深低温冷冻保存同种异体前交叉韧带移植免疫研究  

Immunological Study of Allograft Anterior Cruciate Ligament Graft Transplantation after Deep Hyponthermiap Reservation

作　　者：张培[1] 刘泉[1] 周建生[1] 朱坤[1] 赵皓 项平 ZHANG Pei;LIU Quan;ZHOU Jiansheng(Department of Orthopedics,the First Affiliated Hospital of Bengbu Medical College,Bengbu City,Anhui Province 233000)

机构地区：[1]蚌埠医学院第一附属医院骨科,安徽省蚌埠市233000 [2]组织移植安徽省重点实验室

出　　处：《医学理论与实践》2020年第9期1381-1384,1402,共5页The Journal of Medical Theory and Practice

基　　金：安徽省高校省级自然科学研究重点项目(KJ2016A870);安徽省省级重点实验室开放项目(ZZKF1002)。

摘　　要：目的:探讨同种异体前交叉韧带(Anterior Cruciate Ligament,ACL)重建后移植物的免疫转归。方法:雄性SD大鼠60只,体重250~300g,首先随机选择15只大鼠切取30个ACL,其中随机15个进行深低温冷冻保存,15个非冷冻保存,余45只大鼠随机分三组,每组15只:A组—自体移植,B组—冷冻保存同种异体移植,C组—常温保存同种异体组织移植。观察术后动物一般情况,并于术后12h、24h、48h,每组随机选择5只抽取动物血清及关节局部组织液,ELISA法测定各组织液中肿瘤坏死因子(TNF)-α、白细胞介素(IL)-2、白细胞介素(IL)-6水平,同时测定关节组织液中吞噬细胞吞噬率及吞噬指数,组与组之间对比分析。结果:(1)TNF-α水平比较:血清中(μg/L)术后12h、24h、48h分别为A组:0.515±0.012、0.673±0.012、0.699±0.011,B组:0.506±0.019、0.680±0.019、0.712±0.010,C组:0.431±0.032、0.543±0.018、0.610±0.008,A组与B组差异无显著性(P>0.05),A组、B组均明显高于C组(P<0.01)。关节组织液中(μg/L)A组:24.430±0.385、46.784±2.746、81.988±1.341,B组:24.170±0.483、46.239±2.662、80.537±2.502,C组22.963±1.172、37.933±3.850、56.718±4.890,A组与B组差异无显著性(P>0.05),A组、B组均明显高于C组,术后12h(P<0.05),术后24h、48h(P<0.01)。在纵向比较发现,关节局部组织液中TNF-α水平增幅明显高于血清中(P<0.01)。(2)血清IL-2、IL-6水平比较:在术后每个观察时间点,A组与B组差异无显著性(P>0.05),A组、B组明显高于C组(P<0.01)。(3)关节液吞噬细胞吞噬率、吞噬指数比较:在术后每个观察时间点,A组、B组吞噬细胞吞噬率、吞噬指数基本相同(P>0.05),而每个观察时间点,A组、B组吞噬细胞吞噬率、吞噬指数均明显高于C组(P<0.01)。结论:深低温处理能够有效降低异体移植免疫原性,并抑制移植后排斥反应的发生,其局部与全身的免疫排斥反应有所不同,进而更有效提高交叉韧带的重建效果和生活质量。Objective:To discuss and improve the immune outcome of the allograft anterior cruciate ligament graft transplantation.Methods:60 male SD rats were selected,weight 250~300g,first of all,randomly select 15 rats,cut 30 ACL,among them random select 15 ACL conduct hypothermiap reservation,remaining 15 ACL conduct not hypothermiap reservation.The remaining 45 rats were randomly divided into 3 groups,15 rats each group.Normal autologous transplantation(Group A),transplantation after deep hypothermiap reservation(Group B),transplantation without deep hypothermiap reservation(Group C).Observe the rats general situation after operation,on 12 hours,24 hours,48 hours after operation,each group 5 rats were randomly select,extraction of animal serum and the joint fluid,then assess the level of(TNF)-α,(IL)-2,(IL)-6 through ELISA method,also assess the phagocytic rate and the phagocytic index of the joint fluid,compare and analyze the difference between groups.Results:(1)TNF-αlevel:in serum(μg/L),respectively on 12 hours,24 hours,48 hours after operation,Group A:0.515±0.012,0.673±0.012,0.699±0.011.Group B:0.506±0.019,0.680±0.019,0.712±0.010.Group C:0.431±0.032,0.543±0.018,0.610±0.008.There was no significant difference between Group A and Group B(P>0.05).There was significant difference between Group A,Group B and Group C.The joint fluid(μg/L):Group A:24.430±0.385,46.784±2.746,81.988±1.341.Group B:24.170±0.483,46.239±2.662,80.537±2.502.Group C:22.963±1.172,37.933±3.850,56.718±4.890.There was no significant difference between Group A and Group B(P>0.05),Group A and Group B were significantly higher than that of Group C on 12 hours(P<0.05),24 hours,48 hours(P<0.01).(2)IL-2,IL-6:in serum,respectively on 12 hours,24 hours,48 hours after operation,there was no significant difference between Group A and Group B(P>0.05),there was significant difference between Group A,Group B and Group C(P<0.01).(3)The phagocytic rate and the phagocytic index of the joint fluid:respectively on 12 hours,24 hours,48 hours after oper

关 键 词：前交叉韧带 移植 免疫 转归 大鼠 

分 类 号：R687.4[医药卫生—骨科学] R684[医药卫生—外科学]