不可逆电穿孔消融同种异体肌腱移植对兔肌腱缺损修复的实验研究

Experimental study on repairing rabbit tendon defect by allogenic tendon transplantation pretreated with irreversible electroporation ablation

目的探讨采用不可逆电穿孔法消融对同种异体肌腱移植修复兔肌腱缺损的可行性及消融术后组织转归情况。方法将新西兰大白兔112只随机分为自体肌腱移植组(A组)28只,深低温冷冻同种异体肌腱移植组(B组)28只,不可逆电穿孔消融法(IRE)灭活同种异体肌腱移植组(C组)28只和假手术对照组(D组)28只。各组制备动物后肢第3趾深屈肌腱缺损模型,并采用处理后的肌腱修复缺损;分别于术后1、4、8、12周取材行微血管定量、组织学观察及生物力学测试。结果术后各组移植肌腱直径比较均无统计学意义(P>0.05)。组织学观察提示B组修复再生速度较慢,表现为成纤维细胞爬行替代及微血管再生能力均低于A、C组。微血管灌注观察提示:微血管有效灌注面积在术后1周时A、B、C组显著低于D组(P<0.05),术后4、8、12周时B组显著低于A、C、D组(P<0.05),且8周时A、C组显著高于D组(P<0.05)。生物力学测试提示:B、D组比较移植肌腱最大载荷及硬度仅在术后4周时差异有统计学意义(P<0.05),其余时间点各组间比较差异均无统计学意义(P>0.05)。各时间点各组移植肌腱破损形变比较,差异均无统计学意义(P>0.05)。结论采用IRE灭活的同种异体肌腱修复兔肌腱缺损切实可行,术后移植肌腱组织形态及生物力学性能恢复与自体肌腱移植组无显著差异。

Objective To observe the histological changes of tendon graft pretreated by deep frozen and irreversible electroporation, and assess the feasibility of treatment for tendon defect by using irreversible electroporationpretreated tendon allograft. Methods 112 New Zealand white rabbits were randomly divided into autologous transplantation group, frozen tendon allograft transplantation group, irreversible electroporation ablated tendon allograft transplantation group, and sham operation control group, 28 in each group. The deep flexor tendon defect of the third toe was prepared and the corresponding defect was repaired by treated tendon. Specimen were collected at 1 week, 4 weeks, 8 weeks and 12 weeks post operation for histologic observation, quantitative analysis of microvessel regeneration and biomechanical evaluation.Results There were no significant differences in the tendon diameter between the four groups at each time point （P 〉 005）. Histological observation showed that fibroblast replacement and microvascular regeneration in group B were lower than that in group A and group C. Microvascular perfusion showed that effective perfusion areas in group A, B, C were significantly lower than that in group D at 1 week （P 〈 005）, and effective perfusion areas in group B was significantly lower than that in group A, C, D from 4 week to 12 week （P 〈 005）. At the 12th week, group B was significantly lower than the group A, C ＆ D, the difference was statistically significant （P 〈005）. The biomechanical results showed that there was significant difference in the maximal load and stiffness of tendon between group B and group D at 4 weeks after operation （P 〈 005）. There was no significant difference in damage deformation between the four groups at each time point （P 〉 005）. Conclusion Irreversible electroporation ablated tendon allograft can be used to repair tendon defect, and the morphological and biomechanics property of tendon ablated by irreversible electroporation has no significant difference with autologous transplantation group.