Abstract
The prominent purpose of the study was the evaluation of the in vitro mitogenic effect of three different homologous platelet-rich plasma (PRP) preparations (PRPa, PRPb, PRPc) on three different lines of periodontal ligament (PDL) cells (PDL1,2,3), cultured alone or in combination with a demineralized freeze-dried allograft (DFBA). PDL cell cultures were derived from the mid root of three maxillary caries-free premolars extracted for orthodontic reasons. Cells were grown and reached confluence. To evaluate the mitogenic effect of all exogenous factors (PRPa, PRPb, PRPc and DFBA) on PDL cells, specific number of cells (10.000/well) was cultured in the presence or absence of the above factors. Each PRP preparation (5% v/v) was added in all cell lines, in the absence or presence of 10 mg/ml of DFBA. The cells were also treated with 25 ng/ml bFGF (positive control). The mitogenic effect was evaluated 24 h after incubation, using the Trypan blue exclusion assay. The results revealed that all PRP preparations act as potent mitogens as they significantly induced cell proliferation on PDL1,2,3 lines. All PRP preparations when added alone in the PDL cell cultures, exhibited a significant advantage over the positive control (bFGF). The addition of DFBA to PRP did not influence significantly cell proliferation in all cell lines, comparatively to PRP alone, at the time -period studied. The findings of this study demonstrate the beneficial role of PRP alone or combined with the bone graft on periodontal ligament cells in vitro, suggesting that it may be considered as a potential biological approach in periodontal regeneration.
Similar content being viewed by others
References
Annunziata M, Oliva A, Buonaiuto C, Di Feo A, Di Pasquale R, Passaro I et al (2005) In vitro cell-type specific biological response of human periodontally related cells to platelet-rich plasma. J Periodontal Res 40(6):489–495
Bartold PM, Narayanan AS (2006) Molecular and cell biology of healthy and diseased periodontal tissues. Periodontol 2000 40:29–49
Camargo PM, Lekovic V, Weinlaender M, Divnic-Resnik T, Pavlovic M, Kenney EB (2009) A surgical reentry study on the influence of platelet-rich plasma in enhancing the regenerative effects of bovine porous bone mineral and guided tissue regeneration in the treatment of intrabony defects in humans. J Periodontol 80(6):915–923
Cenni E, Ciapetti G, Pagani S, Perut F, Giunti A, Baldini N (2005) Effects of activated platelet concentrates on human primary cultures of fibroblasts and osteoblasts. J Periodontol 76(3):323–328
Cho MI, Garant PR (2000) Development and general structure of the periodontium. Periodontol 2000 24:9–27
Choi BH, Zhu SJ, Kim BY, Huh JY, Lee SH, Jung JH (2005) Effect of platelet-rich plasma (PRP) concentration on the viability and proliferation of alveolar bone cells: an in vitro study. Int J Oral Maxillofac Surg 34(4):420–424
Christgau M, Moder D, Hiller KA, Dada A, Schmitz G, Schmalz G (2006) Growth factors and cytokines in autologous platelet concentrate and their correlation to periodontal regeneration outcomes. J Clin Periodontol 33(11):837–845
Creeper F, Lichanska AM, Marshall RI, Seymour GJ, Ivanovski S (2009) The effect of platelet-rich plasma on osteoblast and periodontal ligament cell migration, proliferation and differentiation. J Periodontal Res 44(2):258–265
Danesh-Meyer MJ, Filstein MR, Shanaman R (2001) Histological evaluation of sinus augmentation using platelet rich plasma (PRP): a case series. J Int Acad Periodontol 3(2):48–56
Demir B, Sengun D, Berberoglu A (2007) Clinical evaluation of platelet-rich plasma and bioactive glass in the treatment of intra-bony defects. J Clin Periodontol 34(8):709–715
Dereka XE, Markopoulou CE, Mamalis A, Pepelassi E, Vrotsos IA (2006a) Time- and dose-dependent mitogenic effect of basic fibroblast growth factor combined with different bone graft materials: an in vitro study. Clin Oral Implants Res 17(5):554–559
Dereka XE, Markopoulou CE, Vrotsos IA (2006b) Role of growth factors on periodontal repair. Growth Factors 24(4):260–267
Dori F, Huszar T, Nikolidakis D, Arweiler NB, Gera I, Sculean A (2007) Effect of platelet-rich plasma on the healing of intrabony defects treated with an anorganic bovine bone mineral and expanded polytetrafluoroethylene membranes. J Periodontol 78(6):983–990
Dori F, Huszar T, Nikolidakis D, Tihanyi D, Horvath A, Arweiler NB et al (2008) Effect of platelet-rich plasma on the healing of intrabony defects treated with Beta tricalcium phosphate and expanded polytetrafluoroethylene membranes. J Periodontol 79(4):660–669
Ferreira CF, Carriel Gomes MC, Filho JS, Granjeiro JM, Oliveira Simoes CM, Magini Rde S (2005) Platelet-rich plasma influence on human osteoblasts growth. Clin Oral Implants Res 16(4):456–460
Han J, Meng HX, Tang JM, Li SL, Tang Y, Chen ZB (2007) The effect of different platelet-rich plasma concentrations on proliferation and differentiation of human periodontal ligament cells in vitro. Cell Prolif 40(2):241–252
Hanna R, Trejo PM, Weltman RL (2004) Treatment of intrabony defects with bovine-derived xenograft alone and in combination with platelet-rich plasma: a randomized clinical trial. J Periodontol 75(12):1668–1677
Ilgenli T, Dundar N, Kal BI (2007) Demineralized freeze-dried bone allograft and platelet-rich plasma vs platelet-rich plasma alone in infrabony defects: a clinical and radiographic evaluation. Clin Oral Investig 11(1):51–59
Kanno T, Takahashi T, Tsujisawa T, Ariyoshi W, Nishihara T (2005) Platelet-rich plasma enhances human osteoblast-like cell proliferation and differentiation. J Oral Maxillofac Surg 63(3):362–369
Kassolis JD, Rosen PS, Reynolds MA (2000) Alveolar ridge and sinus augmentation utilizing platelet-rich plasma in combination with freeze-dried bone allograft: case series. J Periodontol 71(10):1654–1661
Kawase T, Okuda K, Wolff LF, Yoshie H (2003) Platelet-rich plasma-derived fibrin clot formation stimulates collagen synthesis in periodontal ligament and osteoblastic cells in vitro. J Periodontol 74(6):858–864
Kawase T, Okuda K, Saito Y, Yoshie H (2005) In vitro evidence that the biological effects of platelet-rich plasma on periodontal ligament cells is not mediated solely by constituent transforming-growth factor-beta or platelet-derived growth factor. J Periodontol 76(5):760–767
Klongnoi B, Rupprecht S, Kessler P, Thorwarth M, Wiltfang J, Schlegel KA (2006) Influence of platelet-rich plasma on a bioglass and autogenous bone in sinus augmentation. An explorative study. Clin Oral Implants Res 17(3):312–320
Marcopoulou CE, Vavouraki HN, Dereka XE, Vrotsos IA (2003) Proliferative effect of growth factors TGF-beta1, PDGF-BB and rhBMP-2 on human gingival fibroblasts and periodontal ligament cells. J Int Acad Periodontol 5(3):63–70
Markopoulou CE, Markopoulos P, Dereka XE, Pepelassi E, Vrotsos IA (2009) Effect of homologous PRP on proliferation of human periodontally affected osteoblasts. In vitro preliminary study. Report of a case. J Musculoskelet Neuronal Interact 9(3):167–172
Markou N, Pepelassi E, Vavouraki H, Stamatakis HC, Nikolopoulos G, Vrotsos I et al (2009) Treatment of periodontal endosseous defects with platelet-rich plasma alone or in combination with demineralized freeze-dried bone allograft: a comparative clinical trial. J Periodontol 80(12):1911–1919
Nanci A, Bosshardt DD (2006) Structure of periodontal tissues in health and disease. Periodontol 2000 40:11–28
Okuda K, Kawase T, Momose M, Murata M, Saito Y, Suzuki H et al (2003) Platelet-rich plasma contains high levels of platelet-derived growth factor and transforming growth factor-beta and modulates the proliferation of periodontally related cells in vitro. J Periodontol 74(6):849–857
Okuda K, Tai H, Tanabe K, Suzuki H, Sato T, Kawase T et al (2005) Platelet-rich plasma combined with a porous hydroxyapatite graft for the treatment of intrabony periodontal defects in humans: a comparative controlled clinical study. J Periodontol 76(6):890–898
Papadopoulos CE, Dereka XE, Vavouraki EN, Vrotsos IA (2003) In vitro evaluation of the mitogenic effect of platelet-derived growth factor-BB on human periodontal ligament cells cultured with various bone allografts. J Periodontol 74(4):451–457
Piemontese M, Aspriello SD, Rubini C, Ferrante L, Procaccini M (2008) Treatment of periodontal intrabony defects with demineralized freeze-dried bone allograft in combination with platelet-rich plasma: a comparative clinical trial. J Periodontol 79(5):802–810
Plachokova AS, Nikolidakis D, Mulder J, Jansen JA, Creugers NH (2008) Effect of platelet-rich plasma on bone regeneration in dentistry: a systematic review. Clin Oral Implants Res 19(6):539–545
Pradeep AR, Shetty SK, Garg G, Pai S (2009) Clinical effectiveness of autologous platelet-rich plasma and Peptide-enhanced bone graft in the treatment of intrabony defects. J Periodontol 80(1):62–71
Raja S, Byakod G, Pudakalkatti P (2009) Growth factors in periodontal regeneration. Int J Dent Hyg 7(2):82–89
Sanchez AR, Sheridan PJ, Kupp LI (2003) Is platelet-rich plasma the perfect enhancement factor? A current review. Int J Oral Maxillofac Implants 18(1):93–103
Slapnicka J, Fassmann A, Strasak L, Augustin P, Vanek J (2008) Effects of activated and nonactivated platelet-rich plasma on proliferation of human osteoblasts in vitro. J Oral Maxillofac Surg 66(2):297–301
Somerman MJ, Foster RA, Imm GM, Sauk JJ, Archer SY (1989) Periodontal ligament cells and gingival fibroblasts respond differently to attachment factors in vitro. J Periodontol 60(2):73–77
Terranova VP, Odziemiec C, Tweden KS, Spadone DP (1989) Repopulation of dentin surfaces by periodontal ligament cells and endothelial cells. Effect of basic fibroblast growth factor. J Periodontol 60(6):293–301
Thor A, Wannfors K, Sennerby L, Rasmusson L (2005) Reconstruction of the severely resorbed maxilla with autogenous bone, platelet-rich plasma, and implants: 1-year results of a controlled prospective 5-year study. Clin Implant Dent Relat Res 7(4):209–220
Uggeri J, Belletti S, Guizzardi S, Poli T, Cantarelli S, Scandroglio R et al (2007) Dose-dependent effects of platelet gel releasate on activities of human osteoblasts. J Periodontol 78(10):1985–1991
Weibrich G, Kleis WK, Hafner G (2002) Growth factor levels in the platelet-rich plasma produced by 2 different methods: curasan-type PRP kit versus PCCS PRP system. Int J Oral Maxillofac Implants 17(2):184–190
Weibrich G, Kleis WK, Hitzler WE, Hafner G (2005) Comparison of the platelet concentrate collection system with the plasma-rich-in-growth-factors kit to produce platelet-rich plasma: a technical report. Int J Oral Maxillofac Implants 20(1):118–123
Yamamiya K, Okuda K, Kawase T, Hata K, Wolff LF, Yoshie H (2008) Tissue-engineered cultured periosteum used with platelet-rich plasma and hydroxyapatite in treating human osseous defects. J Periodontol 79(5):811–818
Yilmaz S, Cakar G, Kuru BE, Yildirim B (2007) Platelet-rich plasma in combination with bovine derived xenograft in the treatment of generalized aggressive periodontitis: a case report with re-entry. Platelets 18(7):535–539
Acknowledgments
The authors would like to express their thanks to Prof. M. Koutsilieris (Laboratory of Molecular Endocrinology, Department of Experimental Physiology, Medical School, University of Athens, Greece) for his technical support.
Author information
Authors and Affiliations
Corresponding author
Rights and permissions
About this article
Cite this article
Pantou, A.L., Markopoulou, C.E., Dereka, X.E. et al. The effect of platelet-rich plasma (PRP) combined with a bone allograft on human periodontal ligament (PDL) cells. Cell Tissue Bank 13, 81–88 (2012). https://doi.org/10.1007/s10561-010-9231-3
Received:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s10561-010-9231-3