Evaluation of Platelet-Rich Plasma Effect at different times and concentrations on periodontal ligament fibroblast and osteoblast viability

Adriana Paola Acosta Gómez, Sandra Janeth Gutiérrez Prieto, María Alexandra Bedoya Mejía, Dabeiba Adriana García Robayo, Ximena Edilia Moreno Serrano


ABSTRACT. Background: Growth factors used in health treatments can be obtained from a first-generation source called platelet-rich plasma. The variety of protocols to prepare PRP produces variable results regarding PRP activation time and its effects on cell proliferation and viability. Purpose: To evaluate proliferation and cell viability of periodontal ligament fibroblasts and osteoblasts stimulated with PRP in several concentrations and times after PRP activation. Methods: An in vitro study was carried out using periodontal ligament fibroblast and osteoblast cell cultures. PRP from venous blood of a healthy adult was prepared through centrifugation and activated with 10 % CaCl2. The effect on cell proliferation after application of 1 %, 3 %, and 5% PRP and platelet-poor plasma was evaluated at 0, 12, 24, 48, and 72 hours after activation through MTS. The control group consisted of culture that did not receive any treatment. Data were analyzed using Chi square, Fisher, and McNemar tests. Results: The cell viability assay showed statistically significant differences between the experimental and the control groups. Cell viability increased in cells treated with 5 % PRP 24 hours after activation (p=0.05). Conclusions: Fibroblast and osteoblast cell lines tended to be more viable 24 hours after activation with 5% PRP.


molecular biology; periodontics; cell cultures; cell proliferation; cell viability; in vitro; osteoblast; platelet-rich plasma; periodontal ligament fibroblast

Full Text:

PDF (Español)


Aljehani YA. Risk factors of periodontal disease: review of the literature. Int J Dent. 2014; 2014: 182513.

Armitage GC, Robertson PB. The biology, prevention, diagnosis and treatment of periodontal disease: scientific advances in the United States. J Am Dent Assoc. 2009; 140(Suppl 1): 365-435.

Dentino A, Lee S, Mailhat J, Hefti A. Principles of periodontology. Periodontol 2000. 2013 Feb; 61(1): 16-53.

Anitua E, Troya M, Orive G. An Autologous platelet rich plasma stimulates periodontal ligament regeneration. J Periodontol. 2013 Nov; 84(11): 1556-66.

García-Martinez O, Reyes-Botella C, Díaz-Rodriguez L, Luna-Bertos E, Ramos-Torrecillas J, Vallecillo-Capilla M, Ruiz C. Effect of platelet-rich plasma on growth and antigenic profile of human osteoblasts and its clinical impact. J Oral Maxillofac Surg. 2012; 70: 1558-64.

Rios H, Lin Z, Oh B, Park Ch, Giannobile W. Cell and gene-based therapeutic strategies for periodontal regenerative medicine. J Periodontol. 2011; 82: 1223-37.

Hallman M, Thor A. Bone substitutes and growth factors as an alternative/complement to autogenous bone for grafting in implant dentistry. Periodontol 2000. 2008 Jun; 47(1): 172-92.

Beca T, Hernández G, Morante S, Bascones A. Plasma rico en plaquetas. Una revisión bibliográfica. Avances en Periodoncia. 2007 Abr; 19(1): 39-52.

Hughes FJ, Turner W, Belibasakis G, Martuscelli G. Effects of growth factors and cytokines on osteoblast differentiation. Periodontol 2000. 2006 Jun; 41(1): 48-72.

Polimeni G, Xiropaidis AV, Wikesjö UME. Biology and principles of periodontal wound healing/regeneration. Periodontol 2000. 2006 Jun; 41(1): 30-47.

Cochran DL, Wozney JM. Biological mediators for periodontal regeneration. Periodontol 2000. 1999 Feb; 19(1): 40-58.

Souren J, Van Wijk R. Fibroblastic cell cycling in collagen gels. Cell Proliferation. 1993 Jan; 26(1): 12-23.

Acosta A. Papel del fibroblasto en salud y enfermedad periodontal. En: Gutiérrez S (editora). Fundamentos de ciencias básicas aplicadas a la odontología. 1a edición. Bogotá, Colombia: Pontificia Universidad Javeriana; 2006. pp. 349-68.

Dohan D, Andia I, Zumstein M, Zhang Ch, Pinto N, Bielecki T. Classification of platelet concentrates (Platelet- Rich Plasma- PRP, Platelet- Rich Fibrin- PRF) for topical and infiltrative use in orthopedic and sports medicine: current consensus, clinical implications and perspectives. Muscles, Ligaments and Tendons J. 2014;4 (1): 3-9.

Lee JW, Kwon OH, Kim TK, Cho YK, Choi KY, Chung HY, Cho BC, Yang JD, Shim JH. Platelet-rich plasma: quantitative assessment of growth factor levels and comparative analysis of activated and inactivated groups. Arch Plast Surg. 2013; 40: 530-5.

Carrillo-Mora P, González-Villalta A, Macías-Hernández S, Pineda-Villaseñor C. Plasma rico en plaquetas. Herramienta versátil de la medicina regenerativa? Cir Cir. 2013; 81: 74-82.

Burnouf T, Goubran H, Che TM, Ou KL, El-Ekiaby M, Radosevic M. Blood-derived biomaterials and platelet growth factors in regenerative medicine. Blood Reviews. 2013; 27: 77-89.

Anitua E, Troya M, Orive G. Plasma rich in growth factors promote gingival tissue regeneration by stimulating fibroblast proliferation and migration and by blocking transformin growth factor β-1induced myodifferentiation. J Periodontol. 2012; 83(8): 1028-37.

Dohan D. How to optimize the preparation of leukocyte and platelet- rich fibrin (L-PRF, Choukroun´s technique) clots and membranes: Introducing the PRF Box. Oral Surg Oral Med Oral Pathol Oral Radiol Endod. 2010 Sep; 110(3): 275-8.

Dohan D. Platelet- rich Plasma (PRP) and platelet rich fibrin (PRF) in human cell cultures: Growth factor release and contradictory results. Oral Surg Oral Med Oral Pathol Oral Radiol Endod. 2010; 110(4): 418-21.

He L, Lin Y, Hu X, Zhang Y, Wu H. A comparative study of platelet-rich fibrin (PRF) and platelet- rich plasma (PRP) on the effect of proliferation and differentiation of rat osteoblast in vitro. Oral Surg Oral Med Oral Pathol Oral Radiol Endod. 2009; 108(5): 707-13.

Dohan D, Rasmusson L, Albreksson T. Classification of platelet concentrates: from pure platelet-rich plasma (P-PRP) to leucocyte and platelet-rich fibrin (LPRF). Trends Biotechnol. 2009; 27(3): 158-67.

Nikolidakis D, Jansen J. The biology of platelet- rich plasma and its application in oral surgery: Literature Review. Tissue Eng Part B Rev. 2008; 14(3): 249-58.

Han J, Meng H.X, Tang J.M, Chen Z.B. The effect of different platelet-rich plasma concentration on proliferation of human periodontal ligament cells in vitro. Cell Proliferation. 2007 Apr; 40(2): 241-52.

Dohan D, Choukroun J, Diss A. Platelet-rich fibrin (PRF): a second-generation platelet concentrate. Part I: technological concepts and Evolution. Oral Surg Oral Med Oral Pathol Oral Radiol Endod .2006; 101(3): e37-44.

Zhang N, Wu YP, Quian SJ, Teng Ch, Chen Sh, Li H. Research progress in the mechanism of effect of PRP in bone deficiency healing. Scientific World J. 2013: 1-7.

Saucedo JM, Yaffe MA, Berschback JC, Hsu WK, Kalainov DM. Platelet-rich plasma. J Hand Surg Am. 2012; 37(3): 587-9.

Cervelli V, Scioli M, Gentile P, Doldo E, Bonanno E, Spagnoli L, Orlandi A. Platelet-rich plasma greatly potentiates insulin-induced adipogenic differentiation of human adipose-derived stem cells through a serine threonine kinase akt-dependent mechanism and promotes clinical fat graft maintenance. Stem Cells Transl Med. 2012; 1(3): 206-20.

Gentile P, Bottini D, Spallone D, Curcio B, Cervelli V. Application of PRP in maxillofacial surgery. J Craniofac Surg. 2010; 21: 900-4.

Froum S, Wallace S, Tarnow D, Cho SC. Effect of PRP on bone growth in maxillary sinus grafts. Three bilateral case reports. Int J Periodontics Restorative Dent. 2002; 22: 45-53.

Marx R, Carlson E, Eichstaedt R, Schimmele S, Strauss J, Georgeff K. Platelet-rich plasma- Growth factor enhancement for bone grafts. Oral Surg Oral Med Oral Pathol Oral Radiol Endod. 1998; 85: 638-46.

Chang YC, Zhao JH. Effects of platelet-rich fibrin on human periodontal ligament fibroblasts and application for periodontal infrabony defects. Australian Dental J. 2011; 56: 365-71.

Plachokova A, Nikolidakis D, Mulder J, Jansen J, Creugers N. Effect of Platelet-Rich Plasma on bone regeneration in dentistry: a systematic review. Clin Oral Impl Res. 2008; 19: 539-45.

Mooren R, Hendriks E, van den Beaken J, Merkx M, Meijer G, Jansen J, Stoelinga PJ. The effect of platelet-rich plasma in vitro on primary cells rat osteoblast-like cells and human endotelial cells. Tissue Eng Part A. 2010; 16(10): 3159-72.

Kawase T, Okuda K, Wolff L, Yoshie H. Platelet-rich plasma-derived fibrin clot formation stimulates collagen synthesis in periodontal ligament and osteoblast cells in vitro. J Periodontol 2003; 74: 858-64.

Gonshor A. Technique for producing PRP and platelet concentrate: Background and process. Int J Periodontics Rest Dent. 2002; 22: 547-57.

Slapnicka J, Frassmann A, Strasak L, Augustin P, Vanek J. Effects of activated and nonactivated platelet-rich plasma on proliferation of human osteoblast in vitro. J Oral Maxillofac Surg. 2008; 66: 297-301.

Reyes M, Montero S, Cifuentes J, Zarzar E. Actualización de la técnica de obtención y uso del plasma rico en factores de crecimiento (PRGF). Rev Dental Chile. 2002; 93(2): 25-8.

Sodek J, McKee M. Molecular and cellular biology of alveolar bone. Periodontol 2000. 2000; 24: 99-126.

Demmer RT, Papapanou PN. Epidemiologic patterns of chronic and aggressive periodontitis. Periodontol 2000. 2010 Jun; 53(1): 28-44.

Anitua E, Sanchez M, Zalduendo M, De La Fuente M, Prado R, Orive G, Andia I. Fibroblastic response to treatment with different preparations rich in growth factors. Cell Proliferation. 2009 Apr; 42(2): 162-70.

Acosta A, González O, Pereira E, Sánchez A. Proliferación de fibroblastos gingivales humanos tratados con plasma rico en plaquetas in vitro. Rev Fundación Juan José Carraro (Argentina). 2006 Abr-May; (22): 8-14.

Acosta A, García D, Perdomo S, Gonzalez O. Efecto del plasma rico en plaquetas sobre la proliferación de fibroblastos gingivales humanos in vitro. Rev Fundación Juan José Carraro (Argentina). 2010 Abr-May; (31): 4-13.

Pantou AL, Markopoulo CE, Dereka XE, Vavoraki H, Mamalis A, Vrotsos IA. The effect of platelet-rich plasma (PRP) combined with a bone allograft on human periodontal ligament (PDL) cells. Cell Tissue Bank. 2012; 13: 81-8.

Markopoulou CE, Markopoulou P, Dereka XE, Pepelass E, Vrotsos IA. Effect of homologous PRP on proliferation of human periodontally affected osteoblast. In vitro preliminary study. Report a case. J Musculoskelet Neuronal Interact. 2009; 9(3): 167-72.

Balaram N, Karunakar P, Jayadev M, Marshal R. Role of platelet rich fibrin in wound healing: A critical review. J Conserv Dent. 2013; 16(4): 284-93.

Creeper F, Lichanska AM, Marshall RI, Seymour GJ, Ivanovski S. The effect of platelet-rich plasma on osteoblast and periodontal ligament cell migration, proliferation and differentiation. J Periodont Res. 2009; 44: 258-65.

Fortes-Ferreira C, Carriel M, Filho J, Granjeiro J, Oliveira C, Magini R. Platelet-rich plasma influence on human osteoblasts growth. Clin Oral Impl Res. 2005; 16: 456-60.

Karp JM, Sarraf F, Schoichet MS, Davies JE. Fibrin-filled scaffolds for bone-tissue engineering: an in vivo study. J Biomed Mater Res A. 2004; 71(1): 162-71.

Thorwarth M, Rupprecht S, Falk S, Felszeghy E, Wiltfang J, Schlegel KA. Expression of bone matrix proteins during de novo bone formation using a bovine collagen and platelet-rich plasma (PRP) an immunohistochemical analysis. Biomaterials. 2005; 26: 2575-84.

Tsay RC, Vo J, Burke A. Differential growth factor retention by platelet-rich plasma composites. J Oral Maxillofac Surg. 2005; 63: 521-8.

DOI: http://dx.doi.org/10.11144/Javeriana.uo36-76.eepr

Article Metrics

Metrics Loading ...

Metrics powered by PLOS ALM

Copyright (c) 2017 Adriana Paola Acosta Gómez, Sandra Janeth Gutiérrez Prieto, María Alexandra Bedoya Mejía, Dabeiba Adriana García Robayo, Ximena Edilia Moreno Serrano

Creative Commons
This work is registered under Creative Commons Attribution 4.0 International.
Created from http://revistas.javeriana.edu.co/.