Abstract

Background: VEGF-A expression of human dental pulp stem cells (hDPSCs) can induce the angiogenesis process of dental pulp regeneration. This in vitro study aimed to analyze the effect of various concentrations of Advanced Platelet Rich Fibrin (A-PRF) conditioned media (CM) on the increased expression of vascular endothelial growth factor-A (VEGF-A) of hDPSCs. Methods: hDPSCs were collected from ten third molars extracted from nine healthy donors, cultured, and then harvested at the end of the third passage. The hDPSCs were seeded in four different CM (control group: hDPSCs + DMEM; 1% A-PRF CM group: hDPSCs + 1% A-PRF CM; 5% A-PRF CM group: hDPSCs + 5% A-PRF CM; 10% A-PRF CM group: hDPSCs + 10% A-PRF CM). All of the groups were cultured in biological triplicates (Triplo) and observed for 5, 12, and 24 hours. The VEGF-A protein expression of hDPSCs was measured using human VEGF-A ELISA at a wavelength of 405 nm. Data was analyzed with Kruskal Wallis and post hoc Mann Whitney test with p<0.05. Results: The VEGF-A expression rate of hDPSCs among all groups was statistically significantly different at 5, 12 and 24 hours of observations (p<0.05). Post hoc analysis test showed a statistically significant difference of hDPSCs’s VEGF-A expression between 5% A-PRF groups compared to other groups at 5 and 12 hours of observation (p<0.05). However, there were no statistically significant differences observed of hDPSCs’ VEGF-A expression at 24 hours of observation between 1%, 5% and 10% A-PRF groups (p>0.05). Conclusion: 5% A-PRF CM was superior in increasing VEGF-A expression of hDPSCs at 5, 12 and 24 hours of observations.

Highlights

  • American Association of Endodontists (AAE) approved regenerative endodontics as a potential future therapy in endodontics

  • The International Society for Cellular Therapy (ISCT) stated that 95% of human mesenchymal stem cells (MSCs) can express at least three types of specific surface antigens, CD105, CD73, and CD90/Thy-1 [7, 8]

  • The International Society for Cellular Therapy (ISCT) stated that 95% of human MSCs can express at least three types of specific surface antigens, CD105, CD73, and CD90/Thy-1 (positive cocktail) [7, 8]

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Summary

Introduction

American Association of Endodontists (AAE) approved regenerative endodontics as a potential future therapy in endodontics. Gharaei et al [11] demonstrated that human dental pulp stromal cell conditioned medium can trigger pronounced angiogenic effects in increasing VEGF expression [11]. These results are consistent with that of another study proving that VEGF expression plays an important role in the endothelial differentiation of MSCs derived from human exfoliated deciduous teeth [12]. VEGF-A expression of human dental pulp stem cells (hDPSCs) can induce the angiogenesis process of dental pulp regeneration This in vitro study aimed to analyze the effect of various concentrations of Advanced Platelet Rich Fibrin (A-PRF) conditioned media (CM) on the increased expression of vascular endothelial growth factor-A (VEGF-A) of hDPSCs

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