经骨形态发生蛋白7成骨预处理聚乳酸三维支架的体外培养  

A three-dimensional polylactic acid scaffold pretreated with bone morphogenetic protein 7

作　　者：贝抗胜[1] 刘延晓[2] 熊英辉[1] 吴礼杨[1] 卢林俊[1] 

机构地区：[1]粤北人民医院骨科,广东省韶关市512026 [2]衢州市人民医院骨科,浙江省衢州市324000

出　　处：《中国组织工程研究》2016年第3期346-351,共6页Chinese Journal of Tissue Engineering Research

摘　　要：背景:为改善聚乳酸的细胞亲和性,前期实验在其表面加入骨形态发生蛋白7功能基团,制备了经骨形态发生蛋白成骨预处理的聚乳酸三维支架。目的:验证经骨形态发生蛋白成骨预处理聚乳酸三维支架与骨膜来源细胞的相容性。方法:取传至3代后成骨诱导分化的人骨膜来源细胞,实验组以1×10~9 L^(-1)的细胞浓度接种于含聚乳酸支架的培养板内,聚乳酸支架预先经含骨形态发生蛋白7的成骨诱导培养液浸泡24 h;对照组直接以1×10~9 L^(-1)的细胞浓度接种于培养板内。培养后1,3,5,7,9,11 d,采用CCK-8法检测细胞增殖能力;培养后3,6,9,12 d,电镜下观察细胞-支架复合体。结果与结论:1细胞增殖结果:两组细胞生长曲线均呈"S"型,实验组对数生长期细胞增殖较快,平稳期细胞数目明显下降,对照组平稳期细胞数目继续上升。2透射电镜观察结果:复合培养3-6 d时,支架表面细胞黏附数量逐渐增多,细胞生物状态良好,细胞器旺盛,至第9天,开始出现细胞老化及凋亡。3扫描电镜观察结果:复合培养3-9 d时,支架表面细胞黏附数量逐渐增多,增殖旺盛,至12 d时见凋亡细胞。4结果表明,经骨形态发生蛋白成骨预处理聚乳酸三维支架具有良好的细胞相容性,其与细胞复合培养1周为较佳的体外复合培养时间。BACKGROUND： To improve the cell affinity of polylactic acid, the three-dimensional polylactic acid scaffold is made after pretreatment with bone morphogenetic protein 7（BMP7）. OBJECTIVE： To evaluate the compatibility of BMP7-pretreated three-dimensional polylactic acid scaffold withperiosteum-derived cells. METHODS： Passage 3 human periosteum-derived cells undergoing osteogenic induction at a density of 1×10~9/L were seeded into culture media containing the three-dimensional polylactic acid scaffold（experimental group） or seeded into simple culture media（control group）. In the experimental group, the scaffold was pretreated with BMP7 for 24 hours. At 1, 3, 5, 7, 9, 11 days after culture, cell counting kit-8 method was used to measure cellular proliferation; at 3, 6, 9, 12 days, the cell-scaffold complex was observed under an electron microscope. RESULTS AND CONCLUSION： In the two groups, the cells had an S-shaped growth curve. Compared with the control group, the cell proliferation in the experimental group was faster at the logarithmic phase, but slower at the stationary phase（P 〈0.05）. Under the transmission electron microscope, in the experimental group, the number of adherent cells was increased gradually, and the cells grew well with exuberant organelles at 3-6 days of co-culture, but cell aging and apoptosis were found at 9 days of co-culture. Under the scanning electron microscope, in the experimental group, the number of adherent cells on the scaffold surface gradually increased at 3-9 days of co-culture, and then cell apoptosis was notable at 12 days. These findings indicate that BMP7-pretreated three-dimensional polylactic acid scaffold has good cytocompatibility, and the optimal in vitro co-culture duration is 1 week.

关 键 词：组织支架 骨形态发生蛋白7 骨膜 组织工程 生物材料 骨生物材料 聚乳酸 骨膜来源细胞 细胞培养 超微结构 

分 类 号：R318[医药卫生—生物医学工程]