CAJ | 학술논문

目的：试用两种交联剂对小牛真皮基质来源的支架材料进行交联，比较支架的细胞毒性、结构、生物相容性和细胞贴附的差异，为在体动物试验提供实验依据。方法将脱细胞真皮基质分为两组，分别浸入0.05%戊二醛溶液和0.2%水溶性交联剂进行交联，MTT 法检测细胞毒性和溶血率。将交联后的支架分别植入大鼠皮下，评价生物相容性。以排液法粗测孔隙率，并在电镜下观察支架的结构和孔隙大小。培养间充质干细胞并贴附于两种方法处理的材料表面，电镜下观察贴附情况。结果以戊二醛溶液和水溶性交联剂两种方法处理的支架细胞毒性检测均合格，溶血率分别为4.61%与2.97%，均符合国家标准。经戊二醛交联的支架生物相容性差，炎症反应始终存在，水溶性交联剂处理的真皮基质组织相容性较好，仅有轻微的炎症反应。水溶性交联剂制备的支架材料孔隙率为84.3%±5.0%，戊二醛制备的支架材料孔隙率为79.7%±10.8%，差异不具有统计学意义（P >0.05）。戊二醛交联的支架细胞贴附差，而水溶性交联剂制备的支架细胞贴附良好。结论应用水溶性交联剂处理的支架细胞毒性和溶血率检测均合格，具有良好的生物相容性、孔隙率和细胞贴附性，该法可以作为后期制备软骨细胞移植支架的交联方法。
목적：시용량충교련제대소우진피기질래원적지가재료진행교련，비교지가적세포독성、결구、생물상용성화세포첩부적차이，위재체동물시험제공실험의거。방법장탈세포진피기질분위량조，분별침입0.05%무이철용액화0.2%수용성교련제진행교련，MTT 법검측세포독성화용혈솔。장교련후적지가분별식입대서피하，평개생물상용성。이배액법조측공극솔，병재전경하관찰지가적결구화공극대소。배양간충질간세포병첩부우량충방법처리적재료표면，전경하관찰첩부정황。결과이무이철용액화수용성교련제량충방법처리적지가세포독성검측균합격，용혈솔분별위4.61%여2.97%，균부합국가표준。경무이철교련적지가생물상용성차，염증반응시종존재，수용성교련제처리적진피기질조직상용성교호，부유경미적염증반응。수용성교련제제비적지가재료공극솔위84.3%±5.0%，무이철제비적지가재료공극솔위79.7%±10.8%，차이불구유통계학의의（P >0.05）。무이철교련적지가세포첩부차，이수용성교련제제비적지가세포첩부량호。결론응용수용성교련제처리적지가세포독성화용혈솔검측균합격，구유량호적생물상용성、공극솔화세포첩부성，해법가이작위후기제비연골세포이식지가적교련방법。
Objective Repair of focal articular cartilage defect may be solved by autologous chondrocyte implantation. Preparation of proper scaffold is one of the most important problems to be solved. We are to construct scaffolds for their application in repairing cartilage defect of animal model with two kinds of cross-linking agents. Methods Acellular dermal matrices (ADM) were divided into two groups. One group was treated with 0.05%glutaraldehyde for 2 hours for cross-linking and the other was treated with 0.2%self-made cross-linking agent. Cytotoxity and hemolysis were tested with MTT method. Porosity was examined with liquid discharge method. Mesenchymal stem cells were used for cell adhesion test, and both scaffolds were implanted into SD rats for biocompatibility test. The structure of two kinds of scaffolds was observed under electron microscope. Results Both of the cytotoxicity and hemolysis of two kinds of scaffolds fitted the national standards. And the porocity of scaffold cross-linked with glutaraldehyde and with self-made cross-linking agent were 79.7%± 10.8%and 84.3%± 5.0%, respectively, and there exists no significant difference between the two kinds (P > 0.05). But the biocompatibility of scaffold cross-linked with glutaraldehyde was poorer than that of scaffold cross-linked with self-made agent. According to the results of cell adhesion test, mesenchymal stem cells adhered to the scaffold treated with self-made agent perfectly, but did not adhere to the one treated with glutaraldehyde well. Conclusion The scaffold treated with self-made agent exerts little cytotoxicity, but possesses good biocompatibility, porosity, and cell adhesion. Thus it will be a proper choice for repairing cartilage defect in the future.