Biomechanical evaluation of the effect of a stem cell enhancer on bone implant connection of titanium implants

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Yilmaz-Bozoglan, A. Bozoglan, O. Sunar, O. Polat, B. Tekin This is a preprint; it has not been peer reviewed by a journal. https://doi.org/ 10.21203/rs.3.rs-3790310/v1 This work is licensed under a CC BY 4.0 License Status: Posted Version 1 posted You are reading this latest preprint version Abstract Background This study aimed to conduct a biomechanical investigation of the effects of stem cell enhancer (SCE) application on osseointegration of titanium implants in rat tibias. Methods After surgical implantation of titanium implants in the metaphyseal part of the tibiae of rats, the rats were randomly divided into three equal groups (n = 10): control group, SCE Dosage group 1 (SCE1), and SCE Dosage group 2 (SCE2). Each group consisted of 10 rats. The rats in the SCE1 and SCE2 groups were administered 6 mg and 12 mg SCE (Stemregen®), respectively, every day during the four-week of experimental period after surgery. Implants and surrounding bone tissues were collected for biomechanical bone-implant connection analysis at the end of the experimental procedures. One-way ANOVA was used for statistical analysis. Results There was no significant difference in the biomechanical osseointegration values of the groups; however, the osseointegration of the SCE group was better than that of the control group. Conclusions Stem cell enhancers have the potential to improve the biomechanical bone-implant interface. Biomechanics Bone implant connection Dental implants Osseointegration Stem cell enhancer Figures Figure 1 Figure 2 Background Dental implants replace partially or completely edentulous areas with artificial teeth and function very similar to real teeth [ 1 ]. The success rate of implants exceeds 98%. When osseointegration is achieved, dental implantation is considered a restorative treatment with high long-term predictability [ 2 ]. Osseointegration, with its simplest definition, is known as the mechanical stability of the implant in the bone tissue. Osseointegration can also be defined as the complete interaction between the peri-implant bone niche and the tolerance of the bone marrow after placement of the dental implant. This type of interaction is complex. It also includes heterogeneous cell populations (immune cells, bone, vascular cells, etc.) [ 3 – 5 ]. An abnormal condition that affects bone metabolism or a pathology in the bone marrow may cause the implant to fail [ 6 ]. Factors such as the patient's age and sex, smoking habit, immune system status, and, if any, a chronic disease or drug use, as well as the physical characteristics of the implant, may affect the bone regeneration mechanism and make osseointegration successful or unsuccessful.[ 7 – 8 ]. The body consists of different cells that comprise various organs and tissues. As a natural process of the cell cycle and homeostasis, these organs and tissues lose their cells that need to be regenerated. The rate of this event varies according to the cell/tissue type. For example, the liver is renewed every two–three years, about half of our heart is renewed once in our lifetime, and skin cells are renewed every three–four weeks. In short, our entire body is in a continuous cycle and our health depends on the body's ability to regenerate. To successfully complete this task, the body has a master cell, the stem cell. Stem cells have the ability to multiply indefinitely and transform into cells of almost every organ and tissue in the body. Osteoblasts also develop as a result of a severe process regulated by different signaling pathways from those of mesenchymal stem cells [ 9 ]. During this transformation, the influence of nutrients and food supplements is undeniable. For instance, berberine, which is widely used for musculoskeletal disorders such as osteoporosis, promotes osteogenic differentiation of mesenchymal stem cells via the Wnt/β-catenin signaling pathway [ 10 ]. Ginkgo biloba extract has also been shown to be a promoter in the osteogenic differentiation of stem cells via the same signaling pathwayn [ 11 ]. Another stem cell inducer myricetin, and myricetin stimulates the differentiation of osteoblasts and subsequent bone maturation [ 12 ]. Stem cell-based manipulation may also increase the success of dental implantology and osseointegration. Gli1 + alveolar bone marrow cells lining the alveolar bone marrow vascularity are involved in osseointegration. Due to this, intervention with stem cells or bone marrow with supplemental food, flavonoids, drugs or any other supplement may increase the success of osseointegration. The effect of food supplements on osseointegration, which was previously mainly at the bone level, is now in the dimension of the impact of manipulations that can be performed at the stem cell level of osseointegration [ 13 ]. Stem cell enhancers have been used more frequently to prevent aging and diseases that are higher in the causes of death. The food supplement used in this study contained stem cell enhancers, such as sea buckthorn berry extract, aphanizomenon flos-aquae extract, aloe vera, Fucus vesiculosus extract, Panax notoginseng extract, and beta-glucan. These contents trigger stem cell proliferation, differentiation, and migration from bone marrow [ 14 – 16 ]. This study aimed to biomechanically investigate the effect of a popular stem cell enhancer on the osseointegration levels of titanium implants. Methods Animals and study design Permission for this study was obtained from the Animal Experiments Local Ethics Committee of Firat University (session number:2021/03) and was conducted at the Experimental Research Center of the same university. In the experimental part of the study, care was taken according to the rules of the Declaration of Helsinki. Three groups of animals, each consisting of 10 female Sprague-Dawley rats, were provided by the Firat University Experimental Research Center. Rats weighing approximately 250 g in the same heat period were selected by experts at the center for standardization purposes. The rats were housed in a temperature-controlled room with 55% humidity and 22 ± 2°C, with a 12-hour light and 12-hour dark cycle. After grouping, the rats were placed in pairs in standard cages and fed a regular diet and water ad libitum. Stem cell enhancers (Stemregen®, Texas, USA) were obtained from an importer company in Istanbul, Turkey. Control group (n = 10): The metaphyseal corticocancellous side of the right tibia, one of the lower extremity bones of the rats, was opened. Titanium implants of the size (2.5 mm in diameter and 4 mm in length) that were found to be suitable in previous studies were placed in these sockets [ 6 ]. No additional treatment was applied to the control group during the four-week experimental setup. SCE1 group (n = 10) and SCE2 (n = 10) group: Titanium implants were loaded with proper surgery, as in the control group. The dose of SCE supplement in the rats was calculated using the recommended dose for a 70 kg adult. 6 mg Stemregen® was applied at an SCE dosage of 1 g by oral gavage at the same time each day of the experimental setup, which was planned for four weeks. A double dose of SCE1 was also administered to the SCE2 group in the same manner. After all rats were sacrificed at the end of the scheduled time, the implants were removed along with the surrounding bone tissue and subjected to non-decalcified mechanical testing. Surgical procedures Sterility requirements were compiled for all the surgical procedures. The rats were anesthetized after eight hours of fasting. For general anesthesia, xylazine hydrochloride (Rompun®, Bayer, Germany) and ketamine hydrochloride (Ketasol®, Richter Pharma, Austria) were administered intramuscularly (IM) with an insulin syringe. Mepivacaine hydrochloride (0.3 ml/kg, scandicaine epinephrine 1:100,00 with 2%; Septodont, France) was injected to control wound hemostasis. After shaving the surgical area, it was sterilized with povidone iodide. After making a 15 mm incision over the tibial crest with a scalpel (#15), the proximal tibia was reached using a periosteal elevator. The sockets were opened in the right tibial corticocancellous area [ 5 ] (Fig. 1 A). Resorbable blast material surface titanium implants (Ra:1–2) (Implance, AGS Medical Corporation, Istanbul, Turkey ) were implanted in these sockets (Fig. 1 B). No other surgical procedures were performed until sacrifice during the four-week experiment. After implant loading, the flaps were sutured with absorbable sutures (4/0 Vicryl; Ethicon Inc., Somerville, NJ, USA) for the soft tissues and monofilament sutures (nylon 4.0; Ethicon Inc.) for the skin. After the surgical procedure, the rats were observed daily for signs of pain, detachment, infection, restricted range of motion, and weight loss. Three days postoperatively, antibiotic and analgesic agents were administered IM every day. The subjects in all groups were sacrificed after 4 weeks. The implants loaded at the beginning of the experiment along with the surrounding bone tissues were used for biomechanical analysis. Biomechanical analysis A reverse torque test was applied for biomechanical analysis. The implanted tibial pieces were preserved in special liquid solutions containing formalin. Evaluation was performed as soon as possible to prevent dehydration. All implants were placed in polymethyl methacrylate (PMMA) blocks. A swivel apparatus was used to measure the torque, and a slow and increasing counterclockwise gravitational force was applied manually using a digital torque instrument (Mark10, NY, USA). Rotation was terminated as soon as the implant returned to the bone socket. During the first rotation of the implant in the socket, the highest torque force (N/cm) was recorded using the digital torque device (Fig. 2 ). Biostatistical analysis Data analysis was performed using IBM SPSS v22 software. Quantitative variables are presented as mean ± SD. Statistical significance between groups was analyzed using one-way analysis of variance (ANOVA). p < 0.05 was considered significant. Results Samples whose placement was not at the desired level during reverse torque analysis were not included in the study. The values in the torque analyses did not differ significantly between the groups ( p ˃0.05). However, higher bone-implant attachment values were measured in the SCE groups than in the controls, indicating that osseointegration was achieved at high levels. At the same time, when the SCE 1 and SCE 2 groups were compared, osseointegration increased with the high-dose SCE in the second group (Table 1 ). Table 1 Biomechanic bone implant connection values (Newton/cm) of the groups. Groups N Mean (Newton/cm) SD Min. Max. p* Control 20 19.37 4.95 11.60 28.20 > 0.05 SCE1 12 22.80 10.05 10.40 36.70 SCE2 12 23.60 8.07 12.40 37.60 * One-way ANOVA Test ( p > 0.05). SD: Standard deviation. Discussion When cells need to be replaced, damaged cells send signals to the bone marrow. The bone marrow releases stem cells to circulate throughout the body, and these cells migrate to damaged areas in the tissue. When stem cells come together with damaged cells, they become specialized tissues, which form the building blocks of the body. The last 20 years of research have shown a direct link between stem cells in the cell cycle and the body's natural repair and well-being. In other words, a high number of stem cells in circulation show the participation of stem cells in tissue repair and regeneration [ 9 , 12 ]. As in the entire body, bone tissue is in a state of magnificent and dynamic homeostasis in the form of bone formation by osteoblasts and bone resorption by osteoclasts. Inflammation and oxidative stress, which constitute the pathological basis of almost all diseases, disrupt bone homeostasis. Oxidative stress can result in bone loss, osteoporosis, and fractures. It can even cause many different rheumatological bone diseases such as rheumatoid arthritis and ankylosing spondylitis. Studies have shown that inflammation and oxidative stress inhibit the differentiation of osteoprogenitor cells, which are precursors of bone cells, and osteogenesis of mesenchymal stem cells at the stem cell level [ 17 ]. To prolong human life and prevent aging, the effort and search for human beings are increasing day by day. For this purpose, the effects of various nutrients and supplements, drugs, plants, and even some single-celled organisms on stem cells continue to be studied. For instance, Jensen et al. published an article about the effects of an extract obtained from edible cyanobacteria (Aphanizomenon flos-aquae) enriched with L-selectin in human stem cells. They reported that this extract modulates the expression of CD34 + bone marrow cells and triggers the mobilization of other cells [ 18 ]. In this study, this edible cyanobacteria was identified as a stem cell enhancer. As previously mentioned, Drapeau et al. observed the effects of sea buckthorn fruit extract, rich in proanthocyanidin, a polyphenol, on healthy volunteers [ 16 ]. They reported that blood samples were collected after consumption of 500 mg extract. The mobilization of stem cells, which are involved in regeneration and repair, was increased compared to that of the placebo. Fucoidan is a putative hematopoietic progenitor stem cell-mobilizing agent. Oral ingestion of this agent may contribute to the development of hematopoietic progenitor stem cells by increasing the expression CXCR4 in human CD34 + cells [ 19 ]. Ginseng, which refers to the roots of plants of the genus Panax, is one of the most well-known and used herbal products. In an experimental cerebral infarction model, Xuesaitong capsules containing Panax notoginseng were administered intragastrically for 1 month. At the end of a month, it was observed that the bone marrow mesenchymal stem cell levels and mobilization into the peripheral blood increased [ 20 ]. Aloe vera, which has a history of almost 6000 years, is another frequently used herbal product generally used to heal burns and wounds. In this study, human dental pulp mesenchymal stem cells and mesenchymal stem cells from aloe vera were implanted into a collagen sponge. The effects of aloe vera mesenchymal stem cells on healing non-critical defects of the rat tibia were evaluated. Although the results were not significant, it was observed that A. vera accelerated healing and slowed down the inflammatory process [ 21 ]. The data which is obtained in the current study reveales that the stem cell enhancer stimulates osseointegration biomechanically. Macroclada, a special type of aloe vera, has been used by the local residents of Madagascar to treat a wide variety of clinical conditions. Drapeau et al. investigated the effect of Aloe macroclada on bone marrow stem cells in their study. A. macroclada was prepared in pellet form by Malagasy healers using traditional production methods and was administered to 4 volunteers at conventional doses. A significant increase in circulating stem cells was observed within 2 hours of consumption. These data suggest that Aloe macroclada may be an important mechanism of action for the mobilization of stem cells [ 22 ]. Colostrum contains a mixture of growth factors called transfer factors. They stimulate and organize the activities of various immune cells. However, oral intake of a carbohydrate source, such as immune-enhancing beta-glucan, in combination with colostrum, demonstrates its capacity to influence the endogenous stem cell niche. The transfer factor in the stem cell enhancer in this study also contained growth factors and cytokines that support stem cell functions and enable bone marrow stem cell activation. Webster et al. demonstrated that the production of stem cell-related cytokines such as IL-6, GCSF, and vascular endothelial growth factor can be stimulated by bovine colostrum [ 23 ]. β-Glucans are difficult-to-digest complex carbohydrates that protect and support the bone marrow, triggering the movement of stem cells from the bone marrow [ 24 ]. Maitake beta-glucan are widely used in cancer treatment, especially in Asia. Lin et al. showed for the first time in their study that β-glucan increases hematopoietic stem cell proliferation and induces leukocyte differentiation [ 25 ]. SCE in the current study also contained β-glucan, which is produced by fermentation. This shows that this food supplement is a stem cell enhancer with the content mentioned so far. Cyanobacteria, also known as blue-green algae, are photosynthetic bacteria that occur in freshwater and saltwater environments worldwide. Aphanizomenon flos-aquae is the most common cyanobacteria found naturally in freshwater sources and has been used as a dietary blue-green algae supplement in the United States since the 1980s [ 26 ]. Besides its anti-inflammatory and immunomodulatory properties, aphanizomenon flos-aquae extract has been documented to promote the release of stem cells from the bone marrow and significantly increase the number of circulating stem cells [ 18 ]. According to our literature search, significantly increased osseointegration values were achieved when stem cell-inducing extracts or commercial products were applied to the implant surface [ 27 ]. On the other hand, two years ago, the effect of bone marrow mesenchymal stem cell-derived extracellular matrix sheets on osseointegration was evaluated for the first time. Osseointegration of the stem cell-derived extracellular matrix and implant complex placed in the rat tibia was quite advanced [ 28 ]. Conclusion Based on the limited results of this study, the stem cell enhancer has the potential to alleviate the biomechanical bone-implant interface. Further studies can reveal the relationship between stem cell enhancers and the bone-implant interface. Abbreviations ANOVA analysis of variance cm centimeter g gram GCSF granulocyte colony-stimulating factor IL-6 Interleukin-6 IM intramuscular kg kilogram mg milligram ml milliliter N Newton No number p probability PMMA polymethyl methacrylate SCE stem cell enhancer °C Celsius or centigrade Declarations Acknowledgements Not applicable Authors’ contributions MYB, AB, BT contributed to the study design, the data acquisition, analysis, andinterpretation, and wrote the manuscript draft. BT, OS and OP contributed to the study design, and the interpretation of the results, and revised the manuscript. AB supervised the work. OS and OP provide funding. All authors have read and approved the final manuscript. Funding This work was self-funded by the authors. Availability of data and materials The datasets generated and analyzed during the current study are availablefrom the corresponding author on reasonable request. Ethics approval and consent to participate Permission for this study was obtained from the Animal Experiments Local Ethics Committee of Firat University (session number:2021/03) and performed in accordance with the Declaration of Helsinki. Consent for publication Not applicable. Competing interests The authors declare no competing interests References Jung RE, Zembic A, Pjetursson BE, et al. Systematic review of the survival rate and the incidence of biological, technical, and aesthetic complications of single crowns on implants reported in longitudinal studies with a mean follow-up of 5 years. Clin Oral Implants Res 2012;6:2-21. Lin G, Ye S, Liu F, et al. A retrospective study of 30,959 implants: Risk factors associated with early and late implant loss. J Clin Periodontol 2018;45:733-743. Amengual-Peñafiel L, Brañes-Aroca M, Marchesani-Carrasco F, et al. Coupling between Osseointegration and Mechanotransduction to Maintain Foreign Body Equilibrium in the Long-Term: A Comprehensive Overview. J Clin Med 2019;8:139. Dundar S, Yaman F, Gecor O, et al. Effects of Local and Systemic Zoledronic Acid Application on Titanium Implant Osseointegration: An Experimental Study Conducted on Two Surface Types . J Craniofac Surg 2017;28:935-938. Yu K, Jiang Z, Miao X, et al. circRNA422 enhanced osteogenic differentiation of bone marrow mesenchymal stem cells during early osseointegration through the SP7/LRP5 axis. Mol Ther 2022;30:3226-3240. Ruggiero SL, Mehrotra B, Rosenberg TJ, et al. Osteonecrosis of the jaws associated with the use of bisphosphonates: a review of 63 cases. J Oral Maxil Surg 2004;62,527-553. Talo Yildirim T, Dündar S, Bozoğlan A, et al. Evaluation of the Effects of ß-Adrenergic Receptor-Propranolol on Osseointegration of the Titanium Implants. J Craniofac Surg 2021;32:783-786. Gonzales KAU, Polak L, Matos I, et al. Stem cells expand potency and alter tissue fitness by accumulating diverse epigenetic memories. Science 2021;374(6571):eabh2444. Tao K, Xiao D, Weng J, et al. Berberine promotes bone marrow-derived mesenchymal stem cells osteogenic differentiation via canonical Wnt/β-catenin signaling pathway. Toxicol Lett 2016;240:68-80. Zhang LN, Wang XX, Wang Z, et al. Berberine improves advanced glycation end products‑induced osteogenic differentiation responses in human periodontal ligament stem cells through the canonical Wnt/β‑catenin pathway. Mol Med Rep 2019;19:5440-5452. Gu Q, Chen C, Zhang Z, et al. Ginkgo biloba extract promotes osteogenic differentiation of human bone marrow mesenchymal stem cells in a pathway involving Wnt/β-catenin signaling. Pharmacol Res 2015;97:70-78. Ying X, Chen X, Feng Y, et al. Myricetin enhances osteogenic differentiation through the activation of canonical Wnt/β-catenin signaling in human bone marrow stromal cells. Eur J Pharmacol 2014;738:22-30. Yi Y, Stenberg W, Luo W, et al. Alveolar Bone Marrow Gli1+ Stem Cells Support Implant Osseointegration. J Dent Res 2022;101:73-82. Zhang D, Zhou X, Liu L, et al. Glucomannan from Aloe vera Gel Promotes Intestinal Stem Cell-Mediated Epithelial Regeneration via the Wnt/β-Catenin Pathway. J Agric Food Chem 2021;69:10581-10591. Si Y, Zhu J, Huang X, et al. Effects of Panax notoginseng saponins on proliferation and differentiation of rat embryonic cortical neural stem cells. J Chin Med Assoc 2016;79(5):256-263. Drapeau C, Benson KF, Jensen GS. Rapid and selective mobilization of specific stem cell types after consumption of a polyphenol-rich extract from sea buckthorn berries (Hippophae) in healthy human subjects. Clin Interv Aging 2019;14:253-263. Barbarić Starčević K, Lukač N, Jelić M, et al. Reciprocal Alterations in Osteoprogenitor and Immune Cell Populations in Rheumatoid Synovia. Int J Mol Sci 2022;23:12379. Jensen GS, Hart AN, Zaske LA, et al. Mobilization of human CD34+ CD133+ and CD34+ CD133(-) stem cells in vivo by consumption of an extract from Aphanizomenon flos-aquae--related to modulation of CXCR4 expression by an L-selectin ligand? Cardiovasc Revasc Med 2007;8:189-202. Irhimeh MR, Fitton JH, Lowenthal RM. Fucoidan ingestion increases the expression of CXCR4 on human CD34+ cells. Exp Hematol 2007;35:989-994. Zhang JS, Zhang BX, Du MM, et al. Chinese preparation Xuesaitong promotes the mobilization of bone marrow mesenchymal stem cells in rats with cerebral infarction. Neural Regen Res 2016;11:292-297. Soares IMV, Fernandes GVO, Larissa Cordeiro C, et al. The influence of Aloe vera with mesenchymal stem cells from dental pulp on bone regeneration: characterization and treatment of non-critical defects of the tibia in rats. J Appl Oral Sci 2019;27:e20180103. Drapeau C, Benson KF, James J, et al. Aloe macroclada from Madagascar triggers transient bone marrow stem cell mobilization. J Stem Cell Res Ther 2015;5:2. Webster GA, Lehrke P. Development of a combined bovine colostrum and immune-stimulatory carbohydrate nutraceutical for enhancement of endogenous stem cell activity. The Open Nutraceuticals Journal 2013;6:35-44 Patchen ML, Liang J, Vaudrain T, et al. Mobilization of peripheral blood progenitor cells by Betafectin PGG-Glucan alone and in combination with granulocyte colony-stimulating factor. Stem Cells 1998;16:208-217. Lin H, Cheung SW, Nesin M, et al. Enhancement of umbilical cord blood cell hematopoiesis by maitake beta-glucan is mediated by granulocyte colony-stimulating factor production. Clin Vaccine Immunol 2007;14:21-27. Lyon-Colbert A, Su S, Cude C. A Systematic Literature Review for Evidence of Aphanizomenon flos-aquae Toxigenicity in Recreational Waters and Toxicity of Dietary Supplements: 2000⁻2017. Toxins (Basel) 2018;10:254. Weng CC, Ou KL, Wu CY, et al. Mechanism and Clinical Properties of StemBios Cell Therapy: Induction of Early Osseointegration in Novel Dental Implants. Int J Oral Maxillofac Implants 2017;32:e47-e54. Feng Y , Jiang Z , Zhang Y , et al. Stem-cell-derived ECM sheet-implant complexes for enhancing osseointegration. Biomater Sci 2020;8:6647-56. Additional Declarations No competing interests reported. Cite Share Download PDF Status: Posted Version 1 posted You are reading this latest preprint version Research Square lets you share your work early, gain feedback from the community, and start making changes to your manuscript prior to peer review in a journal. As a division of Research Square Company, we’re committed to making research communication faster, fairer, and more useful. We do this by developing innovative software and high quality services for the global research community. Our growing team is made up of researchers and industry professionals working together to solve the most critical problems facing scientific publishing. Also discoverable on Platform About Our Team In Review Editorial Policies Advisory Board Help Center Resources Author Services Accessibility API Access RSS feed Manage Cookie Preferences © Research Square 2026 | ISSN 2693-5015 (online) Privacy Policy Terms of Service Do Not Sell My Personal Information {"props":{"pageProps":{"initialData":{"identity":"rs-3790310","acceptedTermsAndConditions":true,"allowDirectSubmit":false,"archivedVersions":[],"articleType":"Research Article","associatedPublications":[],"authors":[{"id":262343001,"identity":"54c77996-c296-49b5-a32c-6ab060a9a853","order_by":0,"name":"M. 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(A): Preparation of implant sockets in tibial corticocancellous areas of rats. (B): Implantation of titanium implants into sockets.\u003c/p\u003e","description":"","filename":"Figure1AB.jpg","url":"https://assets-eu.researchsquare.com/files/rs-3790310/v1/8345af26328762b047f2a108.jpg"},{"id":49327087,"identity":"de55e677-8c31-4382-a83d-49734065efae","added_by":"auto","created_at":"2024-01-08 17:35:48","extension":"jpg","order_by":2,"title":"Figure 2","display":"","copyAsset":false,"role":"figure","size":205457,"visible":true,"origin":"","legend":"\u003cp\u003eFixing the samples to apply force on the digital torque device and monitoring the failure values on the digital display.\u003c/p\u003e","description":"","filename":"Figure2.jpg","url":"https://assets-eu.researchsquare.com/files/rs-3790310/v1/143924006624590887eb3a31.jpg"},{"id":49327602,"identity":"4e753c05-c7cd-424a-98a7-9097fc10b989","added_by":"auto","created_at":"2024-01-08 17:43:48","extension":"pdf","order_by":0,"title":"","display":"","copyAsset":false,"role":"manuscript-pdf","size":498759,"visible":true,"origin":"","legend":"","description":"","filename":"manuscript.pdf","url":"https://assets-eu.researchsquare.com/files/rs-3790310/v1/bc1b1958-ceb9-414c-8b3a-2c1ef1d02560.pdf"}],"financialInterests":"No competing interests reported.","formattedTitle":"Biomechanical evaluation of the effect of a stem cell enhancer on bone implant connection of titanium implants","fulltext":[{"header":"Background","content":"\u003cp\u003eDental implants replace partially or completely edentulous areas with artificial teeth and function very similar to real teeth [\u003cspan citationid=\"CR1\" class=\"CitationRef\"\u003e1\u003c/span\u003e]. The success rate of implants exceeds 98%. When osseointegration is achieved, dental implantation is considered a restorative treatment with high long-term predictability [\u003cspan citationid=\"CR2\" class=\"CitationRef\"\u003e2\u003c/span\u003e]. Osseointegration, with its simplest definition, is known as the mechanical stability of the implant in the bone tissue. Osseointegration can also be defined as the complete interaction between the peri-implant bone niche and the tolerance of the bone marrow after placement of the dental implant. This type of interaction is complex. It also includes heterogeneous cell populations (immune cells, bone, vascular cells, etc.) [\u003cspan additionalcitationids=\"CR4\" citationid=\"CR3\" class=\"CitationRef\"\u003e3\u003c/span\u003e\u0026ndash;\u003cspan citationid=\"CR5\" class=\"CitationRef\"\u003e5\u003c/span\u003e]. An abnormal condition that affects bone metabolism or a pathology in the bone marrow may cause the implant to fail [\u003cspan citationid=\"CR6\" class=\"CitationRef\"\u003e6\u003c/span\u003e]. Factors such as the patient's age and sex, smoking habit, immune system status, and, if any, a chronic disease or drug use, as well as the physical characteristics of the implant, may affect the bone regeneration mechanism and make osseointegration successful or unsuccessful.[\u003cspan citationid=\"CR7\" class=\"CitationRef\"\u003e7\u003c/span\u003e\u0026ndash;\u003cspan citationid=\"CR8\" class=\"CitationRef\"\u003e8\u003c/span\u003e].\u003c/p\u003e \u003cp\u003eThe body consists of different cells that comprise various organs and tissues. As a natural process of the cell cycle and homeostasis, these organs and tissues lose their cells that need to be regenerated. The rate of this event varies according to the cell/tissue type. For example, the liver is renewed every two\u0026ndash;three years, about half of our heart is renewed once in our lifetime, and skin cells are renewed every three\u0026ndash;four weeks. In short, our entire body is in a continuous cycle and our health depends on the body's ability to regenerate. To successfully complete this task, the body has a master cell, the stem cell. Stem cells have the ability to multiply indefinitely and transform into cells of almost every organ and tissue in the body. Osteoblasts also develop as a result of a severe process regulated by different signaling pathways from those of mesenchymal stem cells [\u003cspan citationid=\"CR9\" class=\"CitationRef\"\u003e9\u003c/span\u003e]. During this transformation, the influence of nutrients and food supplements is undeniable. For instance, berberine, which is widely used for musculoskeletal disorders such as osteoporosis, promotes osteogenic differentiation of mesenchymal stem cells via the Wnt/β-catenin signaling pathway [\u003cspan citationid=\"CR10\" class=\"CitationRef\"\u003e10\u003c/span\u003e]. Ginkgo biloba extract has also been shown to be a promoter in the osteogenic differentiation of stem cells via the same signaling pathwayn [\u003cspan citationid=\"CR11\" class=\"CitationRef\"\u003e11\u003c/span\u003e]. Another stem cell inducer myricetin, and myricetin stimulates the differentiation of osteoblasts and subsequent bone maturation [\u003cspan citationid=\"CR12\" class=\"CitationRef\"\u003e12\u003c/span\u003e].\u003c/p\u003e \u003cp\u003eStem cell-based manipulation may also increase the success of dental implantology and osseointegration. Gli1\u0026thinsp;+\u0026thinsp;alveolar bone marrow cells lining the alveolar bone marrow vascularity are involved in osseointegration. Due to this, intervention with stem cells or bone marrow with supplemental food, flavonoids, drugs or any other supplement may increase the success of osseointegration. The effect of food supplements on osseointegration, which was previously mainly at the bone level, is now in the dimension of the impact of manipulations that can be performed at the stem cell level of osseointegration [\u003cspan citationid=\"CR13\" class=\"CitationRef\"\u003e13\u003c/span\u003e].\u003c/p\u003e \u003cp\u003eStem cell enhancers have been used more frequently to prevent aging and diseases that are higher in the causes of death. The food supplement used in this study contained stem cell enhancers, such as sea buckthorn berry extract, aphanizomenon flos-aquae extract, aloe vera, Fucus vesiculosus extract, Panax notoginseng extract, and beta-glucan. These contents trigger stem cell proliferation, differentiation, and migration from bone marrow [\u003cspan additionalcitationids=\"CR15\" citationid=\"CR14\" class=\"CitationRef\"\u003e14\u003c/span\u003e\u0026ndash;\u003cspan citationid=\"CR16\" class=\"CitationRef\"\u003e16\u003c/span\u003e].\u003c/p\u003e \u003cp\u003eThis study aimed to biomechanically investigate the effect of a popular stem cell enhancer on the osseointegration levels of titanium implants.\u003c/p\u003e"},{"header":"Methods","content":"\u003cdiv id=\"Sec3\" class=\"Section2\"\u003e \u003ch2\u003eAnimals and study design\u003c/h2\u003e \u003cp\u003e Permission for this study was obtained from the Animal Experiments Local Ethics Committee of Firat University (session number:2021/03) and was conducted at the Experimental Research Center of the same university. In the experimental part of the study, care was taken according to the rules of the Declaration of Helsinki. Three groups of animals, each consisting of 10 female Sprague-Dawley rats, were provided by the Firat University Experimental Research Center. Rats weighing approximately 250 g in the same heat period were selected by experts at the center for standardization purposes. The rats were housed in a temperature-controlled room with 55% humidity and 22\u0026thinsp;\u0026plusmn;\u0026thinsp;2\u0026deg;C, with a 12-hour light and 12-hour dark cycle. After grouping, the rats were placed in pairs in standard cages and fed a regular diet and water ad libitum. Stem cell enhancers (Stemregen\u0026reg;, Texas, USA) were obtained from an importer company in Istanbul, Turkey.\u003c/p\u003e \u003cp\u003eControl group (n\u0026thinsp;=\u0026thinsp;10): The metaphyseal corticocancellous side of the right tibia, one of the lower extremity bones of the rats, was opened. Titanium implants of the size (2.5 mm in diameter and 4 mm in length) that were found to be suitable in previous studies were placed in these sockets [\u003cspan citationid=\"CR6\" class=\"CitationRef\"\u003e6\u003c/span\u003e]. No additional treatment was applied to the control group during the four-week experimental setup.\u003c/p\u003e \u003cp\u003eSCE1 group (n\u0026thinsp;=\u0026thinsp;10) and SCE2 (n\u0026thinsp;=\u0026thinsp;10) group: Titanium implants were loaded with proper surgery, as in the control group. The dose of SCE supplement in the rats was calculated using the recommended dose for a 70 kg adult. 6 mg Stemregen\u0026reg; was applied at an SCE dosage of 1 g by oral gavage at the same time each day of the experimental setup, which was planned for four weeks. A double dose of SCE1 was also administered to the SCE2 group in the same manner.\u003c/p\u003e \u003cp\u003eAfter all rats were sacrificed at the end of the scheduled time, the implants were removed along with the surrounding bone tissue and subjected to non-decalcified mechanical testing.\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec4\" class=\"Section2\"\u003e \u003ch2\u003eSurgical procedures\u003c/h2\u003e \u003cp\u003eSterility requirements were compiled for all the surgical procedures. The rats were anesthetized after eight hours of fasting. For general anesthesia, xylazine hydrochloride (Rompun\u0026reg;, Bayer, Germany) and ketamine hydrochloride (Ketasol\u0026reg;, Richter Pharma, Austria) were administered intramuscularly (IM) with an insulin syringe. Mepivacaine hydrochloride (0.3 ml/kg, scandicaine epinephrine 1:100,00 with 2%; Septodont, France) was injected to control wound hemostasis. After shaving the surgical area, it was sterilized with povidone iodide. After making a 15 mm incision over the tibial crest with a scalpel (#15), the proximal tibia was reached using a periosteal elevator. The sockets were opened in the right tibial corticocancellous area [\u003cspan citationid=\"CR5\" class=\"CitationRef\"\u003e5\u003c/span\u003e] (Fig.\u0026nbsp;\u003cspan refid=\"Fig1\" class=\"InternalRef\"\u003e1\u003c/span\u003eA). Resorbable blast material surface titanium implants (Ra:1\u0026ndash;2) (Implance, AGS Medical Corporation, Istanbul, Turkey ) were implanted in these sockets (Fig.\u0026nbsp;\u003cspan refid=\"Fig1\" class=\"InternalRef\"\u003e1\u003c/span\u003eB). No other surgical procedures were performed until sacrifice during the four-week experiment. After implant loading, the flaps were sutured with absorbable sutures (4/0 Vicryl; Ethicon Inc., Somerville, NJ, USA) for the soft tissues and monofilament sutures (nylon 4.0; Ethicon Inc.) for the skin. After the surgical procedure, the rats were observed daily for signs of pain, detachment, infection, restricted range of motion, and weight loss. Three days postoperatively, antibiotic and analgesic agents were administered IM every day. The subjects in all groups were sacrificed after 4 weeks. The implants loaded at the beginning of the experiment along with the surrounding bone tissues were used for biomechanical analysis.\u003c/p\u003e \u003cp\u003e \u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec5\" class=\"Section2\"\u003e \u003ch2\u003eBiomechanical analysis\u003c/h2\u003e \u003cp\u003eA reverse torque test was applied for biomechanical analysis. The implanted tibial pieces were preserved in special liquid solutions containing formalin. Evaluation was performed as soon as possible to prevent dehydration. All implants were placed in polymethyl methacrylate (PMMA) blocks. A swivel apparatus was used to measure the torque, and a slow and increasing counterclockwise gravitational force was applied manually using a digital torque instrument (Mark10, NY, USA). Rotation was terminated as soon as the implant returned to the bone socket. During the first rotation of the implant in the socket, the highest torque force (N/cm) was recorded using the digital torque device (Fig.\u0026nbsp;\u003cspan refid=\"Fig2\" class=\"InternalRef\"\u003e2\u003c/span\u003e).\u003c/p\u003e \u003cp\u003e \u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec6\" class=\"Section2\"\u003e \u003ch2\u003eBiostatistical analysis\u003c/h2\u003e \u003cp\u003eData analysis was performed using IBM SPSS v22 software. Quantitative variables are presented as mean\u0026thinsp;\u0026plusmn;\u0026thinsp;SD. Statistical significance between groups was analyzed using one-way analysis of variance (ANOVA). \u003cem\u003ep\u003c/em\u003e\u0026thinsp;\u0026lt;\u0026thinsp;0.05 was considered significant.\u003c/p\u003e \u003c/div\u003e"},{"header":"Results","content":"\u003cp\u003eSamples whose placement was not at the desired level during reverse torque analysis were not included in the study. The values in the torque analyses did not differ significantly between the groups (\u003cem\u003ep\u003c/em\u003e˃0.05). However, higher bone-implant attachment values were measured in the SCE groups than in the controls, indicating that osseointegration was achieved at high levels. At the same time, when the SCE 1 and SCE 2 groups were compared, osseointegration increased with the high-dose SCE in the second group (Table\u0026nbsp;\u003cspan refid=\"Tab1\" class=\"InternalRef\"\u003e1\u003c/span\u003e).\u003c/p\u003e \u003cp\u003e \u003cdiv class=\"gridtable\"\u003e\u003ctable float=\"Yes\" id=\"Tab1\" border=\"1\"\u003e \u003ccaption language=\"En\"\u003e \u003cdiv class=\"CaptionNumber\"\u003eTable 1\u003c/div\u003e \u003cdiv class=\"CaptionContent\"\u003e \u003cp\u003eBiomechanic bone implant connection values (Newton/cm) of the groups.\u003c/p\u003e \u003c/div\u003e \u003c/caption\u003e \u003ccolgroup cols=\"7\"\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c1\" colnum=\"1\"\u003e\u003c/div\u003e \u003cdiv align=\"char\" char=\".\" class=\"colspec\" colname=\"c2\" colnum=\"2\"\u003e\u003c/div\u003e \u003cdiv align=\"char\" char=\".\" class=\"colspec\" colname=\"c3\" colnum=\"3\"\u003e\u003c/div\u003e \u003cdiv align=\"char\" char=\".\" class=\"colspec\" colname=\"c4\" colnum=\"4\"\u003e\u003c/div\u003e \u003cdiv align=\"char\" char=\".\" class=\"colspec\" colname=\"c5\" colnum=\"5\"\u003e\u003c/div\u003e \u003cdiv align=\"char\" char=\".\" class=\"colspec\" colname=\"c6\" colnum=\"6\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c7\" colnum=\"7\"\u003e\u003c/div\u003e \u003cthead\u003e \u003ctr\u003e \u003cth align=\"left\" colname=\"c1\"\u003e \u003cp\u003eGroups\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c2\"\u003e \u003cp\u003eN\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c3\"\u003e \u003cp\u003eMean (Newton/cm)\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c4\"\u003e \u003cp\u003eSD\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c5\"\u003e \u003cp\u003eMin.\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c6\"\u003e \u003cp\u003eMax.\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c7\"\u003e \u003cp\u003e\u003cem\u003ep*\u003c/em\u003e\u003c/p\u003e \u003c/th\u003e \u003c/tr\u003e \u003c/thead\u003e \u003ctbody\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eControl\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e20\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e19.37\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e4.95\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e11.60\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c6\"\u003e \u003cp\u003e28.20\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\" morerows=\"2\" rowspan=\"3\"\u003e \u003cp\u003e\u0026gt;\u0026thinsp;0.05\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eSCE1\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e12\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e22.80\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e10.05\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e10.40\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c6\"\u003e \u003cp\u003e36.70\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eSCE2\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e12\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e23.60\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e8.07\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e12.40\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c6\"\u003e \u003cp\u003e37.60\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003c/tbody\u003e \u003c/colgroup\u003e \u003c/table\u003e\u003c/div\u003e \u003c/p\u003e \u003cp\u003e \u003cb\u003e*\u003c/b\u003eOne-way ANOVA Test (\u003cem\u003ep\u003c/em\u003e\u0026thinsp;\u0026gt;\u0026thinsp;0.05). SD: Standard deviation.\u003c/p\u003e"},{"header":"Discussion","content":"\u003cp\u003eWhen cells need to be replaced, damaged cells send signals to the bone marrow. The bone marrow releases stem cells to circulate throughout the body, and these cells migrate to damaged areas in the tissue. When stem cells come together with damaged cells, they become specialized tissues, which form the building blocks of the body. The last 20 years of research have shown a direct link between stem cells in the cell cycle and the body's natural repair and well-being. In other words, a high number of stem cells in circulation show the participation of stem cells in tissue repair and regeneration [\u003cspan citationid=\"CR9\" class=\"CitationRef\"\u003e9\u003c/span\u003e, \u003cspan citationid=\"CR12\" class=\"CitationRef\"\u003e12\u003c/span\u003e].\u003c/p\u003e \u003cp\u003eAs in the entire body, bone tissue is in a state of magnificent and dynamic homeostasis in the form of bone formation by osteoblasts and bone resorption by osteoclasts. Inflammation and oxidative stress, which constitute the pathological basis of almost all diseases, disrupt bone homeostasis. Oxidative stress can result in bone loss, osteoporosis, and fractures. It can even cause many different rheumatological bone diseases such as rheumatoid arthritis and ankylosing spondylitis. Studies have shown that inflammation and oxidative stress inhibit the differentiation of osteoprogenitor cells, which are precursors of bone cells, and osteogenesis of mesenchymal stem cells at the stem cell level [\u003cspan citationid=\"CR17\" class=\"CitationRef\"\u003e17\u003c/span\u003e].\u003c/p\u003e \u003cp\u003eTo prolong human life and prevent aging, the effort and search for human beings are increasing day by day. For this purpose, the effects of various nutrients and supplements, drugs, plants, and even some single-celled organisms on stem cells continue to be studied. For instance, Jensen et al. published an article about the effects of an extract obtained from edible cyanobacteria (Aphanizomenon flos-aquae) enriched with L-selectin in human stem cells. They reported that this extract modulates the expression of CD34\u0026thinsp;+\u0026thinsp;bone marrow cells and triggers the mobilization of other cells [\u003cspan citationid=\"CR18\" class=\"CitationRef\"\u003e18\u003c/span\u003e]. In this study, this edible cyanobacteria was identified as a stem cell enhancer. As previously mentioned, Drapeau et al. observed the effects of sea buckthorn fruit extract, rich in proanthocyanidin, a polyphenol, on healthy volunteers [\u003cspan citationid=\"CR16\" class=\"CitationRef\"\u003e16\u003c/span\u003e]. They reported that blood samples were collected after consumption of 500 mg extract. The mobilization of stem cells, which are involved in regeneration and repair, was increased compared to that of the placebo. Fucoidan is a putative hematopoietic progenitor stem cell-mobilizing agent. Oral ingestion of this agent may contribute to the development of hematopoietic progenitor stem cells by increasing the expression CXCR4 in human CD34\u0026thinsp;+\u0026thinsp;cells [\u003cspan citationid=\"CR19\" class=\"CitationRef\"\u003e19\u003c/span\u003e].\u003c/p\u003e \u003cp\u003eGinseng, which refers to the roots of plants of the genus Panax, is one of the most well-known and used herbal products. In an experimental cerebral infarction model, Xuesaitong capsules containing Panax notoginseng were administered intragastrically for 1 month. At the end of a month, it was observed that the bone marrow mesenchymal stem cell levels and mobilization into the peripheral blood increased [\u003cspan citationid=\"CR20\" class=\"CitationRef\"\u003e20\u003c/span\u003e]. Aloe vera, which has a history of almost 6000 years, is another frequently used herbal product generally used to heal burns and wounds. In this study, human dental pulp mesenchymal stem cells and mesenchymal stem cells from aloe vera were implanted into a collagen sponge. The effects of aloe vera mesenchymal stem cells on healing non-critical defects of the rat tibia were evaluated. Although the results were not significant, it was observed that A. vera accelerated healing and slowed down the inflammatory process [\u003cspan citationid=\"CR21\" class=\"CitationRef\"\u003e21\u003c/span\u003e]. The data which is obtained in the current study reveales that the stem cell enhancer stimulates osseointegration biomechanically. Macroclada, a special type of aloe vera, has been used by the local residents of Madagascar to treat a wide variety of clinical conditions. Drapeau et al. investigated the effect of Aloe macroclada on bone marrow stem cells in their study. A. macroclada was prepared in pellet form by Malagasy healers using traditional production methods and was administered to 4 volunteers at conventional doses. A significant increase in circulating stem cells was observed within 2 hours of consumption. These data suggest that Aloe macroclada may be an important mechanism of action for the mobilization of stem cells [\u003cspan citationid=\"CR22\" class=\"CitationRef\"\u003e22\u003c/span\u003e].\u003c/p\u003e \u003cp\u003eColostrum contains a mixture of growth factors called transfer factors. They stimulate and organize the activities of various immune cells. However, oral intake of a carbohydrate source, such as immune-enhancing beta-glucan, in combination with colostrum, demonstrates its capacity to influence the endogenous stem cell niche. The transfer factor in the stem cell enhancer in this study also contained growth factors and cytokines that support stem cell functions and enable bone marrow stem cell activation. Webster et al. demonstrated that the production of stem cell-related cytokines such as IL-6, GCSF, and vascular endothelial growth factor can be stimulated by bovine colostrum [\u003cspan citationid=\"CR23\" class=\"CitationRef\"\u003e23\u003c/span\u003e]. β-Glucans are difficult-to-digest complex carbohydrates that protect and support the bone marrow, triggering the movement of stem cells from the bone marrow [\u003cspan citationid=\"CR24\" class=\"CitationRef\"\u003e24\u003c/span\u003e]. Maitake beta-glucan are widely used in cancer treatment, especially in Asia. Lin et al. showed for the first time in their study that β-glucan increases hematopoietic stem cell proliferation and induces leukocyte differentiation [\u003cspan citationid=\"CR25\" class=\"CitationRef\"\u003e25\u003c/span\u003e]. SCE in the current study also contained β-glucan, which is produced by fermentation. This shows that this food supplement is a stem cell enhancer with the content mentioned so far. Cyanobacteria, also known as blue-green algae, are photosynthetic bacteria that occur in freshwater and saltwater environments worldwide. Aphanizomenon flos-aquae is the most common cyanobacteria found naturally in freshwater sources and has been used as a dietary blue-green algae supplement in the United States since the 1980s [\u003cspan citationid=\"CR26\" class=\"CitationRef\"\u003e26\u003c/span\u003e]. Besides its anti-inflammatory and immunomodulatory properties, aphanizomenon flos-aquae extract has been documented to promote the release of stem cells from the bone marrow and significantly increase the number of circulating stem cells [\u003cspan citationid=\"CR18\" class=\"CitationRef\"\u003e18\u003c/span\u003e].\u003c/p\u003e \u003cp\u003eAccording to our literature search, significantly increased osseointegration values were achieved when stem cell-inducing extracts or commercial products were applied to the implant surface [\u003cspan citationid=\"CR27\" class=\"CitationRef\"\u003e27\u003c/span\u003e]. On the other hand, two years ago, the effect of bone marrow mesenchymal stem cell-derived extracellular matrix sheets on osseointegration was evaluated for the first time. Osseointegration of the stem cell-derived extracellular matrix and implant complex placed in the rat tibia was quite advanced [\u003cspan citationid=\"CR28\" class=\"CitationRef\"\u003e28\u003c/span\u003e].\u003c/p\u003e"},{"header":"Conclusion","content":"\u003cp\u003eBased on the limited results of this study, the stem cell enhancer has the potential to alleviate the biomechanical bone-implant interface. Further studies can reveal the relationship between stem cell enhancers and the bone-implant interface.\u003c/p\u003e"},{"header":"Abbreviations","content":"\u003cp\u003eANOVA analysis of variance\u003c/p\u003e\n\u003cp\u003ecm centimeter\u003c/p\u003e\n\u003cp\u003eg gram\u003c/p\u003e\n\u003cp\u003eGCSF granulocyte colony-stimulating factor\u003c/p\u003e\n\u003cp\u003eIL-6 Interleukin-6\u003c/p\u003e\n\u003cp\u003eIM intramuscular\u003c/p\u003e\n\u003cp\u003ekg kilogram\u003c/p\u003e\n\u003cp\u003emg milligram\u003c/p\u003e\n\u003cp\u003eml milliliter\u003c/p\u003e\n\u003cp\u003eN Newton\u003c/p\u003e\n\u003cp\u003eNo number\u003c/p\u003e\n\u003cp\u003e\u003cem\u003ep \u003c/em\u003eprobability\u003c/p\u003e\n\u003cp\u003ePMMA polymethyl methacrylate\u003c/p\u003e\n\u003cp\u003eSCE stem cell enhancer\u003c/p\u003e\n\u003cp\u003e\u0026deg;C Celsius or centigrade\u003c/p\u003e"},{"header":"Declarations","content":"\u003cp\u003e\u003cstrong\u003eAcknowledgements\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eNot applicable\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eAuthors\u0026rsquo; contributions\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eMYB, AB, BT contributed to the study design, the data acquisition, analysis, andinterpretation, and wrote the manuscript draft. BT, OS and OP contributed to the study design, and the interpretation of the results, and revised the manuscript. AB supervised the work. OS and OP provide funding. All authors have read and approved the final manuscript.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eFunding\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThis work was self-funded by the authors.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eAvailability of data and materials\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThe datasets generated and analyzed during the current study are availablefrom the corresponding author on reasonable request.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eEthics approval and consent to participate\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003ePermission for this study was obtained from the Animal Experiments Local Ethics Committee of Firat University (session number:2021/03) and performed in accordance with the Declaration of Helsinki.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eConsent for publication\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eNot applicable.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eCompeting interests\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThe authors declare no competing interests\u003c/p\u003e"},{"header":"References","content":"\u003col\u003e\n\u003cli\u003eJung RE, Zembic A, Pjetursson BE, et al. 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Rapid and selective mobilization of specific stem cell types after consumption of a polyphenol-rich extract from sea buckthorn berries (Hippophae) in healthy human subjects. \u003cem\u003eClin Interv Aging\u003c/em\u003e 2019;14:253-263. \u003c/li\u003e\n\u003cli\u003eBarbarić Starčević K, Lukač N, Jelić M, et al. Reciprocal Alterations in Osteoprogenitor and Immune Cell Populations in Rheumatoid Synovia. \u003cem\u003eInt J Mol Sci\u003c/em\u003e 2022;23:12379. \u003c/li\u003e\n\u003cli\u003eJensen GS, Hart AN, Zaske LA, et al. Mobilization of human CD34+ CD133+ and CD34+ CD133(-) stem cells in vivo by consumption of an extract from Aphanizomenon flos-aquae--related to modulation of CXCR4 expression by an L-selectin ligand? \u003cem\u003eCardiovasc Revasc Med\u003c/em\u003e 2007;8:189-202. \u003c/li\u003e\n\u003cli\u003eIrhimeh MR, Fitton JH, Lowenthal RM. 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Aloe macroclada from Madagascar triggers transient bone marrow stem cell mobilization. \u003cem\u003eJ Stem Cell Res Ther\u003c/em\u003e 2015;5:2. \u003c/li\u003e\n\u003cli\u003eWebster GA, Lehrke P. Development of a combined bovine colostrum and immune-stimulatory carbohydrate nutraceutical for enhancement of endogenous stem cell activity. \u003cem\u003eThe Open Nutraceuticals Journal\u003c/em\u003e 2013;6:35-44 \u003c/li\u003e\n\u003cli\u003ePatchen ML, Liang J, Vaudrain T, et al. Mobilization of peripheral blood progenitor cells by Betafectin PGG-Glucan alone and in combination with granulocyte colony-stimulating factor. \u003cem\u003eStem Cells\u003c/em\u003e 1998;16:208-217. \u003c/li\u003e\n\u003cli\u003eLin H, Cheung SW, Nesin M, et al. Enhancement of umbilical cord blood cell hematopoiesis by maitake beta-glucan is mediated by granulocyte colony-stimulating factor production. \u003cem\u003eClin Vaccine Immunol\u003c/em\u003e\u003cem\u003e \u003c/em\u003e2007;14:21-27. \u003c/li\u003e\n\u003cli\u003eLyon-Colbert A, Su S, Cude C. A Systematic Literature Review for Evidence of Aphanizomenon flos-aquae Toxigenicity in Recreational Waters and Toxicity of Dietary Supplements: 2000⁻2017. \u003cem\u003eToxins (Basel)\u003c/em\u003e 2018;10:254. \u003c/li\u003e\n\u003cli\u003eWeng CC, Ou KL, Wu CY, et al. Mechanism and Clinical Properties of StemBios Cell Therapy: Induction of Early Osseointegration in Novel Dental Implants. \u003cem\u003eInt J Oral Maxillofac Implants\u003c/em\u003e 2017;32:e47-e54. \u003c/li\u003e\n\u003cli\u003eFeng Y , Jiang Z , Zhang Y , et al. Stem-cell-derived ECM sheet-implant complexes for enhancing osseointegration. \u003cem\u003eBiomater Sci\u003c/em\u003e 2020;8:6647-56. \u003c/li\u003e\n\u003c/ol\u003e"}],"fulltextSource":"","fullText":"","funders":[],"hasAdminPriorityOnWorkflow":false,"hasManuscriptDocX":true,"hasOptedInToPreprint":true,"hasPassedJournalQc":"","hasAnyPriority":true,"hideJournal":true,"highlight":"","institution":"","isAcceptedByJournal":false,"isAuthorSuppliedPdf":false,"isDeskRejected":"","isHiddenFromSearch":false,"isInQc":false,"isInWorkflow":false,"isPdf":false,"isPdfUpToDate":true,"isWithdrawnOrRetracted":false,"journal":{"display":true,"email":"[email protected]","identity":"researchsquare","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":true,"externalIdentity":"","sideBox":"","snPcode":"","submissionUrl":"/submission","title":"Research Square","twitterHandle":"researchsquare","acdcEnabled":true,"dfaEnabled":false,"editorialSystem":"","reportingPortfolio":"","inReviewEnabled":false,"inReviewRevisionsEnabled":true},"keywords":"Biomechanics, Bone implant connection, Dental implants, Osseointegration, Stem cell enhancer","lastPublishedDoi":"10.21203/rs.3.rs-3790310/v1","lastPublishedDoiUrl":"https://doi.org/10.21203/rs.3.rs-3790310/v1","license":{"name":"CC BY 4.0","url":"https://creativecommons.org/licenses/by/4.0/"},"manuscriptAbstract":"\u003ch2\u003eBackground\u003c/h2\u003e \u003cp\u003eThis study aimed to conduct a biomechanical investigation of the effects of stem cell enhancer (SCE) application on osseointegration of titanium implants in rat tibias.\u003c/p\u003e\u003ch2\u003eMethods\u003c/h2\u003e \u003cp\u003eAfter surgical implantation of titanium implants in the metaphyseal part of the tibiae of rats, the rats were randomly divided into three equal groups (n\u0026thinsp;=\u0026thinsp;10): control group, SCE Dosage group 1 (SCE1), and SCE Dosage group 2 (SCE2). Each group consisted of 10 rats. The rats in the SCE1 and SCE2 groups were administered 6 mg and 12 mg SCE (Stemregen\u0026reg;), respectively, every day during the four-week of experimental period after surgery. Implants and surrounding bone tissues were collected for biomechanical bone-implant connection analysis at the end of the experimental procedures. One-way ANOVA was used for statistical analysis.\u003c/p\u003e\u003ch2\u003eResults\u003c/h2\u003e \u003cp\u003eThere was no significant difference in the biomechanical osseointegration values of the groups; however, the osseointegration of the SCE group was better than that of the control group.\u003c/p\u003e\u003ch2\u003eConclusions\u003c/h2\u003e \u003cp\u003eStem cell enhancers have the potential to improve the biomechanical bone-implant interface.\u003c/p\u003e","manuscriptTitle":"Biomechanical evaluation of the effect of a stem cell enhancer on bone implant connection of titanium implants","msid":"","msnumber":"","nonDraftVersions":[{"code":1,"date":"2024-01-08 17:35:43","doi":"10.21203/rs.3.rs-3790310/v1","editorialEvents":[{"type":"communityComments","content":0}],"status":"published","journal":{"display":true,"email":"[email protected]","identity":"researchsquare","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":true,"externalIdentity":"","sideBox":"","snPcode":"","submissionUrl":"/submission","title":"Research Square","twitterHandle":"researchsquare","acdcEnabled":true,"dfaEnabled":false,"editorialSystem":"","reportingPortfolio":"","inReviewEnabled":false,"inReviewRevisionsEnabled":true}}],"origin":"","ownerIdentity":"be066022-ab9b-4673-8eab-717e5e462844","owner":[],"postedDate":"January 8th, 2024","published":true,"recentEditorialEvents":[],"rejectedJournal":[],"revision":"","amendment":"","status":"posted","subjectAreas":[],"tags":[],"updatedAt":"2024-02-15T23:29:14+00:00","versionOfRecord":[],"versionCreatedAt":"2024-01-08 17:35:43","video":"","vorDoi":"","vorDoiUrl":"","workflowStages":[]},"version":"v1","identity":"rs-3790310","journalConfig":"researchsquare"},"__N_SSP":true},"page":"/article/[identity]/[[...version]]","query":{"redirect":"/article/rs-3790310","identity":"rs-3790310","version":["v1"]},"buildId":"J0_U0BvcaRcwD8yVFaRlm","isFallback":false,"isExperimentalCompile":false,"dynamicIds":[84888],"gssp":true,"scriptLoader":[]}