摘要
目的:研究中药提取物菟丝子总多糖局部运用修复兔关节软骨全层缺损的可行性和部分机理。
方法:将72只成年健康日本大耳白兔随机分为菟丝子多糖组(A组)、人骨形态发生蛋白/明胶复合物组(B组)、空白组(C组),每组24只,48个关节。在膝关节股骨内髁滑车面上,用手摇钻造成直径3mm、深约3mm的软骨全层缺损区,直至软骨下骨渗血为准。冲洗后分别给予A组菟丝子总多糖凝胶、B组骨形态发生蛋白/明胶复合物填充,C组只做钻孔处理,空白对照。术后于2、4、8周分批处死实验动物,取标本行大体观察、HE和甲苯胺蓝染色、改良Pinsda法组织学评分、Ⅱ型胶原免疫组化和原位杂交的方法进行评估。
结果:菟丝子总多糖和rhBMP/明胶复合物在2、4、8周缺损区填充均优于空白组,为乳白色半透明质软类透明软骨组织;组织学评分与空白组存在极显著性差异(p＜0.01),用药组之间没有统计学意义(p＞0.05)。关节软骨缺损区修复组织经Ⅱ型胶原免疫组化、原位杂交检测定性为Ⅱ型胶原。用药组Ⅱ型胶原的表达在术后2、4、8周与空白组比较,均有极显著性差异(p＜0.01),用药组之间比较没有统计学意义(p＞0.05)。
结论:菟丝子总多糖局部运用可以修复家兔关节软骨缺损,可能是通过促进Ⅱ型胶原mRNA表达等方式实现的,经组织学观察修复组织以透明软骨为主,接近正常关节软骨。其修复关节软骨缺损是可行的,具有很好的实用价值。
OBJECTIVE: To study the feasibility and the mechanism of the total polysaccharide from Cuscuta for repair of articular cartilage defects in rabbits.
METHODS: 72 healthy adult Japanese White rabbits were randomly divided into the total polysaccharide from Cuscuta(A group), the compounds of bone morphogenetic protein and gelatin(B group), the blank control group(C group).There were 24 rabbits including 48 knee joints in every group. Full thickness articular cartilage defects with 3mm in diameter by 3mm depth were created over the trochlear surface of the intercondyle femur, in depth until had punctate hemorrhage. After washing, A group with the total polysaccharide from Cuscuta and B group with the compounds of bone morphogenetic protein and gelatin were filled in the defects, C group was left only without any treatment. The rabbits of group A,B,C were sacrificed postoperatively respectively at the end of 2,4,8 weeks. The repair tissues in the defects were performed as macroscopically observation,HE staining,toluidine blue staining, histology score, immunohistochemistry and in situ hybridization essay with collagen typeⅡ.
RESULTS: The therapeutic groups could promote the repair of the defects by ivory white translucent cartilage. The group A and group B got a better histological score than the blank control group (p<0.01) .There was no statistical significance (p>0.05) between the group A and group B .The newly cartilaginous tissue collagen was typeⅡ.The therapeutic groups on the expression of collagen typeⅡhad significant difference (p<0.01) compared with C group after 2,4,8 weeks postoperatively,but the medication groups had no difference (p>0.05) .
CONCLUSION: The total polysaccharide from Cuscuta can promote the expression of collagen typeⅡmRNA in the repair of the defects. Transparent cartilage was main,nearly to normal articular cartilage after histology observation. TCM for repair of articular cartilage defects is feasible and economical.
引文
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