负载软骨细胞的透明质酸可注射材料修复软骨缺损

Injectable hyaluronic acid carrying autologous chondrocytes repairs cartilage defects

背景:在软骨组织工程材料中,以透明质酸为代表的多聚糖材料因具备良好的材料细胞作用界面,有利于软骨细胞的生长,成为近年来的研究热点。目的:探究负载软骨细胞的透明质酸可注射材料修复大鼠软骨缺损的可行性。方法:取90只SD大鼠,制备软骨缺损模型,造模后第2天随机分3组干预,实验组关节腔内注射负载软骨细胞的透明质酸水凝胶,对照组关节腔内注射透明质酸水凝胶,空白对照组不进行干预。注射后1,3,6周取修复部位软骨组织,进行苏木精-伊红染色、Masson染色、扫描电镜观察及血红素氧合酶表达、胶原水平检测。结果与结论:①苏木精-伊红染色:注射后6周,空白对照组以肉芽组织填充修复区;对照修复部位被黄白色修复组织填满,与正常软骨边界明显,表面不平整,淋巴细胞较3周时减少;实验组修复组织呈半透明状,与正常软骨边界模糊,淋巴细胞较3周时明显减少;②Masson染色:注射后6周,实验组修复区胶原纤维合成状况最佳,其次为对照组,均强于空白对照组;③扫描电镜观察:注射后6周,空白对照组修复区胶原纤维排列不规则,部分断裂;对照组修复区胶原纤维排列较整齐,仍可见部分断裂;实验组实验部位胶原纤维排列整齐,与正常软骨边缘分界不明;④血红素氧合酶表达与胶原水平:注射后6周,实验组血红素氧合酶表达高于对照组、空白对照组(P<0.05);实验组注射后1,3,6周的胶原水平高于对照组、空白对照组(P<0.05);⑤结果表明:负载软骨细胞的透明质酸可注射材料可促进大鼠软骨缺损的修复。

BACKGROUND:In the cartilage tissue engineering materials,hyaluronic acid as the representative of polysaccharide materials has good material-cell interface that is beneficial to the growth of chondrocytes,which has become a hot research topic in recent years.OBJECTIVE:To explore the feasibility of injectable hyaluronic acid material loaded with chondrocytes to repair cartilage defects in rats.METHODS:Ninety Sprague-Dawley rats were selected to prepare a cartilage defect model,and they were randomly divided into three groups at the2nd day after modeling.The experimental group was injected hyaluronic acid hydrogel loaded with chondrocytes into the articular cavity,the control group was injected with hyaluronic acid hydrogel into the articular cavity,and the blank control group received no intervention.At1,3and6weeks after injection,the repaired cartilage tissues were taken out for hematoxylin eosin staining,Masson staining,and scanning electron microscope observation,and the expression of heme oxygenase and level of collagen were detected.RESULTS AND CONCLUSION:(1)Hematoxylin eosin staining:at6weeks after injection,granulation tissues filled the repair area in the blank control group.The control group was full of yellowish white tissues in the repair area,with distinct boundary with the normal cartilage,the surface was not smooth and the lymphocytes were reduced compared with those at3weeks.Repair tissue of the experimental group was semi-transparent and showed a fuzzy boundary with the normal cartilage,and moreover,lymphocyte was significantly reduced compared with those at3weeks.(2)Masson staining:at6weeks after injection,collagen fiber synthesis in the repair area in the experimental group was the best,successively followed by the control group and the blank control group.(3)Scanning electron microscope observation:at6weeks after injection,collagen fiber arrangement in the repair area was irregular and partially broken in the blank control group,and the arrangement became more orderly in the control group but still partially broken.The collagen fibers in the experimental group were arranged orderly,and the boundary with normal cartilage was unclear.(4)Expression of heme oxygenase and level of collagen:at6weeks after injection,the expression of heme oxygenase in the experimental group was higher than that in the control group and blank control group(P<0.05).The levels of collagen in the experimental group at1,3and6weeks after injection were higher than those in the control group and the blank control group(P<0.05).To conclude,hyaluronic acid injectable material loaded with chondrocytes can promote the repair of cartilage defects in rats.