','.js?sv='); search file_upload Submit your research search menu close search Browse Gateways & Collections How to Publish Submit your Research My Submissions Article Guidelines Article Guidelines (New Versions) Open Data, Software and Code Guidelines Open Data and Accessible Source Materials Guidelines (HSS) Open Data, Software and Code Guidelines (PSE) Prepublication Checks Production Process Posters and Slides Guidelines Document Guidelines Article Processing Charges Peer Review Finding Article Reviewers About How it Works For Reviewers Our Advisors Policies Glossary FAQs For Developers Newsroom Contact My Research Submissions Content and Tracking Alerts My Details Sign In file_upload Submit your research { "@context": "https://schema.org", "@type": "ScholarlyArticle", "mainEntityOfPage": { "@type": "WebPage", "@id": "https://f1000research.com/articles/12-447" }, "headline": "Enhanced human periodontal ligament stem cell viability and osteogenic differentiation on two implant...", "datePublished": "2023-04-26T11:46:25", "dateModified": "2023-08-08T10:32:55", "author": [ { "@type": "Person", "name": "Sara F. El Shafei" }, { "@type": "Person", "name": "Shereen N. Raafat" }, { "@type": "Person", "name": "Engy A. Farag" } ], "publisher": { "@type": "Organization", "name": "F1000Research", "logo": { "@type": "ImageObject", "url": "https://f1000research.com/img/AMP/F1000Research_image.png", "height": 480, "width": 60 } }, "image": { "@type": "ImageObject", "url": "https://f1000research.com/img/AMP/F1000Research_image.png", "height": 1200, "width": 150 }, "description": "Background: Surface roughness of dental implants impacts the survival of adult periodontal stem cells and rate of differentiation. This research was conducted to test how human periodontal ligament stem cells behaved on yttria stabilized tetragonal zirconia polycrystals and polyetheretherketone (PEEK) discs with different surface topographies. Methods: Discs roughening was prepared by sandblasting. Stem cells were cultivated on zirconia discs with a polished surface, PEEK discs with a polished surface, sandblasted zirconia discs and sandblasted PEEK discs. Cells viability was assessed after 24, 48, 72 hours. Scanning electron microscopy was used to examine the adherence and attachment of cells. Osteoblastic differentiation capacity was studied by checking the mineralization clusters development through alizarin red S staining and alkaline phosphatase assay. ANOVA and the Tukey post hoc test were used for the statistical analysis. Results: Polished PEEK discs showed lower cell viability, whereas roughened sandblasted zirconia and PEEK discs showed the highest proliferation rates and cell viability percent. The osteogenic differentiation was enhanced for rough surfaces in comparison to polished surfaces. Sandblasted zirconia and PEEK discs showed a markedly increased mineralized nodule development and ALP enzyme activity compared to the polished surface and control. Conclusions: Micro- topographies creation on the PEEK implant surface enhances stem cell attachment, viability, and osteogenic differentiation." } { "@context": "http://schema.org", "@type": "BreadcrumbList", "itemListElement": [ { "@type": "ListItem", "position": "1", "item": { "@id": "https://f1000research.com/", "name": "Home" } }, { "@type": "ListItem", "position": "2", "item": { "@id": "https://f1000research.com/browse/articles", "name": "Browse" } }, { "@type": "ListItem", "position": "3", "item": { "@id": "https://f1000research.com/articles/12-447", "name": "Enhanced human periodontal ligament stem cell viability and osteogenic..." } } ] } Home Browse Enhanced human periodontal ligament stem cell viability and osteogenic... ALL Metrics - Views Downloads Get PDF Get XML Cite How to cite this article El Shafei SF, Raafat SN and Farag EA. Enhanced human periodontal ligament stem cell viability and osteogenic differentiation on two implant materials: An experimental in vitro study [version 2; peer review: 2 approved] . F1000Research 2023, 12 :447 ( https://doi.org/10.12688/f1000research.129562.2 ) NOTE: If applicable, it is important to ensure the information in square brackets after the title is included in all citations of this article. Close Copy Citation Details Export Export Citation Sciwheel EndNote Ref. Manager Bibtex ProCite Sente EXPORT Select a format first Track Share ▬ ✚ Research Article Revised Enhanced human periodontal ligament stem cell viability and osteogenic differentiation on two implant materials: An experimental in vitro study [version 2; peer review: 2 approved] Sara F. El Shafei https://orcid.org/0000-0002-5060-8941 1 , Shereen N. Raafat 2 , Engy A. Farag 3 Sara F. El Shafei https://orcid.org/0000-0002-5060-8941 1 , Shereen N. Raafat 2 , Engy A. Farag 3 PUBLISHED 08 Aug 2023 Author details Author details 1 Removable Prosthodontics, Faculty of Dentistry, The British University in Egypt, Cairo, Egypt 2 Department of Pharmacology, Director of Stem Cell and Tissue Culture Hub, Centre of Innovative Dental Sciences (CIDS), Faculty of Dentistry,, The British University in Egypt, Cairo, Egypt 3 Fixed Prosthodontics, Faculty of Dentistry, The British University in Egypt, Cairo, Egypt Sara F. El Shafei Roles: Conceptualization, Data Curation, Methodology, Writing – Review & Editing Shereen N. Raafat Roles: Conceptualization, Data Curation, Formal Analysis, Investigation, Methodology, Writing – Review & Editing Engy A. Farag Roles: Conceptualization, Data Curation, Methodology, Writing – Original Draft Preparation, Writing – Review & Editing OPEN PEER REVIEW DETAILS REVIEWER STATUS This article is included in the Cell & Molecular Biology gateway. Abstract Background: Surface roughness of dental implants impacts the survival of adult periodontal stem cells and rate of differentiation. This research was conducted to test how human periodontal ligament stem cells behaved on yttria stabilized tetragonal zirconia polycrystals and polyetheretherketone (PEEK) discs with different surface topographies. Methods: Discs roughening was prepared by sandblasting. Stem cells were cultivated on zirconia discs with a polished surface, PEEK discs with a polished surface, sandblasted zirconia discs and sandblasted PEEK discs. Cells viability was assessed after 24, 48, 72 hours. Scanning electron microscopy was used to examine the adherence and attachment of cells. Osteoblastic differentiation capacity was studied by checking the mineralization clusters development through alizarin red S staining and alkaline phosphatase assay. ANOVA and the Tukey post hoc test were used for the statistical analysis. Results: Polished PEEK discs showed lower cell viability, whereas roughened sandblasted zirconia and PEEK discs showed the highest proliferation rates and cell viability percent. The osteogenic differentiation was enhanced for rough surfaces in comparison to polished surfaces. Sandblasted zirconia and PEEK discs showed a markedly increased mineralized nodule development and ALP enzyme activity compared to the polished surface and control. Conclusions: Micro- topographies creation on the PEEK implant surface enhances stem cell attachment, viability, and osteogenic differentiation. READ ALL READ LESS Keywords PEEK, PLDSCs, Sandblasting ,Zirconia Corresponding Author(s) Sara F. El Shafei ( [email protected] ) Close Corresponding author: Sara F. El Shafei Competing interests: No competing interests were disclosed. Grant information: The author(s) declared that no grants were involved in supporting this work. Copyright: © 2023 El Shafei SF et al . This is an open access article distributed under the terms of the Creative Commons Attribution License , which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited. How to cite: El Shafei SF, Raafat SN and Farag EA. Enhanced human periodontal ligament stem cell viability and osteogenic differentiation on two implant materials: An experimental in vitro study [version 2; peer review: 2 approved] . F1000Research 2023, 12 :447 ( https://doi.org/10.12688/f1000research.129562.2 ) First published: 26 Apr 2023, 12 :447 ( https://doi.org/10.12688/f1000research.129562.1 ) Latest published: 08 Aug 2023, 12 :447 ( https://doi.org/10.12688/f1000research.129562.2 ) Revised Amendments from Version 1 Abbreviated words in abstract have been extended to full form. The referencing style has not been changed since this style is in accordance with the journal submission regulations. The sentences that were unclear have been rephrased. PEEK abbreviations have been clarified as Polyetheretherketone. The antibiotic dose used was clarified as added once in the culture medium bottle (500 ml) which is used in the whole study. Repetition of RRID:SCR_002798 was corrected. The Resolution of Figures 2a and 3 has been improved. Figures 2b and 5 have been edited and relabelled correctly. The Mean values of absorbance of the study group were added in the manuscript. In the discussion, the sentence A strategy that has the potential to improve PLDSC attachment …. has been rephrased to clarify the meaning. The label of figure 2b has been corrected to Chondrogenesis. The kind cell/material binding has been added to the discussion section. Opposing opinions have been added to the discussion section of the manuscript to enrich the study details. Abbreviated words in abstract have been extended to full form. The referencing style has not been changed since this style is in accordance with the journal submission regulations. The sentences that were unclear have been rephrased. PEEK abbreviations have been clarified as Polyetheretherketone. The antibiotic dose used was clarified as added once in the culture medium bottle (500 ml) which is used in the whole study. Repetition of RRID:SCR_002798 was corrected. The Resolution of Figures 2a and 3 has been improved. Figures 2b and 5 have been edited and relabelled correctly. The Mean values of absorbance of the study group were added in the manuscript. In the discussion, the sentence A strategy that has the potential to improve PLDSC attachment …. has been rephrased to clarify the meaning. The label of figure 2b has been corrected to Chondrogenesis. The kind cell/material binding has been added to the discussion section. Opposing opinions have been added to the discussion section of the manuscript to enrich the study details. See the authors' detailed response to the review by Samar Darwish See the authors' detailed response to the review by Sahar El Refai READ REVIEWER RESPONSES Introduction The biological interface between a dental implant and the host tissue in which it is embedded is a critical determinant of clinical outcome. 1 To enable osteointegration of the implant, osteoconductive material and coatings are usually used. 2 However, in case of significant loss of bone volume (and the unsuitability of bone autografts), the use of osteoconductive materials only will not suffice. The use of osteoinductive material should provide an improved clinical outcome. These materials should stimulate ontogenesis, increase vascularization and hence improve mechanical stability of the implant. 3 Human periodontal ligament stem cells (hPDLS) have mesenchymal stem cells characteristics, may regenerate, and develop into a variety of cells, including osteoblasts. Furthermore, their angiogenic potential has been demonstrated via the release of vascular endothelial growth factor (VEGF), indicating that they play an important role in implant osseointegration. 4 The biocompatibility of titanium with bone tissue makes it a popular material for bone implants. Unfortunately, due to its dark color, titanium implants may become obvious through the gingiva. 5 , 6 Tetragonal zirconia polycrystals (TZP) on the other hand show outstanding esthetic performance. Additionally, due to their biocompatibility and favorable mechanical properties, TZP has become a pillar material for dental restorations. 7 However, from an osteointegration perspective, titanium still shows superiority. 8 PEEK is a synthetic polymer which shows potential to be used as an aesthetic dental implant material given its tooth-like color. It possesses exceptional chemical tolerance and biomechanical characteristics making it well-suited for biomedical applications. Notwithstanding, PEEK's bioactivity and osseointegration are also debatable due to shortage of evidence and lack of comprehensive assessment of osseointegration. 9 Surface topography modification by lithography, pattern transfer, sandblasting, acid-etching, or plasma immersion ion implantation has been shown to promote cellular adhesion and proliferation. Micro-topographical modifications also create areas in which the bone infiltrates and grows, enhancing implant fixation and stability into bone tissue. 10 Sandblasting is one of the most versatile techniques used for implant surface modification. Increased surface area of implants to promote osseointegration is its main benefit. The rougher surfaces also improve osteoblast adhesion and proliferation. 11 , 12 In comparison to titanium implants, the significance of surface alterations in cell proliferation and the development of osteogenic tissue on zirconia and PEEK implants is much less understood. This study aimed to ascertain the effect of implant material (TZP and PEEK) and surface characteristics (rough versus smooth) on human periodontal ligament stem cell growth and osteogenic differentiation. Methods This study was conducted from 1 st of February 2022 to 1 st of November 2022. This study is an experimental in vitro study investigating hPDLS behavior using different implant materials with different topographies. Disc preparation Yttria stabilized tetragonal zirconia polycrystals (ZrO 2 balanced, Y 2 O 3 < 5.15 mass %, HfO 3 < 3 mass%, Al 2 O 3 < 0.5 mass%, SiO 2 < 0.02 mass %, Fe 2 O 3 < 0.01, Na 2 O < 0.04) (Bruxzir Shaded, Glidewell) and polyether ether ketone polyetheretherketone (PEEK) blocks were used. The blocks were cut into discs of 1 mm thick and 10 mm in diameter. Four experimental groups of discs were formed according to the type of material and surface treatment: experimental zirconia discs with polished surface (Z); experimental zirconia discs with a sandblasted surface (ZS); experimental PEEK discs with smooth surface (P) and experimental PEEK discs with a sandblasted surface. Using a polishing apparatus (Ecomet 3, Buehler), polished TZP samples were first ground on the two sides with diamond discs (70 m, then 45 m). They were then polished with a polishing cloth, diamond particles measuring 3 and 9 microns, and colloidal silica measuring 0.6 microns. 13 PEEK polished samples, however, were produced by first polishing the PEEK disc with 1000 and 2000 grit silicon carbide sandpapers, then finishing with 8000 grit alumina lapping film to offer a smooth surface. 14 At 50 mm perpendicular distance on all sides, particles of 110 m alumina (Korox 110, BEGO) were used to sandblast specimens of TZP and PEEK at 5 bar air pressure using a sandblaster (Basic eco, Renfert). All discs were sandblasted for 10 seconds. 15 All samples underwent a 10 minutes ultrasonic cleaning procedure using acetone and distilled water before being used in cell culture experiments, followed by a half an hour UV disinfection lamp application on each side of each disc. In-vitro stem cell isolation and characterization Periodontal ligament (PDL) cells isolation and culture Under the guidance of the British University in Egypt's Ethical Committee (approval # 22-003) and with patients' (n=3) full agreement and signing an informed consent, three human permanent teeth were gathered from healthy donors at the oral and maxillofacial dental department. The teeth had been extracted for orthodontic purposes. The patients were informed about the study privately and the patient’s data confidentiality was ensured. The patients’ inclusion criteria were; not suffering from any chronic diseases, and average age between 20–40 years. The patients were not further included in the study and were considered as teeth donors only. The selected teeth were free from any carious lesions. The extracted teeth were kept in Dulbecco’s modified Eagle’s medium (DMEM; Sigma, St. Louis, MO), which received antibiotic doses (300 U/ml penicillin and 300 mg/ml streptomycin; Sigma). Within the first 24 hours from teeth extraction, primary cell culture was performed. Sterile phosphate-buffered saline (PBS, Sigma) was used to disinfect the PDL tissues removed from healthy third molars root surfaces. PBS was then added with antibiotics at escalating doses to provide further protection. Stem cells were isolated from these PDL tissues by using the outgrowth method. 16 Smaller pieces of PDL were put onto 35 mm plates (NUNC, Roskilde) with 1 mL of culture media, which was composed of Dulbecco's modified Eagle's medium/nutrient combination F-12 Ham medium (DMEM/F12, Sigma) with 10% fetal bovine serum (FBS, Gibco) added as a supplement and 1% penicillin/streptomycin. 37°C incubation was conducted with 5% CO 2 in a humid atmosphere. An inverted microscope was used to monitor cell growth and morphology. The cells were trypsinized and sub-cultured once they had reached 80% confluence. Every three days, the medium was changed. All tests were carried out on cells acquired during the fourth passage ( Figure 1a , b ). Figure 1. hPLDSCs growth observation under microscope. (a) Cell aggregates at passage 0; (b) Confluent cells at passage 4 (Mag. 40×). Identification of isolated PDL cells as mesenchymal stem cells (MSCs) Cell surface marker identification. The expression of specific surface markers on the extracted PDLSCs was identified by fluorescence-activated cell sorting (FACS). The cells were collected, moved, and then fixed for 15 minutes in 4% paraformaldehyde. First, 3% bovine serum albumin (BSA; Sigma-Aldrich) was used to incubate the cells before being incubated for one hour with primary antibodies (eBioscience, Thermo Fisher Scientific), produced against CD105, CD90, CD73, CD34, CD45, and HLA-DR for 1 hour. The secondary antibody was applied and left on the cells for 45 minutes at room temperature after being rinsed with wash buffer. After three rounds of washing, a flow cytometer (FACSCalibur, BD Biosciences) was used to evaluate the cells. 17 Multilineage differentiation. To evaluate the in vitro potential differentiation capacity of hPDLSCs, 5×10 4 cells/ml from the fourth passage were cultured in 24-well plate with OsteoDiff media, AdipoDiff media, and CondroDiff media (Human mesenchymal stem cell functional identification kit, R&D Systems) for three weeks to achieve osteogenic, adipogenic, and chondrogenic differentiation, respectively. Alizarin red staining (Sigma-Aldrich) was used to assess mineralization, which demonstrates osteogenesis. Oil red O solution (Sigma-Aldrich) was utilized to find lipid droplet aggregation, which demonstrates adipogenesis. Alcan blue staining (Sigma-Aldrich) was used to identify glycosaminoglycans verifying chondrogenic differentiation. Assessment of cell viability, morphology and osteogenic differentiation capacity upon culture on disks Cell viability. Six discs of each group (Z, ZS, P and PS) were put at the bottom of a 6-well plate. Cells were then put to the wells at a density of 30×104 cell for each well. The culturing of cells was on the discs for the following 24, 48, and 72 hours in which cellular viability was evaluated, utilizing methylthiazolyldiphenyl-tetrazolium bromide (MTT) assay (Sigma Aldrich), relative to control cells being cultured in 6-well plates without discs. Absorbance was recorded using a benchtop microplate reader (Thermo Scientific Multiscan). 18 The findings were presented as mean viability % relative to controls cultured in the absence of disks. Cell morphology. Cells were seeded in 6-well plates on Z, ZS, P and PS discs as detailed earlier and 72 hours were spent cultivating. Subsequently, 4% glutaraldehyde was used to fix the cells for 2 hours at 4°C. 19 For critical point drying, samples dehydration was done using increasing ethanol concentrations from 25%,50%, 75%, 95%, to 100%, 5 minutes in each concentration. Samples were subjected to sputter-coating with gold using 15 mA for 4 min (HUMMER 8.0, ANATECH). Discs without cells and those with cultured cells were analyzed by scanning electron microscopy (SEM, Leo Supra 55). Osteogenic differentiation assay. For osteogenic differentiation assay, cells were implanted in a 6-well plate at a density of (30 × 104 cells/well) on disks. The medium was changed the following day to osteogenic differentiation medium, which contains MEM-Alpha, 10% FBS, dexamethasone 100 nm (Sigma Aldrich, Steinheim), ascorbic acid-2-phosphate 200 uM (Sigma Aldrich), and beta glycerophosphate 10 mM (Merck, Darmstadt). 20 Differentiation was performed for 14 days. A positive control was achieved using cells cultivated in osteogenic media without discs, whereas a negative control used cells cultured in normal medium (DMEM). Both osteogenic induction medium and basal culture medium were altered twice per week. After 14 days of cultivation, the osteogenic distinction capacity was assessed by the calcium deposition mineralization assay using alizarin red staining and alkaline phosphatase (ALP) activity. Alizarin red S assay. Once calcified, mineralized nodules were observed after 14 days of induction, alizarin red S staining was conducted. In more detail, the medium was aspirated and the cells were fixed with 10% formaldehyde (Sigma, v/v) at room temperature for 15 minutes. After three PBS washes to remove non-attached cells, the plates were stained for 30 minutes at room temperature with 40 mm alizarin red S (Sigma, pH 4.2). To eliminate excess stain, cells were washed 3 more times with PBS before being examined using a light microscope. Additionally, at 405 nm, the absorbance was measured after staining was solubilized in a 10% glacial acetic acid (Sigma-Aldrich) solution. 21 Alkaline phosphatase (ALP) activity assay. ALP activity of PDLSCs on every specimen was assayed after differentiation for 14 days. Briefly, monolayers were cleaned twice with PBS and then once with 1 mL alkaline phosphatase buffer (ALPB). One mL of ALPB was added to each well and equal volumes of para-Nitrophenylphosphate (p-NPP) (Sigma Aldrich) equilibrated to 4°C were added. Immediately, 50 ul were removed and mixed with same volume of NaOH to stop the reaction. The previous step was repeated every minute for each well till 10 minutes had passed. The rate of accumulation of p-nitrophenolate was plotted for each well and the initial rate of the reaction, a way to express the rate of the reaction, was calculated by obtaining the gradient at the linear phase for each group. Monolayers were then cleaned in PBS and kept at -20°C for total protein quantification. 22 Total protein amounts were detected utilizing enhanced BCA protein assay kit (Pierce) based on the manufacturer’s procedures. 23 The absorbance at 405 nm was detected by spectrophotometry. The initial rate of the reaction in each group (slope) was divided by protein concentrations (mg). Statistical analysis Each in-vitro assay was carried out in triplicate and analyzed across three distinct studies. Graph-Pad Prism v8.1.0 (RRID:SCR 002798) was used to conduct ANOVA test, followed by a pair-wise Tukey's post hoc test, after validating the homogeneity of variance and normal distribution of the data. Results PLDSCs identification According to flow cytometry findings, isolated PLDSCs were CD73 (98.76%), CD90 (95.67%), and CD105 (95.52%) positive; and less than 5% for CD34 (4.44%), CD45 (4.48%), and HLA-DR (2.66%) ( Figure 2a ). The rates are provided in respect to the presence of mesenchymal stem cell surface markers. Additionally, hPDLSCs were found to be capable of differentiating into osteoblasts, chondrocytes, and adipocytes ( Figure 2b ). PLDSCs differentiated into osteoblasts showed positive red staining of the mineralized matrix using alizarin red S stain, cells differentiated into chondrocytes showed positive staining of the produced glycosaminoglycans via alcian blue stain and cells differentiated into adipocytes showed positive staining of the oil droplets using oil red staining. Figure 2. PLDSCs identification (a) Flowcytometry Figures (b) In vitro multilineage cell differentiation (Mag. 100×). Cytotoxicity assay An MTT assay was utilized to determine the viability of the hPLDSCs ( Figure 3 ). Data are presented after 24,48, and 72 hours of exposure of the PLDSCs to discs and were normalized to the control (cells + medium). Polished PEEK surface discs showed a notably reduced cell viability % in comparison to the three other groups at 24,48 and 72 hours (p<0.05). The sandblasted roughened zirconia and PEEK discs showed increased cell viability % throughout the whole experiment, the roughened PEEK surface possessed significant higher cell viability % compared to the same respective group of smooth polished surface (p<0.001), and the roughened Zirconia discs also showed higher cell viability % compared to the same respective group of smooth polished surface (p<0.05). However, the percentage of cell viability was not significantly different between the roughened zirconia and PEEK discs. Figure 3. MTT assay. The data are presented as absorbance values (570 nm) at 24, 48, and 72 h of the PLDSCs being exposed to discs. *p significant difference of the respective group in comparison to the Z group, # p significant difference of the respective group in comparison to the P group, p values <0.05 are statistically significant. For each material's surface properties, each experimental condition was run in triplicate. Cell adhesion and morphology The discs’ surface microstructure and stem cell morphology are shown in Figure 4 . Cells with extending cytoplasmic processes were frequent and well-observed in smooth polished zirconia discs while no well-defined cells were seen attached to the polished PEEK discs. Cell adhesion and structure of hPLDSCs on roughened sandblasted specimen surfaces revealed the presence of numerous well-attached-type cells with extending stretched cytoplasmic processes. Figure 4. Representative SEM micrographs illustrate (a) polished smooth zirconia surface, (b) cell adhesion on polished smooth zirconia surface, (c) sandblasted roughened zirconia surface, (d) cell adhesion on sandblasted roughened zirconia surface, (e) polished smooth PEEK surface, (f) cell adhesion on polished smooth PEEK surface, (g) sandblasted roughened PEEK surface, (h) cell adhesion on sandblasted roughened PEEK surface, (i) cell adhesion on sandblasted roughened PEEK higher resolution. Mineralization assay calcium deposition On day 14, extracellular matrix rich in calcium was assessed as a byproduct of the osteogenic differentiation process when stained with alizarin red S ( Figure 5 ). There were notable variations in between the groups examined; the negative control (cells+normal medium) displayed fewest mineralized nodules amounts; the positive control (osteogenic medium only) exhibited higher amount of calcium deposition (p<0.001) than polished zirconia and PEEK groups ( Figure 5 ). When compared to the other experimental groups, roughened sandblasted zirconia and PEEK groups possessed greatest grades of mineralized nodules (p<0.001). Average mean values of absorbance in the control, Osteo, Z, P, ZS, and PS groups are 0.048±0.01, 0.24±0.01, 0.1784±0.01, 0.11±0.02, 0.33±0.01, 0.32±0.01, respectively. Figure 5. Osteogenic differentiation and alizarin assay: On day 14, as a byproduct of the differentiation of osteoblasts, calcium deposition was examined using alizarin red S staining. (Mag. 40×). (a) Control; (b) Osteogenic medium; (c) Smooth zirconia surface; (d) Smooth PEEK surface; (e) Roughened zirconia surface; (f) Roughened PEEK surface. Every experimental condition was carried out in triplicate for every material surface characteristics. (g) Alizarin red staining data from three independent experiments were statistically analyzed and given as mean standard deviation, *p significant difference of the respective group compared to the control, # p significant difference of the respective group in comparison to the osteo group. $ p significant difference of the respective group in comparison to the Z group. p values <0.05 is statistically significant. ALP (Alkaline phosphatase) assay results The ALP assay kinetic profile demonstrated the aggregation of the yellow p-nitrophenolate (p-NP) as a growing product among the various groups (shown in Figure 6a ). The control group possessed the lowest rate of accumulation, the polished zirconia and PEEK showed lower rate of p-NP accumulation than the roughened sandblasted disks of same respective groups. Statistical analysis was done by calculating the slope of each curve and divided by the total amount of protein in each well. The results showed that the ALP activity at day 14 in cells cultured on the polished zirconia and PEEK discs was significantly lower than the positive control (p<0.05); polished PEEK discs possessed the lowest ALP activity compared to the polished zirconia discs and positive control (p<0.001). Roughened sandblasted Zirconia and PEEK discs indicated most ALP activity in comparison to the polished discs (p<0.001) ( Figure 6b ). Regarding ALP activity, no discernible change was found in cells cultured on the roughened sandblasted zirconia discs, PEEK discs and the positive control group, suggesting that roughened sandblasted surfaces can stimulate the osteogenic potential of PDLSCs. Mean values of the rate of ALP reaction (slope) of the control, Osteo, Z, P, ZS and PS groups are 0.0038±0.0002, 0.0125±0.001, 0.0107±0.001, 0.009±0.004, 0.0131±0.001, 0.0135±0.0003, respectively. Figure 6. ALP (Alkaline phosphatase) activity after 14 days of osteogenic induction. a) An illustration of an ALP assay's kinetic curve showing how different groups over time accumulated the yellow p-nitrophenolate product. b) The initial rate of the reaction in each group by calculating the slope, values were scaled back to the quantity of total protein. The values revealed as mean±SD of three independent experiments, *p significant difference of the respective group in comparison to the control (cells in normal medium), # p significant difference of the respective group in comparison to the osteo group (osteogenic medium), $ p significant difference of the respective group compared to the Z group. p values <0.05 is statistically significant. Discussion Titanium and zirconia are commonly used as dental implant materials, whereas PEEK is widely used in orthopedic research. PEEK, on the other hand, has little biological activity when utilized as an implant, making it unable to successfully osseointegrate with adjacent bone tissue. These flaws significantly restrict the osteogenic properties of PEEK, which is a crucial factor for implants long-term stability. As a result, recent years have seen a surge in research into various methods of modification that might improve PEEK's biological activities, such as cell compatibility, osteogenic activity, and antibacterial capability. 24 By enhancing bone-to-implant connection and peri-implant bone density, implant surface modification aimed to accelerate the early phases of osteointegration. 25 The surface pattern can be altered by acid etching and airborne particle abrasion, or the surface's physiochemical characteristics can be altered through coating application to enhance the surface's bioactive and osteoconductive qualities. 26 Meanwhile, MSCs are capable of self-renewal and cell lineage differentiation into a wide range of cell types, including osteoblasts, chondrocytes, adipocytes, neurocytes, and myocytes. hPDLSCs have been used in this study for the advantages of MSCs as self-renewal, multipotency and immunomodulation, in addition to being a cell population that is conveniently accessible and has the required properties for future clinical use in human studies. 27 This study assessed the in vitro behavior of hPLDSCs on TZP and PEEK with varying surface topography. The sandblast method was used to micro-roughen the surface; this approach has been utilized for metal implants and is straightforward and affordable. 28 To assess the behavior of hPLDSCs on different substrates, we employed different in vitro assays including cell viability, morphology and osteogenic potential. Our results showed that polished TZP had better hPLDSCs cell attachment and viability than polished PEEK, whereas sandblasted TZP and PEEK had the highest significant hPLDSCs viability and osteogenic differentiation. Cell viability was higher on the rough TZP surface than on the smooth surface. Different surface topography presumably modifies the shape and arrangement of the cells' cytoskeletons influencing cell attachment proteins secretion, hence enhanced cell viability and proliferation. 29 The viability of hPLDSCs on sandblasted PEEK was significantly higher than on smooth PEEK. Sandblasting produced peaks and valleys on PEEK surfaces resulting in increased PEEK's surface roughness and increased contact angle. 30 Danen et al. 31 proposed that cell adhesion proteins have an arginine-glycine-aspartate peptide sequence that serves as the integrin-binding domain, a protein known to play a role in the adhesion of osteoblasts to biomaterials, and is essential for initial cell attachment, morphology, and proliferation. Because roughened PEEK has a larger surface area, more proteins can be adsorbed on it, resulting in better initial cell attachment and proliferation. Our findings agree with those of Sunarso et al., 14 suggesting that roughened PEEK could achieve cell-cell interactions faster than mirror smooth PEEK. In the current study, the smooth surface specimens of TZP had micro-topography. It displayed advanced cell attachment, a well-adhered cell morphology, and some fibres running between cells, indicating that cells aggregated together. This could be due to the presence of micro-scale granules in the smooth-surfaced TZP. The observed filamentous scaffolding was previously reported to be important in cell adhesion and spread. 32 The SEM observations revealed that the cells in the roughened groups exhibited multiple aggregates of cells. Cell attachment samples revealed to be dependent on the finger-like cytoplasmic extensions formation, which was clearly observed on the roughened PEEK specimens in contrast to the polished smooth PEEK. These structures appeared to serve as anchors for the cells to the underlying surface. 8 These findings imply that cell rearrangement might result from the sandblasted structure's conversion of the material's physical signals into intracellular biological signals. 33 The alizarin red staining and ALP activity assays were used to detect the impact of the tested specimens on the osteogenic potential of hPLDSCs. Our findings revealed that calcium-rich extracellular matrix and ALP activity were significantly higher on sandblasted surfaces than on smooth specimens. All samples showed evidence of calcium nodule development, with differences in aggregation and dispersion based on the topographical type. The mineralization tendency was confirmed by the distribution and density of the nodules. These findings are consistent with Sima et al . 34 who proved that increased surface granulation, particularly on sandblasted surfaces, indicates that roughened surfaces act as mineralization nucleation centres. The enhanced osteogenic activity of hPLDSCs could be due to the synergistic effect of micro-topography which may provide a biomimetic surface and influences hPLDSC proliferation and osteogenic differentiation. 2 A previous study by Taniguchi et al . 35 examined how surface roughness affected cultured osteoblast-like cell morphology, proliferation, differentiation and reported that a roughened surface has a greater chance to speed up the calcification process and boost cells' osteogenic ability. Furthermore, the study showed that surface roughness promoted osseointegration of the peri-implant area after application on induced bone defects of rats’ tibiae. Jung et al. illustrated that the kind of cell/material binding seems to be regulated by the implant material. It proved that in osteoblasts, the cell/material binding occur in 60–70% via fibronectin and in 40–50% via laminin. They postulated binding via laminin as a second type of binding and as an enhancement of binding via fibronectin. Fibronectin expression increases over time and with the strongest alteration and surface topography alteration of implant surface. 36 Sandblasted zirconia and PEEK surfaces with microscale roughness may provide a suitable niche for hPLDSC proliferation and osteogenic differentiation. A study by Zhu et al. 37 discovered groove-ridge patterns on the surface of polystyrene that resembled collagen fibrillar architecture more closely in profile and size and that it could be used as a scaffold to provide directed physical cues for osteoblast-like cell orientation, spreading, and directional mineralization. In contrast with the findings of our study Fukuda et al. demonstrated that sandblast treatment alone did not increase osteogenesis in rat-derived MSCs although a slight increase in the osteocalcin level was observed. The study highlighted the importance of combined chemical modifications in addition to surface roughness. 38 The non-significant effect of surface roughness in this study may be attributed to the different type of stem cells used and the different sample preparation procedures. Surface roughness has an impact on cell attachment, proliferation, differentiation, and mineralized matrix synthesis. Our results are in line with those of Lincks et al., 39 who found that roughness is an important variable in promoting osteogenic differentiation and appears to affect cell orientation. Furthermore, as surface roughness increases, so does bone development rates and the percentage of surface area in close interaction with bone. In addition, Schneider et al. 40 studied the influence of implant surface microtopography on the expression of transcription factors regulating osteoblast differentiation. It was observed that there is a relation between cell attachment and surface architecture through gene expression regulation. To summarize the current study findings, we found that roughened PEEK exhibited high PLDSCs attachment and viability, close to that of roughened zirconia and higher in comparison to the polished PEEK. In addition, higher level of mineralized matrix and ALP activity were observed on the roughened TZP and PEEK. Sandblasted PEEK showed improved osteogenic potential through elevation of the responses of hPLDSCs. The development of micro-topographies on the PEEK surface is a strategy that has the potential to improve PLDSC attachment, viability and osteogenic differentiation. Future efforts should be directed at detailed understanding of the development of more efficient and the study of PEEK modification techniques to enhance accelerated osteogenic differentiation. Data availability All data underlying the results are available as part of the article and no additional source data are required. Extended data Figshare: https://figshare.com/articles/dataset/Raw_data_xlsx/21724385.v1 . 41 This project contains the following extended data ‐ Supplementary raw data for and statistical analysis of all in vitro assays ‐ Supplementary data of the control figures of flow-cytometry in stem cells characterization ‐ All in vitro assays protocols and procedures were done in accordance to; Mesenchymal stem cells assays and applications. https://link.springer.com/protocol/10.1007/978-1-60761-999-4_17 Data are available under the terms of the creative commons attribution 4.0 international license CC BY 4.0. Acknowledgements The authors would like to thank all members in the Stem cells and Tissue Culture Hub, Centre of Innovative Dental Sciences, Faculty of Dentistry, The British University in Egypt (BUE), for their assistance in the study. References 1. Parithimarkalaignan S, Padmanabhan TV: Osseointegration: An Update. J. Indian Prosthodont. Soc. 2013; 13 (1): 2–6. PubMed Abstract | Publisher Full Text | Free Full Text 2. Hirano T, Sasaki H, Honma S, et al. : Proliferation and osteogenic differentiation of human mesenchymal stem cells on zirconia and titanium with different surface topography. Dent. Mater. J. 2015; 34 (6): 872–880. PubMed Abstract | Publisher Full Text 3. Kazimierczak P, Przekora A: Osteoconductive and Osteoinductive Surface Modifications of Biomaterials for Bone Regeneration: A Concise Review. Coatings. 2020; 10 (10): 971. Publisher Full Text 4. Marconi GD, Fonticoli L, Della Rocca Y, et al. : Human Periodontal Ligament Stem Cells Response to Titanium Implant Surface: Extracellular Matrix Deposition. Biology (Basel). 2021; 10 (9): 931. Publisher Full Text 5. Sicilia A, Cuesta S, Coma G, et al. : Titanium allergy in dental implant patients: a clinical study on 1500 consecutive patients. Clin. Oral Implants Res. 2008; 19 (8): 823–835. PubMed Abstract | Publisher Full Text 6. Egusa H, Ko N, Shimazu T, et al. : Suspected association of an allergic reaction with titanium dental implants: a clinical report. J. Prosthet. Dent. 2008; 100 (5): 344–347. PubMed Abstract | Publisher Full Text 7. Depprich R, Zipprich H, Ommerborn M, et al. : Osseointegration of zirconia implants compared with titanium: an in vivo study. Head Face Med. 2008; 4 (1): 1–8. Publisher Full Text 8. Piconi C, Maccauro G: Zirconia as a ceramic biomaterial. Biomaterials. 1999; 20 (1): 1–25. Publisher Full Text 9. Schwitalla A, Müller WD: PEEK dental implants: a review of the literature. J. Oral Implantol. 2013; 39 (6): 743–749. PubMed Abstract | Publisher Full Text 10. Sunarso TA, Toita R, et al. : Enhanced Osseointegration Capability of Poly (ether ether ketone) via Combined Phosphate and Calcium Surface-Functionalization. Int. J. Mol. Sci. 2019; 21 (1): 198. PubMed Abstract | Publisher Full Text | Free Full Text 11. Fischer J, Schott A, Märtin S: Surface micro-structuring of zirconia dental implants. Clin. Oral Implants Res. 2016; 27 (2): 162–166. PubMed Abstract | Publisher Full Text 12. Brett PM, Harle J, Salih V, et al. : Roughness response genes in osteoblasts. Bone. 2004; 35 (1): 124–133. Publisher Full Text 13. Ito H, Sasaki H, Saito K, et al. : Response of osteoblast-like cells to zirconia with different surface topography. Dent. Mater. J. 2013; 32 (1): 122–129. PubMed Abstract | Publisher Full Text 14. Sunarso TA, Fukuda N, et al. : Effect of micro-roughening of poly (ether ether ketone) on bone marrow derived stem cell and macrophage responses, and osseointegration. J. Biomater. Sci. Polym. Ed. 2018; 29 (12): 1375–1388. PubMed Abstract | Publisher Full Text 15. El Awadly TA, Wu G, Ayad M, et al. : A histomorphometric study on treated and untreated ceramic filled PEEK implants versus titanium implants: Preclinical in vivo study. Clin. Oral Implants Res. 2020; 31 (3): 246–254. PubMed Abstract | Publisher Full Text | Free Full Text 16. Tran HLB, Doan VN, Le HTN, et al. : Various methods for isolation of multipotent human periodontal ligament cells for regenerative medicine. In Vitro Cell. Dev. Biol. Anim. 2014; 50 (7): 597–602. PubMed Abstract | Publisher Full Text 17. Dominici M, Le Blanc K, Mueller I, et al. : Minimal criteria for defining multipotent mesenchymal stromal cells. The International Society for Cellular Therapy position statement. Cytotherapy. 2006; 8 (4): 315–317. PubMed Abstract | Publisher Full Text 18. Sequeira DB, Seabra CM, Palma PJ, et al. : Effects of a New Bioceramic Material on Human Apical Papilla Cells. J. Funct. Biomater. 2018; 9 (4). PubMed Abstract | Publisher Full Text | Free Full Text 19. Al Shehadat S, Gorduysus MO, Hamid SSA, et al. : Optimization of scanning electron microscope technique for amniotic membrane investigation: A preliminary study. Eur. J. Dent. 2018; 12 (4): 574–578. PubMed Abstract | Publisher Full Text 20. Hong L, Sultana H, Paulius K, et al. : Steroid regulation of proliferation and osteogenic differentiation of bone marrow stromal cells: a gender difference. J. Steroid Biochem. Mol. Biol. 2009; 114 (3–5): 180–185. PubMed Abstract | Publisher Full Text | Free Full Text 21. Rodríguez-Lozano FJ, López-García S, García-Bernal D, et al. : Cytocompatibility and bioactive properties of the new dual-curing resin-modified calcium silicate-based material for vital pulp therapy. Clin. Oral Investig. 2021; 25 (8): 5009–5024. PubMed Abstract | Publisher Full Text 22. Krause U, Seckinger A, Gregory CA: Assays of osteogenic differentiation by cultured human mesenchymal stem cells. Methods Mol. Biol. 2011; 698 : 215–230. Publisher Full Text 23. Li J, Zhang F, Zhang N, et al. : Osteogenic capacity and cytotherapeutic potential of periodontal ligament cells for periodontal regeneration in vitro and in vivo. PeerJ. 2019; 7 : e6589. PubMed Abstract | Publisher Full Text | Free Full Text 24. Ma Z, Zhao X, Zhao J, et al. : Biologically Modified Polyether Ether Ketone as Dental Implant Material. Front. Bioeng. Biotechnol. 2020; 8 : 620537. PubMed Abstract | Publisher Full Text | Free Full Text 25. Van Oirschot BAJA, Alghamdi HS, Närhi TO, et al. : In vivo evaluation of bioactive glass-based coatings on dental implants in a dog implantation model. Clin. Oral Implants Res. 2014; 25 (1): 21–28. PubMed Abstract | Publisher Full Text 26. Majhi R, Majhi RK, Garhnayak L, et al. : Comparative evaluation of surface-modified zirconia for the growth of bone cells and early osseointegration. J. Prosthet. Dent. 2021; 126 (1): 92.e1–92.e8. Publisher Full Text 27. Marconi GD, Fonticoli L, Della RY, et al. : Enhanced Extracellular Matrix Deposition on Titanium Implant Surfaces: Cellular and Molecular Evidences. Biomedicines. 2021; 9 (11): 1710. PubMed Abstract | Publisher Full Text | Free Full Text 28. Le Guéhennec L, Soueidan A, Layrolle P, et al. : Surface treatments of titanium dental implants for rapid osseointegration. Dent. Mater. 2007; 23 (7): 844–854. Publisher Full Text 29. Boyan BD, Lohmann CH, Dean DD, et al. : Mechanisms involved in osteoblast response to implant surface morphology. Annu. Rev. Mater. Sci. 2001; 31 : 357–371. Publisher Full Text 30. Ourahmoune R, Salvia M, Mathia TG, et al. : Surface morphology and wettability of sandblasted PEEK and its composites. Scanning. 2014; 36 (1): 64–75. PubMed Abstract | Publisher Full Text 31. Danen EHJ, Yamada KM: Fibronectin, integrins, and growth control. J. Cell. Physiol. 2001; 189 (1): 1–13. Publisher Full Text 32. Wei C, Gong T, Pow EHN, et al. : Adhesive and oxidative response of stem cell and pre-osteoblasts on titanium and zirconia surfaces in vitro. J. Investig. Clin. Dent. 2019; 10 (3): e12407. PubMed Abstract | Publisher Full Text 33. Sun Y, Sun J, Wu X, et al. : Mechanism of zirconia microgroove surface structure for osseointegration. Mater. Today Adv. 2021; 12 : 100159. Publisher Full Text 34. Sima LE, Bonciu A, Baciu M, et al. : Bioinstructive Micro-Nanotextured Zirconia Ceramic Interfaces for Guiding and Stimulating an Osteogenic Response In Vitro. Nanomater. 2020; 10 (12): 1–25. 35. Taniguchi Y, Kakura K, Yamamoto K, et al. : Accelerated Osteogenic Differentiation and Bone Formation on Zirconia with Surface Grooves Created with Fiber Laser Irradiation. Clin. Implant. Dent. Relat. Res. 2016; 18 (5): 883–894. PubMed Abstract | Publisher Full Text 36. Jung S, Bohner L, Hanisch M, et al. : Influence of Implant Material and Surface on Mode and Strength of Cell/Matrix Attachment of Human Adipose Derived Stromal Cell. Int. J. Mol. Sci. 2020; 21 (11): 4110. 37. Zhu B, Lu Q, Yin J, et al. : Alignment of osteoblast-like cells and cell-produced collagen matrix induced by nanogrooves. Tissue Eng. 2005; 11 (5–6): 825–834. PubMed Abstract | Publisher Full Text 38. Fukuda N, Kanazawa M, Tsuru K, et al. : Synergistic effect of surface phosphorylation and micro-roughness on enhanced osseointegration ability of poly (ether ether ketone) in the rabbit tibia. Sci. Rep. 2018; 8 (1): 16887. PubMed Abstract | Publisher Full Text | Free Full Text 39. Lincks J, Boyan BD, Blanchard CR, et al. : Response of MG63 osteoblast-like cells to titanium and titanium alloy is dependent on surface roughness and composition. Biomaterials. 1998; 19 (23): 2219–2232. PubMed Abstract | Publisher Full Text 40. Schneider GB, Perinpanayagam H, Clegg M, et al. : Implant Surface Roughness Affects Osteoblast Gene Expression. J. Dent. Res. 2003; 82 (5): 372–376. Publisher Full Text 41. El Shafei S, Raafat SN, Farag EA: Enhanced human periodontal ligament stem cell viability and osteogenic differentiation on two implant materials: An experimental in vitro study, Raw data. figshare. 2022. Publisher Full Text Comments on this article Comments (0) Version 2 VERSION 2 PUBLISHED 26 Apr 2023 ADD YOUR COMMENT Comment Author details Author details 1 Removable Prosthodontics, Faculty of Dentistry, The British University in Egypt, Cairo, Egypt 2 Department of Pharmacology, Director of Stem Cell and Tissue Culture Hub, Centre of Innovative Dental Sciences (CIDS), Faculty of Dentistry,, The British University in Egypt, Cairo, Egypt 3 Fixed Prosthodontics, Faculty of Dentistry, The British University in Egypt, Cairo, Egypt Sara F. El Shafei Roles: Conceptualization, Data Curation, Methodology, Writing – Review & Editing Shereen N. Raafat Roles: Conceptualization, Data Curation, Formal Analysis, Investigation, Methodology, Writing – Review & Editing Engy A. Farag Roles: Conceptualization, Data Curation, Methodology, Writing – Original Draft Preparation, Writing – Review & Editing Competing interests No competing interests were disclosed. Grant information The author(s) declared that no grants were involved in supporting this work. Article Versions (2) version 2 Revised Published: 08 Aug 2023, 12:447 https://doi.org/10.12688/f1000research.129562.2 version 1 Published: 26 Apr 2023, 12:447 https://doi.org/10.12688/f1000research.129562.1 Copyright © 2023 El Shafei SF et al . This is an open access article distributed under the terms of the Creative Commons Attribution License , which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited. Download Export To Sciwheel Bibtex EndNote ProCite Ref. Manager (RIS) Sente metrics Views Downloads F1000Research - - PubMed Central info_outline Data from PMC are received and updated monthly. - - Citations open_in_new 0 open_in_new 0 open_in_new SEE MORE DETAILS CITE how to cite this article El Shafei SF, Raafat SN and Farag EA. Enhanced human periodontal ligament stem cell viability and osteogenic differentiation on two implant materials: An experimental in vitro study [version 2; peer review: 2 approved] . F1000Research 2023, 12 :447 ( https://doi.org/10.12688/f1000research.129562.2 ) NOTE: If applicable, it is important to ensure the information in square brackets after the title is included in all citations of this article. COPY CITATION DETAILS track receive updates on this article Track an article to receive email alerts on any updates to this article. TRACK THIS ARTICLE Share Open Peer Review Current Reviewer Status: ? Key to Reviewer Statuses VIEW HIDE Approved The paper is scientifically sound in its current form and only minor, if any, improvements are suggested Approved with reservations A number of small changes, sometimes more significant revisions are required to address specific details and improve the papers academic merit. Not approved Fundamental flaws in the paper seriously undermine the findings and conclusions Version 2 VERSION 2 PUBLISHED 08 Aug 2023 Revised Views 0 Cite How to cite this report: El Refai S. Reviewer Report For: Enhanced human periodontal ligament stem cell viability and osteogenic differentiation on two implant materials: An experimental in vitro study [version 2; peer review: 2 approved] . F1000Research 2023, 12 :447 ( https://doi.org/10.5256/f1000research.154099.r194731 ) The direct URL for this report is: https://f1000research.com/articles/12-447/v2#referee-response-194731 NOTE: it is important to ensure the information in square brackets after the title is included in this citation. Close Copy Citation Details Reviewer Report 21 Aug 2023 Sahar El Refai , Princess Nourah bint Abdulrahman University, Riyadh, Riyadh Province, Saudi Arabia Approved VIEWS 0 https://doi.org/10.5256/f1000research.154099.r194731 I have no further comments ... Continue reading READ ALL I have no further comments than the ones the authors addressed Competing Interests: No competing interests were disclosed. I confirm that I have read this submission and believe that I have an appropriate level of expertise to confirm that it is of an acceptable scientific standard. Close READ LESS CITE CITE HOW TO CITE THIS REPORT El Refai S. Reviewer Report For: Enhanced human periodontal ligament stem cell viability and osteogenic differentiation on two implant materials: An experimental in vitro study [version 2; peer review: 2 approved] . F1000Research 2023, 12 :447 ( https://doi.org/10.5256/f1000research.154099.r194731 ) The direct URL for this report is: https://f1000research.com/articles/12-447/v2#referee-response-194731 NOTE: it is important to ensure the information in square brackets after the title is included in all citations of this article. COPY CITATION DETAILS Report a concern Respond or Comment COMMENT ON THIS REPORT Views 0 Cite How to cite this report: Darwish S. Reviewer Report For: Enhanced human periodontal ligament stem cell viability and osteogenic differentiation on two implant materials: An experimental in vitro study [version 2; peer review: 2 approved] . F1000Research 2023, 12 :447 ( https://doi.org/10.5256/f1000research.154099.r194730 ) The direct URL for this report is: https://f1000research.com/articles/12-447/v2#referee-response-194730 NOTE: it is important to ensure the information in square brackets after the title is included in this citation. Close Copy Citation Details Reviewer Report 10 Aug 2023 Samar Darwish , Pharmacology and Toxicology Department, Badr University in Cairo, Badr City, Cairo Governorate, Egypt Approved VIEWS 0 https://doi.org/10.5256/f1000research.154099.r194730 No further ... Continue reading READ ALL No further comments to make. Competing Interests: No competing interests were disclosed. Reviewer Expertise: In vivo studies, inflammation, hepatology, endocrinology, cancer, oxidative stress, kidney, ulcerative colitis I confirm that I have read this submission and believe that I have an appropriate level of expertise to confirm that it is of an acceptable scientific standard. Close READ LESS CITE CITE HOW TO CITE THIS REPORT Darwish S. Reviewer Report For: Enhanced human periodontal ligament stem cell viability and osteogenic differentiation on two implant materials: An experimental in vitro study [version 2; peer review: 2 approved] . F1000Research 2023, 12 :447 ( https://doi.org/10.5256/f1000research.154099.r194730 ) The direct URL for this report is: https://f1000research.com/articles/12-447/v2#referee-response-194730 NOTE: it is important to ensure the information in square brackets after the title is included in all citations of this article. COPY CITATION DETAILS Report a concern Respond or Comment COMMENT ON THIS REPORT Version 1 VERSION 1 PUBLISHED 26 Apr 2023 Views 0 Cite How to cite this report: El Refai S. Reviewer Report For: Enhanced human periodontal ligament stem cell viability and osteogenic differentiation on two implant materials: An experimental in vitro study [version 2; peer review: 2 approved] . F1000Research 2023, 12 :447 ( https://doi.org/10.5256/f1000research.142252.r171165 ) The direct URL for this report is: https://f1000research.com/articles/12-447/v1#referee-response-171165 NOTE: it is important to ensure the information in square brackets after the title is included in this citation. Close Copy Citation Details Reviewer Report 25 Jul 2023 Sahar El Refai , Princess Nourah bint Abdulrahman University, Riyadh, Riyadh Province, Saudi Arabia Approved with Reservations VIEWS 0 https://doi.org/10.5256/f1000research.142252.r171165 This article is clearly written covering all the details of the material and methods, I acknowledged the standardization in the used technique and the clarity of the results which mostly verified the aim despite figure 2b, (the most right image ... Continue reading READ ALL This article is clearly written covering all the details of the material and methods, I acknowledged the standardization in the used technique and the clarity of the results which mostly verified the aim despite figure 2b, (the most right image should have been titled chondrogenesis instead of the 2 images having the osteogenesis title. In regards to the discussion, it was sequential and consistent, however would have preferred if the author highlighted the kind of cell/material binding being regulated by cell type and implant material. Finally, the addition of opposing opinions to the results addressed in the present research would have enriched the current study outcomes and elucidated all reported literature views. Is the work clearly and accurately presented and does it cite the current literature? Yes Is the study design appropriate and is the work technically sound? Yes Are sufficient details of methods and analysis provided to allow replication by others? Yes If applicable, is the statistical analysis and its interpretation appropriate? Yes Are all the source data underlying the results available to ensure full reproducibility? Yes Are the conclusions drawn adequately supported by the results? Yes References 1. Jung S, Bohner L, Hanisch M, Kleinheinz J, et al.: Influence of Implant Material and Surface on Mode and Strength of Cell/Matrix Attachment of Human Adipose Derived Stromal Cell. Int J Mol Sci . 2020; 21 (11). PubMed Abstract | Publisher Full Text Competing Interests: No competing interests were disclosed. Reviewer Expertise: Oral cancer I confirm that I have read this submission and believe that I have an appropriate level of expertise to confirm that it is of an acceptable scientific standard, however I have significant reservations, as outlined above. Close READ LESS CITE CITE HOW TO CITE THIS REPORT El Refai S. Reviewer Report For: Enhanced human periodontal ligament stem cell viability and osteogenic differentiation on two implant materials: An experimental in vitro study [version 2; peer review: 2 approved] . F1000Research 2023, 12 :447 ( https://doi.org/10.5256/f1000research.142252.r171165 ) The direct URL for this report is: https://f1000research.com/articles/12-447/v1#referee-response-171165 NOTE: it is important to ensure the information in square brackets after the title is included in all citations of this article. COPY CITATION DETAILS Report a concern Author Response 08 Aug 2023 Sara El Shafei , Removable Prosthodontics, Faculty of Dentistry, The British University in Egypt, Cairo, Egypt 08 Aug 2023 Author Response Thank you for your efforts in reviewing our manuscript. All the comments have been addressed. 1-despite figure 2b, (the most right image should have been titled chondrogenesis instead of the ... Continue reading Thank you for your efforts in reviewing our manuscript. All the comments have been addressed. 1-despite figure 2b, (the most right image should have been titled chondrogenesis instead of the 2 images having the osteogenesis title -Done and corrected. 2- preferred if the author highlighted the kind of cell/material binding being regulated by cell type and implant material Added in the manuscript, discussion section and highlighted. 3- the addition of opposing opinions to the results addressed in the present research would have enriched the current study outcomes and elucidated all reported literature views Added in the manuscript, discussion section and highlighted. Thank you for your efforts in reviewing our manuscript. All the comments have been addressed. 1-despite figure 2b, (the most right image should have been titled chondrogenesis instead of the 2 images having the osteogenesis title -Done and corrected. 2- preferred if the author highlighted the kind of cell/material binding being regulated by cell type and implant material Added in the manuscript, discussion section and highlighted. 3- the addition of opposing opinions to the results addressed in the present research would have enriched the current study outcomes and elucidated all reported literature views Added in the manuscript, discussion section and highlighted. Competing Interests: No competing interests Close Report a concern Respond or Comment COMMENTS ON THIS REPORT Author Response 08 Aug 2023 Sara El Shafei , Removable Prosthodontics, Faculty of Dentistry, The British University in Egypt, Cairo, Egypt 08 Aug 2023 Author Response Thank you for your efforts in reviewing our manuscript. All the comments have been addressed. 1-despite figure 2b, (the most right image should have been titled chondrogenesis instead of the ... Continue reading Thank you for your efforts in reviewing our manuscript. All the comments have been addressed. 1-despite figure 2b, (the most right image should have been titled chondrogenesis instead of the 2 images having the osteogenesis title -Done and corrected. 2- preferred if the author highlighted the kind of cell/material binding being regulated by cell type and implant material Added in the manuscript, discussion section and highlighted. 3- the addition of opposing opinions to the results addressed in the present research would have enriched the current study outcomes and elucidated all reported literature views Added in the manuscript, discussion section and highlighted. Thank you for your efforts in reviewing our manuscript. All the comments have been addressed. 1-despite figure 2b, (the most right image should have been titled chondrogenesis instead of the 2 images having the osteogenesis title -Done and corrected. 2- preferred if the author highlighted the kind of cell/material binding being regulated by cell type and implant material Added in the manuscript, discussion section and highlighted. 3- the addition of opposing opinions to the results addressed in the present research would have enriched the current study outcomes and elucidated all reported literature views Added in the manuscript, discussion section and highlighted. Competing Interests: No competing interests Close Report a concern COMMENT ON THIS REPORT Views 0 Cite How to cite this report: Darwish S. Reviewer Report For: Enhanced human periodontal ligament stem cell viability and osteogenic differentiation on two implant materials: An experimental in vitro study [version 2; peer review: 2 approved] . F1000Research 2023, 12 :447 ( https://doi.org/10.5256/f1000research.142252.r171166 ) The direct URL for this report is: https://f1000research.com/articles/12-447/v1#referee-response-171166 NOTE: it is important to ensure the information in square brackets after the title is included in this citation. Close Copy Citation Details Reviewer Report 14 Jul 2023 Samar Darwish , Pharmacology and Toxicology Department, Badr University in Cairo, Badr City, Cairo Governorate, Egypt Approved with Reservations VIEWS 0 https://doi.org/10.5256/f1000research.142252.r171166 In this manuscript, the scientists examined and tested how human periodontal ligament stem cells responded to polyetheretherketone (PEEK) discs with various surface topographies and yttria-stabilized tetragonal zirconia polycrystals. The study clarifies how forming micro-topographies on the surface of PEEK implants ... Continue reading READ ALL In this manuscript, the scientists examined and tested how human periodontal ligament stem cells responded to polyetheretherketone (PEEK) discs with various surface topographies and yttria-stabilized tetragonal zirconia polycrystals. The study clarifies how forming micro-topographies on the surface of PEEK implants improves stem cell adhesion, vitality, and osteogenic differentiation. However, there are few minor revisions that I advise the authors to take them in concern : Abstract section : It is preferred not to write the keywords in their abbreviated forms, such as PEEK and PLDSCs. Introduction section : It is observed that all reference numbers are written after the full stops of the sentences in the whole manuscript. It is preferred to write the reference numbers before not after. 3 rd paragraph, 2 nd Line: What is meant by “ Given their color... ”, please rephrase the sentence to give a clear idea. 4 th paragraph, 1 st line: “ PEEK is a synthetic polymer which shows potential...", there is a missing word I think, please revise. 4 th paragraph, 3 rd line: the authors mentioned that PEEK's bioactivity and osseointegration are also debatable, please write in more detail what you mean by debatable to clarify your idea. Methods section : PEEK abbreviation refers to which word? Polyetheretherketone or polyether-ether-ketone? Please check. In 'Disc preparation section': please write more details about how yttria stabilized tetragonal zirconia polycrystals were prepared or utilized. In 'Periodontal ligament (PDL) cells isolation and culture': the authors mentioned that " The extracted teeth were kept in Dulbecco’s modified Eagle’s medium (DMEM; Sigma, St. Louis, MO), which received antibiotic doses (300 U/ml penicillin and 300 mg/ml streptomycin; Sigma). " Please mention how many doses were given and the dosing frequency. In 'Statistical analysis': " (RRID:SCR_002798) " is repeated, mention only once. Results section : Figure 2a: if possible, it is preferred to increase the resolution to make the numbers more clear. Figure 2b: put labels on the 6 different pictures to identify them clearly. Figure 3: again the resolution needs to be increased. Figure 5: there are two rows of pictures, (a, b , c….). What is the difference? Is it the magnification? Please clarify and label them because it is only mentioned that " (Mag. 40x) ". In the comment on 'Mineralization assay calcium deposition' results, mention the percentage or folds difference calculated between the groups as you can. Discussion : the discussion was well described and covered the study points in detail In the last paragraph: what did the authors mean by this sentence, " A strategy that has the potential to improve PLDSC attachment is the development of micro-topographies on the PEEK surface, viability and osteogenic differentiation "? Please rephrase to clarify your idea. Reference section : Some references are preferred to be renewed such as: no 8, 29, 31, 37 Is the work clearly and accurately presented and does it cite the current literature? Yes Is the study design appropriate and is the work technically sound? Yes Are sufficient details of methods and analysis provided to allow replication by others? Yes If applicable, is the statistical analysis and its interpretation appropriate? Yes Are all the source data underlying the results available to ensure full reproducibility? Yes Are the conclusions drawn adequately supported by the results? Yes Competing Interests: No competing interests were disclosed. Reviewer Expertise: In vivo studies, inflammation, hepatology, endocrinology, cancer, oxidative stress I confirm that I have read this submission and believe that I have an appropriate level of expertise to confirm that it is of an acceptable scientific standard, however I have significant reservations, as outlined above. Close READ LESS CITE CITE HOW TO CITE THIS REPORT Darwish S. Reviewer Report For: Enhanced human periodontal ligament stem cell viability and osteogenic differentiation on two implant materials: An experimental in vitro study [version 2; peer review: 2 approved] . F1000Research 2023, 12 :447 ( https://doi.org/10.5256/f1000research.142252.r171166 ) The direct URL for this report is: https://f1000research.com/articles/12-447/v1#referee-response-171166 NOTE: it is important to ensure the information in square brackets after the title is included in all citations of this article. COPY CITATION DETAILS Report a concern Author Response 08 Aug 2023 Sara El Shafei , Removable Prosthodontics, Faculty of Dentistry, The British University in Egypt, Cairo, Egypt 08 Aug 2023 Author Response Thank you for your efforts in reviewing our manuscript. All the comments have been addressed. Reviewer 1: Abstract section : It is preferred not to write the keywords ... Continue reading Thank you for your efforts in reviewing our manuscript. All the comments have been addressed. Reviewer 1: Abstract section : It is preferred not to write the keywords in their abbreviated forms, such as PEEK and PLDSCs. -Corrected PEEK in abstract and keywords highlighted -Corrected hPLDSCs in keywords and highlighted Introduction section : 1-It is observed that all reference numbers are written after the full stops of the sentences in the whole manuscript. It is preferred to write the reference numbers before not after. -The followed reference numbering style in text is according to the journal submission regulations 2-3 rd paragraph, 2 nd Line: What is meant by “ Given their color... ”, please rephrase the sentence to give a clear idea. -Rephrased and highlighted 3-4 th paragraph, 1 st line: “PEEK is a synthetic polymer which shows potential..." , there is a missing word I think, please revise -Added missing word and highlighted 4-4 th paragraph, 3 rd line: the authors mentioned that PEEK's bioactivity and osseointegration are also debatable, please write in more detail what you mean by debatable to clarify your idea -Clarified the sentence and highlighted Methods section : 1-PEEK abbreviation refers to which word? Polyetheretherketone or polyether-ether-ketone? Please check. -Clarified the word and highlighted 2-In 'Disc preparation section': please write more details about how yttria stabilized tetragonal zirconia polycrystals were prepared or utilized -Rephrased to clarify the idea and highlighted 3- In 'Periodontal ligament (PDL) cells isolation and culture': the authors mentioned that " The extracted teeth were kept in Dulbecco’s modified Eagle’s medium (DMEM; Sigma, St. Louis, MO), which received antibiotic doses (300 U/ml penicillin and 300 mg/ml streptomycin; Sigma). " Please mention how many doses were given and the dosing frequency. Antibiotic-antimycotic drug is added once in the culture medium bottle (500 ml) which is used in the whole study; changing medium for the cells is maximum 2-5 ml / well or flask twice per week. 4- In 'Statistical analysis': " (RRID:SCR_002798) " is repeated, mention only once. -Removed and mentioned only once. Results section : 1-Figure 2a: if possible, it is preferred to increase the resolution to make the numbers more clear. 2-Figure 2b: put labels on the 6 different pictures to identify them clearly -Figure 2a. Edited and added in the manuscript -Figure 2b. Edited and added in the manuscript 3-Figure 3: again the resolution needs to be increased -Edited and added in the manuscript Figure 5: there are two rows of pictures, (a, b , c….). What is the difference? Is it the magnification? Please clarify and label them because it is only mentioned that " (Mag. 40x) ". - Done and labeled In the comment on 'Mineralization assay calcium deposition' results, mention the percentage or folds difference calculated between the groups as you can - Mean values of absorbance of the study group are added in the manuscript and highlighted. Discussion : 1-In the last paragraph: what did the authors mean by this sentence, " A strategy that has the potential to improve PLDSC attachment is the development of micro-topographies on the PEEK surface, viability and osteogenic differentiation "? Please rephrase to clarify your idea. - Rephrased and highlighted Thank you for your efforts in reviewing our manuscript. All the comments have been addressed. Reviewer 1: Abstract section : It is preferred not to write the keywords in their abbreviated forms, such as PEEK and PLDSCs. -Corrected PEEK in abstract and keywords highlighted -Corrected hPLDSCs in keywords and highlighted Introduction section : 1-It is observed that all reference numbers are written after the full stops of the sentences in the whole manuscript. It is preferred to write the reference numbers before not after. -The followed reference numbering style in text is according to the journal submission regulations 2-3 rd paragraph, 2 nd Line: What is meant by “ Given their color... ”, please rephrase the sentence to give a clear idea. -Rephrased and highlighted 3-4 th paragraph, 1 st line: “PEEK is a synthetic polymer which shows potential..." , there is a missing word I think, please revise -Added missing word and highlighted 4-4 th paragraph, 3 rd line: the authors mentioned that PEEK's bioactivity and osseointegration are also debatable, please write in more detail what you mean by debatable to clarify your idea -Clarified the sentence and highlighted Methods section : 1-PEEK abbreviation refers to which word? Polyetheretherketone or polyether-ether-ketone? Please check. -Clarified the word and highlighted 2-In 'Disc preparation section': please write more details about how yttria stabilized tetragonal zirconia polycrystals were prepared or utilized -Rephrased to clarify the idea and highlighted 3- In 'Periodontal ligament (PDL) cells isolation and culture': the authors mentioned that " The extracted teeth were kept in Dulbecco’s modified Eagle’s medium (DMEM; Sigma, St. Louis, MO), which received antibiotic doses (300 U/ml penicillin and 300 mg/ml streptomycin; Sigma). " Please mention how many doses were given and the dosing frequency. Antibiotic-antimycotic drug is added once in the culture medium bottle (500 ml) which is used in the whole study; changing medium for the cells is maximum 2-5 ml / well or flask twice per week. 4- In 'Statistical analysis': " (RRID:SCR_002798) " is repeated, mention only once. -Removed and mentioned only once. Results section : 1-Figure 2a: if possible, it is preferred to increase the resolution to make the numbers more clear. 2-Figure 2b: put labels on the 6 different pictures to identify them clearly -Figure 2a. Edited and added in the manuscript -Figure 2b. Edited and added in the manuscript 3-Figure 3: again the resolution needs to be increased -Edited and added in the manuscript Figure 5: there are two rows of pictures, (a, b , c….). What is the difference? Is it the magnification? Please clarify and label them because it is only mentioned that " (Mag. 40x) ". - Done and labeled In the comment on 'Mineralization assay calcium deposition' results, mention the percentage or folds difference calculated between the groups as you can - Mean values of absorbance of the study group are added in the manuscript and highlighted. Discussion : 1-In the last paragraph: what did the authors mean by this sentence, " A strategy that has the potential to improve PLDSC attachment is the development of micro-topographies on the PEEK surface, viability and osteogenic differentiation "? Please rephrase to clarify your idea. - Rephrased and highlighted Competing Interests: No competing interests Close Report a concern Respond or Comment COMMENTS ON THIS REPORT Author Response 08 Aug 2023 Sara El Shafei , Removable Prosthodontics, Faculty of Dentistry, The British University in Egypt, Cairo, Egypt 08 Aug 2023 Author Response Thank you for your efforts in reviewing our manuscript. All the comments have been addressed. Reviewer 1: Abstract section : It is preferred not to write the keywords ... Continue reading Thank you for your efforts in reviewing our manuscript. All the comments have been addressed. Reviewer 1: Abstract section : It is preferred not to write the keywords in their abbreviated forms, such as PEEK and PLDSCs. -Corrected PEEK in abstract and keywords highlighted -Corrected hPLDSCs in keywords and highlighted Introduction section : 1-It is observed that all reference numbers are written after the full stops of the sentences in the whole manuscript. It is preferred to write the reference numbers before not after. -The followed reference numbering style in text is according to the journal submission regulations 2-3 rd paragraph, 2 nd Line: What is meant by “ Given their color... ”, please rephrase the sentence to give a clear idea. -Rephrased and highlighted 3-4 th paragraph, 1 st line: “PEEK is a synthetic polymer which shows potential..." , there is a missing word I think, please revise -Added missing word and highlighted 4-4 th paragraph, 3 rd line: the authors mentioned that PEEK's bioactivity and osseointegration are also debatable, please write in more detail what you mean by debatable to clarify your idea -Clarified the sentence and highlighted Methods section : 1-PEEK abbreviation refers to which word? Polyetheretherketone or polyether-ether-ketone? Please check. -Clarified the word and highlighted 2-In 'Disc preparation section': please write more details about how yttria stabilized tetragonal zirconia polycrystals were prepared or utilized -Rephrased to clarify the idea and highlighted 3- In 'Periodontal ligament (PDL) cells isolation and culture': the authors mentioned that " The extracted teeth were kept in Dulbecco’s modified Eagle’s medium (DMEM; Sigma, St. Louis, MO), which received antibiotic doses (300 U/ml penicillin and 300 mg/ml streptomycin; Sigma). " Please mention how many doses were given and the dosing frequency. Antibiotic-antimycotic drug is added once in the culture medium bottle (500 ml) which is used in the whole study; changing medium for the cells is maximum 2-5 ml / well or flask twice per week. 4- In 'Statistical analysis': " (RRID:SCR_002798) " is repeated, mention only once. -Removed and mentioned only once. Results section : 1-Figure 2a: if possible, it is preferred to increase the resolution to make the numbers more clear. 2-Figure 2b: put labels on the 6 different pictures to identify them clearly -Figure 2a. Edited and added in the manuscript -Figure 2b. Edited and added in the manuscript 3-Figure 3: again the resolution needs to be increased -Edited and added in the manuscript Figure 5: there are two rows of pictures, (a, b , c….). What is the difference? Is it the magnification? Please clarify and label them because it is only mentioned that " (Mag. 40x) ". - Done and labeled In the comment on 'Mineralization assay calcium deposition' results, mention the percentage or folds difference calculated between the groups as you can - Mean values of absorbance of the study group are added in the manuscript and highlighted. Discussion : 1-In the last paragraph: what did the authors mean by this sentence, " A strategy that has the potential to improve PLDSC attachment is the development of micro-topographies on the PEEK surface, viability and osteogenic differentiation "? Please rephrase to clarify your idea. - Rephrased and highlighted Thank you for your efforts in reviewing our manuscript. All the comments have been addressed. Reviewer 1: Abstract section : It is preferred not to write the keywords in their abbreviated forms, such as PEEK and PLDSCs. -Corrected PEEK in abstract and keywords highlighted -Corrected hPLDSCs in keywords and highlighted Introduction section : 1-It is observed that all reference numbers are written after the full stops of the sentences in the whole manuscript. It is preferred to write the reference numbers before not after. -The followed reference numbering style in text is according to the journal submission regulations 2-3 rd paragraph, 2 nd Line: What is meant by “ Given their color... ”, please rephrase the sentence to give a clear idea. -Rephrased and highlighted 3-4 th paragraph, 1 st line: “PEEK is a synthetic polymer which shows potential..." , there is a missing word I think, please revise -Added missing word and highlighted 4-4 th paragraph, 3 rd line: the authors mentioned that PEEK's bioactivity and osseointegration are also debatable, please write in more detail what you mean by debatable to clarify your idea -Clarified the sentence and highlighted Methods section : 1-PEEK abbreviation refers to which word? Polyetheretherketone or polyether-ether-ketone? Please check. -Clarified the word and highlighted 2-In 'Disc preparation section': please write more details about how yttria stabilized tetragonal zirconia polycrystals were prepared or utilized -Rephrased to clarify the idea and highlighted 3- In 'Periodontal ligament (PDL) cells isolation and culture': the authors mentioned that " The extracted teeth were kept in Dulbecco’s modified Eagle’s medium (DMEM; Sigma, St. Louis, MO), which received antibiotic doses (300 U/ml penicillin and 300 mg/ml streptomycin; Sigma). " Please mention how many doses were given and the dosing frequency. Antibiotic-antimycotic drug is added once in the culture medium bottle (500 ml) which is used in the whole study; changing medium for the cells is maximum 2-5 ml / well or flask twice per week. 4- In 'Statistical analysis': " (RRID:SCR_002798) " is repeated, mention only once. -Removed and mentioned only once. Results section : 1-Figure 2a: if possible, it is preferred to increase the resolution to make the numbers more clear. 2-Figure 2b: put labels on the 6 different pictures to identify them clearly -Figure 2a. Edited and added in the manuscript -Figure 2b. Edited and added in the manuscript 3-Figure 3: again the resolution needs to be increased -Edited and added in the manuscript Figure 5: there are two rows of pictures, (a, b , c….). What is the difference? Is it the magnification? Please clarify and label them because it is only mentioned that " (Mag. 40x) ". - Done and labeled In the comment on 'Mineralization assay calcium deposition' results, mention the percentage or folds difference calculated between the groups as you can - Mean values of absorbance of the study group are added in the manuscript and highlighted. Discussion : 1-In the last paragraph: what did the authors mean by this sentence, " A strategy that has the potential to improve PLDSC attachment is the development of micro-topographies on the PEEK surface, viability and osteogenic differentiation "? Please rephrase to clarify your idea. - Rephrased and highlighted Competing Interests: No competing interests Close Report a concern COMMENT ON THIS REPORT Comments on this article Comments (0) Version 2 VERSION 2 PUBLISHED 26 Apr 2023 ADD YOUR COMMENT Comment keyboard_arrow_left keyboard_arrow_right Open Peer Review Reviewer Status info_outline Alongside their report, reviewers assign a status to the article: Approved The paper is scientifically sound in its current form and only minor, if any, improvements are suggested Approved with reservations A number of small changes, sometimes more significant revisions are required to address specific details and improve the papers academic merit. Not approved Fundamental flaws in the paper seriously undermine the findings and conclusions Reviewer Reports Invited Reviewers 1 2 Version 2 (revision) 08 Aug 23 read read Version 1 26 Apr 23 read read Samar Darwish , Badr University in Cairo, Badr City, Egypt Sahar El Refai , Princess Nourah bint Abdulrahman University, Riyadh, Saudi Arabia Comments on this article All Comments (0) Add a comment Sign up for content alerts Sign Up You are now signed up to receive this alert Browse by related subjects keyboard_arrow_left Back to all reports Reviewer Report 0 Views copyright © 2023 El Refai S. This is an open access peer review report distributed under the terms of the Creative Commons Attribution License , which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited. 21 Aug 2023 | for Version 2 Sahar El Refai , Princess Nourah bint Abdulrahman University, Riyadh, Riyadh Province, Saudi Arabia 0 Views copyright © 2023 El Refai S. This is an open access peer review report distributed under the terms of the Creative Commons Attribution License , which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited. format_quote Cite this report speaker_notes Responses (0) Approved info_outline Alongside their report, reviewers assign a status to the article: Approved The paper is scientifically sound in its current form and only minor, if any, improvements are suggested Approved with reservations A number of small changes, sometimes more significant revisions are required to address specific details and improve the papers academic merit. Not approved Fundamental flaws in the paper seriously undermine the findings and conclusions I have no further comments than the ones the authors addressed Competing Interests No competing interests were disclosed. I confirm that I have read this submission and believe that I have an appropriate level of expertise to confirm that it is of an acceptable scientific standard. reply Respond to this report Responses (0) El Refai S. Peer Review Report For: Enhanced human periodontal ligament stem cell viability and osteogenic differentiation on two implant materials: An experimental in vitro study [version 2; peer review: 2 approved] . F1000Research 2023, 12 :447 ( https://doi.org/10.5256/f1000research.154099.r194731) NOTE: it is important to ensure the information in square brackets after the title is included in this citation. The direct URL for this report is: https://f1000research.com/articles/12-447/v2#referee-response-194731 keyboard_arrow_left Back to all reports Reviewer Report 0 Views copyright © 2023 Darwish S. This is an open access peer review report distributed under the terms of the Creative Commons Attribution License , which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited. 10 Aug 2023 | for Version 2 Samar Darwish , Pharmacology and Toxicology Department, Badr University in Cairo, Badr City, Cairo Governorate, Egypt 0 Views copyright © 2023 Darwish S. This is an open access peer review report distributed under the terms of the Creative Commons Attribution License , which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited. format_quote Cite this report speaker_notes Responses (0) Approved info_outline Alongside their report, reviewers assign a status to the article: Approved The paper is scientifically sound in its current form and only minor, if any, improvements are suggested Approved with reservations A number of small changes, sometimes more significant revisions are required to address specific details and improve the papers academic merit. Not approved Fundamental flaws in the paper seriously undermine the findings and conclusions No further comments to make. Competing Interests No competing interests were disclosed. Reviewer Expertise In vivo studies, inflammation, hepatology, endocrinology, cancer, oxidative stress, kidney, ulcerative colitis I confirm that I have read this submission and believe that I have an appropriate level of expertise to confirm that it is of an acceptable scientific standard. reply Respond to this report Responses (0) Darwish S. Peer Review Report For: Enhanced human periodontal ligament stem cell viability and osteogenic differentiation on two implant materials: An experimental in vitro study [version 2; peer review: 2 approved] . F1000Research 2023, 12 :447 ( https://doi.org/10.5256/f1000research.154099.r194730) NOTE: it is important to ensure the information in square brackets after the title is included in this citation. The direct URL for this report is: https://f1000research.com/articles/12-447/v2#referee-response-194730 keyboard_arrow_left Back to all reports Reviewer Report 0 Views copyright © 2023 El Refai S. This is an open access peer review report distributed under the terms of the Creative Commons Attribution License , which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited. 25 Jul 2023 | for Version 1 Sahar El Refai , Princess Nourah bint Abdulrahman University, Riyadh, Riyadh Province, Saudi Arabia 0 Views copyright © 2023 El Refai S. This is an open access peer review report distributed under the terms of the Creative Commons Attribution License , which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited. format_quote Cite this report speaker_notes Responses (1) Approved With Reservations info_outline Alongside their report, reviewers assign a status to the article: Approved The paper is scientifically sound in its current form and only minor, if any, improvements are suggested Approved with reservations A number of small changes, sometimes more significant revisions are required to address specific details and improve the papers academic merit. Not approved Fundamental flaws in the paper seriously undermine the findings and conclusions This article is clearly written covering all the details of the material and methods, I acknowledged the standardization in the used technique and the clarity of the results which mostly verified the aim despite figure 2b, (the most right image should have been titled chondrogenesis instead of the 2 images having the osteogenesis title. In regards to the discussion, it was sequential and consistent, however would have preferred if the author highlighted the kind of cell/material binding being regulated by cell type and implant material. Finally, the addition of opposing opinions to the results addressed in the present research would have enriched the current study outcomes and elucidated all reported literature views. Is the work clearly and accurately presented and does it cite the current literature? Yes Is the study design appropriate and is the work technically sound? Yes Are sufficient details of methods and analysis provided to allow replication by others? Yes If applicable, is the statistical analysis and its interpretation appropriate? Yes Are all the source data underlying the results available to ensure full reproducibility? Yes Are the conclusions drawn adequately supported by the results? Yes References 1. Jung S, Bohner L, Hanisch M, Kleinheinz J, et al.: Influence of Implant Material and Surface on Mode and Strength of Cell/Matrix Attachment of Human Adipose Derived Stromal Cell. Int J Mol Sci . 2020; 21 (11). PubMed Abstract | Publisher Full Text Competing Interests No competing interests were disclosed. Reviewer Expertise Oral cancer I confirm that I have read this submission and believe that I have an appropriate level of expertise to confirm that it is of an acceptable scientific standard, however I have significant reservations, as outlined above. reply Respond to this report Responses (1) Author Response 08 Aug 2023 Sara El Shafei, Removable Prosthodontics, Faculty of Dentistry, The British University in Egypt, Cairo, Egypt Thank you for your efforts in reviewing our manuscript. All the comments have been addressed. 1-despite figure 2b, (the most right image should have been titled chondrogenesis instead of the 2 images having the osteogenesis title -Done and corrected. 2- preferred if the author highlighted the kind of cell/material binding being regulated by cell type and implant material Added in the manuscript, discussion section and highlighted. 3- the addition of opposing opinions to the results addressed in the present research would have enriched the current study outcomes and elucidated all reported literature views Added in the manuscript, discussion section and highlighted. View more View less Competing Interests No competing interests reply Respond Report a concern El Refai S. Peer Review Report For: Enhanced human periodontal ligament stem cell viability and osteogenic differentiation on two implant materials: An experimental in vitro study [version 2; peer review: 2 approved] . F1000Research 2023, 12 :447 ( https://doi.org/10.5256/f1000research.142252.r171165) NOTE: it is important to ensure the information in square brackets after the title is included in this citation. The direct URL for this report is: https://f1000research.com/articles/12-447/v1#referee-response-171165 keyboard_arrow_left Back to all reports Reviewer Report 0 Views copyright © 2023 Darwish S. This is an open access peer review report distributed under the terms of the Creative Commons Attribution License , which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited. 14 Jul 2023 | for Version 1 Samar Darwish , Pharmacology and Toxicology Department, Badr University in Cairo, Badr City, Cairo Governorate, Egypt 0 Views copyright © 2023 Darwish S. This is an open access peer review report distributed under the terms of the Creative Commons Attribution License , which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited. format_quote Cite this report speaker_notes Responses (1) Approved With Reservations info_outline Alongside their report, reviewers assign a status to the article: Approved The paper is scientifically sound in its current form and only minor, if any, improvements are suggested Approved with reservations A number of small changes, sometimes more significant revisions are required to address specific details and improve the papers academic merit. Not approved Fundamental flaws in the paper seriously undermine the findings and conclusions In this manuscript, the scientists examined and tested how human periodontal ligament stem cells responded to polyetheretherketone (PEEK) discs with various surface topographies and yttria-stabilized tetragonal zirconia polycrystals. The study clarifies how forming micro-topographies on the surface of PEEK implants improves stem cell adhesion, vitality, and osteogenic differentiation. However, there are few minor revisions that I advise the authors to take them in concern : Abstract section : It is preferred not to write the keywords in their abbreviated forms, such as PEEK and PLDSCs. Introduction section : It is observed that all reference numbers are written after the full stops of the sentences in the whole manuscript. It is preferred to write the reference numbers before not after. 3 rd paragraph, 2 nd Line: What is meant by “ Given their color... ”, please rephrase the sentence to give a clear idea. 4 th paragraph, 1 st line: “ PEEK is a synthetic polymer which shows potential...", there is a missing word I think, please revise. 4 th paragraph, 3 rd line: the authors mentioned that PEEK's bioactivity and osseointegration are also debatable, please write in more detail what you mean by debatable to clarify your idea. Methods section : PEEK abbreviation refers to which word? Polyetheretherketone or polyether-ether-ketone? Please check. In 'Disc preparation section': please write more details about how yttria stabilized tetragonal zirconia polycrystals were prepared or utilized. In 'Periodontal ligament (PDL) cells isolation and culture': the authors mentioned that " The extracted teeth were kept in Dulbecco’s modified Eagle’s medium (DMEM; Sigma, St. Louis, MO), which received antibiotic doses (300 U/ml penicillin and 300 mg/ml streptomycin; Sigma). " Please mention how many doses were given and the dosing frequency. In 'Statistical analysis': " (RRID:SCR_002798) " is repeated, mention only once. Results section : Figure 2a: if possible, it is preferred to increase the resolution to make the numbers more clear. Figure 2b: put labels on the 6 different pictures to identify them clearly. Figure 3: again the resolution needs to be increased. Figure 5: there are two rows of pictures, (a, b , c….). What is the difference? Is it the magnification? Please clarify and label them because it is only mentioned that " (Mag. 40x) ". In the comment on 'Mineralization assay calcium deposition' results, mention the percentage or folds difference calculated between the groups as you can. Discussion : the discussion was well described and covered the study points in detail In the last paragraph: what did the authors mean by this sentence, " A strategy that has the potential to improve PLDSC attachment is the development of micro-topographies on the PEEK surface, viability and osteogenic differentiation "? Please rephrase to clarify your idea. Reference section : Some references are preferred to be renewed such as: no 8, 29, 31, 37 Is the work clearly and accurately presented and does it cite the current literature? Yes Is the study design appropriate and is the work technically sound? Yes Are sufficient details of methods and analysis provided to allow replication by others? Yes If applicable, is the statistical analysis and its interpretation appropriate? Yes Are all the source data underlying the results available to ensure full reproducibility? Yes Are the conclusions drawn adequately supported by the results? Yes Competing Interests No competing interests were disclosed. Reviewer Expertise In vivo studies, inflammation, hepatology, endocrinology, cancer, oxidative stress I confirm that I have read this submission and believe that I have an appropriate level of expertise to confirm that it is of an acceptable scientific standard, however I have significant reservations, as outlined above. reply Respond to this report Responses (1) Author Response 08 Aug 2023 Sara El Shafei, Removable Prosthodontics, Faculty of Dentistry, The British University in Egypt, Cairo, Egypt Thank you for your efforts in reviewing our manuscript. All the comments have been addressed. Reviewer 1: Abstract section : It is preferred not to write the keywords in their abbreviated forms, such as PEEK and PLDSCs. -Corrected PEEK in abstract and keywords highlighted -Corrected hPLDSCs in keywords and highlighted Introduction section : 1-It is observed that all reference numbers are written after the full stops of the sentences in the whole manuscript. It is preferred to write the reference numbers before not after. -The followed reference numbering style in text is according to the journal submission regulations 2-3 rd paragraph, 2 nd Line: What is meant by “ Given their color... ”, please rephrase the sentence to give a clear idea. -Rephrased and highlighted 3-4 th paragraph, 1 st line: “PEEK is a synthetic polymer which shows potential..." , there is a missing word I think, please revise -Added missing word and highlighted 4-4 th paragraph, 3 rd line: the authors mentioned that PEEK's bioactivity and osseointegration are also debatable, please write in more detail what you mean by debatable to clarify your idea -Clarified the sentence and highlighted Methods section : 1-PEEK abbreviation refers to which word? Polyetheretherketone or polyether-ether-ketone? Please check. -Clarified the word and highlighted 2-In 'Disc preparation section': please write more details about how yttria stabilized tetragonal zirconia polycrystals were prepared or utilized -Rephrased to clarify the idea and highlighted 3- In 'Periodontal ligament (PDL) cells isolation and culture': the authors mentioned that " The extracted teeth were kept in Dulbecco’s modified Eagle’s medium (DMEM; Sigma, St. Louis, MO), which received antibiotic doses (300 U/ml penicillin and 300 mg/ml streptomycin; Sigma). " Please mention how many doses were given and the dosing frequency. Antibiotic-antimycotic drug is added once in the culture medium bottle (500 ml) which is used in the whole study; changing medium for the cells is maximum 2-5 ml / well or flask twice per week. 4- In 'Statistical analysis': " (RRID:SCR_002798) " is repeated, mention only once. -Removed and mentioned only once. Results section : 1-Figure 2a: if possible, it is preferred to increase the resolution to make the numbers more clear. 2-Figure 2b: put labels on the 6 different pictures to identify them clearly -Figure 2a. Edited and added in the manuscript -Figure 2b. Edited and added in the manuscript 3-Figure 3: again the resolution needs to be increased -Edited and added in the manuscript Figure 5: there are two rows of pictures, (a, b , c….). What is the difference? Is it the magnification? Please clarify and label them because it is only mentioned that " (Mag. 40x) ". - Done and labeled In the comment on 'Mineralization assay calcium deposition' results, mention the percentage or folds difference calculated between the groups as you can - Mean values of absorbance of the study group are added in the manuscript and highlighted. Discussion : 1-In the last paragraph: what did the authors mean by this sentence, " A strategy that has the potential to improve PLDSC attachment is the development of micro-topographies on the PEEK surface, viability and osteogenic differentiation "? Please rephrase to clarify your idea. - Rephrased and highlighted View more View less Competing Interests No competing interests reply Respond Report a concern Darwish S. Peer Review Report For: Enhanced human periodontal ligament stem cell viability and osteogenic differentiation on two implant materials: An experimental in vitro study [version 2; peer review: 2 approved] . F1000Research 2023, 12 :447 ( https://doi.org/10.5256/f1000research.142252.r171166) NOTE: it is important to ensure the information in square brackets after the title is included in this citation. The direct URL for this report is: https://f1000research.com/articles/12-447/v1#referee-response-171166 Alongside their report, reviewers assign a status to the article: Approved - the paper is scientifically sound in its current form and only minor, if any, improvements are suggested Approved with reservations - A number of small changes, sometimes more significant revisions are required to address specific details and improve the papers academic merit. Not approved - fundamental flaws in the paper seriously undermine the findings and conclusions Adjust parameters to alter display View on desktop for interactive features Includes Interactive Elements View on desktop for interactive features Competing Interests Policy Provide sufficient details of any financial or non-financial competing interests to enable users to assess whether your comments might lead a reasonable person to question your impartiality. Consider the following examples, but note that this is not an exhaustive list: Examples of 'Non-Financial Competing Interests' Within the past 4 years, you have held joint grants, published or collaborated with any of the authors of the selected paper. You have a close personal relationship (e.g. parent, spouse, sibling, or domestic partner) with any of the authors. You are a close professional associate of any of the authors (e.g. scientific mentor, recent student). You work at the same institute as any of the authors. You hope/expect to benefit (e.g. favour or employment) as a result of your submission. You are an Editor for the journal in which the article is published. Examples of 'Financial Competing Interests' You expect to receive, or in the past 4 years have received, any of the following from any commercial organisation that may gain financially from your submission: a salary, fees, funding, reimbursements. You expect to receive, or in the past 4 years have received, shared grant support or other funding with any of the authors. You hold, or are currently applying for, any patents or significant stocks/shares relating to the subject matter of the paper you are commenting on. Stay Updated Sign up for content alerts and receive a weekly or monthly email with all newly published articles Register with F1000Research Already registered? Sign in Not now, thanks close PLEASE NOTE If you are an AUTHOR of this article, please check that you signed in with the account associated with this article otherwise we cannot automatically identify your role as an author and your comment will be labelled as a “User Comment”. If you are a REVIEWER of this article, please check that you have signed in with the account associated with this article and then go to your account to submit your report, please do not post your review here. If you do not have access to your original account, please contact us . All commenters must hold a formal affiliation as per our Policies . The information that you give us will be displayed next to your comment. User comments must be in English, comprehensible and relevant to the article under discussion. We reserve the right to remove any comments that we consider to be inappropriate, offensive or otherwise in breach of the User Comment Terms and Conditions . Commenters must not use a comment for personal attacks. When criticisms of the article are based on unpublished data, the data should be made available. I accept the User Comment Terms and Conditions Please confirm that you accept the User Comment Terms and Conditions. Affiliation ✕ refresh Please enter your institution. Note: To add your institution or organisation, start typing the name and then select the correct name from the list. Where applicable, the name will appear in both the original language and in English. Do not paste in the name. If the name does not appear in the drop-down list, we will display the information you have entered. ✕ refresh Country/Region * USA UK Canada China France Germany Afghanistan Aland Islands Albania Algeria American Samoa Andorra Angola Anguilla Antarctica Antigua and Barbuda Argentina Armenia Aruba Australia Austria Azerbaijan Bahamas Bahrain Bangladesh Barbados Belarus Belgium Belize Benin Bermuda Bhutan Bolivia Bosnia and Herzegovina Botswana Bouvet Island Brazil British Indian Ocean Territory British Virgin Islands Brunei Bulgaria Burkina Faso Burundi Cambodia Cameroon Canada Cape Verde Cayman Islands Central African Republic Chad Chile China Christmas Island Cocos (Keeling) Islands Colombia Comoros Congo Cook Islands Costa Rica Cote d'Ivoire Croatia Cuba Cyprus Czech Republic Democratic Republic of the Congo Denmark Djibouti Dominica Dominican Republic Ecuador Egypt El Salvador Equatorial Guinea Eritrea Estonia Ethiopia Falkland Islands Faroe Islands Federated States of Micronesia Fiji Finland France French Guiana French Polynesia French Southern Territories Gabon Georgia Germany Ghana Gibraltar Greece Greenland Grenada Guadeloupe Guam Guatemala Guernsey Guinea Guinea-Bissau Guyana Haiti Heard Island and Mcdonald Islands Holy See (Vatican City State) Honduras Hong Kong Hungary Iceland India Indonesia Iran Iraq Ireland Israel Italy Jamaica Japan Jersey Jordan Kazakhstan Kenya Kiribati Kosovo (Serbia and Montenegro) Kuwait Kyrgyzstan Lao People's Democratic Republic Latvia Lebanon Lesotho Liberia Libya Liechtenstein Lithuania Luxembourg Macao Madagascar Malawi Malaysia Maldives Mali Malta Marshall Islands Martinique Mauritania Mauritius Mayotte Mexico Minor Outlying Islands of the United States Moldova Monaco Mongolia Montenegro Montserrat Morocco Mozambique Myanmar Namibia Nauru Nepal Netherlands Antilles New Caledonia New Zealand Nicaragua Niger Nigeria Niue Norfolk Island North Korea North Macedonia Northern Mariana Islands Norway Oman Pakistan Palau Palestinian Territory Panama Papua New Guinea Paraguay Peru Philippines Pitcairn Poland Portugal Puerto Rico Qatar Reunion Romania Russian Federation Rwanda Saint Helena Saint Kitts and Nevis Saint Lucia Saint Pierre and Miquelon Saint Vincent and the Grenadines Samoa San Marino Sao Tome and Principe Saudi Arabia Senegal Serbia Seychelles Sierra Leone Singapore Slovakia Slovenia Solomon Islands Somalia South Africa South Georgia and the South Sandwich Is South Korea South Sudan Spain Sri Lanka Sudan Suriname Svalbard and Jan Mayen Swaziland Sweden Switzerland Syria Taiwan Tajikistan Tanzania Thailand The Gambia The Netherlands Timor-Leste Togo Tokelau Tonga Trinidad and Tobago Tunisia Turkey Turkmenistan Turks and Caicos Islands Tuvalu UK USA Uganda Ukraine United Arab Emirates United States Virgin Islands Uruguay Uzbekistan Vanuatu Venezuela Vietnam Wallis and Futuna West Bank and Gaza Strip Western Sahara Yemen Zambia Zimbabwe Please select your country/region. You must enter a comment. Competing Interests Please disclose any competing interests that might be construed to influence your judgment of the article's or peer review report's validity or importance. Competing Interests Policy Provide sufficient details of any financial or non-financial competing interests to enable users to assess whether your comments might lead a reasonable person to question your impartiality. Consider the following examples, but note that this is not an exhaustive list: Examples of 'Non-Financial Competing Interests' Within the past 4 years, you have held joint grants, published or collaborated with any of the authors of the selected paper. You have a close personal relationship (e.g. parent, spouse, sibling, or domestic partner) with any of the authors. You are a close professional associate of any of the authors (e.g. scientific mentor, recent student). You work at the same institute as any of the authors. You hope/expect to benefit (e.g. favour or employment) as a result of your submission. You are an Editor for the journal in which the article is published. Examples of 'Financial Competing Interests' You expect to receive, or in the past 4 years have received, any of the following from any commercial organisation that may gain financially from your submission: a salary, fees, funding, reimbursements. You expect to receive, or in the past 4 years have received, shared grant support or other funding with any of the authors. You hold, or are currently applying for, any patents or significant stocks/shares relating to the subject matter of the paper you are commenting on. Please state your competing interests The comment has been saved. An error has occurred. Please try again. Cancel Post var lTitle = "Enhanced human periodontal ligament stem...".replace("'", ''); var linkedInUrl = "http://www.linkedin.com/shareArticle?url=https://f1000research.com/articles/12-447/v2" + "&title=" + encodeURIComponent(lTitle) + "&summary=" + encodeURIComponent('Read the article by '); var deliciousUrl = "https://del.icio.us/post?url=https://f1000research.com/articles/12-447/v2&title=" + encodeURIComponent(lTitle); var redditUrl = "http://reddit.com/submit?url=https://f1000research.com/articles/12-447/v2" + "&title=" + encodeURIComponent(lTitle); linkedInUrl += encodeURIComponent('El Shafei SF et al.'); var offsetTop = /chrome/i.test( navigator.userAgent ) ? 4 : -10; var addthis_config = { ui_offset_top: offsetTop, services_compact : "facebook,twitter,www.linkedin.com,www.mendeley.com,reddit.com", services_expanded : "facebook,twitter,www.linkedin.com,www.mendeley.com,reddit.com", services_custom : [ { name: "LinkedIn", url: linkedInUrl, icon:"/img/icon/at_linkedin.svg" }, { name: "Mendeley", url: "http://www.mendeley.com/import/?url=https://f1000research.com/articles/12-447/v2/mendeley", icon:"/img/icon/at_mendeley.svg" }, { name: "Reddit", url: redditUrl, icon:"/img/icon/at_reddit.svg" }, ] }; var addthis_share = { url: "https://f1000research.com/articles/12-447", templates : { twitter : "Enhanced human periodontal ligament stem cell viability and osteogenic.... El Shafei SF et al., published by " + "@F1000Research" + ", https://f1000research.com/articles/12-447/v2" } }; if (typeof(addthis) != "undefined"){ addthis.addEventListener('addthis.ready', checkCount); addthis.addEventListener('addthis.menu.share', checkCount); } $(".f1r-shares-twitter").attr("href", "https://twitter.com/intent/tweet?text=" + addthis_share.templates.twitter); $(".f1r-shares-facebook").attr("href", "https://www.facebook.com/sharer/sharer.php?u=" + addthis_share.url); $(".f1r-shares-linkedin").attr("href", addthis_config.services_custom[0].url); $(".f1r-shares-reddit").attr("href", addthis_config.services_custom[2].url); $(".f1r-shares-mendelay").attr("href", addthis_config.services_custom[1].url); function checkCount(){ setTimeout(function(){ $(".addthis_button_expanded").each(function(){ var count = $(this).text(); if (count !== "" && count != "0") $(this).removeClass("is-hidden"); else $(this).addClass("is-hidden"); }); }, 1000); } close How to cite this report {{reportCitation}} Cancel Copy Citation Details $(function(){R.ui.buttonDropdowns('.dropdown-for-downloads');}); $(function(){R.ui.toolbarDropdowns('.toolbar-dropdown-for-downloads');}); $.get("/articles/acj/129562/154099") new F1000.Clipboard(); new F1000.ThesaurusTermsDisplay("articles", "article", "154099"); $(document).ready(function() { $( "#frame1" ).on('load', function() { var mydiv = $(this).contents().find("div"); var h = mydiv.height(); console.log(h) }); var tooltipLivingFigure = jQuery(".interactive-living-figure-label .icon-more-info"), titleLivingFigure = tooltipLivingFigure.attr("title"); tooltipLivingFigure.simpletip({ fixed: true, position: ["-115", "30"], baseClass: 'small-tooltip', content:titleLivingFigure + " " }); tooltipLivingFigure.removeAttr("title"); $("body").on("click", ".cite-living-figure", function(e) { e.preventDefault(); var ref = $(this).attr("data-ref"); $(this).closest(".living-figure-list-container").find("#" + ref).fadeIn(200); }); $("body").on("click", ".close-cite-living-figure", function(e) { e.preventDefault(); $(this).closest(".popup-window-wrapper").fadeOut(200); }); $(document).on("mouseup", function(e) { var metricsContainer = $(".article-metrics-popover-wrapper"); if (!metricsContainer.is(e.target) && metricsContainer.has(e.target).length === 0) { $(".article-metrics-close-button").click(); } }); var articleId = $('#articleId').val(); if($("#main-article-count-box").attachArticleMetrics) { $("#main-article-count-box").attachArticleMetrics(articleId, { articleMetricsView: true }); } }); var figshareWidget = $(".new_figshare_widget"); if (figshareWidget.length > 0) { window.figshare.load("f1000", function(Widget) { // Select a tag/tags defined in your page. In this tag we will place the widget. _.map(figshareWidget, function(el){ var widget = new Widget({ articleId: $(el).attr("figshare_articleId") //height:300 // this is the height of the viewer part. [Default: 550] }); widget.initialize(); // initialize the widget widget.mount(el); // mount it in a tag that's on your page // this will save the widget on the global scope for later use from // your JS scripts. This line is optional. //window.widget = widget; }); }); } close Error Close Add Reset F1000.MICROSERVICES.AFFILIATION = ''; $(document).ready(function () { $('.js-affiliations-form').each((index, form) => { new AffiliationForm({ formId: form.id, institutionErrorSelector: '.comment-enter-institution', departmentErrorSelector: '.comment-enter-department', placeSelector: '.js-add-comment-place', stateSelector: '.js-add-comment-state', zipCodeSelector: '.js-add-comment-zipcode', countrySelector: '.js-add-comment-country', countryErrorSelector: '.comment-enter-country', }); }); }); $(document).ready(function () { var reportIds = { "171168": 0, "186886": 0, "186884": 0, "194730": 6, "186858": 0, "186890": 0, "194731": 4, "186859": 0, "186888": 0, "186857": 0, "186860": 0, "186875": 0, "171166": 10, "171167": 0, "186877": 0, "171165": 12, }; $(".referee-response-container,.js-referee-report").each(function(index, el) { var reportId = $(el).attr("data-reportid"), reportCount = reportIds[reportId] || 0; $(el).find(".comments-count-container,.js-referee-report-views").html(reportCount); }); var uuidInput = $("#article_uuid"), oldUUId = uuidInput.val(), newUUId = "a3e52a66-ad34-4128-9602-4dd8c8e03160"; uuidInput.val(newUUId); $("a[href*='article_uuid=']").each(function(index, el) { var newHref = $(el).attr("href").replace(oldUUId, newUUId); $(el).attr("href", newHref); }); }); An innovative open access publishing platform offering rapid publication and open peer review, whilst supporting data deposition and sharing. Browse Gateways Collections How it Works Contact For Developers Cookie Notice Privacy Notice RSS Submit Your Research Follow us © 2012-2026 F1000 Research Ltd. ISSN 2046-1402 | Legal | Partner of Research4Life • CrossRef • ORCID • FAIRSharing R.templateTests.simpleTemplate = R.template(' $text $text $text $text $text '); R.templateTests.runTests(); var F1000platform = new F1000.Platform({ name: "f1000research", displayName: "F1000Research", hostName: "f1000research.com", id: "1", editorialEmail: "[email protected]", infoEmail: "[email protected]", usePmcStats: true }); $(function(){R.ui.dropdowns('.dropdown-for-authors, .dropdown-for-about, .dropdown-for-myresearch');}); // $(function(){R.ui.dropdowns('.dropdown-for-referees');}); $(document).ready(function () { if ($(".cookie-warning").is(":visible")) { $(".sticky").css("margin-bottom", "35px"); $(".devices").addClass("devices-and-cookie-warning"); } $(".cookie-warning .close-button").click(function (e) { $(".devices").removeClass("devices-and-cookie-warning"); $(".sticky").css("margin-bottom", "0"); }); $("#tweeter-feed .tweet-message").each(function (i, message) { var self = $(message); self.html(linkify(self.html())); }); $(".partner").on("mouseenter mouseleave", function() { $(this).find(".gray-scale, .colour").toggleClass("is-hidden"); }); }); Sign In Remember me Forgotten your password? Sign In Cancel Email or password not correct. Please try again Please wait... $(function(){ // Note: All the setup needs to run against a name attribute and *not* the id due the clonish // nature of facebox... $("a[id=googleSignInButton]").click(function(event){ event.preventDefault(); $("input[id=oAuthSystem]").val("GOOGLE"); $("form[id=oAuthForm]").submit(); }); $("a[id=facebookSignInButton]").click(function(event){ event.preventDefault(); $("input[id=oAuthSystem]").val("FACEBOOK"); $("form[id=oAuthForm]").submit(); }); $("a[id=orcidSignInButton]").click(function(event){ event.preventDefault(); $("input[id=oAuthSystem]").val("ORCID"); $("form[id=oAuthForm]").submit(); }); }); If you've forgotten your password, please enter your email address below and we'll send you instructions on how to reset your password. The email address should be the one you originally registered with F1000. Email address not valid, please try again You registered with F1000 via Google, so we cannot reset your password. To sign in, please click here . If you still need help with your Google account password, please click here . You registered with F1000 via Facebook, so we cannot reset your password. To sign in, please click here . If you still need help with your Facebook account password, please click here . Code not correct, please try again Reset password Cancel Email us for further assistance. Server error, please try again. If your email address is registered with us, we will email you instructions to reset your password. If you think you should have received this email but it has not arrived, please check your spam filters and/or contact for further assistance. Please wait... Register $(document).ready(function () { signIn.createSignInAsRow($("#sign-in-form-gfb-popup")); $(".target-field").each(function () { var uris = $(this).val().split("/"); if (uris.pop() === "login") { $(this).val(uris.toString().replace(",","/")); } }); });