转染CDMP-2基因的成肌细胞与PLGA材料的相容性
发布时间：2018-10-10 14:35
【摘要】：目的 探讨转染CDMP-2(软骨源性形态发生蛋白2)基因后的成肌细胞与PLGA（聚乳酸/乙醇酸共聚物）材料的相容性。 方法 取新生5d的SD大鼠骨骼肌，采用混合酶一步消化分离获取成肌细胞，将差速贴壁法和胰酶消化法相结合进行纯化，观察形态学特点，生长状况,并检测desmin表达。采用脂质体转染方法将CDMP2基因转染P3代成肌细胞，，将成肌细胞和转染CDMP-2基因的成肌细胞分别接种于PLGA支架上。分4组：A成肌细胞;B转染CDMP-2基因的成肌细胞;C成肌细胞+PLGA支架;D转染CDMP-2基因的成肌细胞+PLGA支架。采用MTT法检测各组成肌细胞的生物活性，绘制生长曲线，在扫描电镜下观察细胞形态。 结果 细胞接种于PLGA支架后早期，倒置相差显微镜视野内见不到细胞在支架表面生长、附着，只能看到纵横交错的纤维网状结构。72小时后可见细胞增殖，细胞数量显著增多。MTT示各组成肌细胞在492nm波长时分别为0.046414,0.057665,0.038945,0.049911；在629nm波长时分别为0.132158，0.178811，0.108284，0.173810，ABCD各组在统计学上都无统计学意义（P0.05）。 结论 成肌细胞能够在PLGA支架上生长良好，说明PLGA材料对成肌细胞无细胞毒性，具有优异的生物相容性，可用于组织修复，为进一步研究软骨损伤的修复提供了新的思路和方法。
[Abstract]:Objective to investigate the compatibility of CDMP-2 (chondrogenic morphogenetic protein-2) gene transfected myoblasts with PLGA (poly (lactic acid / glycolic acid) material. Methods the skeletal muscle of 5 days old SD rats was isolated and isolated by one step digestion of mixed enzymes. The differential adherent method and trypsin digestion method were combined to purify the myoblasts. The morphological characteristics, growth status and expression of desmin were observed. The CDMP2 gene was transfected into P3 generation myoblast by liposome transfection. The myoblast and the CDMP-2 gene transfected myoblast were inoculated on the PLGA scaffold respectively. Four groups were divided into four groups: myoblast A, myoblast transfected with CDMP-2 gene B, PLGA scaffold with C myoblast and PLGA scaffold transfected with CDMP-2 gene. The biological activity of muscle cells was measured by MTT method, and the growth curve was drawn. The morphology of the cells was observed by scanning electron microscope (SEM). Results at the early stage after inoculation with PLGA scaffold, the cells could not grow and attach on the scaffold surface in inverted phase contrast microscope, only the crisscross fibrous reticular structure could be seen. After 72 hours, cell proliferation could be seen. MTT showed that the number of muscle cells was 0.0464140.0576650.038945and 0.1321580.1788110.1082840.1738101010 at the wavelength of 492nm, respectively, and there was no statistical significance in each group (P0.05). Conclusion myoblast can grow well on PLGA scaffold, which indicates that PLGA has no cytotoxicity to myoblasts and has excellent biocompatibility, and can be used for tissue repair. It provides a new idea and method for the further study of cartilage repair.
【学位授予单位】：辽宁医学院
【学位级别】：硕士
【学位授予年份】：2012
【分类号】：R318.08;R687
[Abstract]:Objective to investigate the compatibility of CDMP-2 (chondrogenic morphogenetic protein-2) gene transfected myoblasts with PLGA (poly (lactic acid / glycolic acid) material. Methods the skeletal muscle of 5 days old SD rats was isolated and isolated by one step digestion of mixed enzymes. The differential adherent method and trypsin digestion method were combined to purify the myoblasts. The morphological characteristics, growth status and expression of desmin were observed. The CDMP2 gene was transfected into P3 generation myoblast by liposome transfection. The myoblast and the CDMP-2 gene transfected myoblast were inoculated on the PLGA scaffold respectively. Four groups were divided into four groups: myoblast A, myoblast transfected with CDMP-2 gene B, PLGA scaffold with C myoblast and PLGA scaffold transfected with CDMP-2 gene. The biological activity of muscle cells was measured by MTT method, and the growth curve was drawn. The morphology of the cells was observed by scanning electron microscope (SEM). Results at the early stage after inoculation with PLGA scaffold, the cells could not grow and attach on the scaffold surface in inverted phase contrast microscope, only the crisscross fibrous reticular structure could be seen. After 72 hours, cell proliferation could be seen. MTT showed that the number of muscle cells was 0.0464140.0576650.038945and 0.1321580.1788110.1082840.1738101010 at the wavelength of 492nm, respectively, and there was no statistical significance in each group (P0.05). Conclusion myoblast can grow well on PLGA scaffold, which indicates that PLGA has no cytotoxicity to myoblasts and has excellent biocompatibility, and can be used for tissue repair. It provides a new idea and method for the further study of cartilage repair.
【学位授予单位】：辽宁医学院
【学位级别】：硕士
【学位授予年份】：2012
【分类号】：R318.08;R687
【参考文献】
相关期刊论文 前10条
1 李全双;梅晰凡;郭占鹏;王滢丽;秦书俭;;成肌干细胞在透明质酸凝胶载体中诱导分化为软骨细胞的研究[J];解放军医学杂志;2010年07期
2 张力;范明;王伟;陈晓萍;刘淑红;孙亮;;改良法体外培养大鼠成肌细胞的实验研究[J];军事医学科学院院刊;2007年01期
3 李志华,宋锦
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