低强度脉冲超声对同种异体软骨细胞-藻酸钙凝胶复合物修复兔膝关节软骨缺损的影响  

EFFECT OF ALLOGENEIC CHONDROCYTES-CALCIUM ALGINATE GEL COMPOSITE UNDER INTERVENTION OF LOW INTENSIVE PULSED ULTRASOUND FOR REPAIRING RABBIT KNEE ARTICULAR CARTILAGE DEFECT

作　　者：郭杨[1] 马勇[1,2] 董睿[1] 刘尚仑[1] 屠娟[3] 

机构地区：[1]南京中医药大学骨伤研究所,南京210023 [2]南京中医药大学附属医院骨伤科 [3]南京大学声学研究所

出　　处：《中国修复重建外科杂志》2013年第8期928-934,共7页Chinese Journal of Reparative and Reconstructive Surgery

基　　金：江苏省"六大人才高峰"资助项目(2010-WS-082);江苏省自然基金资助项目(BK2011812)

摘　　要：目的探讨低强度脉冲超声(low intensive pulsed ultrasound,LIPUS)干预后的同种异体软骨细胞-藻酸钙凝胶复合物修复兔膝关节软骨缺损的有效性。方法取8只2周龄新西兰白兔,采用机械法联合Ⅱ型胶原酶消化法分离双侧膝关节软骨细胞。取第3代细胞与1.2%藻酸钠溶液制成细胞悬液,调整细胞密度为5×106个/mL,制备软骨细胞-藻酸钙凝胶复合物,并给予超声干预,参数为:中心频率1 MHz,脉冲重复频率1 kHz,超声强度60 mW/cm2,作用周期50,作用时间20 min,每天1次,共干预3 d。取28～35周龄健康新西兰白兔18只,体重2.1～2.8 kg,随机分为超声凝胶组、普通凝胶组及模型组,每组6只。于3组实验动物双侧膝关节制备直径约3 mm、深3 mm的膝关节软骨缺损模型后,超声凝胶组、普通凝胶组分别于缺损区对应植入经超声干预及未行干预的软骨细胞-藻酸钙凝胶复合物,模型组不作处理。术后观察实验动物一般情况,并于术后8、12周处死动物分别行大体观察、组织学观察及Wakitani组织学评分、Ⅱ型胶原免疫组织化学染色观察。结果大体观察见超声凝胶组与普通凝胶组缺损区均逐渐被新生组织填充,但前者为透明软骨样组织,后者为灰白色软骨样组织,且前者表面平整度及组织整合程度均优于后者;模型组缺损区修复缓慢且新生组织弹性差。术后8、12周修复区组织学观察及Wakitani组织学评分示超声凝胶组优于普通凝胶组,两凝胶组均优于模型组,差异有统计学意义(P<0.05);两凝胶组组内8周与12周间比较,差异均有统计学意义(P<0.05)。术后8、12周,超声凝胶组及普通凝胶组Ⅱ型胶原染色阳性表达面积、吸光度(A)值均显著高于模型组(P<0.05),两凝胶组术后12周时比较差异亦有统计学意义(P<0.05)。超声凝胶组组内8周与12周间比较,差异有统计学意义(P<0.05);普通凝胶组及模型组两时间点间比较,差异无统计学意义(P>0.05)�Objective To investigate the effect of allogeneic chondrocytes-calcium alginate gel composite under the intervention of low intensive pulsed ultrasound（LIPUS） for repairing rabbit articular cartilage defects. Methods Bilateral knee articular cartilage were harvested from 8 2-week-old New Zealand white rabbits to separate the chondrocytes by mechanical-collagen type II enzyme digestion.The 3rd passage chondrocytes were diluted by 1.2% sodium alginate to 5 × 106 cells/mL,then mixed with CaCl2 solution to prepare chondrocytes-calcium alginate gel composite,which was treated with LIPUS for 3 days（F0： 1 MHz;PRF： 1 kHz;Amp： 60 mW/cm2;Cycle： 50;Time： 20 minutes）.An articular cartilage defect of 3 mm in diameter and 3 mm in thickness was established in both knees of 18 New Zealand white rabbits（aged 28-35 weeks;weighing,2.1-2.8 kg）,and divided into 3 groups randomly,6 rabbits in each group： LIPUS group,common group,and model group.Defect was repaired with LIPUS-intervention gel composite,non LIPUS-intervention gel composite in LIPUS group and common group,respectively;defect was not treated in the model group.The general condition of rabbits was observed after operation.The repair effect was evaluated by gross and histological observations,immunohistochemical staining,and Wakitani score at 8 and 12 weeks after operation. Results Defect was filled with hyaline chondroid tissue and white chondroid tissue in LIPUS and common groups,respectively.LIPUS group was better than common group in the surface smooth degree and the degree of integration with surrounding tissue.Defect was repaired slowly,and the new tissue had poor elasticity in model group.Histological observation and Wakitani score showed that LIPUS group had better repair than common group at 8 and 12 weeks after operation;the repair effect of the 2 groups was significantly better than that of model group（P 0.05）;and significant differences in repair effect were found between at 8 and 12 weeks in LIPUS and common groups（P 0.05）.The c

关 键 词：组织工程软骨 软骨细胞 藻酸钙凝胶 低强度脉冲超声 软骨缺损 兔 

分 类 号：R684[医药卫生—骨科学]