为了观察壳聚糖膜对转染载有骨形成蛋白基因（bone morphogenetic protein,BMPDNA）的成骨样细胞（MG 63）粘附及增殖的影响,本研究构建重组BMP-2质粒并转染入MG 63,分别经酶切、PCR、Western、免疫组化鉴定;制备壳聚糖膜（chitosan,CS）,然后将转染细胞与该材料分别培养1、3、5天,扫描电镜观察细胞在材料表面的粘附形态;24、48、72、96小时后MTS检测细胞的增殖情况。结果发现转染了BMP-2 DNA MG63在壳聚糖膜材料上形态生长良好,有利于细胞的粘附行为;96小时后与未转染细胞组相比无明显差异。结果表明虽然体外转染BMP-2的MG63的增殖没有明显促进作用,但有利于细胞与材料间的粘附。结论：BMP-2转染成骨细胞增加细胞壳聚糖膜间的粘附,这将有助于了解壳聚糖细胞复合材料临床骨再生的应用前景。
To evaluate the adhesion and proliferation of bone morphogenetic protein-2（BMP-2）-transfected MG 63 cells,recombinant bone morphogenetic protein-2（BMP-2） plasmids were constructed and transfected into MG 63 cells.The transfection was analyzed and confirmed by pulimerase chain reaction（PCR）,Western bloting,and cell immunohistochemisty.Chitosan membranes（CSs） were prepared,on which BMP-2-transfected MG 63 cells were cultured.The adhesion between the cells and the membranes was analyzed with scanning electron microscopy（SEM） in d1,d3,and d5,and proliferation of the cells was assessed with（3-（4,5-dimethylthiazol-2-yl）-5-（3-carboxymethoxyphenyl）-2-（4-sulfophenyl）-2H-tetrazolium）,（MTS） assay in 24h,48h,72h,and 96h.The recombinant BMP-2 plasmid DNA was successfully constructed and transfected into the MG63 cells.SEM analysis indicated that significant increase of adhesion occurred in d1,d3,and d5 compared with the control,while MTS showed no significant difference in cell proliferation.BMP-2-transfected MG63 cells significantly increased their adhesion to chitosan membranes,which may have clinical application in bone rebuilding.
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