LAUSR

AIM To investigate the proliferation and differentiation potential of human dental pulp stem cells (DPSCs) in a three-dimensional culture model (TDM) by incorporation of VEGF and BMP-2. METHODOLOGY TDM was established using fibrin gel (fg) as a soft tissue matrix and demineralized dentine disc (dd) as a hard tissue matrix. DPSCs and vascular endothelial growth factor (VEGF) were encapsulated in fibrin gel (fg-VEGF) and then inserted into bone morphogenetic protein (BMP-2)-coated demineralized dentine discs (dd-BMP-2). DPSCs were incubated for 28 days in various fg/dd combinations in the absence or presence of VEGF and BMP-2. Proliferation and morphology of DPSCs in fibrin gel were analysed using MTT and Live&Dead assays. Release profiles of VEGF and BMP-2 from fibrin gel and dentine discs were quantified using ELISA, and the expressions of angiogenic and odontogenic differentiation markers were determined with RT-qPCR analysis. Data were analysed statistically using Wilcoxon signed rank tests, Kruskal-Wallis tests with Mann-Whitney U tests and Bonferroni adjustment. The level of significance was set at P < 0.05. RESULTS DPSCs were able to proliferate and showed interconnected cellular elongations in fibrin gel depending on fibrinogen concentration whilst monolayer control group showed typical fibroblast-like cell morphology. Encapsulating of VEGF in fibrin gel and BMP-2 in gelatin that was used to coat dentine discs allowed the controlled releases of growth factors, which induced angiogenic and odontogenic gene expressions by DPSCs. Higher expressions of PECAM as an angiogenic factor, and BSP, DMP-1, OCN and CBFA as odontogenic factors, were observed in TDM as compared to the other fg/dd combinations and the monolayer control group (P < 0.05). CONCLUSIONS TDM consisting of fibrin gel and dentine matrix allowed cell-cell interactions. TDM was highly effective in delivering both VEGF and BMP-2 that enhanced the angiogenic and odontogenic potential of DPSCs.