两种方法冷冻保存异体髌腱重建膝关节交叉韧带比较

Comparative study on two freezing methods of treating patellar tendon allograft for anterior cruciate ligament reconstruction

目的观察程序冷冻液氮保存和-80℃深低温保存方法处理同种异体髌腱后重建膝关节交叉韧带的愈合过程并比较其差异。方法将兔的1/2骨-髌腱-骨复合体经程序冷冻液氮保存和-80℃深低温保存2周后,观察冻存变化差异并行同种异体移植重建前交叉韧带,分别于术后3周和8周观察细胞毒反应、细胞活性、最大载荷和形态学变化等指标进行比较并与自体移植组对照。结果①经程序冷冻液氮保存方法处理后,髌腱的最大载荷无明显下降,细胞活性得到了较好的保存,组织学观察冷冻损伤较-80℃深低温保存方法轻微。②程序冷冻液氮保存处理的移植物在术后未表现明显的排斥反应,且免疫反应随时间的推移而下降。③移植后3周,各组移植物的最大载荷差异无显著性,移植后8周,程序冷冻液氮保存组移植物的最大载荷优于-80℃深低温保存组,但和自体移植组相近。④和-80℃深低温保存组相比,程序冷冻液氮保存组的移植物在术后的愈合过程和组织学行为更接近于自体移植组。结论①经程序冷冻液氮保存和-80℃深低温保存方法冻存的兔异体髌腱重建膝关节交叉韧带后愈合过程和自体韧带移植重建过程相似。②程序冷冻液氮保存法优于传统的-80℃深低温保存法。

Objective To study the difference of healing process of reconstructing the knee cruciate ligament with patellar allograft tendons which were treated by program freezing and - 80℃ deep-frozen. Methods Bone-patellar tendonbone （ BPTB ） specimens of rabbits were undergone liquid nitrogen deep-frozen for 2 weeks after program freezing or - 80℃ deep-frozen. Reconstruction the anterior cruciate ligament（ACL） of the knee with the BPTB was performed and the ceils poison response, ceils activity,the maximum load and morphologic changes of the grafts were observed and compared between the two allograft groups with the autograft group. Results （1） After treated by program freezing,the maximum load of the allograft have no evident decrease and the activity of allograft was still good, and the hurt after program freezing was more slight than - 80℃ deep-frozen. （2） No evident reject reaction can be found during the healing process after program freezing and the reject reaction declined as the time pass by. （3） Three weeks after operation,there was no significant difference between the 2 allograft groups in maximum load（P 〉0. 05）. Eight weeks later,the maximum load of the group that was preserved by liquid nitrogen after program freezing（76. 60 ±4. 67） was similar with the autograft group （ 76. 72 ± 2. 55 ） and better than the group that was preserved by - 80℃ deep-frozen （ 75.99 ± 4. 20）. （4）The healing process and histological behavior of the group that was preserved by liquid nitrogen after program freezing was more similar with the autograft group than the group that was preserved by - 80℃ deep-frozen. Conclusions （1） Healing process of the groups that was preserved by program freezing or - 80℃ deep-frozen is similar with the autograft group. （2） The program freezing method is better than the traditional -80℃ deep-frozen method in this experiment.