CAJ | 학술논문

背景:脱细胞膀胱黏膜下层是一种天然的细胞外基质生物材料,主要由Ⅰ、Ⅲ型纤维胶原蛋白构成,是一种理想的生物支架材料.目的:脱细胞猪膀胱组织作为组织工程支架材料的生物学评价.方法:取适量健康猪膀胱置于PBS和叠氮钠的混合溶液,浸泡过夜,刮去膀胱黏膜层.用低渗、-80℃反复冻融、DNase和RNase混合液消化和NaOH裂解连续方法制备脱细胞膀胱黏膜下层.通过观察其组织学结构特征、DNA残留、细胞毒性、细胞黏附效果及皮下炎症反应等来综合评价脱细胞膀胱黏膜下层的生物相容性.结果与结论:正常猪的膀胱经改进的脱细胞方法处理后,绝大部分细胞组分被去处,细胞外基质结构保持完好.细胞毒性试验结果显示脱细胞膀胱黏膜下层细胞毒性为1级,DNA提取结果中未见有DNA残留,细胞黏附结果显示猪平滑肌细胞能在脱细胞支架上较好的贴附生长,SD大鼠皮下植入试验显示无明显的炎症反应.结果证实所制备的脱细胞膀胱黏膜下层结构保存完好,有良好的组织相容性,可能成为组织工程修复的替代材料.
배경:탈세포방광점막하층시일충천연적세포외기질생물재료,주요유Ⅰ、Ⅲ형섬유효원단백구성,시일충이상적생물지가재료.목적:탈세포저방광조직작위조직공정지가재료적생물학평개.방법:취괄량건강저방광치우PBS화첩담납적혼합용액,침포과야,괄거방광점막층.용저삼、-80℃반복동융、DNase화RNase혼합액소화화NaOH렬해련속방법제비탈세포방광점막하층.통과관찰기조직학결구특정、DNA잔류、세포독성、세포점부효과급피하염증반응등래종합평개탈세포방광점막하층적생물상용성.결과여결론:정상저적방광경개진적탈세포방법처리후,절대부분세포조분피거처,세포외기질결구보지완호.세포독성시험결과현시탈세포방광점막하층세포독성위1급,DNA제취결과중미견유DNA잔류,세포점부결과현시저평활기세포능재탈세포지가상교호적첩부생장,SD대서피하식입시험현시무명현적염증반응.결과증실소제비적탈세포방광점막하층결구보존완호,유량호적조직상용성,가능성위조직공정수복적체대재료.
BACKGROUND: Acellular bladder submucosa is a natural extracellular matrix, which is mainly composed of collagen Ⅰ and Ⅲ. It is regarded as an ideal biological scaffold material. OBJECTIVE: To evaluate the biocompatibility of acellular bladder submucosa as a tissue engineered scaffold material. METHODS: Pig urinary bladder was immersed in the solution of PBS and sodium azide for a night, and the mucosa was removed. Acellular bladder submucosa was prepared using continuous hypotension, freeze-thawed treatment and NaOH spallation. The biocompatibility of acellular bladder submucosa was evaluated through histologic structure, DNA residual, cytotoxicity, cell adhesion, as well as subcutaneous inflammatory reactions. RESULTS AND CONCLUSION: The cell components were completely eliminated after deoellularization treatment, while the extracellular matrix was remained intact as normal bladder:According to MTT results, cytotoxicity of acellular bladder matrix was assigned to be the first grade. No DNA signal was observed after extraction, and the matrix also supported porcine smooth muscle cell attachment and proliferation. Subcutaneous implantation of the matrix indicated that the acellular bladder submucosa trigger no immunologic rejection reaction obviously. The results demonstrated that: the acellular bladder submucosa prepared here exhibits excellent biocompatibility, which can be used as substitution in tissue-engineering field.