| Objective: To investigate the effect and the mechanism of transforming growth factor-β3(TGF-β3)on the osteogenic differentiation of rabbit dental pulp stem cells(rDPSCs).Methods:rDPSCs in third passage were divided into the experimental group(rDPSCs+80ng/mL TGF-β3),control group(rDPSCs+ osteogenic medium)and blank group(conventional culture medium);For each group after7,14 days,ALP activity was measured,RT-qPCR was carried out to measure the expression of ALP,COL-1A1,Runx-2,OCN,OPN,BSP and Smad4;After 3,5,7,14 days,immunohistochemical staining for COL-IA1,OCN and Runx-2 was performed;And after 7,14,21 days,Alizarin Red S staining was carried out to testify mineralized matrix productivity.Results: rDPSCs was isolated successfully.ALP activity showed significant difference between each two groups(P<0.05).Immunohistochemical staining on the experimental group showed strong expression of COL-1A1,Runx-2 and OCN than the other groups.RT—qPCR results revealed that experimental group showed higher expression of COL-1A1,Runx-2,OPN,BSP and Smad4 than the blank group(P<0.05)and higher expression of COL-1A1 and Smad4 than the control group(P<0.05).The average number of mineralized nodules in the experimental group was higher than the other groups.Conclusion: TGF-β3 can improve the osteogenic differentiation of rDPSCs and this process maybe through activating Smad pathway. |
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