转化生长因子-β1和甲壳胺联合作用对老年人牙周膜细胞生长分化功能的影响

Effects of chitosan associating with TGF-β1 on proliferation and differentiation of elderly people's periodontal ligament cells

目的探讨转化生长因子(TGF)-β1和甲壳胺(chitosan,Chi)联合作用对老年人牙周膜细胞增殖和碱性磷酸酶(ALP)活性的影响。方法用原代培养的老年人牙周膜细胞,采用四甲基偶氮唑蓝(MTT)法和酶动力学方法检测Chi 0.1 g/L、TGF-β1 1μg/L和Chi(0.1 g/L)加TGF-β1(1μg/L)对老年人牙周膜细胞增殖和ALP的作用。结果①与对照组比较,3 d和5 d时Chi加TGF-β1组和TGF-β1组及7d时Chi加TGF-β1组和Chi组均能明显增强老年人牙周膜细胞的增殖能力(P<0.05,P<0.01),Chi加TGF-β1组的促增殖能力明显强于Chi组和TGF-β1组;②3个实验组的细胞裂解液和Chi加TGF-β1组和TGF-β1组细胞培养液中牙周膜细胞ALP活性均高于对照组(P<0.05,P<0.01),以Chi加TGF-β1组最高(P<0.01)。结论Chi和TGF-β1单独都能增加老年人牙周膜细胞增殖的能力和ALP活性,Chi和TGF-β1联合使用增强老年人牙周膜细胞增殖和ALP活性的能力强于单独使用Chi和TGF-β1。

Objective To observe effects of chitosan associating with TGF-β1 on proliferation and alkaline phosphatase activity of elderly people＇s periodontal ligament cells（PDLCs）. Methods The elderly people＇s PDLCs were isolated and cultured. The effects of chitosan （0.1 g/L）, TGF-β1 （ 1 μg/L） and chitosan（0.1 g/L） plus TGF-β1 （ 1 μg/L） on the proliferative ability of elderly people＇s PDLCs were evaluated with MTF method and alkaline phosphatase activity of elderly people＇s PDLCs were measured with spectrophotometric assay. Results ① Compared with control group, the proliferative ability of elderly people＇s PDLCs in Chi（0.1 g/L） plus TGF-β1 （ 1 μg/L）group and TGF-β1 （ 1 μg/L） group on day 3,5 and Chi（0. 1 g/L） plus TGF-β1（ 1 μg/L） group and Chi（0. 1 g/L） group on day 7 was considerably increased （ P 〈 0.05, P 〈 0.01 ） .The effect of Chi（0. 1 g/L） plus TGF-β1（ 1 μg/L） group was the strongest among three groups. ②Compared with control group, alkaline phosphatase activities of elderly people＇s PDLCs of all three groups in cell cracked liquid, and Chi（0. 1 g/L） plus TGF-β1（ 1 μg/L）group and TGF-β1（ 1 μg/L） group in cell culture liquid were significantly enhanced （P 〈 0.05, P 〈 0.01 ）. The alkaline phosphatase activities of elderly people＇s PDLCs of Chi（0. 1 g/L） plus TGF-β1（ 1 μg/L）group were the highest among three groups. Conclusion These findings suggested that chitosan and TGF-β1 may be capable of producing an accelerative effect on proliferation and differentiation functions of human PDLCs. The effects of chitosan associating with TGF-β1 are considerably stronger than those of chitosan or TGF-β1.