Comparison of the level of growth factors in injectable platelet-rich fibrin obtained from healthy individuals and patients with chronic periodontitis

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Methods Venous blood samples were obtained from patients diagnosed with chronic periodontitis (test group) and people with healthy periodontal conditions (control group). The I-PRF was acquired from blood that had been centrifuged. The growth factors released from i-PRFs were compared among groups using the ELISA test. Furthermore, the amounts of white blood cells (WBCs) and platelets were also compared. Results Comparison was made between the i-PRFs obtained from the test and control groups. No discernible disparity was detected. The test group had a markedly elevated white blood cell (WBC) count compared to the control group (P = 0.025). Nevertheless, the data obtained from the whole-blood analysis, including the platelet count, did not show any statistically significant differences between the groups (P = 0.387). Furthermore, there was no notable link observed between the white blood cells (WBCs) and growth factors in either of the groups (P > 0.05). Conclusions The levels of growth factors (VEGF, IGF-1, TGF-β1, PDGF-BB, EGF) in i-PRFs varied significantly among people. Nevertheless, there was no substantial disparity observed between the two groups. This suggests that the levels of growth factors may be unaffected by the periodontal disease state of the participants. These findings provide evidence for the practical application of i-PRFs as a method for delivering autologous growth factors. Injectable platelet rich fibrin Growth factors White blood cells Platelets Chronic periodontitis Figures Figure 1 Figure 2 Figure 3 Figure 4 Figure 5 Background Periodontitis is a condition characterized by the gradual deterioration of the periodontal ligament and alveolar bone. This deterioration is caused by inflammation resulting from the presence of harmful microorganisms around the tooth and the body's immune response to them [ 1 ]. The main objective of periodontal therapy is to halt the progression of periodontal disease and restore the structures that have been destroyed as a result of the illness [ 2 ]. During the typical wound healing process, several clusters of cells, particularly platelets, travel towards the affected region. Platelet cells initiate and establish a durable blood clot in the area. After the development of a blood clot, the growth factors contained in the granules within the platelets are released into the surrounding area, promoting the healing process [ 3 ]. The presence of significant quantities of platelet-derived growth factor (PDGF) and transforming growth factor-beta (TGF-β) in the α granules of platelets, and their subsequent release into the surrounding environment during the initial stages of wound healing, serves as compelling evidence for the crucial role played by these cells in the wound healing process [ 4 ]. Platelet-rich fibrin (PRF), a type of platelet concentrate from the second generation, is acquired by ensnaring platelets, abundant leukocytes, and cytokines within the fibrin network. Some platelet concentrations include PDGF, insulin-like growth factor-1 (IGF-1), TGF-β, fibroblast growth factor, and vascular endothelial growth factor (VEGF). It enhances the healing process of both soft and hard tissues by elevating the levels of growth factors, such as epidermal growth factor (EGF). Since PRF is derived from the patient's own body, there is no chance of disease transmission or immune response [ 5 – 9 ]. Over the past few years, different platelet concentrates have been created based on the method used, each with different centrifugation processes. Joseph Choukron created Injectable Platelet-Rich Fibrin (i-PRF) in 2014. The i-PRF is a fluid and injectable form, distinct from other PRF kinds acquired through alternative methods. It comprises not only leukocytes and thrombocytes but also hematopoietic stem cells and endothelial cells. Due to this rationale, it is regarded not only as a platelet concentrate but also as a "blood concentrate" [ 10 ]. I-PRF is acquired by collecting blood samples in anticoagulant-free tubes and subjecting the tubes to centrifugation at 700 rpm for 3 minutes (60 g) at room temperature. Following centrifugation, a distinct orange-colored structure (i-PRF) is created on the upper portion of the tube, while a red-colored structure including other components of the blood is generated at the bottom of the tube [ 10 ]. Injecting leukocyte-rich i-PRF, which is produced during a 3-minute centrifugation process, into soft tissue promotes vascularization. Additionally, it can be utilized by blending it with graft materials. The solid and inflexible structure that develops after a waiting period of 5–10 minutes for polymerization is referred to as "sticky bone". It can be conveniently used in rebuilding techniques [ 11 ]. Research has demonstrated that platelet-rich factors have an impact on both the amount and the characteristics of PRF [ 12 ]. Nevertheless, limited research has assessed the patient-specific variables that influence the quantity of growth factors derived from platelet-rich fibrin (PRF). No literature study has investigated the potential impact of the inflammatory condition of patients with chronic periodontitis or their platelet count on the levels of growth factors present and released by i-PRF. Hence, the primary objective of this study was to assess and contrast the concentrations of growth factors in i-PRF samples obtained from both healthy individuals and patients diagnosed with chronic periodontitis. Additionally, the study aimed to examine the correlation between the levels of growth factors in i-PRF and the count of white blood cells (WBCs) and platelets. Methods Collaborative Learning Groups The study was conducted at Alanya Alaaddin Keykubat University, namely in the Faculty of Dentistry's Department of Periodontology and the Faculty of Medicine's Department of Biochemistry. The study was commenced after obtaining approval from the Akdeniz University Faculty of Medicine Clinical Research Ethics Committee (dated 05.12.2018 and protocol# 859). The participants were chosen from the pool of patients who sought treatment at the periodontology clinic throughout the period from September 2019 to January 2020. The study's objectives and extent were thoroughly explained to the volunteers, along with comprehensive information regarding potential complications. A signed informed consent form was obtained from all participants who volunteered. The subjects were then categorized into two groups based on their periodontal health status. [ 13 ]. Control group: Individuals without periodontal disease Test group: Patients diagnosed with stage II and stage III, grade B chronic periodontitis Furthermore, severe generalized chronic periodontitis was categorized as individuals experiencing bleeding from over 30% of their teeth on probing, with probing depth equal to or greater than 7 mm, clinical attachment loss equal to or greater than 5 mm, and radiographic bone loss above 30%. The control group consisted of individuals who had periodontal health. The eligibility requirements encompassed those aged 30–65 years, who were free from systemic illness or had well-managed systemic illness. The determination of systemic disease relied on patient self-disclosure. The exclusion criteria included patients with an advanced and uncontrolled systemic disease (HbA1c ≥ 6.5%); patients who were morbidly obese (body mass index < 18.5 or ≥ 40 kg/m2); pregnant patients; patients using immunosuppressed drugs; patients who smoked or consumed alcohol; patients who had received chemotherapy and radiotherapy; patients who had used corticosteroids, anticoagulants, antiplatelet, or non-steroid anti-inflammatory drugs; patients who were drug abusers; patients who had received antibiotics treatment in the last 6 months; and patients who had received active periodontal treatment in the last 2 years [ 14 ]. Preparation for PRF Upon completing the collection of demographic data and conducting a periodontal examination. The patients were provided with follow-up numbers, and the tubes used to collect the i-PRF samples were labeled with tracking numbers. As part of the study, two blood samples were collected from the forearm area (specifically the antecubital vein) of each subject. One tube was utilized to prepare i-PRF, while the other tube was designated for whole blood, containing both white blood cells (WBC) and platelets. The duration it took to fill the tubes with blood was measured using a timer when the patient's blood was collected. The blood extraction procedure from the subjects was completed at a maximum duration of 25 seconds. The blood samples were promptly deposited in centrifuge equipment without delay (within 60 s). The blood was transferred to i-PRF tubes that are made of plastic and do not contain any anticoagulant. The sample was subjected to fixed-angle centrifugation in a tabletop centrifuge (ElektroMag M 415P, Istanbul, TR) at a maximum relative centrifugal force (RCF) of 700 for a duration of 3 minutes. Following centrifugation, the blood was divided into two distinct layers. The lower stratum comprised red blood cells, whereas the top stratum comprised plasma, platelets, and coagulation factors. The plasma and platelets that were divided were of a pale yellow color. Above the point where the two layers meet, the uppermost layer was meticulously extracted using a Pasteur pipette. Aspirate refers to a partially active injectable type of platelet-rich fibrin (PRF). The i-PRF samples were transferred to Eppendorf tubes. The sample was kept at a temperature of -80°C until the day of analysis. Evaluation of the complete blood count Automated hematology analyzers are used to measure the complete blood count (CBC), which quantifies the quantity of various blood cells within a specified volume of blood. We performed a calculation to determine the white blood cell (WBC) and platelet levels. Analysis of biological substances at a molecular level The levels of TGF-β, human IGF-1, human PDGF-BB, human EGF, and human VEGF-A were analyzed in duplicate using the ELISA method provided by Elabscience, located in TX, USA [ 15 , 18 ]. Following the completion of the reactions, absorbance measurements were conducted using a microplate reader instrument (Biotek Synergy H1, VT, USA). A concentration vs optical density plot was generated using the seven-level standard for each parameter. The concentrations of TGF-β, human IGF-1, human PDGF-BB, human EGF, and human VEGF-A in the samples were determined by utilizing this graph. The kit specifications for each parameter, including sensitivity, evaluation range, specificity, and repeatability, were as follows: TGF-β: 0.1 ng/mL; 1.16–10 ng/mL, 100% specificity, and coefficient of variation (CV) less than 10%; human IGF-1: 0.94 ng/mL, 1.56–100 ng/mL, 100% specificity, and CV less than 10%; human PDGF-BB: 18.75 pg/mL; 31.2–2000 pg/mL, 100% specificity, and CV less than 10%; human EGF: 2.35 pg/mL, 3.91–250 pg/mL, 100% specificity, and CV less than 10%; human VEGF-A: 18.75 pg/mL; 31.2–2000 pg/mL, 100% specificity, and CV less than 10%. Data analysis The statistical analyses were conducted using SPSS 19.0 (IBM Inc., IL, USA). The normality distribution was assessed using the Kolmogorov-Smirnov and Shapiro-Wilk tests. A significance level of 0.05 was employed while discussing the findings. The "independent t test" was employed to compare the groups because the data adhered to a normal distribution. Prior to the investigation, the reliability of the significance values was assessed by conducting power analysis. The calculation was based on measurements of growth factors. The power value for comparing the two groups in terms of growth factor mean and standard deviation values was 95%. Results Demographic findings The study involved a cohort of 60 participants, comprising 23 males and 37 females. The average age of the volunteers was 48.93 ± 3.66. The study did not find a statistically significant difference in the average age between the groups. The test group had an average age of 52.5 ± 3.46 years, whereas the control group had an average age of 45.36 ± 3.73 years (P = 0.332). The gender distribution between the two groups does not show any notable disparity. The demographic information of the volunteers is displayed in Table 1 . Table 1 Demographic characteristics of the patients Control Test N (%) P Sex Male 12 (40.0) 11 (36.7) 0.560 Female 18 (60.0) 19 (63.3) Mean ± standard deviation Age 45.36 ± 3.73 52.50 ± 3.46 0.332 Levels of growth factors The amounts of growth factors in i-PRF are shown in Table 2 and illustrated in Figs. 1 – 5 . The levels of VEGF (P = 0.084), IGF-1 (P = 0.268), TGF-β1 (P = 0.121), PDGF-BB (P = 0.388), and EGF (P = 0.069) in the i-PRF of the test and control groups did not show any significant difference. The concentrations of VEGF, IGF-1, TGF-β1, PDGF-BB, and EGF were elevated in the test group in comparison to the control group. However, the disparity between the groups did not reach statistical significance (P > 0.05). Table 2 The distribution of Growth factor levels in i-PRF between groups n Mean Standard deviation Min Max Median Q1 Q3 t P IGF1 (ng/mL) Control 30 38.82 14.99 10.08 66.22 41.18 27.77 60.43 -1.117 0.268 Test 30 42.46 9.69 22.30 58.60 42.15 34.30 57.27 PDGF-BB (pg/mL) Control 30 414.25 107.19 120.74 745.22 419.65 213.60 725.11 0.869 0.388 Test 30 466.28 102 105.21 836,41 419.24 231.46 822.45 VEGF (pg/mL) Control 30 375.69 76.10 196.25 622.52 369.36 250.54 620.66 1.737 0.084 Test 30 412.18 68.71 144.51 650.50 396.66 294.06 748.35 TGF-β1 (ng/mL) Control 30 21.50 8.11 14.41 44.62 20.40 15.45 30.16 1.478 0.121 Test 30 26.21 7.67 13.94 46.42 25.82 22.95 42.15 EGF (pg/mL) Control 30 138.62 22.39 69.36 181.23 155.00 1008.43 177.12 1.854 0.069 Test 30 154.82 40.97 81.66 196.32 157.59 145.26 192.75 Comparative analysis of white blood cells (WBC) and platelets The test group had a markedly elevated white blood cell (WBC) count compared to the control group (P = 0.025). Nevertheless, the data obtained by analyzing whole-blood samples, which included the platelet count, did not show any statistically significant differences between the groups (P = 0.387) (Table 3 ). Furthermore, there was no correlation observed between white blood cells (WBCs) and the growth factors examined in both the test and control groups (P > 0.05). Table 3 WBC and platelet values between groups Median (Q1–Q3) Control Test P WBC (cells/mm 3 ) 6.60 (5.25–7.55) 8.80 (7.38–9.78) 0.025 * Thrombocyte (cells/mm 3 ) 242.00 (216–287.75) 262.50 (230.75–298) 0.387 Q1 25 percentile, Q3 75 percentile. * The test group had a significantly higher WBC value than the control group ( P < 0.05). Discussions The findings of our investigation revealed that the levels of growth factors (IGF-1, EGF, VEGF, PDGF-BB, and TGF-1) released from i-PRFs varied significantly among individuals. Nevertheless, there was no notable disparity observed between the control and test groups. This investigation revealed that the levels of growth factors were not influenced by the periodontal disease condition of the participants. These findings provided support for the clinical application of i-PRF as a method of delivering autologous growth factors. No correlation was observed between the white blood cell (WBC) counts and the levels of growth factors across the groups, despite the fact that the data indicated higher WBC levels in the test group. The presence of elevated leukocytes indicated the expected occurrence of an inflammatory condition. The test group consisted of patients diagnosed with widespread severe and mild chronic periodontitis [ 20 ]. No significant differences were identified in the findings of the whole-blood analysis between the groups. These results were expected as all patients had either managed systemic disorders or were in a systemically healthy state. Multiple PRF preparation protocols are accessible. The PRF centrifuge procedure employed in this work effectively ensnared leukocytes and thrombocytes within the fibrin clot [ 20 ]. Nevertheless, TGF-β1, EGF, VEGF, IGF-1, and PDGF-BB primarily originate from platelets rather than leukocytes [ 21 ]. Therefore, the absence of a link between white blood cell count (WBC) and the levels of growth factors derived from platelet-rich fibrin (PRF) is to be expected. There is currently no available information on the impact of whole-blood analysis and the presence of chronic periodontitis on the quantity or release of growth factors from i-PRF. In 2015, Maurao et al. achieved a distinct platelet concentration of i-PRF using horizontal centrifugation using a B-40 centrifuge (RDE, Brazil) for a duration of 2 minutes at 3300 rpm. The tubes used in this process contained 9 mL of venous blood and were not treated with any anticoagulants (11). Miron et al. obtained i-PRF by centrifuging 10 mL of whole blood without anticoagulant in tubes at a speed of 700 rpm for 3 minutes (equivalent to 60g) at room temperature using a Duo Centrifuge, a device specifically designed for obtaining PRF, manufactured in Nice, France (10). It is widely accepted that i-PRF contains not just leukocytes and thrombocytes, but also mesenchymal stem cells and endothelial cells. Thus, it is considered a "blood concentrate" rather than solely a platelet concentrate [ 22 ]. Chang et al. conducted a comparison of the release of growth factors, white blood cell count, and thrombocyte amount in the PRF membrane and exudate of both periodontally healthy persons (control group) and patients with chronic periodontitis (test group). No significant disparity in the quantity of growth factor was observed between the test and control groups. The test group exhibited a markedly elevated WBC count. No association was found between white blood cell count (WBC), platelet count, and growth factors [ 14 ]. The findings of the study conducted by Chang et al. were in line with the results of the current investigation. Kobayashi et al. examined the growth factors present in platelet rich plasma (PRP), PRF, and advanced-platelet rich fibrin (A-PRF) derived from individuals who are in good health. The quantities and release of growth factors, including TGF-β, IGF, VEGF, EGF, PDGF-AA, PDGF-BB, and PDGF-AB, were assessed from platelet concentrates using ELISA over a duration of 10 days. The authors proposed that factors connected to the patient could influence the release of growth factors. As a result, there may be a significant variation in the quantities of growth factors released [ 23 ]. Miron et al. conducted a comparison of the growth factors present in platelet-rich plasma (PRP) and injectable platelet-rich fibrin (i-PRF). According to their statement, PRP produced a greater amount of growth factors initially, but i-PRF released higher levels of PDGF-AA, PDGF-AB, EGF, and IGF over a longer duration [ 10 ]. In their study comparing i-PRF with PRP in gingival fibroblasts on titanium-implant surfaces, Wang et al. discovered higher levels of PDGF and TGF-β in the i-PRF group. Furthermore, the i-PRF group had a higher level of cellular migration and collagen-1, as evidenced by the detection of these factors [ 24 ]. In their investigation on the release of leukocytes, platelets, and growth factors in liquid PRF products, Choukroun et al. found that low-speed centrifugation resulted in an increase in these components in PRF-based matrices [ 25 ]. Varela et al. assessed the liberation of growth factors in i-PRF, the gene expression of type I collagen, the morphological characteristics, and the cellular composition. According to their statement, i-PRF, which includes platelets, growth factors, type I collagen, leukocytes, and osteocalcin, may be an effective method for promoting the healing of both hard and soft tissues [ 26 ]. The release of growth factors from PRF varied among patients. However, the levels of growth factors derived from the i-PRF did not exhibit a statistically significant disparity between the test and control groups. The findings of the whole-blood analysis indicated that the test group exhibited a statistically significant increase in white blood cell count (P < 0.05). However, there was no substantial correlation observed between the white blood cell counts and the levels of the growth factors in i-PRF. The current investigation is limited by its modest and straightforward nature. Additional constraints of the study include the exclusive acquisition of i-PRF and the absence of evaluation for other types of PRF. Furthermore, the determination of participants' systemic status relied solely on their self-reported remarks. Conclusions Based on the constraints of this investigation, it was determined that i-PRF can be utilized as a self-derived source of growth factors that remain unaffected by the periodontal condition and white blood cell counts of healthy individuals. Declarations Acknowledgements Not applicable Authors contributions: Study concept and design: BK. Data Acquisition: BK. Analysis and interpretation of data: BK, HBS. Writing - original draft preparation: BK. Writing - review and editing: BK, HBS. All authors read and approved the final manuscript. Funding This study was supported by scientific research projects (ALKU BAP, grant 2019-06-01-MAP02). Data Availability All data generated or analyzed during this study are included in this published article & supporting material. Ethics approval and consent to participate The study was initiated following the approval of the Akdeniz University Faculty of Medicine Clinical Research Ethics Committee (dated 05.12.2018 and protocol# 859). All procedures performed in studies involving human participants were in accordance with the ethical standards of the institutional research committee. Informed consent term was obtained from every individual participant included in this study. Consent for publication Not applicable Competing interests The authors declare that they have no competing interests Author details 1 Alanya Alaaddin Keykubat University, Faculty of Dentistry, Department of Periodontology, Antalya, Turkey 2 Mardin Artuklu University, Faculty of Medicine, Department of Biochemistry, Mardin, Turkey References Ohlrich EJ, Cullinan MP, Seymour GJ. The immunopathogenesis of periodontal disease. Aust Dent J. 2009;54(Suppl):2. https://doi.org/10.1111/j.1834-7819.2009.01139.x . S-10S. Gotlow J, Nyman S, Karring T. 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Cite Share Download PDF Status: Published Journal Publication published 03 May, 2024 Read the published version in BMC Oral Health → Version 1 posted Editorial decision: Revision requested 11 Mar, 2024 Reviews received at journal 22 Feb, 2024 Reviewers agreed at journal 13 Feb, 2024 Reviewers agreed at journal 11 Feb, 2024 Reviewers agreed at journal 11 Feb, 2024 Reviewers invited by journal 11 Feb, 2024 Editor assigned by journal 05 Feb, 2024 Editor invited by journal 05 Feb, 2024 Submission checks completed at journal 05 Feb, 2024 First submitted to journal 10 Jan, 2024 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. 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Karci","email":"data:image/png;base64,iVBORw0KGgoAAAANSUhEUgAAAZAAAAAyAQMAAABI0h/eAAAABlBMVEX///8AAABVwtN+AAAACXBIWXMAAA7EAAAOxAGVKw4bAAAA50lEQVRIie3PsQrCMBCA4QuBZIl2LSj4Ci0FRSj2YQSn4Kw4KBQ69QWK79HZksGl7nVTu7gIurkIXrq5NI6C+YdCyn0kB2Cz/WAepRv8umLA0+aH6JoJQbII+35aAuyQMCMBTcpZCJVsCBjJiJP4+kiUINmtqB9y0mdAz5eqhYxjkowzJLw3n3pFPsWHsSCQbQ9TJAk6+patHLpFTpEI1jOSFxI4lpqsvyJxjesLqIQmykz0LiRdhMJPZeAd8r1g1LDLyFHq/vTcaMBL/7TMV5HD43PdRjDmfp5p+3gzcjfP2Gw221/3BkRFRslZUO5mAAAAAElFTkSuQmCC","orcid":"","institution":"Alanya Alaaddin Keykubat University","correspondingAuthor":true,"prefix":"","firstName":"Bilge","middleName":"","lastName":"Karci","suffix":""},{"id":271337605,"identity":"0d4114e6-ef40-44a3-92f5-5b1f6ef5f0e1","order_by":1,"name":"Hasan Basri Savas","email":"","orcid":"","institution":"Mardin Artuklu University","correspondingAuthor":false,"prefix":"","firstName":"Hasan","middleName":"Basri","lastName":"Savas","suffix":""}],"badges":[],"createdAt":"2024-01-10 12:46:34","currentVersionCode":1,"declarations":"","doi":"10.21203/rs.3.rs-3850305/v1","doiUrl":"https://doi.org/10.21203/rs.3.rs-3850305/v1","draftVersion":[],"editorialEvents":[{"content":"https://doi.org/10.1186/s12903-024-04301-x","type":"published","date":"2024-05-03T19:57:54+00:00"}],"editorialNote":"","failedWorkflow":false,"files":[{"id":50817500,"identity":"2df463ad-deab-41cc-9824-aa76f57378c0","added_by":"auto","created_at":"2024-02-07 20:10:14","extension":"jpg","order_by":1,"title":"Figure 1","display":"","copyAsset":false,"role":"figure","size":580459,"visible":true,"origin":"","legend":"\u003cp\u003eComparison of insulin-like growth factor (IGF)1 levels between the groups.\u003c/p\u003e","description":"","filename":"Figure1..jpg","url":"https://assets-eu.researchsquare.com/files/rs-3850305/v1/1aaa5598587ca684f7323cfd.jpg"},{"id":50817499,"identity":"0e7b9e69-6a5f-42d3-97d8-42bf7f28c840","added_by":"auto","created_at":"2024-02-07 20:10:14","extension":"jpg","order_by":2,"title":"Figure 2","display":"","copyAsset":false,"role":"figure","size":592387,"visible":true,"origin":"","legend":"\u003cp\u003eComparison of vascular endothelial growth factor (VEGF) levels between the groups.\u003c/p\u003e","description":"","filename":"Figure2..jpg","url":"https://assets-eu.researchsquare.com/files/rs-3850305/v1/be9788fe6922eaf10740cfd4.jpg"},{"id":50817501,"identity":"89f51ed0-6adb-4c4e-a636-34fab063339f","added_by":"auto","created_at":"2024-02-07 20:10:14","extension":"jpg","order_by":3,"title":"Figure 3","display":"","copyAsset":false,"role":"figure","size":594019,"visible":true,"origin":"","legend":"\u003cp\u003eComparison of transforming growth factor (TGF)-β1 levels between the groups.\u003c/p\u003e","description":"","filename":"Figure3..jpg","url":"https://assets-eu.researchsquare.com/files/rs-3850305/v1/0d48759e2338f8219ca9fe21.jpg"},{"id":50817502,"identity":"7dbffef5-5223-486e-8004-53ff34e9e25b","added_by":"auto","created_at":"2024-02-07 20:10:14","extension":"jpg","order_by":4,"title":"Figure 4","display":"","copyAsset":false,"role":"figure","size":579701,"visible":true,"origin":"","legend":"\u003cp\u003eComparison of endothelial growth factor (EGF) levels between the groups.\u003c/p\u003e","description":"","filename":"Figure4..jpg","url":"https://assets-eu.researchsquare.com/files/rs-3850305/v1/c49d5a50107162ad5cb49636.jpg"},{"id":50817503,"identity":"96d9b237-e636-4325-ae42-a13088d34451","added_by":"auto","created_at":"2024-02-07 20:10:14","extension":"jpg","order_by":5,"title":"Figure 5","display":"","copyAsset":false,"role":"figure","size":579127,"visible":true,"origin":"","legend":"\u003cp\u003eComparison of platelet-derived growth factor (PDGF)-BB levels between the groups.\u003c/p\u003e","description":"","filename":"Figure5..jpg","url":"https://assets-eu.researchsquare.com/files/rs-3850305/v1/1e563ac2c2176ee1198b981f.jpg"},{"id":56043176,"identity":"f8fdd3cf-b87c-4a80-aec7-d501eaf9e044","added_by":"auto","created_at":"2024-05-07 20:10:36","extension":"pdf","order_by":0,"title":"","display":"","copyAsset":false,"role":"manuscript-pdf","size":705953,"visible":true,"origin":"","legend":"","description":"","filename":"manuscript.pdf","url":"https://assets-eu.researchsquare.com/files/rs-3850305/v1/df806d0d-5eef-4c17-b91b-03015173de95.pdf"}],"financialInterests":"No competing interests reported.","formattedTitle":"Comparison of the level of growth factors in injectable platelet-rich fibrin obtained from healthy individuals and patients with chronic periodontitis","fulltext":[{"header":"Background","content":"\u003cp\u003ePeriodontitis is a condition characterized by the gradual deterioration of the periodontal ligament and alveolar bone. This deterioration is caused by inflammation resulting from the presence of harmful microorganisms around the tooth and the body's immune response to them [\u003cspan citationid=\"CR1\" class=\"CitationRef\"\u003e1\u003c/span\u003e]. The main objective of periodontal therapy is to halt the progression of periodontal disease and restore the structures that have been destroyed as a result of the illness [\u003cspan citationid=\"CR2\" class=\"CitationRef\"\u003e2\u003c/span\u003e].\u003c/p\u003e \u003cp\u003eDuring the typical wound healing process, several clusters of cells, particularly platelets, travel towards the affected region. Platelet cells initiate and establish a durable blood clot in the area. After the development of a blood clot, the growth factors contained in the granules within the platelets are released into the surrounding area, promoting the healing process [\u003cspan citationid=\"CR3\" class=\"CitationRef\"\u003e3\u003c/span\u003e]. The presence of significant quantities of platelet-derived growth factor (PDGF) and transforming growth factor-beta (TGF-β) in the α granules of platelets, and their subsequent release into the surrounding environment during the initial stages of wound healing, serves as compelling evidence for the crucial role played by these cells in the wound healing process [\u003cspan citationid=\"CR4\" class=\"CitationRef\"\u003e4\u003c/span\u003e].\u003c/p\u003e \u003cp\u003ePlatelet-rich fibrin (PRF), a type of platelet concentrate from the second generation, is acquired by ensnaring platelets, abundant leukocytes, and cytokines within the fibrin network. Some platelet concentrations include PDGF, insulin-like growth factor-1 (IGF-1), TGF-β, fibroblast growth factor, and vascular endothelial growth factor (VEGF). It enhances the healing process of both soft and hard tissues by elevating the levels of growth factors, such as epidermal growth factor (EGF). Since PRF is derived from the patient's own body, there is no chance of disease transmission or immune response [\u003cspan additionalcitationids=\"CR6 CR7 CR8\" citationid=\"CR5\" class=\"CitationRef\"\u003e5\u003c/span\u003e\u0026ndash;\u003cspan citationid=\"CR9\" class=\"CitationRef\"\u003e9\u003c/span\u003e].\u003c/p\u003e \u003cp\u003eOver the past few years, different platelet concentrates have been created based on the method used, each with different centrifugation processes. Joseph Choukron created Injectable Platelet-Rich Fibrin (i-PRF) in 2014. The i-PRF is a fluid and injectable form, distinct from other PRF kinds acquired through alternative methods. It comprises not only leukocytes and thrombocytes but also hematopoietic stem cells and endothelial cells. Due to this rationale, it is regarded not only as a platelet concentrate but also as a \"blood concentrate\" [\u003cspan citationid=\"CR10\" class=\"CitationRef\"\u003e10\u003c/span\u003e].\u003c/p\u003e \u003cp\u003eI-PRF is acquired by collecting blood samples in anticoagulant-free tubes and subjecting the tubes to centrifugation at 700 rpm for 3 minutes (60 g) at room temperature. Following centrifugation, a distinct orange-colored structure (i-PRF) is created on the upper portion of the tube, while a red-colored structure including other components of the blood is generated at the bottom of the tube [\u003cspan citationid=\"CR10\" class=\"CitationRef\"\u003e10\u003c/span\u003e].\u003c/p\u003e \u003cp\u003eInjecting leukocyte-rich i-PRF, which is produced during a 3-minute centrifugation process, into soft tissue promotes vascularization. Additionally, it can be utilized by blending it with graft materials. The solid and inflexible structure that develops after a waiting period of 5\u0026ndash;10 minutes for polymerization is referred to as \"sticky bone\". It can be conveniently used in rebuilding techniques [\u003cspan citationid=\"CR11\" class=\"CitationRef\"\u003e11\u003c/span\u003e].\u003c/p\u003e \u003cp\u003eResearch has demonstrated that platelet-rich factors have an impact on both the amount and the characteristics of PRF [\u003cspan citationid=\"CR12\" class=\"CitationRef\"\u003e12\u003c/span\u003e]. Nevertheless, limited research has assessed the patient-specific variables that influence the quantity of growth factors derived from platelet-rich fibrin (PRF). No literature study has investigated the potential impact of the inflammatory condition of patients with chronic periodontitis or their platelet count on the levels of growth factors present and released by i-PRF.\u003c/p\u003e \u003cp\u003eHence, the primary objective of this study was to assess and contrast the concentrations of growth factors in i-PRF samples obtained from both healthy individuals and patients diagnosed with chronic periodontitis. Additionally, the study aimed to examine the correlation between the levels of growth factors in i-PRF and the count of white blood cells (WBCs) and platelets.\u003c/p\u003e"},{"header":"Methods","content":"\u003cdiv id=\"Sec3\" class=\"Section2\"\u003e \u003ch2\u003eCollaborative Learning Groups\u003c/h2\u003e \u003cp\u003eThe study was conducted at Alanya Alaaddin Keykubat University, namely in the Faculty of Dentistry's Department of Periodontology and the Faculty of Medicine's Department of Biochemistry. The study was commenced after obtaining approval from the Akdeniz University Faculty of Medicine Clinical Research Ethics Committee (dated 05.12.2018 and protocol# 859). The participants were chosen from the pool of patients who sought treatment at the periodontology clinic throughout the period from September 2019 to January 2020. The study's objectives and extent were thoroughly explained to the volunteers, along with comprehensive information regarding potential complications. A signed informed consent form was obtained from all participants who volunteered. The subjects were then categorized into two groups based on their periodontal health status. [\u003cspan citationid=\"CR13\" class=\"CitationRef\"\u003e13\u003c/span\u003e].\u003c/p\u003e \u003cp\u003eControl group: Individuals without periodontal disease\u003c/p\u003e \u003cp\u003eTest group: Patients diagnosed with stage II and stage III, grade B chronic periodontitis\u003c/p\u003e \u003cp\u003eFurthermore, severe generalized chronic periodontitis was categorized as individuals experiencing bleeding from over 30% of their teeth on probing, with probing depth equal to or greater than 7 mm, clinical attachment loss equal to or greater than 5 mm, and radiographic bone loss above 30%. The control group consisted of individuals who had periodontal health. The eligibility requirements encompassed those aged 30\u0026ndash;65 years, who were free from systemic illness or had well-managed systemic illness. The determination of systemic disease relied on patient self-disclosure. The exclusion criteria included patients with an advanced and uncontrolled systemic disease (HbA1c\u0026thinsp;\u0026ge;\u0026thinsp;6.5%); patients who were morbidly obese (body mass index\u0026thinsp;\u0026lt;\u0026thinsp;18.5 or \u0026ge;\u0026thinsp;40 kg/m2); pregnant patients; patients using immunosuppressed drugs; patients who smoked or consumed alcohol; patients who had received chemotherapy and radiotherapy; patients who had used corticosteroids, anticoagulants, antiplatelet, or non-steroid anti-inflammatory drugs; patients who were drug abusers; patients who had received antibiotics treatment in the last 6 months; and patients who had received active periodontal treatment in the last 2 years [\u003cspan citationid=\"CR14\" class=\"CitationRef\"\u003e14\u003c/span\u003e].\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec4\" class=\"Section2\"\u003e \u003ch2\u003ePreparation for PRF\u003c/h2\u003e \u003cp\u003eUpon completing the collection of demographic data and conducting a periodontal examination. The patients were provided with follow-up numbers, and the tubes used to collect the i-PRF samples were labeled with tracking numbers. As part of the study, two blood samples were collected from the forearm area (specifically the antecubital vein) of each subject. One tube was utilized to prepare i-PRF, while the other tube was designated for whole blood, containing both white blood cells (WBC) and platelets.\u003c/p\u003e \u003cp\u003eThe duration it took to fill the tubes with blood was measured using a timer when the patient's blood was collected. The blood extraction procedure from the subjects was completed at a maximum duration of 25 seconds. The blood samples were promptly deposited in centrifuge equipment without delay (within 60 s).\u003c/p\u003e \u003cp\u003eThe blood was transferred to i-PRF tubes that are made of plastic and do not contain any anticoagulant. The sample was subjected to fixed-angle centrifugation in a tabletop centrifuge (ElektroMag M 415P, Istanbul, TR) at a maximum relative centrifugal force (RCF) of 700 for a duration of 3 minutes. Following centrifugation, the blood was divided into two distinct layers. The lower stratum comprised red blood cells, whereas the top stratum comprised plasma, platelets, and coagulation factors. The plasma and platelets that were divided were of a pale yellow color. Above the point where the two layers meet, the uppermost layer was meticulously extracted using a Pasteur pipette. Aspirate refers to a partially active injectable type of platelet-rich fibrin (PRF). The i-PRF samples were transferred to Eppendorf tubes. The sample was kept at a temperature of -80\u0026deg;C until the day of analysis.\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec5\" class=\"Section2\"\u003e \u003ch2\u003eEvaluation of the complete blood count\u003c/h2\u003e \u003cp\u003eAutomated hematology analyzers are used to measure the complete blood count (CBC), which quantifies the quantity of various blood cells within a specified volume of blood. We performed a calculation to determine the white blood cell (WBC) and platelet levels.\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec6\" class=\"Section2\"\u003e \u003ch2\u003eAnalysis of biological substances at a molecular level\u003c/h2\u003e \u003cp\u003eThe levels of TGF-β, human IGF-1, human PDGF-BB, human EGF, and human VEGF-A were analyzed in duplicate using the ELISA method provided by Elabscience, located in TX, USA [\u003cspan citationid=\"CR15\" class=\"CitationRef\"\u003e15\u003c/span\u003e, \u003cspan citationid=\"CR18\" class=\"CitationRef\"\u003e18\u003c/span\u003e]. Following the completion of the reactions, absorbance measurements were conducted using a microplate reader instrument (Biotek Synergy H1, VT, USA). A concentration vs optical density plot was generated using the seven-level standard for each parameter. The concentrations of TGF-β, human IGF-1, human PDGF-BB, human EGF, and human VEGF-A in the samples were determined by utilizing this graph. The kit specifications for each parameter, including sensitivity, evaluation range, specificity, and repeatability, were as follows: TGF-β: 0.1 ng/mL; 1.16\u0026ndash;10 ng/mL, 100% specificity, and coefficient of variation (CV) less than 10%; human IGF-1: 0.94 ng/mL, 1.56\u0026ndash;100 ng/mL, 100% specificity, and CV less than 10%; human PDGF-BB: 18.75 pg/mL; 31.2\u0026ndash;2000 pg/mL, 100% specificity, and CV less than 10%; human EGF: 2.35 pg/mL, 3.91\u0026ndash;250 pg/mL, 100% specificity, and CV less than 10%; human VEGF-A: 18.75 pg/mL; 31.2\u0026ndash;2000 pg/mL, 100% specificity, and CV less than 10%.\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec7\" class=\"Section2\"\u003e \u003ch2\u003eData analysis\u003c/h2\u003e \u003cp\u003eThe statistical analyses were conducted using SPSS 19.0 (IBM Inc., IL, USA). The normality distribution was assessed using the Kolmogorov-Smirnov and Shapiro-Wilk tests. A significance level of 0.05 was employed while discussing the findings.\u003c/p\u003e \u003cp\u003eThe \"independent t test\" was employed to compare the groups because the data adhered to a normal distribution.\u003c/p\u003e \u003cp\u003ePrior to the investigation, the reliability of the significance values was assessed by conducting power analysis. The calculation was based on measurements of growth factors. The power value for comparing the two groups in terms of growth factor mean and standard deviation values was 95%.\u003c/p\u003e \u003c/div\u003e"},{"header":"Results","content":"\u003cdiv id=\"Sec9\" class=\"Section2\"\u003e \u003ch2\u003eDemographic findings\u003c/h2\u003e \u003cp\u003eThe study involved a cohort of 60 participants, comprising 23 males and 37 females. The average age of the volunteers was 48.93\u0026thinsp;\u0026plusmn;\u0026thinsp;3.66. The study did not find a statistically significant difference in the average age between the groups. The test group had an average age of 52.5\u0026thinsp;\u0026plusmn;\u0026thinsp;3.46 years, whereas the control group had an average age of 45.36\u0026thinsp;\u0026plusmn;\u0026thinsp;3.73 years (P\u0026thinsp;=\u0026thinsp;0.332). The gender distribution between the two groups does not show any notable disparity. The demographic information of the volunteers is displayed in 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\u003eDemographic characteristics of the patients\u003c/p\u003e \u003c/div\u003e \u003c/caption\u003e \u003ccolgroup cols=\"6\"\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c1\" colnum=\"1\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c2\" colnum=\"2\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c3\" colnum=\"3\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c4\" colnum=\"4\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c5\" colnum=\"5\"\u003e\u003c/div\u003e \u003cdiv align=\"char\" char=\".\" class=\"colspec\" colname=\"c6\" colnum=\"6\"\u003e\u003c/div\u003e \u003cthead\u003e \u003ctr\u003e \u003cth align=\"left\" colname=\"c1\"\u003e\u0026nbsp;\u003c/th\u003e \u003cth align=\"left\" colname=\"c2\"\u003e\u0026nbsp;\u003c/th\u003e \u003cth align=\"left\" colname=\"c3\"\u003e \u003cp\u003eControl\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c4\"\u003e \u003cp\u003eTest\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c5\"\u003e\u0026nbsp;\u003c/th\u003e \u003cth align=\"left\" colname=\"c6\"\u003e\u0026nbsp;\u003c/th\u003e \u003c/tr\u003e \u003ctr\u003e \u003cth align=\"left\" colname=\"c1\"\u003e\u0026nbsp;\u003c/th\u003e \u003cth align=\"left\" colname=\"c2\"\u003e\u0026nbsp;\u003c/th\u003e \u003cth align=\"left\" colspan=\"3\" nameend=\"c5\" namest=\"c3\"\u003e \u003cp\u003e\u003cem\u003eN\u003c/em\u003e (%)\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c6\"\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\u003eSex\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eMale\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e12 (40.0)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e11 (36.7)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c6\"\u003e \u003cp\u003e0.560\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eFemale\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e18 (60.0)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e19 (63.3)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e\u0026nbsp;\u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colspan=\"3\" nameend=\"c5\" namest=\"c3\"\u003e \u003cp\u003e\u003cb\u003eMean\u0026thinsp;\u0026plusmn;\u0026thinsp;standard deviation\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e\u0026nbsp;\u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eAge\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e45.36\u0026thinsp;\u0026plusmn;\u0026thinsp;3.73\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e52.50\u0026thinsp;\u0026plusmn;\u0026thinsp;3.46\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c6\"\u003e \u003cp\u003e0.332\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003c/tbody\u003e \u003c/colgroup\u003e \u003c/table\u003e\u003c/div\u003e \u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec10\" class=\"Section2\"\u003e \u003ch2\u003eLevels of growth factors\u003c/h2\u003e \u003cp\u003eThe amounts of growth factors in i-PRF are shown in Table\u0026nbsp;\u003cspan refid=\"Tab2\" class=\"InternalRef\"\u003e2\u003c/span\u003e and illustrated in Figs.\u0026nbsp;\u003cspan refid=\"Fig1\" class=\"InternalRef\"\u003e1\u003c/span\u003e\u0026ndash;\u003cspan refid=\"Fig5\" class=\"InternalRef\"\u003e5\u003c/span\u003e. The levels of VEGF (P\u0026thinsp;=\u0026thinsp;0.084), IGF-1 (P\u0026thinsp;=\u0026thinsp;0.268), TGF-β1 (P\u0026thinsp;=\u0026thinsp;0.121), PDGF-BB (P\u0026thinsp;=\u0026thinsp;0.388), and EGF (P\u0026thinsp;=\u0026thinsp;0.069) in the i-PRF of the test and control groups did not show any significant difference. The concentrations of VEGF, IGF-1, TGF-β1, PDGF-BB, and EGF were elevated in the test group in comparison to the control group. However, the disparity between the groups did not reach statistical significance (P\u0026thinsp;\u0026gt;\u0026thinsp;0.05).\u003c/p\u003e \u003cp\u003e \u003cdiv class=\"gridtable\"\u003e\u003ctable float=\"Yes\" id=\"Tab2\" border=\"1\"\u003e \u003ccaption language=\"En\"\u003e \u003cdiv class=\"CaptionNumber\"\u003eTable 2\u003c/div\u003e \u003cdiv class=\"CaptionContent\"\u003e \u003cp\u003eThe distribution of Growth factor levels in i-PRF between groups\u003c/p\u003e \u003c/div\u003e \u003c/caption\u003e \u003ccolgroup cols=\"13\"\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c1\" colnum=\"1\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c2\" colnum=\"2\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c3\" colnum=\"3\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c4\" colnum=\"4\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c5\" colnum=\"5\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c6\" colnum=\"6\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c7\" colnum=\"7\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c8\" colnum=\"8\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c9\" colnum=\"9\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c10\" colnum=\"10\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c11\" colnum=\"11\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c12\" colnum=\"12\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c13\" colnum=\"13\"\u003e\u003c/div\u003e \u003cthead\u003e \u003ctr\u003e \u003cth align=\"left\" colspan=\"2\" nameend=\"c2\" namest=\"c1\"\u003e\u0026nbsp;\u003c/th\u003e \u003cth align=\"left\" colname=\"c3\"\u003e \u003cp\u003e\u003cem\u003en\u003c/em\u003e\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c4\"\u003e \u003cp\u003eMean\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c5\"\u003e \u003cp\u003eStandard deviation\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c6\"\u003e \u003cp\u003eMin\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c7\"\u003e \u003cp\u003eMax\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colspan=\"2\" nameend=\"c9\" namest=\"c8\"\u003e \u003cp\u003eMedian\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c10\"\u003e \u003cp\u003eQ1\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c11\"\u003e \u003cp\u003eQ3\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c12\"\u003e \u003cp\u003e\u003cem\u003et\u003c/em\u003e\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c13\"\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\" morerows=\"1\" rowspan=\"2\"\u003e \u003cp\u003eIGF1 (ng/mL)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eControl\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e30\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e38.82\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e14.99\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e10.08\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e66.22\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003e41.18\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colspan=\"2\" nameend=\"c10\" namest=\"c9\"\u003e \u003cp\u003e27.77\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c11\"\u003e \u003cp\u003e60.43\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c12\" morerows=\"1\" rowspan=\"2\"\u003e \u003cp\u003e-1.117\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c13\" morerows=\"1\" rowspan=\"2\"\u003e \u003cp\u003e0.268\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eTest\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e30\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e42.46\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e9.69\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e22.30\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e58.60\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003e42.15\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colspan=\"2\" nameend=\"c10\" namest=\"c9\"\u003e \u003cp\u003e34.30\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c11\"\u003e \u003cp\u003e57.27\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\" morerows=\"1\" rowspan=\"2\"\u003e \u003cp\u003ePDGF-BB (pg/mL)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eControl\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e30\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e414.25\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e107.19\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e120.74\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e745.22\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003e419.65\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colspan=\"2\" nameend=\"c10\" namest=\"c9\"\u003e \u003cp\u003e213.60\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c11\"\u003e \u003cp\u003e725.11\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c12\" morerows=\"1\" rowspan=\"2\"\u003e \u003cp\u003e0.869\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c13\" morerows=\"1\" rowspan=\"2\"\u003e \u003cp\u003e0.388\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eTest\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e30\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e466.28\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e102\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e105.21\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e836,41\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003e419.24\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colspan=\"2\" nameend=\"c10\" namest=\"c9\"\u003e \u003cp\u003e231.46\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c11\"\u003e \u003cp\u003e822.45\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\" morerows=\"1\" rowspan=\"2\"\u003e \u003cp\u003eVEGF (pg/mL)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eControl\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e30\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e375.69\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e76.10\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e196.25\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e622.52\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003e369.36\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colspan=\"2\" nameend=\"c10\" namest=\"c9\"\u003e \u003cp\u003e250.54\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c11\"\u003e \u003cp\u003e620.66\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c12\" morerows=\"1\" rowspan=\"2\"\u003e \u003cp\u003e1.737\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c13\" morerows=\"1\" rowspan=\"2\"\u003e \u003cp\u003e0.084\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eTest\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e30\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e412.18\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e68.71\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e144.51\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e650.50\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003e396.66\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colspan=\"2\" nameend=\"c10\" namest=\"c9\"\u003e \u003cp\u003e294.06\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c11\"\u003e \u003cp\u003e748.35\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\" morerows=\"1\" rowspan=\"2\"\u003e \u003cp\u003eTGF-β1 (ng/mL)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eControl\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e30\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e21.50\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e8.11\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e14.41\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e44.62\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003e20.40\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colspan=\"2\" nameend=\"c10\" namest=\"c9\"\u003e \u003cp\u003e15.45\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c11\"\u003e \u003cp\u003e30.16\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c12\" morerows=\"1\" rowspan=\"2\"\u003e \u003cp\u003e1.478\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c13\" morerows=\"1\" rowspan=\"2\"\u003e \u003cp\u003e0.121\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eTest\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e30\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e26.21\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e7.67\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e13.94\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e46.42\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003e25.82\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colspan=\"2\" nameend=\"c10\" namest=\"c9\"\u003e \u003cp\u003e22.95\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c11\"\u003e \u003cp\u003e42.15\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\" morerows=\"1\" rowspan=\"2\"\u003e \u003cp\u003eEGF (pg/mL)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eControl\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e30\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e138.62\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e22.39\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e69.36\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e181.23\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003e155.00\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colspan=\"2\" nameend=\"c10\" namest=\"c9\"\u003e \u003cp\u003e1008.43\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c11\"\u003e \u003cp\u003e177.12\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c12\" morerows=\"1\" rowspan=\"2\"\u003e \u003cp\u003e1.854\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c13\" morerows=\"1\" rowspan=\"2\"\u003e \u003cp\u003e0.069\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eTest\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e30\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e154.82\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e40.97\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e81.66\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e196.32\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003e157.59\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colspan=\"2\" nameend=\"c10\" namest=\"c9\"\u003e \u003cp\u003e145.26\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c11\"\u003e \u003cp\u003e192.75\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 \u003c/p\u003e \u003cp\u003e \u003c/p\u003e \u003cp\u003e \u003c/p\u003e \u003cp\u003e \u003c/p\u003e \u003cp\u003e \u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec11\" class=\"Section2\"\u003e \u003ch2\u003eComparative analysis of white blood cells (WBC) and platelets\u003c/h2\u003e \u003cp\u003eThe test group had a markedly elevated white blood cell (WBC) count compared to the control group (P\u0026thinsp;=\u0026thinsp;0.025). Nevertheless, the data obtained by analyzing whole-blood samples, which included the platelet count, did not show any statistically significant differences between the groups (P\u0026thinsp;=\u0026thinsp;0.387) (Table\u0026nbsp;\u003cspan refid=\"Tab3\" class=\"InternalRef\"\u003e3\u003c/span\u003e). Furthermore, there was no correlation observed between white blood cells (WBCs) and the growth factors examined in both the test and control groups (P\u0026thinsp;\u0026gt;\u0026thinsp;0.05).\u003c/p\u003e \u003cp\u003e \u003cdiv class=\"gridtable\"\u003e\u003ctable float=\"Yes\" id=\"Tab3\" border=\"1\"\u003e \u003ccaption language=\"En\"\u003e \u003cdiv class=\"CaptionNumber\"\u003eTable 3\u003c/div\u003e \u003cdiv class=\"CaptionContent\"\u003e \u003cp\u003eWBC and platelet values between groups\u003c/p\u003e \u003c/div\u003e \u003c/caption\u003e \u003ccolgroup cols=\"4\"\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c1\" colnum=\"1\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c2\" colnum=\"2\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c3\" colnum=\"3\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c4\" colnum=\"4\"\u003e\u003c/div\u003e \u003cthead\u003e \u003ctr\u003e \u003cth align=\"left\" colname=\"c1\"\u003e\u0026nbsp;\u003c/th\u003e \u003cth align=\"left\" colname=\"c2\"\u003e \u003cp\u003eMedian (Q1\u0026ndash;Q3)\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c3\"\u003e\u0026nbsp;\u003c/th\u003e \u003cth align=\"left\" colname=\"c4\"\u003e\u0026nbsp;\u003c/th\u003e \u003c/tr\u003e \u003ctr\u003e \u003cth align=\"left\" colname=\"c1\"\u003e\u0026nbsp;\u003c/th\u003e \u003cth align=\"left\" colname=\"c2\"\u003e \u003cp\u003eControl\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c3\"\u003e \u003cp\u003eTest\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c4\"\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\u003e\u003cb\u003eWBC (cells/mm\u003c/b\u003e\u003csup\u003e\u003cb\u003e3\u003c/b\u003e\u003c/sup\u003e\u003cb\u003e)\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e6.60 (5.25\u0026ndash;7.55)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e8.80 (7.38\u0026ndash;9.78)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e0.025\u003csup\u003e*\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e\u003cb\u003eThrombocyte (cells/mm\u003c/b\u003e\u003csup\u003e\u003cb\u003e3\u003c/b\u003e\u003c/sup\u003e\u003cb\u003e)\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e242.00 (216\u0026ndash;287.75)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e262.50 (230.75\u0026ndash;298)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e0.387\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003c/tbody\u003e \u003c/colgroup\u003e \u003ctfoot\u003e \u003ctr\u003e\u003ctd colspan=\"4\"\u003eQ1 25 percentile, Q3 75 percentile.\u003c/td\u003e\u003c/tr\u003e \u003ctr\u003e\u003ctd colspan=\"4\"\u003e\u003csup\u003e*\u003c/sup\u003eThe test group had a significantly higher WBC value than the control group (\u003cem\u003eP\u003c/em\u003e\u0026thinsp;\u0026lt;\u0026thinsp;0.05).\u003c/td\u003e\u003c/tr\u003e \u003c/tfoot\u003e \u003c/table\u003e\u003c/div\u003e \u003c/p\u003e \u003c/div\u003e"},{"header":"Discussions","content":"\u003cp\u003eThe findings of our investigation revealed that the levels of growth factors (IGF-1, EGF, VEGF, PDGF-BB, and TGF-1) released from i-PRFs varied significantly among individuals. Nevertheless, there was no notable disparity observed between the control and test groups. This investigation revealed that the levels of growth factors were not influenced by the periodontal disease condition of the participants. These findings provided support for the clinical application of i-PRF as a method of delivering autologous growth factors.\u003c/p\u003e \u003cp\u003eNo correlation was observed between the white blood cell (WBC) counts and the levels of growth factors across the groups, despite the fact that the data indicated higher WBC levels in the test group. The presence of elevated leukocytes indicated the expected occurrence of an inflammatory condition. The test group consisted of patients diagnosed with widespread severe and mild chronic periodontitis [\u003cspan citationid=\"CR20\" class=\"CitationRef\"\u003e20\u003c/span\u003e]. No significant differences were identified in the findings of the whole-blood analysis between the groups. These results were expected as all patients had either managed systemic disorders or were in a systemically healthy state.\u003c/p\u003e \u003cp\u003eMultiple PRF preparation protocols are accessible. The PRF centrifuge procedure employed in this work effectively ensnared leukocytes and thrombocytes within the fibrin clot [\u003cspan citationid=\"CR20\" class=\"CitationRef\"\u003e20\u003c/span\u003e]. Nevertheless, TGF-β1, EGF, VEGF, IGF-1, and PDGF-BB primarily originate from platelets rather than leukocytes [\u003cspan citationid=\"CR21\" class=\"CitationRef\"\u003e21\u003c/span\u003e]. Therefore, the absence of a link between white blood cell count (WBC) and the levels of growth factors derived from platelet-rich fibrin (PRF) is to be expected. There is currently no available information on the impact of whole-blood analysis and the presence of chronic periodontitis on the quantity or release of growth factors from i-PRF.\u003c/p\u003e \u003cp\u003eIn 2015, Maurao et al. achieved a distinct platelet concentration of i-PRF using horizontal centrifugation using a B-40 centrifuge (RDE, Brazil) for a duration of 2 minutes at 3300 rpm. The tubes used in this process contained 9 mL of venous blood and were not treated with any anticoagulants (11). Miron et al. obtained i-PRF by centrifuging 10 mL of whole blood without anticoagulant in tubes at a speed of 700 rpm for 3 minutes (equivalent to 60g) at room temperature using a Duo Centrifuge, a device specifically designed for obtaining PRF, manufactured in Nice, France (10). It is widely accepted that i-PRF contains not just leukocytes and thrombocytes, but also mesenchymal stem cells and endothelial cells. Thus, it is considered a \"blood concentrate\" rather than solely a platelet concentrate [\u003cspan citationid=\"CR22\" class=\"CitationRef\"\u003e22\u003c/span\u003e].\u003c/p\u003e \u003cp\u003eChang et al. conducted a comparison of the release of growth factors, white blood cell count, and thrombocyte amount in the PRF membrane and exudate of both periodontally healthy persons (control group) and patients with chronic periodontitis (test group). No significant disparity in the quantity of growth factor was observed between the test and control groups. The test group exhibited a markedly elevated WBC count. No association was found between white blood cell count (WBC), platelet count, and growth factors [\u003cspan citationid=\"CR14\" class=\"CitationRef\"\u003e14\u003c/span\u003e]. The findings of the study conducted by Chang et al. were in line with the results of the current investigation.\u003c/p\u003e \u003cp\u003eKobayashi et al. examined the growth factors present in platelet rich plasma (PRP), PRF, and advanced-platelet rich fibrin (A-PRF) derived from individuals who are in good health. The quantities and release of growth factors, including TGF-β, IGF, VEGF, EGF, PDGF-AA, PDGF-BB, and PDGF-AB, were assessed from platelet concentrates using ELISA over a duration of 10 days. The authors proposed that factors connected to the patient could influence the release of growth factors. As a result, there may be a significant variation in the quantities of growth factors released [\u003cspan citationid=\"CR23\" class=\"CitationRef\"\u003e23\u003c/span\u003e].\u003c/p\u003e \u003cp\u003eMiron et al. conducted a comparison of the growth factors present in platelet-rich plasma (PRP) and injectable platelet-rich fibrin (i-PRF). According to their statement, PRP produced a greater amount of growth factors initially, but i-PRF released higher levels of PDGF-AA, PDGF-AB, EGF, and IGF over a longer duration [\u003cspan citationid=\"CR10\" class=\"CitationRef\"\u003e10\u003c/span\u003e].\u003c/p\u003e \u003cp\u003eIn their study comparing i-PRF with PRP in gingival fibroblasts on titanium-implant surfaces, Wang et al. discovered higher levels of PDGF and TGF-β in the i-PRF group. Furthermore, the i-PRF group had a higher level of cellular migration and collagen-1, as evidenced by the detection of these factors [\u003cspan citationid=\"CR24\" class=\"CitationRef\"\u003e24\u003c/span\u003e].\u003c/p\u003e \u003cp\u003eIn their investigation on the release of leukocytes, platelets, and growth factors in liquid PRF products, Choukroun et al. found that low-speed centrifugation resulted in an increase in these components in PRF-based matrices [\u003cspan citationid=\"CR25\" class=\"CitationRef\"\u003e25\u003c/span\u003e].\u003c/p\u003e \u003cp\u003eVarela et al. assessed the liberation of growth factors in i-PRF, the gene expression of type I collagen, the morphological characteristics, and the cellular composition. According to their statement, i-PRF, which includes platelets, growth factors, type I collagen, leukocytes, and osteocalcin, may be an effective method for promoting the healing of both hard and soft tissues [\u003cspan citationid=\"CR26\" class=\"CitationRef\"\u003e26\u003c/span\u003e].\u003c/p\u003e \u003cp\u003eThe release of growth factors from PRF varied among patients. However, the levels of growth factors derived from the i-PRF did not exhibit a statistically significant disparity between the test and control groups. The findings of the whole-blood analysis indicated that the test group exhibited a statistically significant increase in white blood cell count (P\u0026thinsp;\u0026lt;\u0026thinsp;0.05). However, there was no substantial correlation observed between the white blood cell counts and the levels of the growth factors in i-PRF.\u003c/p\u003e \u003cp\u003eThe current investigation is limited by its modest and straightforward nature. Additional constraints of the study include the exclusive acquisition of i-PRF and the absence of evaluation for other types of PRF. Furthermore, the determination of participants' systemic status relied solely on their self-reported remarks.\u003c/p\u003e"},{"header":"Conclusions","content":"\u003cp\u003eBased on the constraints of this investigation, it was determined that i-PRF can be utilized as a self-derived source of growth factors that remain unaffected by the periodontal condition and white blood cell counts of healthy individuals.\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 contributions:\u0026nbsp;\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eStudy concept and design: BK. Data Acquisition: BK. Analysis and interpretation of data: BK, HBS. Writing - original draft preparation: BK. Writing - review and editing: BK, HBS. All authors read and approved the final manuscript.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eFunding\u003c/strong\u003e\u0026nbsp;\u003c/p\u003e\n\u003cp\u003eThis study was supported by scientific research projects (ALKU BAP, grant 2019-06-01-MAP02).\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eData Availability\u0026nbsp;\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eAll data generated or analyzed during this study are included in this published article \u0026amp; supporting material.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eEthics approval and consent to participate\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThe study was initiated following the approval of the Akdeniz University Faculty of Medicine Clinical Research Ethics Committee (dated 05.12.2018 and protocol# 859).\u0026nbsp;All procedures performed in studies involving human participants were in accordance with the ethical standards of the institutional research committee. Informed consent term was obtained from every individual participant included in this study.\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 that they have no competing interests\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eAuthor details\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003e\u003csup\u003e1\u0026nbsp;\u003c/sup\u003eAlanya Alaaddin Keykubat University, Faculty of Dentistry, Department of Periodontology, Antalya, Turkey\u003c/p\u003e\n\u003cp\u003e\u003csup\u003e2\u0026nbsp;\u003c/sup\u003eMardin Artuklu University, Faculty of Medicine,\u0026nbsp;Department of\u0026nbsp;Biochemistry, Mardin, Turkey\u003c/p\u003e"},{"header":"References","content":"\u003col\u003e\u003cli\u003e\u003cspan\u003eOhlrich EJ, Cullinan MP, Seymour GJ. 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Clin Oral Investig. 2019;23(3):1309\u0026ndash;18. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1007/s00784-018-2555-2\u003c/span\u003e\u003cspan address=\"10.1007/s00784-018-2555-2\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e.\u003c/span\u003e\u003c/li\u003e\u003c/ol\u003e"}],"fulltextSource":"","fullText":"","funders":[],"hasAdminPriorityOnWorkflow":false,"hasManuscriptDocX":true,"hasOptedInToPreprint":true,"hasPassedJournalQc":"","hasAnyPriority":false,"hideJournal":false,"highlight":"","institution":"","isAcceptedByJournal":true,"isAuthorSuppliedPdf":false,"isDeskRejected":"","isHiddenFromSearch":false,"isInQc":false,"isInWorkflow":false,"isPdf":false,"isPdfUpToDate":true,"isWithdrawnOrRetracted":false,"journal":{"display":true,"email":"[email protected]","identity":"bmc-oral-health","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":false,"externalIdentity":"ohea","sideBox":"Learn more about [BMC Oral Health](http://bmcoralhealth.biomedcentral.com/)","snPcode":"","submissionUrl":"https://www.editorialmanager.com/ohea/default.aspx","title":"BMC Oral Health","twitterHandle":"BMC_series","acdcEnabled":true,"dfaEnabled":false,"editorialSystem":"em","reportingPortfolio":"BMC Series","inReviewEnabled":true,"inReviewRevisionsEnabled":true},"keywords":"Injectable platelet rich fibrin, Growth factors, White blood cells, Platelets, Chronic periodontitis","lastPublishedDoi":"10.21203/rs.3.rs-3850305/v1","lastPublishedDoiUrl":"https://doi.org/10.21203/rs.3.rs-3850305/v1","license":{"name":"CC BY 4.0","url":"https://creativecommons.org/licenses/by/4.0/"},"manuscriptAbstract":"\u003ch2\u003eBackground\u003c/h2\u003e \u003cp\u003eThis study purposed to assess and compare the concentrations of growth factors, white blood cells (WBCs), and platelets in injectable platelet-rich fibrin (i-PRF) derived from persons with healthy periodontal conditions and those with chronic periodontitis.\u003c/p\u003e\u003ch2\u003eMethods\u003c/h2\u003e \u003cp\u003eVenous blood samples were obtained from patients diagnosed with chronic periodontitis (test group) and people with healthy periodontal conditions (control group). The I-PRF was acquired from blood that had been centrifuged. The growth factors released from i-PRFs were compared among groups using the ELISA test. Furthermore, the amounts of white blood cells (WBCs) and platelets were also compared.\u003c/p\u003e\u003ch2\u003eResults\u003c/h2\u003e \u003cp\u003eComparison was made between the i-PRFs obtained from the test and control groups. No discernible disparity was detected. The test group had a markedly elevated white blood cell (WBC) count compared to the control group (P\u0026thinsp;=\u0026thinsp;0.025). Nevertheless, the data obtained from the whole-blood analysis, including the platelet count, did not show any statistically significant differences between the groups (P\u0026thinsp;=\u0026thinsp;0.387). Furthermore, there was no notable link observed between the white blood cells (WBCs) and growth factors in either of the groups (P\u0026thinsp;\u0026gt;\u0026thinsp;0.05).\u003c/p\u003e\u003ch2\u003eConclusions\u003c/h2\u003e \u003cp\u003eThe levels of growth factors (VEGF, IGF-1, TGF-β1, PDGF-BB, EGF) in i-PRFs varied significantly among people. Nevertheless, there was no substantial disparity observed between the two groups. This suggests that the levels of growth factors may be unaffected by the periodontal disease state of the participants. These findings provide evidence for the practical application of i-PRFs as a method for delivering autologous growth factors.\u003c/p\u003e","manuscriptTitle":"Comparison of the level of growth factors in injectable platelet-rich fibrin obtained from healthy individuals and patients with chronic periodontitis","msid":"","msnumber":"","nonDraftVersions":[{"code":1,"date":"2024-02-07 20:10:09","doi":"10.21203/rs.3.rs-3850305/v1","editorialEvents":[{"type":"communityComments","content":0},{"type":"decision","content":"Revision requested","date":"2024-03-11T08:31:42+00:00","index":"","fulltext":""},{"type":"editorInvitedReview","content":"","date":"2024-02-22T16:24:02+00:00","index":"hide","fulltext":""},{"type":"reviewerAgreed","content":"8fac170d-467e-4108-8a94-23aa16c45ed3","date":"2024-02-13T14:19:14+00:00","index":"hide","fulltext":""},{"type":"reviewerAgreed","content":"6438e212-a6cb-488a-9ad9-0a4cc747feb3","date":"2024-02-11T13:14:53+00:00","index":"hide","fulltext":""},{"type":"reviewerAgreed","content":"29bcc17b-a4cb-459d-9877-550dedf992e1","date":"2024-02-11T09:47:37+00:00","index":"hide","fulltext":""},{"type":"reviewersInvited","content":"","date":"2024-02-11T08:36:52+00:00","index":"","fulltext":""},{"type":"editorAssigned","content":"","date":"2024-02-05T11:22:56+00:00","index":"","fulltext":""},{"type":"editorInvited","content":"","date":"2024-02-05T11:06:20+00:00","index":"","fulltext":""},{"type":"checksComplete","content":"","date":"2024-02-05T11:04:08+00:00","index":"","fulltext":""},{"type":"submitted","content":"BMC Oral Health","date":"2024-01-10T12:45:28+00:00","index":"","fulltext":""}],"status":"published","journal":{"display":true,"email":"[email protected]","identity":"bmc-oral-health","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":false,"externalIdentity":"ohea","sideBox":"Learn more about [BMC Oral Health](http://bmcoralhealth.biomedcentral.com/)","snPcode":"","submissionUrl":"https://www.editorialmanager.com/ohea/default.aspx","title":"BMC Oral Health","twitterHandle":"BMC_series","acdcEnabled":true,"dfaEnabled":false,"editorialSystem":"em","reportingPortfolio":"BMC Series","inReviewEnabled":true,"inReviewRevisionsEnabled":true}}],"origin":"","ownerIdentity":"0da3c244-8dc2-4847-8d9e-91e585290b60","owner":[],"postedDate":"February 7th, 2024","published":true,"recentEditorialEvents":[],"rejectedJournal":[],"revision":"","amendment":"","status":"published-in-journal","subjectAreas":[],"tags":[],"updatedAt":"2024-05-07T20:08:34+00:00","versionOfRecord":{"articleIdentity":"rs-3850305","link":"https://doi.org/10.1186/s12903-024-04301-x","journal":{"identity":"bmc-oral-health","isVorOnly":false,"title":"BMC Oral Health"},"publishedOn":"2024-05-03 19:57:54","publishedOnDateReadable":"May 3rd, 2024"},"versionCreatedAt":"2024-02-07 20:10:09","video":"","vorDoi":"10.1186/s12903-024-04301-x","vorDoiUrl":"https://doi.org/10.1186/s12903-024-04301-x","workflowStages":[]},"version":"v1","identity":"rs-3850305","journalConfig":"researchsquare"},"__N_SSP":true},"page":"/article/[identity]/[[...version]]","query":{"redirect":"/article/rs-3850305","identity":"rs-3850305","version":["v1"]},"buildId":"qtupq5eGEP_6zYnWcrvyt","isFallback":false,"isExperimentalCompile":false,"dynamicIds":[84888],"gssp":true,"scriptLoader":[]}