Ⅰ型胶原修饰纯钛片促进人脂肪间充质干细胞增殖  

Type I collagen modified titanium sheet improves the proliferation of human adipose-derived mesenchymal stem cells

作　　者：叶川[1] 马敏先[2] 张弢[3] 汤晋[1] 孙博[1] 杨龙[1] 任厚相 孙琦[1] 杨静[2] 张俊标 

机构地区：[1]贵阳医学院附属医院骨科,贵州省贵阳市550004 [2]贵阳医学院附属医院口腔修复科,贵州省贵阳市550004 [3]解放军第117医院骨科,浙江省杭州市310013

出　　处：《中国组织工程研究》2014年第25期4032-4037,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);贵州省科学技术基金(黔科合J字20102177);贵州省科技厅资助项目(LG2012008)~~

摘　　要：背景:钛及钛合金在人工关节、骨折固定、口腔种植等临床领域应用最多,但在这些领域都存在骨量不足的复杂病例,干细胞研究的深入为骨缺损提供了促进新骨生成的解决方案,钛和干细胞的生物相容性以及钛表面改性优化等问题已引起重视。目的:观察Ⅰ型胶原修饰纯钛片后能否改善其与人脂肪间充质干细胞的生物相容性。方法:实验分为2组,修饰组用Ⅰ型胶原修饰纯钛片,未修饰组为未用Ⅰ型胶原修饰的纯钛片,分别将第6代人脂肪间充质干细胞种植于两组钛片上。分别计算细胞种植于两组钛片上1,2,4 h贴壁细胞的数量,比较两组细胞的黏附率。MTT比色法比较细胞种植于两组钛片上2,4,6,8 d细胞在钛片上的增殖情况。比较细胞种植于两组钛片上3,6,9 d细胞的DNA和蛋白含量。将人脂肪间充质干细胞分别接种于修饰组和未修饰组钛片,细胞种植于两组钛片上6 d时,电镜扫描观察细胞在两组钛片上的生长情况。结果与结论:修饰组的细胞黏附率在培养1,2 h时高于未修饰组(P<0.05)。MTT比色法显示,修饰组和未修饰组细胞的吸光度值均随培养时间延长而增高,培养第4,6,8天修饰组吸光度值较未修饰组明显升高(P<0.05)。培养第6,9天,修饰组的DNA和蛋白含量均高于未修饰组(P<0.05)。电镜扫描观察显示,培养第6天,修饰组在细胞贴壁数量、贴壁细胞基质分泌情况明显好于未修饰组。结果证实,Ⅰ型胶原修饰纯钛片的表面活性及生物相容性较好,可促进人脂肪间充质干细胞增殖。BACKGROUND：Titanium and titanium aloy are used mostly in artificial joints, fracture fixation, and oral transplantation, while there are complex cases of insufficient bone mass in these areas. The deepened research of stem cels offers a solution for bone injury to promote new bone formation. The biocompatibility of titanium and stem cels and optimization of titanium surface modification have aroused people＆#39;s attention. OBJECTIVE：To investigate whether the biocompatibility of titanium and human adipose-derived mesenchymal stem cels can be improved by type I colagen modification of titanium sheets. METHODS：The experiment was divided into two groups. Modification group： titanium sheet was modified with type I colagen; control group： titanium sheet was not modified with type I colagen. Human adipose-derived mesenchymal stem cels at passage 6 were implanted into titanium sheet in two groups. Then we calculated the number of adherent cels in two groups at 1, 2 and 4 hours after implantation, and compared the celladhesion rate. MTT assay was used to observe the proliferation of cels on titanium sheet at 2, 4, 6 and 8 days after implantation. DNA and protein content of cels were detected at 3, 6, 9 days after implantation. The growth of human adipose-derived mesenchymal stem cels seeded upon the titanium sheets was observed under scanning electron microscope at 6 days. RESULTS AND CONCLUSION：When the cels were cultured for 1 hour and 2 hours, the number of adherent cels in the modification group was higher than in the control group （P 〈 0.05）. The absorbance of cels in two groups was increased as the culture time, as detected by MTT assay. The modification group had a significantly higher absorbance value than the control group at 4, 6, 8 days （P 〈 0.05）. DNA and protein contents of the cels in the modification group were higher than that in control group at 6 and 9 days （P 〈 0.05）. At 6 days, the number of adherent cels and secretion of adherent stromal cellmatrix in the modification

关 键 词：生物材料 材料相容性 Ⅰ型胶原 钛 生物相容性 生物材料 人脂肪间充质干细胞 细胞贴壁率 流式 细胞增殖 体外 DNA含量 蛋白含量 国家自然科学基金 

分 类 号：R318[医药卫生—生物医学工程]