目的:探讨兔膝关节前交叉韧带(ACL)重建后,金黄色葡萄球菌滤液制剂(金葡素)对移植肌腱与骨隧道愈合的作用.方法:将3月龄新西兰大白兔24只(体重平均2.56 kg,雌雄不限)随机分为实验组和空白对照组,每组12只.采用兔自体趾长伸肌腱建立膝前交叉韧带重建后的腱骨愈合模型.实验组分别于术中及术后第2天向胫骨端腱骨界面处缓慢均匀注射金葡素0.1ml,对照组以相同时间和方法注射等量0.9％ NaCl注射液.术后双膝不予制动,以青霉素注射液80万U/d连续肌肉注射3d.分别于术后4、8和12周各处死动物8只,完整获取腱骨愈合标本并分别进行组织学分析,肉眼观察判断ACL重建大体情况,苏木精-伊红染色进行Yamakado腱骨界面组织形态分型评价,苦味酸-天狼猩红染色观察腱骨界面胶原纤维情况,血管内皮生长因子免疫组化染色定量评价腱骨界面再血管化情况,甲苯胺蓝染色定量评价腱骨界面新骨生成情况.结果:大体观察显示全部动物术后伤口Ⅰ期愈合,关节功能恢复良好,未见感染、死亡及金葡素注射不良反应.腱骨界面形态学分型结果显示术后各时间点实验组腱骨界面愈合质量均优于对照组(P＜0.05)).苦味酸-天狼猩红染色显示实验组胶原纤维术后4周生成少,8周大量生成,12周Sharpey纤维有序排列,集结成束,各时间点胶原纤维生成情况均优于对照组.基于血管内皮生长因子免疫组化染色和甲苯胺蓝染色的定量分析显示实验组术后4、8和12周腱骨界面新生血管面积和新骨生成面积均高于对照组(P＜0.05).结论:金葡素能够在葡萄球菌肠毒素C等多种明确成分的协同作用下,通过局部启动无菌性炎症反应,加快腱骨界面血管生成和血液供应,激活骨细胞和纤维细胞大量增殖,有效促进兔ACL重建后腱骨愈合,有望成为促进ACL重建后腱骨愈合的新的临床方法.%Objective:To explore highly agglutinative staphylococcin (HAS) pomoting on tendon-bone healing after reconstruction of anterior cruciate ligament (ACL) in rabbits.Methods:Animal model of ACL reconstruction in 24 mature New Zealand white rabbits (3 months,2.56 kg on average,either gender) were established using autologous digital long extensor tendon and randomly classified into 2 groups (HAS and control group),12 rabbits for each group.HAS group was separately injected 0.1ml highly agglutinative staphylococcin immediately into tendon-bone interface during the operation and 2 days after operation.Control group was injected with the same dose of physiological saline for 3 days.All animals were sacrificed at 4,8,and 12 weeks after operation for histological examinations.The specimens were stained with hematoxylin-eosin,picric acid-sirius red,VEGF immunohistochemistry stain,and toluidine blue to histologically analysis the total pathology changes of the tendon-bone healing tissue,the tendon bone interface morphology classification,hyperplasia and arrangement of collagen fiber,vascularization and new bone formation,respectively.Results:The Yamakado morphological interface results showed that the tissue healing at tendon-bone interface of the HAS group was better than that of the control group.The histological examination revealed that on the 4th week after operation,the tendon-bone interface of HAS group was filled with fibrous connective tissue.The proliferated fibroblasts,chondroblasts and the angiogenesis were rich.On the 8th week after operation,the healing tissue at the bone-tendon interface had developed into dense connective tissue,the neo-vessels were very rich,the collagen fibers w,ere formed abundantly,some Sharpey's fibers spanning parts of the tendon-bone interface.On the 12th week after operation,the transition zones were full of Sharpey's fibers;the neo-vessels were not as much as the 8th weeks,but new bone formation was further increased and immature fibrocartilage appeared.For quantitative histological analysis at 4,8 and 12 weeks after operation,the proportion of neo-vessel area and the area of now bone formation of the HAS group were all significantly higher than those of the control group (P＜0.05).Conclusion:Under the synergy of staphylococcal enterotoxin C and other active ? ingredients,Highly Agglutinative Staphylococcin can significantly improve the tendon-bone healing after ACL reconstruction in rabbit knees,which is expected to be a new method to improve the clinical results of ACL reconstruction.
展开▼
