Effect of bFGF on the Co-breeding of Periodontal Ligament Fibroblasts and Bone Marrow Mesenchymal Stem Cells in Vitro
Objective To investigate the effect of basic fibroblast growth factor(bFGF)on the differentiation of bone marrow mesenchymal stem cells(BMSCs)into periodontal ligament fibroblasts(PDLFs)in the co-breeding condition of BMSCs and PDLFs.Methods BMSCs and PDLFs were cultured in vitro and identified.BMSCs/PDLFs indirect co-culture model was estab-lished in Millicell hanging cell co-culture room.The experiment was divided into 5 groups:negative control group(single BMSCs group);positive control group(PDLFs group alone);experimental group:0 ng/mL bFGF+BMSCs/PDLFs group,1 ng/mL bFGF+ BMSCs/PDLFs group and10 ng/mL bFGF+BMSCs/PDLFs group.ALP activity was measured on days3,5 and 7 of cell co-breed-ing.After cultured to 2 w,the co-breeding was stopped,and mRNA contents of osteocalcin(OCN),collagen typeⅢ(COLⅢ),osteopontin(OPN)and metalloproteinase 1(ADAMTS1)were monitored by RT-PCR.SPSS 17.0 software was used for statistical analysis of the data.Results The ALP activity test showed that there were statistically significant differences between the experimen-tal group and the negative control group at each time point after 3 d,5 d and 7 d co-culture(P<0.05);compared with positive con-trol group,the difference was not statistically significant(P>0.05);there was no statistical significance among all experimental groups(P>0.05).RT-PCR detection results showed that the mRNA expression of OCN and OPN between the negative control group and the other 4 groups had statistical significance(P<0.05),but there was no statistical significance between the latter 4 groups(P>0.05);the mRNA expressions of COLⅢand ADAMTS1 in the single BMSCs group were significantly different from those in the oth-er 4 groups(P<0.05),among which the mRNA expressions of COLⅢand ADAMTS1 in the 10 ng/mL bFGF+BMSCs/PDLFs group were higher than those in the other 4 groups.The difference was statistically significant(P<0.05).Conclusion bFGF can induce BMSCs to differentiate into PDLFs,and the induction effect of bFGF in high concentration group(10 ng/mL)is better than that in low concentration group(1 ng/mL).
万方数据
维普
