温固化水凝胶复合细胞外基质与膀胱平滑肌细胞的相容性  

Biocompatibility of the compound pattern of extracellular matrix and temperature dependent hydrogel with bladder smooth muscle cells

作　　者：姚颐[1] 张杰[1] 方立[1] 叶达夫[1] 彭建平[1] 李强[1] 

机构地区：[1]武汉大学人民医院泌尿外科,湖北省武汉市430060

出　　处：《中国组织工程研究与临床康复》2007年第26期5090-5093,共4页Journal of Clinical Rehabilitative Tissue Engineering Research

基　　金：国家自然科学基金资助项目(30471734)~~

摘　　要：目的:选用膀胱平滑肌细胞为实验细胞,评估细胞外基质/温度敏感性水凝胶复合支架材料的生物相容性。方法:实验于2005-02/05在武汉大学人民医院泌尿外科研究室完成。①实验分组:实验分为4组。细胞外基质/温度敏感性水凝胶复合支架组:细胞外基质/温度敏感性水凝胶复合支架种植平滑肌细胞;单纯细胞外基质组:单纯细胞外基质种植平滑肌细胞;阴性对照组:单纯培养平滑肌细胞;空白对照组:无细胞培养液。②实验操作:种植细胞前先用培养液将支架材料预湿,取第3代兔膀胱平滑肌细胞悬液(1×108L-1)50μL缓慢接种于材料上。③实验评估:培养2d后倒置相差显微镜下观察细胞黏附、生长情况以及扫描电镜下观察细胞在材料上的空间生长情况;培养5,10d取细胞外基质/温度敏感性水凝胶复合支架组、单纯细胞外基质组材料,进行细胞计数,观察细胞黏附数量;培养1,3,5,7d时MTT法检测细胞的活力。结果:①相差显微镜下可见细胞外基质/温度敏感性水凝胶复合材料为红染的网状结构,纤维较粗,网孔直径较小,未见到细胞碎片;种植平滑肌细胞后6h,可见细胞紧密黏附于材料表面;种植平滑肌细胞后12h,可见细胞已伸展,有多个突起;培养3d时,贴附于材料的细胞数量增加,细胞增殖明显。②扫描电镜下可见发育良好的细胞附于材料上,伪足沿纤维伸展,附着牢固,细胞间连接紧密,可见到细胞外基质分泌。③膀胱平滑肌细胞在单纯细胞外基质上黏附性良好,黏附率为87.6%,复合温度敏感性水凝胶后的细胞外基质表面黏附能力较之增强,黏附率为96.7%。④细胞活力:细胞外基质/温度敏感性水凝胶复合支架组、阴性对照组细胞活力(A)高于单纯细胞外基质组,差异有显著性意义(P<0.05,0.01),培养1,3,5d细胞外基质/温度敏感性水凝胶复合支架组细胞活力(A)低于阴性对照组,培养7d高于阴性对照组,差异�AIM：To evaluate the biocompatibility of compound pattem of extracellular matrix （ECM） and temperature-sensitive hydrogel （HG） in vitro to the bladder smooth muscle cells （SMCs）. METHODS： The study was finished in the Laboratory of Urology in the Renmin Hospital of Wuhan University from February to May in 2005. ①There were 4 groups in this experiment： SMCs were cultured in ECM （group A）, in the ECM/HG compounds materials （group B） and on the plastic surfaces as a negative control （group C） respectively, There was no cell culture fluid in group D, which played as a blank control.②The compound materials were moistened with culture fluid and inoculated with 50 p.L rabbit bladder SMC suspension （1×10^8L^-1） at third passage.③Two days after culturing, cells and the compound materials were observed by phase contrast microscopy and scanning electron microscope to assess cell adhesion, proliferation, and growth, At the 5th and 10th days, the cells of group A and group B were counted to observe the cell adhesion. Cell activity was detected with MTT assay at 1, 3, 5 and 7 days after culture. RESULTS： ①Histologically, the ECM/HG compounds appeared as a net-like and red-stained construction with enough meshes and without any cellular fragments, Six hours after implantation, cells were observed to adhere on the compounds and extend spuriously along the fibers 12 hours later, Three days later, the adhesive cells were increased in number and proliferated obviously, ②Under scanning electron microscope, the proliferated cells adhered closely on materials, and pseudopodia extended along fibers, even some ECMs were secreted.③The SMC adhesion was notable on the ECM and the compound materials as well, the adhesion rate was 87,6% and 96,7%, respectively, ④The cell activity in group B and group C was significantly higher than that of group A （P 〈 0,05, 0.01）. At 1, 3, 5 days after culture, the cell activity of group B was lower than that of group C and higher at 7 days, with the s

关 键 词：水凝胶 细胞外基质 复合支架材料 生物相容性 

分 类 号：R318.08[医药卫生—生物医学工程]