摘要
目的制备兔膀胱细胞外基质材料,检测其生物相容性,探讨其作为组织工程泌尿道修复重建支架材料的可行性。方法联合应用渗透溶液法-酶消化法-去垢剂洗涤法制备兔膀胱细胞外基质,将体外培养的兔膀胱移行上皮细胞与其复合培养,观察细胞在材料表面的黏附、生长、增殖情况;MTT法检测材料浸提液对膀胱移行上皮细胞的细胞毒性;将材料植入兔背部皮下检测其组织相容性。结果兔膀胱细胞外基质冻干后呈白色半透明薄膜状,扫描电镜观察材料一面呈纤维网状结构,另一面呈平坦致密结构,两面均无细胞残留。膀胱移行上皮细胞能够在材料表面正常黏附、生长、增殖,细胞形态良好。MTT法检测材料细胞毒性分级为0级或1级。皮下植入实验中动物无异常反应,材料能在体内逐渐降解并引导新生组织长入。结论本研究制备的兔膀胱细胞外基质已完全除去组织中的细胞成份,同时保留了细胞外基质的纤维网状结构,具有良好的细胞相容性和组织相容性,可以作为组织工程泌尿道修复重建的支架材料。
Objective To investigate the biocompatibility of rabbit bladder extracellular matrix （BECM） and evaluate the feasibility of using BECM as scaffold for reconstruction of tissue engineering urinary tract. Methods By the application of associating the solution diosmosis, enzymatic digestion with chemical detergent, the rabbit bladder was deeellularized to prepare the BECM, on which rabbit bladder transitional epithelial cells were cultured and seeded in vitro, of which the adhesion and proliferation were observed. MTT method was used to evaluate the eytotoxieity of BECM, which was implanted into the rabbit back for evaluating its histoeompatibility meant that the toxicity, degradation and local inflammatory response were studied through gross observation and HE staining. Results The prepared rabbit BECMs were freeze-drying, and looked like thin semitransparent membrane. By electron microscope examination, there were no residues of cells found on BECM membrane, of which one side had the reticular fibrous structure, and the other side had the compact structure. The co-culture of BECM membrane with bladder transitional epithelial cells indicated that BECM had a good eytoeompatibility. The eytotoxieity score tested by MTT method was number 0 and 1. The rabbits in the implant test had no abnormal response. The BECMs degraded gradually in vivo, and growth of peripheral tissue could be seen in the materials after eight weeks. Conclusion The BECMs prepared by our process are cell-free under scanning electron microscopy. Meanwhile, the reticular structure of the tissue matrix is well preserved. The BECMs have the good eytoeompatibility with transitional epithelial cells but without eytotoxieity. The BECMs can degrade gradually in vivo with good histoeompatibility. BECM is a good bio-derived material of tissue engineering as scaffold for urinary tract reconstruction.
出处
《四川大学学报（医学版）》
CAS
CSCD
北大核心
2007年第6期1009-1012,1032,共5页
Journal of Sichuan University(Medical Sciences)
关键词
膀胱
细胞外基质
生物相容性
Bladder Extraeellular matrix Bioeompatibility
