CAJ | 학술논문

目的评价猪来源关节软骨脱细胞支架的生物安全性.方法取猪四肢关节软骨,分别制备脱细胞和未脱细胞软骨支架.在人软骨细胞中分别加入浓度为100%、50%、25%的脱细胞软骨支架浸提液[无血清DMEM培养液与支架表面积按1 ml:(3～6)cm~2比例混合,37℃静置72h],计算相对增殖率,判定其体外细胞毒性.0.9%NaCl与支架表面积按1 ml:(3～6)cm~2比例混合,37℃静置72 h制备脱细胞软骨支架浸提液,分别用于全身急性毒性实验(小白鼠)、溶血实验、热原检测实验、皮内刺激实验(大白兔),分别观察其体内毒性、溶血程度、热原和刺激情况.在大白兔体内分别埋植脱细胞软骨支架与未脱细胞软骨支架,观察炎症反应和细胞免疫情况.结果猪脱细胞软骨支架浸提液对细胞生长无抑制作用,无细胞毒性.注射支架浸提液后小白鼠一般情况良好,无全身急性毒性反应.支架浸提液溶血程度为1.3%,无溶血作用.支架浸提液无热原存在且原发刺激指数为0分,皮内刺激实验阴性.脱细胞软骨支架与未脱细胞软骨支架相比免疫原性小,无炎症反应和淋巴细胞浸润.结论猪来源关节软骨脱细胞支架具有良好的生物相容性,将有可能成为理想的天然生物材料,具有良好的应用前景.
목적 평개저래원관절연골탈세포지가적생물안전성.방법 취저사지관절연골,분별제비탈세포화미탈세포연골지가.재인연골세포중분별가입농도위100%、50%、25%적탈세포연골지가침제액[무혈청DMEM배양액여지가표면적안1 ml:(3～6)cm~2비례혼합,37℃정치72h],계산상대증식솔,판정기체외세포독성.0.9%NaCl여지가표면적안1 ml:(3～6)cm~2비례혼합,37℃정치72 h제비탈세포연골지가침제액,분별용우전신급성독성실험(소백서)、용혈실험、열원검측실험、피내자격실험(대백토),분별관찰기체내독성、용혈정도、열원화자격정황.재대백토체내분별매식탈세포연골지가여미탈세포연골지가,관찰염증반응화세포면역정황.결과 저탈세포연골지가침제액대세포생장무억제작용,무세포독성.주사지가침제액후소백서일반정황량호,무전신급성독성반응.지가침제액용혈정도위1.3%,무용혈작용.지가침제액무열원존재차원발자격지수위0분,피내자격실험음성.탈세포연골지가여미탈세포연골지가상비면역원성소,무염증반응화림파세포침윤.결론 저래원관절연골탈세포지가구유량호적생물상용성,장유가능성위이상적천연생물재료,구유량호적응용전경.
Objective Evaluate the biological safety of porcine articular acellular cartilage scaffold. Methods Porcine articular cartilage was used to prepare acellular and non-acellular cartilage scaffold, Add 100%, 50% and 25% acellular cartilage scaffold leaching fluid [DMEM culture solution and surface area of scaffold were mixed according to 1 ml: (3 - 6) cm~2, 37 ℃, 72 h], RGR was valued to determine its vitro cytotoxicity. After acellular cartilage scaffold leaching fluid were prepared [0.9% NaCl and surface area of scaffold were mixed according to 1 ml: (3 - 6) cm~2, 37 ℃, 72 h], the biocompatility was evaluated both in vivo and in vitro, including long term subcutaneous implantation test, cytotoxic test, acute systemic toxicity test, hemolysis test, intradermal test, pyrogenic test. Results Porcine acellular cartilage scaffold leaching liquid has no inhibitory effect on cell growth. After the injection of leaching fluid, rats showed no acute systemic toxicity. The leaching fluid has no hemolysis, the degree of hemolysis was 1.3%. Biologic assays were pyrogen free and primary stimulation index was 0. Compared with non-acellular cartilage scaffold, acellular cartilage scaffold showed low immunogenicity, no inflammatory response and no lymphocyte infiltration. Conclusions Porcine acellular cartilage scaffold had good biocompatibility. It will he an ideal natural biological material and has good application prospects.