摘要
目的探讨不同浓度比例的骨保护素(osteoprotegerin,OPG)与脱蛋白骨(deproteinized bone,DPB)凝胶复合物对前交叉韧带(anterior cruciate ligament,ACL)重建术后骨隧道的影响。方法取市售新鲜新生小牛股骨骺端,采用H2O2-甲醇/氯仿方法制备异种DPB,将其分别浸泡于浓度为30、60、100μg/m L的OPG溶液中,制成3种浓度比例(30%、60%、100%)的凝胶复合物。取健康新西兰大白兔60只,雌雄不限,体质量(2.7±0.4)kg,随机分为4组(n=15),分别为空白对照组(A组)及30%、60%、100%OPG/DPB凝胶复合物组(B、C、D组)。各组建立自体跟腱重建ACL模型后,将不同浓度比例OPG/DPB凝胶复合物分别植入B、C、D组股骨及胫骨骨隧道内,A组不作处理。术后观察动物一般情况,于术后4、8、12周取材行大体观察(包括股骨隧道扩大率计算)、组织学观察以及生物力学检测。结果术后A、D组各1只,B、C组各2只动物因感染死亡并补充。大体观察示,4周时各组股骨及胫骨隧道内口均有少量瘢痕组织覆盖;8周时股骨及胫骨隧道内口周围组织呈白色软骨样变,其中C、D组较明显;12周时A、B、C组股骨及胫骨隧道内口扩大,D组股骨及胫骨隧道内口完全封闭。各时间点B、C、D组股骨隧道扩大率均显著低于A组,D组低于B、C组(P<0.05);除8周时C组股骨隧道扩大率显著低于B组(P<0.05)外,4、12周时B、C组间比较差异无统计学意义(P>0.05)。组织学观察示,4周时各组可见新生纤维结缔组织生长;8周时可见排列不一的Sharpey纤维,D组偶见不典型的4层结构;12周时Sharpey纤维排列有序,除A组外其余组均可见典型的4层结构,形成不规则的"潮线",尤以D组显著。生物力学测试示,除C组12周和D组8、12周时均为韧带实质断裂外,其余各时间点主要是从骨隧道拔出,其中以从股骨隧道拔出多见。术后4周时,D组最大抗拉力负荷显著大于A、B组(P<0.05);A、B、C组间以及C、D组间比较,差异均无统计学意义(P>0.05)。8周时,C、D组大于A组,D组大于B组(P<0.05);其余组间比较差异均无统计学意义(P>0.05)。12周时,C、D组大于A、B组,D组大于C组(P<0.05);A、B组间比较差异无统计学意义(P>0.05)。结论不同浓度比例OPG/DPB凝胶复合物对ACL重建术后骨隧道影响程度不同,100%OPG/DPB凝胶复合物预防骨隧道扩大、促进腱-骨愈合作用最显著。
Objective To investigate the effects of different concentrations of osteoprotegerin（OPG） combined with deproteinized bone（DPB） on the bone tunnel after the anterior cruciate ligament（ACL） reconstruction. Methods The femoral epiphyseal side was harvested from newborn calf, and allogenic DPB were prepared by hydrogen peroxide-chloroform/methanol method. Then, DPB were immersed in 3 concentrations levels of OPG（30, 60, 100 μg/ m L） and 3 concentration ratios（30%, 60%, 100%） of the gel complex were prepared. Sixty healthy New Zealand white rabbits, male or female, weighing（2.7±0.4） kg, were divided randomly into 4 groups（n=15）： control group（group A）, 30%（group B）, 60%（group C）, and 100%（group D） OPG/DPB gel complex. The ACL reconstruction models were established by autologous Achilles tendon. Different ratios of OPG/DPB gel complex were implanted in the femoral and tibial bone tunnel of groups B, C, and D, but group A was not treated. The pathology observation（including the percentage of the femoral bone tunnel enlargement） and histological observation were performed and the biomechanical properties were measured at 4, 8, and 12 weeks after operation. Results One rabbit died of infection in groups A and D, 2 rabbits in groups B and C respectively, and were added. General pathology observation showed that the internal orifices of the femoral and tibia tunnels were covered by a little of scar tissue at 4 weeks in all groups. At 8 weeks, white chondroid tissues were observed around the internal orifices of the femoral and tibia tunnels, especially in groups C and D. At 12 weeks, the internal orifices of the femoral and tibia tunnels enlarged in groups A, B, and C, but it was completely closed in group D. At each time point, the rates of the femoral bone tunnel enlargement in groups B, C, and D were significantly lower than that in group A, and group D was significantly lower than groups B and C（P〈0.05）; group C was significantly lower than group B at 8 weeks, but no significant difference was found at 4 and 12 weeks（P〈0.05）. Hisological observation showed that fresh fibrous connective tissue was observed in 4 groups at 4 weeks; there was various arrangements of Sharpey fiber in all groups at 8 weeks and the atypical 4-layer structure of bone was seen in group D; at 12 weeks, Sharpey fiber arranged regularly in all groups, with typical 4-layer structure of bone in groups B, C, and D, and an irregular ＂tidal line＂ formed, especially in group D. Biomechanics measurement showed that the maximum tensile load in group D was significantly higher than that in groups A and B at 4 weeks（P〈0.05）, but no significant difference was shown among groups A, B, and C, and between groups C and D（P〈0.05）; at 8 weeks, it was significantly higher in groups C and group D than group A, and in group D than group B（P〈0.05）, but there was no significant difference between groups A, C and group B（P〈0.05）; at 12 weeks, it was significantly higher in groups C and D than groups A and B, and in group D than group C（P〈0.05）, but difference was not significant between groups A and B（P〈0.05）. Conclusion Different concentrations ratios of OPG/DPB gel complexes have different effects on the bone tunnel after ACL reconstruction. 100% OPG/DPB gel complex has significant effects to prevent the enlargement of bone tunnel and to enhance tendon bone healing.
出处
《中国修复重建外科杂志》
CAS
CSCD
北大核心
2015年第11期1369-1375,共7页
Chinese Journal of Reparative and Reconstructive Surgery
基金
广西壮族自治区自然科学基金资助项目(2014GXNSFAA118239)~~
关键词
前交叉韧带
骨保护素
脱蛋白骨
骨隧道
生物力学
兔
Anterior cruciate ligament
Osteoprotegerin
Deproteinized bone
Bone tunnel
Biomechanics
Rabbit
