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

作　　者：张弢[1,2] 何志旭[3,4] 叶川[3,5] 刘杰麟[3,6] 马敏先[7,8] 孙博[5] 王梅[7] 杨华[5] 

机构地区：[1]贵阳医学院,贵州省贵阳市550004 [2]解放军117医院骨科,浙江省杭州市310013 [3]贵阳医学院组织工程与干细胞实验中心,贵州省贵阳市550004 [4]贵阳医学院儿科教研室,贵州省贵阳市550004 [5]贵阳医学院附属医院骨科,贵州省贵阳市550004 [6]贵阳医学院免疫学教研室,贵州省贵阳市550004 [7]贵阳医学院口腔医学院,贵州省贵阳市550004 [8]贵阳医学院附属医院口腔科,贵州省贵阳市550004

出　　处：《中国组织工程研究》2014年第12期1920-1925,共6页Chinese Journal of Tissue Engineering Research

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)~~

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。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 cel s in vitro. METHODS：Human bone marrow mesenchymal stem cel s at passage 3 were seeded upon PHBV membrane as experimental group and upon conventional culture plates as control group. Then we calculated the adherent cel number of two groups at 1, 2 and 4 hours and got the cel adherent rate. 3-（4,5-Dimethylthiazol-2-yl）-2,5-diphenyltetrazoliumbromide assay was used at days 2, 4, 6, 8 to observe the cel proliferation of two groups. Fluorimetric method with the fluorescent dye Hoechst 33258 was used to detect the DNA content of cel s at days 3, 6, 9 and 12 in both groups. After cel s were seeded upon PHBV membrane for 5 days, the cel growth upon the＆amp;nbsp;material was examined under a scanning electron microscope. RESULTS AND CONCLUSION：When the cel s 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 cel proliferation and the DNA content between the two groups. Human bone marrow mesenchymal stem cel s seeded upon PHBV membrane for 5 days grew wel with spindle morphology and the intercel ular connections were tight and more extracel ular matrices were observed by scanning electron microscopy. Taken together, PHBV membrane material shows a good biocompatibility with human bone marrow mesenchymal stem cells.

关 键 词：生物材料 材料生物相容性 聚羟基丁酸 戊酸酯共聚物 人骨髓间充质干细胞 生物相容性 组织工程 细胞贴壁率 细胞增殖 体外 DNA含量 高分子 材料活性 国家自然科学基金 

分 类 号：R394.2[医药卫生—医学遗传学]