Effect of tumor necrosis factor-α on the proliferation and multi-directional differentiation of stem cells from rat apical papilla / 重庆医学

ABSTRACT

Objective To evaluate the biological effect of tumor necrosis factor-α(TNF-α) on the proliferation and multi-directional differentiation of stem cells from rat apical papilla(SCAP).Methods SCAP was extracted by combining enzyme digestion method with tissue block method.The cells were divided into control group(TNF-α 0 ng/mL) and experimental group(TNF-α 5,10,20,50 ng/mL).The ability of proliferation of SCAP was measured by MTT method.The ability of osteogenic/dentinogenic differentiation of SCAP was measured by alizarin red staining and quantitative real-time PCR.The ability of adipogenic of SCAP was measured by oil red O staining.The expression of vascular related genes of SCAP was measured by quantitative real-time PCR.Results SCAP was consistent with the characteristics of mesenchymal stem cells and possessed the ability of multi-directional differentiation.The MTT results showed that experimental group promoted the proliferation of SCAP in comparison with the control group.The difference was statistically significant(P<0.05),and 10 ng/mL was the optimum concentration.The results of alizarin red staining showed that with the increase of the concentration of TNF-α,the mineralized nodules in the experimental group gradually became smaller,and the number of the formation decreased gradually.The results of quantitative real-time PCR showed that the expression of OC,DMP-1 and DSPP in the experimental group was significantly lower than that of the control group at 3 and 7 days,in which the expression of OC was statistically significant different(P<0.05);at 14 days,the expression of OC,DMP-1 in the experimental group was significantly lower than that of the control group(P<0.05).The result of Oil red O staining showed that with the increase of the concentration of TNF-α,the lipid droplets formation in the experimental group gradually decreased.The result of quantitative real-time PCR showed that the expression of ANGPT1,VEGFA,PECAM-1 in the experimental group was significantly lower than that of the control group(P<0.05).Conclusion TNF-α might promote the proliferation and inhibit the multi-directional differentiation of SCAP.