hBMP2基因转染对人牙龈成纤维细胞生物学特性的影响

目的分析hBMP2基因转染人牙龈成纤维细胞的生物学特性.方法用脂质体转染法将hBMP2基因转入人牙龈成纤维细胞内,经G418筛选后,获得阳性克隆.以原位杂交和免疫组化对转染细胞进行鉴定;进一步观察细胞形态、生长特性、碱性磷酸酶活性、骨钙素合成以及体外形成矿化结节的能力.结果hBMP2基因转染后,人牙龈成纤维细胞有hBMP2 mRNA的转录和蛋白表达;部分细胞由原来的长梭形转化为多角形,碱性磷酸酶活性和骨钙素合成均明显高于对照组(两组均P＜0.01);在矿化液作用下,能形成体外矿化结节.但细胞的增殖特性无明显变化.结论外源性hBMP2基因能够在人牙龈成纤维细胞表达,并促进其向成骨细胞方向转化,而对细胞的生长特性无明显影响.

Effects of human BMP2 gene transfection on human gingival fibroblast in vitro

AIM:To study the effects of human BMP_2 gene transfection on human gingival fibroblasts.METHODS: hBMP_2 gene was transfected into human gingival fibroblasts by using LipofectAMINE.After positive clones selected with G-418,the expression of hBMP_2 gene were determined by in situ hybridization and immunohistochemistry.We also observed the shape,proliferation,alkaline phosphatase (ALP) activity,osteocalcin(OC) and mineralization of the transfected gingival fibroblasts.RESULTS:The results showed that the hBMP_2 mRNA and protein were synthesized in the gingival fibroblasts. It was also found that the shape of parts of gingival fibroblast changed from fusiform to polygonal,gene transfection enhanced both ALP activity and OC synthsis ( P <0.01),and mineral nodules developed in mineralization culture medium,after gene transfected.But,there was no obvious difference in doubling time and proliferative ability compared with parental cells.CONCLUSION:Those findings seem to indicate that the exogenous hBMP_2 gene could be expressed in human gingival fibroblasts and induce them differentiated into osteoblast-like cell,without obvious effect in their proliferation.