兔骨髓基质细胞在去抗原牛松质骨载体上生长状态的观察  

The Observation of Growth of MSC in Rabbit on the Surface of Antigen-cleared Bovine Cancellous Bone in Vitro

作　　者：刘丹平[1] 李丹[2] 栗刚[1] 董明岩[1] 张元和[1] 胡蕴玉[2] 于洋[1] 

机构地区：[1]锦州医学院附属第一医院骨科,辽宁锦州121001 [2]第四军医大学西京医院全军骨科研究所,陕西西安710033

出　　处：《锦州医学院学报》2005年第1期1-4,共4页Journal of Jinzhou Medical College

基　　金：国家自然基金课题资助 (编号 :3 0 0 70 75 7)

摘　　要：目的　牛松质骨具有天然的多孔结构 ,微孔规则 ,相互贯通 ,是骨组织工程载体的选择之一。本研究观察了体外状态下表面不加修饰的去抗原牛松质骨载体的生物相容性 ,为骨组织工程研究寻找新的载体提供实验依据。方法　将牛四肢骨干骺端松质骨锯成 0 6～ 0 8cm3 的松质骨块 ,经过反复水洗 (40～ 5 0℃ )、脱脂、超声振荡、离心、脱蛋白过程 ,清除牛松质骨内的细胞成份、油脂、骨髓组织及残留血液。选择孔隙直径约为 30 0～70 0 μm ,孔隙均匀的骨块 ,风干后再经砂轮仔细打磨精修 ,制成约 0 5cm3 的去抗原牛松质骨骨块。用兔骨髓基质细胞与载体复合 ,体外培养 7、 14d行扫描电镜观察了解细胞的生长情况。结果　 1 去抗原牛松质骨载体表面干涩不平 ,洞沟遍布 ;2 培养第一周的载体壁上 ,细胞较少 ,形态不规则 ,大小不一 ,散在于孔壁上 ,细胞分布不均 ,无突起伸展贴壁 ,并可见有细胞死亡后崩解的碎片散在于细胞之间。培养第二周载体孔壁上细胞数量更少 ,提示有细胞死亡并脱落。结论　体外状态下表面不加修饰的去抗原牛松质骨载体不适于MSC贴壁生长。Objective Bovine cancellous bone (BCB) has natural regular porosity in which the holes are approximately even in diameter and linked each other and for this reason antigen-cleared BCB can be taken as a carrier for bone tiscue engineering. In the present study we observed the biocompatibility of antigen-cleared BCB without surface modification in vitro to provide experiment data of this new carrier for bone tissue engineering. Methods The metaphyses of bovine extremities bone were sawed into 0.6～0.8 cm3 cancellous blocks and the cells, oil, marrow and the trace of blood in the blocks were cleared by repeatedly rinsing(40～50℃ water ),defatting in chloroform methanol, removing proteins in 30% peroxide solution, supersonic oscillation and centrifuge.The blocks with regular porosities 300～700μm in diameter were chosen and dried in wind. After rubbed and trimmed carefully with an emery wheel, the 0.5cm3 blocks of antigen-cleared BCB were made as carrier.The blocks were composited with marrow stromal cells(MSC) in vitro and cultured.The growth of MSC in the blocks were observed with a scaning electron microscope(SEM) at 7,14 days respectively. Results (1) Under the SEM, the surfaces of antigen-cleared BCB are dry and unsmooth. Ridges, cracks and holes disperse all the walls of the block.(2)After cultured for 7 days, there is no MSC stretching at the wall and proliferation.Cells with irregular morphosis distribute anywhere.Some debris of the dead cells intersperse among the MSC. After cultured for 14 days, less cells and more debris are seen on the surface of the blocks than those at the 7th days, demonstrating that the majority of the MSC die and exfoliate from the wall of the carrier. Conclusions Antigen-cleared BCB with no surface modified does not fit for the growth of MSC in vitro.

关 键 词：兔骨髓基质细胞 去抗原牛松质骨 生长状态 

分 类 号：R329.2[医药卫生—人体解剖和组织胚胎学]