Effects of transforming growth factor beta 3 on the proliferation and mineralization of dental pulp stem cells from human deciduous teeth

Ren Fei; Liu Jian-ping; Lin Jin-mei; Zhou Hui; Chen Xiao-chun; Xu Ping-ping; Yang Qin

doi:10.3969/j.issn.2095-4344.2014.28.019

Abstract：BACKGROUND:The role of transforming growth factorβsuperfamily has been reported in bone mineralization of various types of stem cells, but the effects of transforming growth factorβ3 (TGF-β3) combined with heparin on proliferation and mineralization of dental pulp stem cells from human deciduous teeth remains to be studied. OBJECTIVE:To evaluate the effects of TGF-β3 on the proliferation and mineralization of dental pulp stem cells from human deciduous teeth. METHODS:The enzyme digestion method was utilized to separately culture dental pulp stem cells from human deciduous teeth. The cellcolony forming efficiency was determined. Flow cytometry was utilized to identify cellsurface marker CD146. Immunochemistry for Vimentin and STRO1 was performed to measure dental pulp stem cells from human deciduous teeth. The third passage dental pulp stem cells from human deciduous teeth cultured in vitro were intervened with heparin and TGF-β3 of 1, 5, 25μg/L mass concentration. The MTS method was applied to measure cellgrowth curves. Alizarin red staining was carried out. The changes in alkaline phosphatase activity were determined with alkaline phosphatase kit. RESULTS AND CONCLUSION:The cellcolony forming efficiency was high. cells were positive for CD146, and strongly positive for Vimentin and STRO1. Dental pulp stem cells from human deciduous teeth were identified. MTS assay indicated that there was no obvious effect on promoting proliferation of dental pulp stem cells from human deciduous teeth after stimulation of TGF-β3. Detection results of alkaline phosphatase activity demonstrated that the combination of TGF-β3 and heparin could strengthen the alkaline phosphatase activity of dental pulp stem cells from human deciduous teeth with increased concentration. Alkaline phosphatase activities were significantly higher in the TGF-β3+heparin group, TGF-β3 group and heparin group than in the control group (P<0.01). Alizarin red staining was positive in the TGF-β3+heparin group, and the staining was strongest in the 5μg/L TGF-β3+heparin group. Results indicated that TGF-β3 combined with heparin promoted mineralization of dental pulp stem cells from human deciduous teeth.