退变大鼠椎间盘器官培养模型的建立及其意义  

Development and significance of an in vitro organ culture model of degenerative intervertebral disc in rats

作　　者：徐宏光[1] 章平治[1] 宋俊兴[1] 程加峰[1] 卢林明[2] 何雷[2] 周珏[2] 

机构地区：[1]皖南医学院附属弋矶山医院脊柱外科,安徽芜湖241001 [2]皖南医学院附属弋矶山医院病理科,安徽芜湖241001

出　　处：《中国骨与关节外科》2012年第3期233-237,197,共6页Chinese Journal of Bone and Joint Surgery

基　　金：国家自然科学基金(30973025);卫生部行业基金(201002018);安徽省教育厅自然科学基金(KJ2010A320)

摘　　要：背景:椎间盘退变(intervertebral disc degeneration,IDD)是慢性腰背疼和脊柱功能损害的主要原因,建立退变的椎间盘器官培养模型能为IDD机制的研究提供一个更好的试验平台。目的:建立退变大鼠椎间盘器官培养模型,观察正常和退变大鼠椎间盘的组织形态及细胞活性,以评价培养结果。方法:选取SD大鼠30只,老年组15只,青年组15只。反转录-PCR(reverse transcription-polymerase chain reaction,RT-PCR)法分别检测2组椎间盘蛋白多糖(aggrecan,AGN)和Ⅱ型胶原(typeⅡcollagen,COLⅡ)的基因表达。无菌条件下完整取出两组大鼠腰段椎间盘,在培养液中经过不同时段培养后HE染色观察椎间盘组织学变化,氯化硝基四氮唑蓝(NBT)和4’,6-二眯基-2-苯基吲哚(DAPI)复染检测椎间盘细胞存活率。结果:老年组大鼠椎间盘较青年组发生明显退变,可视为退变椎间盘组织。两组大鼠椎间盘器官在体外培养4周,其组织学特点和细胞存活率无明显变化。结论:成功建立退变大鼠椎间盘器官培养模型,为研究椎间盘退变机制提供理想的实验模型。Background： Intervertebral disc degeneration （IDD） is the main reason of low back pain and the damage of spine. The estab- lishment of an in vitro organ culture model of degenerative intervertebral disc can provide a preferable experimental plat- form for studying the mechanism of IDD. Objective： The purpose of the present study is to develop an organ culture model of IDD in rats and to observe the tissue structure and cellular activity so as to access the culture results. Methods： Totally 30 SD rats were divided into old group （n=15） with 6 weeks old and young group （n=15） with 15 months old. RT-PCR was used to detect the mRNA expression of aggrecan and type II collagen of chondroeytes in all rat interverte- bral discs. Intervertebral discs were taken out completely by asepsis and cultured in medium for different periods. The change of histomorphology in intervertebral disc tissue was assessed by HE staining and the cell viability was investigated by counterstain with NBT, DAPI. Results： IDD was more obvious in old rats than in young ones. Rat intervertebral disc organs cultured for up to 4 weeks showed minimal changes in histomorphology and cell viability. Conclusions： A degenerative organ culture model of intervertebral disc has been established successfully in vitro in rats. It may offer a better experimental model for exploring the mechanism of IDD.

关 键 词：椎间盘 器官培养 退变模型 

分 类 号：R681.53[医药卫生—骨科学]