不同方法深低温保存兔肢体再植后血管组织的病理学变化李波1，2，何建平3，张树明2，朱泽兴2，乔林2，乔雅楠1，21辽宁医学院研究生学院，辽宁省锦州市121000；2解放军第二炮兵总医院骨科，北京市100088；3北京市平谷区人民医院，北京市101200PathologicalchangesofvasculartissueafterrabbitlimbreplantationwithdifferentmethodsofcryopreservationLiBo1,2,HeJian-ping3,ZhangShu-ming2,ZhuZe-xing2,QiaoLin2,QiaoYa-nan1,21GraduateSchool,LiaoningMedicalUniversity,Jinzhou121000,LiaoningProvince,China;2DepartmentofOrthopedics,theSecondArtilleryGeneralHospitalofPLA,Beijing100088,China;3PingguDistrictPeople’sHospital,Beijing101200,China背景：虽然对于单一组织的冷冻保存获得了空前的成果，并且已逐渐应用于临床，但是对于复合组织的冷冻保存及应用还鲜有研究。目的：通过实验研究深低温冷冻不同复温方法下兔肢体再植后血管形态学变化，找出一种对复合组织中血管损伤最小的复温方法，从而为深低温处理后断肢再植可行性提供理论依据。方法：30只健康新西兰大白兔随机数字表法均分为对照组、慢速冷冻-慢速复温组、慢速冷冻-快速复温组，均给予大白兔右后肢自膝上1cm处离断。慢速冷冻两组复温后均行断肢再植，再植肢体成活6h后给予再次离断右后下肢。3组兔断肢均取股血管组织采用染色光镜及电镜进行形态学观察及大体观察，光镜病理计分结果用显著性分析。结果与结论：慢速冷冻-慢速复温组、慢速冷冻-快速复温组兔肢体再植6h后的病理变化(大体标本、光镜、电镜)均较对照组差，但慢速冷冻-慢速复温组与慢速冷冻-快速复温组相比血管内皮细胞完整性较好，细胞器破坏较少。证实，经过深低温冷冻-复温处理后兔断肢再植6h，离断肢体血管组织能保持一定的结构完整性，再植后6h兔肢体获得成活，且慢速冷冻-慢速复温组更为适合离断肢体的保存，为深低温处理后离断肢体行断肢再植远期成活可行性提供了依据。中国组织工程研究杂志出版内容重点：组织构建；骨细胞；软骨细胞；细胞培养；成纤维细胞；血管内皮细胞；骨质疏松；组织工程全文链接：关键词：组织构建；组织工程；深低温处理；断肢再植；血管；内皮细胞；组织学；Abstract：BACKGROUND:Thecryopreservationofsingletissuehasachievedgreatadvancementandisgraduallyappliedinclinics.However,thecryopreservationofcomplextissueisrarelyreported.OBJECTIVE:Toinvestigatethemorphologicalchangeinrabbitlimbtissueafterreplantationthroughdifferentrewarmingmethods,findthebestrewarmingmethodsofcompoundtexturedbloodvessels,andprovidetheoreticalbasisforthefeasibilityoflimbreplantationafterlong-termcryopreservation.METHODS:ThirtyNewZealandwhiterabbitswererandomlydividedintocontrolgroup,slowfreezing-slowthawinggroup,andslowfreezing-rapidthawinggroup.Therightposteriorlimbsofalltherabbitswerecutoff1cmabovethekneejoint.Exceptcontrolgroup,thelattertwogroupsweregivenlimbreplantationafterthawing,andthentherightposteriorlimbwasagaincutoffafterthereplantedlimbsweresurvivedfor6hours.Forallgroups,thehistologicalchangesandgrossobservationinaortatissuewereobservedbylightmicroscopyandtransmissionelectronmicroscope,andtheresultswereanalyzedwithstatisticalmethods.RESULTSANDCONCLUSION:Intheslowfreezing-slowthawing,slowfreezing-rapidthawinggroups,thepathologicalchanges(grossspecimen,lightmicroscope,electronmicroscope)ofrabbitlimbs6hoursafterreplantationwereworsethanthoseincontrolgroup.Comparedwithslowfreezing-rapidthawinggroup,betterintegrityofendothelialcellsandlessdamageoftheorganelleswerefoundinslowfreezing-slowthawinggroup.Throughdeepcryogenicfreezing-thawingprocess,rabbitlimbbloodvesselscanmaintainthestructuralintegrityafterreplantationandsurvivedat6hours.Slowfreezing-slowthawingisbetterthanslowfreezing-rapidthawingforthepreservationofseveredlimbs,providingevidencesforthelong-termsurvivalfollowingadeepcryogenictreatmentaftertheseveredlimbreplantation.中国组织工程研究杂志出版内容重点：组织构建；骨细胞；软骨细胞；细胞培养；成纤维细胞；血管内皮细胞；骨质疏松；组织工程全文链接：Keywords：cryopreservation;hindlimb;transplantation;disarticulation;bloodvessels;endothelialcells;中图分类号:R318通讯作者:张树明，男，汉族，教授、硕士生导师，解放军第二炮兵总医院骨科，北京市100088作者简介:李波，男，1986年生，内蒙自治区古包头市人，汉族，辽宁医学院在读硕士。引用本文:李波,何建平,张树明等.不同方法深低温保存兔肢体再植后血管组织的病理学变化[J].中国组织工程研究,2014,18(15):2357-2362.
