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聚羟基丁酸/戊酸酯共聚物膜与人骨髓间充质干细胞的体外生物相容性

In vitro biocompatibility of polyhydroxybutyrate-co-volerate membrane with human bone marrow mesenchymal stem cells

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【作者】 张弢何志旭叶川刘杰麟马敏先孙博王梅杨华

【Author】 Zhang Tao;He Zhi-xu;Ye Chuan;Liu Jie-lin;Ma Min-xian;Sun Bo;Wang Mei;Yang Hua;Guiyang Medical University;Department of Orthopedics, the117th Hospital of PLA;the Center of Stem Cells and Tissue Engineering Research,Guiyang Medical University;Department of Pediatrics,Guiyang Medical University;Department of Orthopedics,Affiliated Hospital of Guiyang Medical University;Department of Immunology, Guiyang Medical University;School of Stomatology, Guiyang Medical University;Department of Stomatology,Affiliated Hospital of Guiyang Medical University;

【机构】 贵阳医学院解放军117医院骨科贵阳医学院组织工程与干细胞实验中心贵阳医学院儿科教研室贵阳医学院附属医院骨科贵阳医学院免疫学教研室贵阳医学院口腔医学院贵阳医学院附属医院口腔科

【摘要】 背景:聚羟基丁酸/戊酸酯共聚物是近年来受到重视的聚羟基脂肪酸族组织工程支架材料,具有免疫排斥反应低、生物相容性好和降解产物无毒副作用的优点。目的:观察聚羟基丁酸/戊酸酯共聚物膜材料与人骨髓间充质干细胞的体外生物相容性。方法:将第3代人骨髓间充质干细胞种植于聚羟基丁酸/戊酸酯共聚物膜上作为实验组,培养板单纯培养细胞作为对照组,计算1,2,4 h两组贴壁细胞的数量,得出细胞贴壁率。MTT比色法观察2,4,6,8 d两组细胞的增殖情况。采用Hoechst33258荧光法,检测3,6,9,12 d两组细胞内的DNA含量。将人骨髓间充质干细胞接种聚羟基丁酸/戊酸酯共聚物膜材料上5 d后,电镜扫描观察细胞在材料上的生长情况。结果与结论:共培养1 h时,实验组的细胞贴壁率低于对照组;其他时间段两组之间细胞贴壁率差异无显著意义。两组各时点间的细胞增殖差异无显著意义。两组各时间点细胞内DNA含量差异无显著意义。扫描电镜观察人骨髓间充质干细胞在聚羟基丁酸/戊酸酯共聚物膜上生长良好,形态呈梭形,细胞间连接紧密,分泌较多细胞基质。证明聚羟基丁酸/戊酸酯共聚物膜材料与人骨髓间充质干细胞有良好的生物相容性。

【Abstract】 BACKGROUND: Polyhydroxybutyrate-co-volerate(PHBV) is a noticeable tissue engineering material of polyhydroxyalkanoates family. It has the properties of low immune rejection response and good biocompatibility, and its degradation products are non-toxic. OBJECTIVE: To investigate the biocompatibility of PHBV membrane material and human bone marrow mesenchymal stem cells in vitro. METHODS: Human bone marrow mesenchymal stem cells at passage 3 were seeded upon PHBV membrane as experimental group and upon conventional culture plates as control group. Then we calculated the adherent cell number of two groups at 1, 2 and 4 hours and got the cell adherent rate. 3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazoliumbromide assay was used at days 2, 4, 6, 8 to observe the cell proliferation of two groups. Fluorimetric method with the fluorescent dye Hoechst 33258 was used to detect the DNA content of cells at days 3, 6, 9 and 12 in both groups. After cells were seeded upon PHBV membrane for 5 days, the cell growth upon thematerial was examined under a scanning electron microscope. RESULTS AND CONCLUSION: When the cells were cultured for 1 hour, the adherent rate in the experimental group was lower than that in the control group; but there were no significant differences between two groups at the other two periods. No difference was found in the cell proliferation and the DNA content between the two groups. Human bone marrow mesenchymal stem cells seeded upon PHBV membrane for 5 days grew well with spindle morphology and the intercellular connections were tight and more extracellular matrices were observed by scanning electron microscopy. Taken together, PHBV membrane material shows a good biocompatibility with human bone marrow mesenchymal stem cells.

【基金】 国家自然科学基金资助项目(81360232);贵州省卫生厅资助项目(gzwkj2010-1-042);贵州省国际科技合作项目(黔科合外G字2010-7);贵州省科技厅社发联合基金项目(黔科合20103166)~~
  • 【文献出处】 中国组织工程研究 ,Chinese Journal of Tissue Engineering Research , 编辑部邮箱 ,2014年12期
  • 【分类号】R318.08
  • 【被引频次】2
  • 【下载频次】127
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