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[目的]对比分析复合软骨细胞的海藻酸钠凝胶球在体内外的降解特点。[方法]取2个月龄新西兰兔5只,酶解消化获取膝关节软骨细胞,与海藻酸钠混合制成细胞密度为5.8×106/ml的混悬液,制成体积为25μl软骨细胞-海藻酸钠凝胶球,在6孔板中进行体外培养对照;体内组取4个月龄新西兰兔36只,双侧股骨滑车行直径4 mm的全层软骨缺损,并将软骨细胞-海藻酸钠凝胶球置入缺损处并经MRI评估缺损模型。分别于1、2、4、6周,通过倒置显微镜观察体外对照海藻酸钠降解情况;通过MRI、大体显微镜和番红O染色观察体内组中海藻酸钠凝胶球体内降解过程。[结果]大体显微观察体外对照组中1周海藻酸钠凝胶球边界清楚,4周凝胶球边界模糊不清,6周藻酸钠边界消失且成糊状。体内组中Roberts MRI评价修复效果显示4周(2.25±0.500)与1周(1.00±0.817)相比差异有统计学意义(t=2.611 P=0.0401),与2周(1.25±0.500)相比差异有统计学意义(t=2.828,P=0.0300),与6周(2.75±0.500)相比差异无统计学意义(t=1.416,P=0.2070);大体观察1周时凝胶球开始液化,4周时海藻酸钠完全消失,且随着凝胶球的降解,缺损以组织修复;番红O染色示实验组修复1、2、4、6周后分别表现为无修复、部分纤维修复、大部分修复、完全纤维修复。[结论]与体外对比,复合软骨细胞的海藻酸钠凝胶球在兔膝关节内生物相容性较好,降解时间明显缩短,大约4周吸收完全,可以为临床应用提供理论依据。
[Objective] To comparatively analyze the degradation characteristics of composite alginate gelatin balls in vitro and in vivo. [Method] Five New Zealand rabbits of 2 months old were taken and digested by enzymolysis to obtain the chondrocytes of knee joint. The suspension was mixed with sodium alginate to make a cell density of 5.8 × 106 / ml, and the volume of 25μl chondrocytes - Alginate gelatin balls were cultured in a 6-well plate in vitro. Thirty-six New Zealand rabbits, 4 months old, were performed in vivo. The bilateral femoral trochlear cartilage was excised as 4 mm diameter cartilage defects. Chondrocytes, Sodium sodium gel balls were placed into the defect and the defect model was evaluated by MRI. The degradation of alginate in vitro was observed by inverted microscope at 1, 2, 4 and 6 weeks respectively. The degradation of alginate gel in vivo was observed by MRI, gross microscope and Safranin O staining. [Results] The microscopic observation showed that the boundary of alginate gel gel in one week was clear, the boundary of gel gel in four weeks was blurred, and the border of alginate disappeared in six weeks and became a paste. The results of Roberts MRI evaluation in vivo showed that the difference was statistically significant at 4 weeks (2.25 ± 0.500) vs 1 week (1.00 ± 0.817) (t = 2.611 P = 0.0401), compared with 2 weeks (1.25 ± 0.500) The difference was statistically significant (t = 2.828, P = 0.0300), and there was no significant difference compared with 6 weeks (2.75 ± 0.500) (t = 1.416, P = 0.2070) Week alginate disappeared completely, and with the degradation of the gel ball, the defect was repaired by tissue. The safranin O staining showed that the experimental group showed no repair, some fiber repair, Partial repair, complete fiber repair. [Conclusion] In comparison with in vitro, the chondrocytes of alginate gels had better biocompatibility in rabbit knee joints and significantly shortened the degradation time. The absorption was complete in about 4 weeks, which could provide a theoretical basis for clinical application.