Ideal Concentration of Advanced-Platelet Rich Fibrin (A-PRF) Conditioned Media for Human Dental Pulp Stem Cells Differentiation

Dini Asrianti Bagio, Indah Julianto, Endang Suprastiwi, Anggraini Margono


Objective:To discover the ideal concentration of Advanced Platelet Rich Fibrin (A-PRF) as modification of PRF, for human Dental Pulp Stem Cells (hDPSCs) differentiation. Material and Methods:hDPSCs were devided into five experimental groups: Group I (control group) consist of hDPSCs cultured in 10% FBS, Group II consist of hDPSCs cultured in 1% A-PRF, Group III consist of hDPSCs cultured in 5% A-PRF, Group IV consist of hDPSCs cultured in 10% A-PRF and Group V consist of hDPSCs cultured in 25% A-APRF. All group have been observed for 7 and 14 days and each group had three biological replicates (triplo). Formation of the mineralized nodules was detected after 7 days by Alizarin red-based assay and Dentin Sialophosphoprotein (DSPP) expression after 7 and 14 days quantified by ELISA reader. Statistical analysis was proven with Kruskal-Wallis and post hoc Mann-Whitney test. Results:The differentiation of hDPSCs in all A-PRF groups was significantly different on day-7 (p<0.05) compare to control group (Group I). There were no significant differences between all groups on day-14 (p>0.05). Significantly differences between Group II (1% A-PRF) and Group I (control), Group II (1% A-PRF) and Group III (5% A-PRF), also Group II (1% A-PRF) and Group V (25% A-PRF) was found from post hoc test analysis. Conclusion:The ideal conditioned media concentration for differentiation of human dental pulp stem cells was on 1% up to 5% A-PRF group.


Platelet-Rich Fibrin; Dental Pulp; Cell Differentiation.

Full Text:



Hargreaves KM, Diogenes A, Teixeira FB. Treatment options: Biological basis of regenerative endodontic procedures. J Endod 2013; 39(3 Suppl):S30-S43.

Murray PE. Regenerative endodontics: A Review of current status and a call for action. J Endod 2007; 33(4):377-90.

Pawitan JA. Prospect of stem cell conditioned medium in regenerative medicine. Biomed Res Int 2014; 2014:965849.

Hotwani K, Sharma K. Platelet rich fibrin - A novel acumen into regenerative endodontic therapy. Restor Dent Endod 2014; 39(1):1-6.

Dohan DM, Choukroun J, Diss A, Dohan SL, Dohan AJ, Mouhyi J, et al. 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.

Ledesma-Martínez E, Mendoza-Núñez VM, Santiago-Osorio E. Mesenchymal stem cells derived from dental pulp: A review. Stem Cells Int 2016; 2016:4709572.

Gronthos S1, Mankani M, Brahim J, Robey PG, Shi S. Postnatal human dental pulp stem cells (DPSC) in vitro and in vivo. Proc Natl Acad Sci USA 2000; 97(25):13625-30.

Huang FM, Yang SF, Zhao JH, Chang YC. Platelet-rich fibrin increases proliferation and differentiation of human dental pulp cells. J Endod 2010; 36(10):1628-32.

Diananda, Kamizar, Margono A, Asrianti D, Meydiawati R. The efficacy of Advanced Platelet Rich Fibrin (A-PRF) of fibroblast cell regeneration. J Int Dent Med Res 2017; 10(Sp. Issue):789-92.

Masuki H, Okudera T, Watanebe T, Suzuki M, Nishiyama K, Okudera H, et al. Growth factor and pro-inflammatory cytokine contents in platelet-rich plasma (PRP), plasma rich in growth factors (PRGF), advanced platelet-rich fibrin (A-PRF), and concentrated growth factors (CGF). Int J Implant Dent 2016; 2(1):19.

Saeed MA, El-Rahman MA, Helal ME, Zaher AR, Grawish ME. Efficacy of human platelet rich fibrin exudate vs fetal bovine serum on proliferation and differentiation of dental pulp stem cells. Int J Stem Cells 2017; 10(1):38-47.

Vahabi S, Vaziri S, Torshabi M, Esfahrood ZR. Effects of plasma rich in growth factors and platelet rich fibrin on proliferation and viability of human gingival fibroblasts. J Dent 2015; 12(7):504-12.

Wirohadidjojo YW, Budiyanto A, Soebono H. Platelet-Rich fibrin lysate can ameliorate dysfunction of chronically uva-irradiated human dermal fibroblasts. Yonsei Med J 2016; 57(5):1282-5.

Margono DA, Bagio DA, Nursasongko B, Nazar K, Yulianto I, Gayatri E, et al. Comparison of Advanced Platelet Rich Fibrin (A-PRF) and Culture Media Conditioned Warton's Jelly (CMCWJ) on fibroblast cells proliferation. Pesqui Bras Odontopediatria Clín Integr 2018; 18(1):e4085.

Asri RS, Asrianti D, Margono A, Usman M, Yulianto I. Optimum concentration of platelet-rich fibrin lysate for human dental pulp stem cells culture medium. J Int Dent Med Res 2019; 12(1)105-10.