Modulation of the TGF-β1-related response of human dental pulp stem cells by PRF membranes with and without bone allograft

Introduction To our knowledge, no previous study has directly compared the TGF-β1-related responses of human dental pulp stem cells (hDPSCs) to conditioned media derived from A-PRF+, H-PRF, Gel A-PRF+, and e-PRF, with and without bone allograft. Aim To evaluate the TGF-β1-related cellular response of hDPSCs following exposure to conditioned media derived from different PRF preparations, used alone or combined with bone allograft. Materials and methods Four PRF preparations (A-PRF+, H-PRF, Gel A-PRF+, and e-PRF) were investigated with and without bone allograft. Each experimental group included 10 independent biological samples (n = 10). Conditioned media were collected after 24 h and on day 14. hDPSCs cultured in unconditioned medium served as controls. After 48 h of treatment, TGF-β1-related cellular immunoreactivity was assessed using Cell ELISA and normalized to glyceraldehyde-3-phosphate dehydrogenase (GAPDH). Statistical analysis included the Kruskal–Wallis test with Dunn’s post hoc test and Bonferroni correction. Direct paired comparisons were performed using the Wilcoxon signed-rank test with Bonferroni correction. Results PRF preparations without allograft showed no significant increases in the normalized TGF-β1/GAPDH signal compared with controls. After 24 h of conditioning, significant increases were observed for H-PRF, Gel A-PRF+, and e-PRF combined with allograft compared with controls. After 14 days, significant increases were observed for Gel A-PRF+ and e-PRF with allograft. Direct paired comparisons demonstrated significantly higher signals for Gel A-PRF+ and e-PRF with allograft after 14 days compared with the corresponding preparations without allograft, whereas no significant differences were detected after 24 h. Conclusion The TGF-β1-related response of hDPSCs varied according to PRF preparation, allograft supplementation, and conditioning period. The significant effects observed for Gel A-PRF+ and e-PRF combined with allograft support further investigation of these constructs as potential bioactive materials for vital pulp therapy.