聚氨酯-脱细胞基质复合支架的制备及体内外生物相容性研究  

FABRICATION AND BIOCOMPATIBILITY EVALUATION OF POLYURETHANEACELLULAR MATRIX COMPOSITE SCAFFOLD IN VITRO AND IN VIVO

作　　者：肖原[1,2] 张杰[2] 陆燕蓉[2] 员海超[3] 柏琳[2] 蒋霞[2] 程惊秋[2] 

机构地区：[1]四川大学生命科学学院成都,610065 [2]四川大学华西医院再生医学研究中心 [3]四川大学华西医院泌尿外科

出　　处：《中国修复重建外科杂志》2015年第8期1016-1021,共6页Chinese Journal of Reparative and Reconstructive Surgery

基　　金：国家自然科学基金青年基金资助项目(51203100);中国博士后科学基金资助项目(2013M531967)~~

摘　　要：目的构建用于膀胱修复与重建的聚氨酯(polyurethane,PU)-膀胱脱细胞基质(bladder acellular matrix,BACM)复合支架,并对其力学性能和生物相容性进行体内外评价。方法取10只雄性新西兰大白兔膀胱,经1%SDS及1%Triton X-100脱细胞处理后,用PU乳液对所得BACM进行表面涂覆,获得PU-BACM复合支架;用力学试验仪测试BACM与PU-BACM支架的拉伸强度及断裂伸长率。将人膀胱平滑肌细胞(human bladder smooth muscle cells,HBSMC)分别与BACM、PU、PU-BACM支架共培养,通过细胞计数试剂盒8法比较各组材料对细胞增殖的影响。以DMEM培养基制得上述3种材料浸提液,并以DMEM培养基作对照,与HBSMC共培养,流式细胞仪测定细胞周期。取12只雄性新西兰大白兔膀胱剪开5 mm小口,将PU-BACM支架以锁边法缝合于膀胱切口部位以建立膀胱损伤修复模型,术后10、20、40、60 d取材行HE染色观察炎性浸润、材料降解和上皮再生情况。结果 BACM和PU-BACM的拉伸强度分别为(5.78±0.85)N和(11.88±3.21)N,断裂伸长率分别为14.46%±3.21%和23.14%±1.32%,差异均有统计学意义(t=3.182,P=0.034;t=4.332,P=0.012)。细胞增殖实验显示,体外培养5、7 d后,PU-BACM支架组细胞增殖显著高于BACM组(P<0.05);细胞周期实验中,对照组及PU、BACM、PU-BACM组的细胞增殖率分别为36.78%±1.21%、30.49%±0.89%、18.92%±0.84%和22.42%±1.55%,PUBACM组显著高于BACM组(P<0.05)。兔膀胱修复实验中,术后10 d移植部位出现单核炎性细胞浸润情况;20 d炎性反应逐渐减弱、材料逐步降解;40 d观测到上皮再生;60 d移植部位形成与正常组织类似结构,修复基本完成。结论 PU-BACM复合支架较BACM力学性能及生物相容性均有显著提升;在兔膀胱修复实验中,复合支架未导致强烈的免疫反应,并在移植部位形成新生膀胱组织,可为进一步研究提供基础数据。Objective To prepare a composite scaffold using bladder acellular matrix （BACM） and polyurethane （PU） for bladder repair and regeneration, and to evaluate its mechanical properties and biocompatibility. Methods Fresh bladder tissues were obtained from New Zealand rabbits and then treated with 1%SDS and 1%Triton X-100 to obtain BACM. The BACM was combined with PU to fabricate PU-BACM composite scaffold. The tensile strength and elongation at break of BACM and PU-BACM scaffolds were tested. Scaffolds and extracts of scaffolds were prepared to evaluate the biocompatibility. For cell-proliferation analysis, cell counting kit 8 method was used at 1, 3, 5, and 7 days after co-culture of human bladder smooth muscle cell （HBSMC） and scaffolds. The cell cycle was tested by flow cytometry after HBSMC co-cultured with extracts of scaffolds and DMEM culture medium （control group） for 24 hours. Finally, 12 New Zealand rabbits were used to establish the model of bladder repair and regeneration. Incision of 5 mm was made on the bladder, and PU-BACM scaffold was sutured with the incision. The rabbits were sacrificed at 10, 20, 40, and 60 days after surgery to observe the inflammatory cell infiltration, new tissues formation, and regeneration of epithelium by HE staining. Results The tensile strength of BACM and PU-BACM composite scaffold was （5.78 ±0.85） N and （11.88 ± 3.21） N, and elongation at break was 14.46%±3.21% and 23.14%±1.32% respectively, all showing significant diffeence （t=3.182, P=0.034; t=4.332, P=0.012）. The cell-proliferation rates of controls, PU, BACM, and PU-BACM were 36.78%＋ 1.21%, 30.49%＋0.89%, 18.92%＋0.84%, and 22.42%±1.55%, it was significantly higher in PU-BACM than BACM （P〈0.05）. In the bladder repair and regeneration experiment, inflammatory cell infiltration was observed at 10 days after operation, and reduced at 20 days after implantation. In the meanwhile, the degradation of scaffolds was observed in vivo. The regeneration of epithelium could be observed a

关 键 词：脱细胞基质 聚氨酯 膀胱修复 组织工程 兔 

分 类 号：R318.08[医药卫生—生物医学工程]