碳化二亚胺交联同种异体肌腱修复兔前交叉韧带损伤生物力学实验研究

Reconstruction of anterior cruciate ligament with rabbit tendon allograft cross-linked and heparinized by EDC/NHS

目的探讨利用碳化二亚胺(1-3-(dimethylamino)propyl-3-ethylcarbodiimide methiodide,EDC)交联并肝素化的方法对同种异体肌腱进行预处理改善其修复韧带损伤效果,为临床应用提供实验基础和理论依据。方法采用冷冻干燥法获得兔同种异体肌腱,应用EDC交联并肝素化方法进行再处理;自体肌腱移植组、未交联同种异体肌腱移植组和EDC交联同种异体肌腱移植组分别对兔前交叉韧带损伤进行修复,于1、3、6个月生物力学测试检测各时间点肌腱最大拉伸强度,其结果进行统计学分析;交联组和未交联组肌腱取材做光镜和扫描电镜观察,对比1、3、6个月不同时间段腱骨愈合情况。结果术后1个月最大拉伸强度自体肌腱移植组、EDC交联肌腱移植组、未交联肌腱移植组两两比较,差异有统计学意义(P<0.05);术后3、6个月进行比较,EDC交联肌腱移植组和自体肌腱移植组比较无显著性差异(P>0.05),与未交联肌腱移植组相比较有显著性差异(P<0.05)。组织学观察EDC交联并肝素化的同种异体肌腱在修复ACL损伤过程中的腱骨愈合能力比未交联的同种异体肌腱强,Sharpey’s纤维出现早。结论经碳化二亚胺交联并肝素化处理的同种异体肌腱,其稳定性明显增强,免疫原性明显降低,有利于肌腱与骨的愈合。

Objective To evaluate the anterior cruciate ligament reconstruction effect of rabbit tendon allograft cross-linked and heparinized by EDC/NHS. Methods The rabbit＇ s tendon allograft was obtained by lyophilization, then cross-linked and heparinized by EDC/NHS; the rabbit anterior cruciate ligaments were reconstructed with cross-linked and non-crosslinked liga- ment allograft. After 1, 3, 6 months, the tendons were harvested, the maximum tensile strength was compared; the tendon-bone junction were harvested and evaluated by microscopy and scanning electric microscopy. Results The maximum tensile strength of the cross-linked and heparinized tendon allograft was higher than that of the non-crosslinked allograft and autolo- gous tendon; the union at the bone-tendon junction was faster in cross-linked and hepranized group and the sharpey＇ s fiber appeared earlier. Conclusion Cross-linking and heparinizing process by EDC/NHS may improve the biocompatibility of rabbit tendon allograft, facilitate the union of tendon-bone junction.