组织工程骨-80℃低温保存的初步研究.pdfVIP

中国骨伤2008年1月第21卷第1期 China J Orthop＆Trauma，Jan．2008，Vo1．21，No．1 · 49· · 基 础 研 究 · 组织工程骨一8O CIC低温保存的初步研究 蓝旭，文益民，葛宝丰，刘雪梅 (解放军兰州军区总医院脊柱外科，甘肃 兰州 730050) 【摘要】 目的：了解不同冻存方法对组织工程骨生物活性的影响。方法：以骨髓基质干细胞(marrow stromal cells， MSCs)复合部分脱蛋白骨培养制备组织工程骨，实验分为：A组，组织工程骨用添加冻存保护荆的保存液保存；B组，组 织工程骨用不添加冻存保护荆的保存液保存；C组，组织工程骨未行低温保存；D组。单纯MSCs培养。A组和B组的组 织工程骨于-80 oC深低温保存3个月，3个月后复温冻存的组织工程骨。扫描电镜观察MSCs的黏附和分布情况，M1Tr 法检测细胞活力，对硝基苯磷酸法检测碱性磷酸酶(alkalinephosphatase，AI )活性，流式细胞仪分析细胞周期。结果： MSCs在材料表面和孔隙内均可黏附和分布，黏附于材料的细胞活力大小依次为C组A组B组 (P0．01，P0．05)．黏 附于材料的细胞ALP活性大小依次为C组A组B组(P0．01)。各组细胞周期未见明显变化，未见异倍体细胞。结 论：选择适宜的冻存保护荆对组织工程骨的生物活性有一定的保护作用。 【关键词】组织工程； 骨；低温保存 Preliminary study on cryopreservation oftissue engineered bone at-80℃ LAN Xu，WEN Yi-min，GE Boo-feng，LIU Xue-mei．Department of ine Surgery，the General Ho~ital ofLanzhou Milhary Command，Lanzhou 730050，Gansu，China ABSTRACT Objective：To study the effects of various methods of cryopreservation on the bioactivity of tissue engineered bone．Methods：MSCs were cocultured with partialy deproteinised bone to produce tissue engineered bone．The experiment was divided into A，B，C and D group．Group A：Tissue engineered bone was stored in preservation solution with cryopreservation medium．Group B：Tissue engineered bone was stored in preservation solution without cryopreservation medium．Group C：Tis— sue engineered bone was stored witl1out cryopreservation．Group D：MSCs were cultured without c~opreservation．The tissue engineered bone of group A and B had been c~opreserved at-80℃for three months and thawed three months later．The elec— tronic scanning microscope wag used to evaluate the adhesion and distribution of MSCs，cell viability WaS meaSured by MTr， AIJP activity WaS detected by P-nitrophosphate，cell cycle WaS analysed by flow c~ometry．Results：MSCs could adhere to the surf