力学刺激促进软骨细胞与骨髓基质干细胞共培养体外软骨分化  

Mechanical Stimulation to Promote in Vitro Chondrogenesis by Co-culture of Bone Marrow Stromal Cells and Chondrocytes

作　　者：田甜[1] 章庆国 

机构地区：[1]东南大学附属中大医院整形外科,江苏南京210009 [2]中国医学科学院整形外科医院,北京100144

出　　处：《中国美容医学》2009年第5期658-662,共5页Chinese Journal of Aesthetic Medicine

摘　　要：目的:研究不同的应力刺激对软骨细胞与骨髓基质干细胞(BMSCs)共培养体外构建组织工程化软骨的影响。方法:分离、培养、扩传兔MSCs及软骨细胞,二者按7∶3比例混和,以5.0×107/ml的细胞密度接种于聚羟基乙酸(PGA)支架上,一周后根据不同的施加力分为4组:离心组、摇床组、搅拌组,静止培养作为对照组。6周后取材行相关检测。结果:三受力组形成的细胞材料复合物基本保持原来的体积与外形。HE染色结果显示大量成熟软骨陷窝形成,细胞外基质沉积均匀;Safranin-O及甲苯胺兰染色显示有大量的GAG形成,免疫组化检测Ⅱ型胶原表达强阳性。三受力组标本组织湿重、体积、GAG含量等指标均优于对照组。结论:力学刺激有利于促进少量软骨细胞与BMSCs共培养体外软骨分化;并在三维支架材料上构建组织工程化软骨。Objective To evaluate the influence of different mechanical stresses on in vitro cultured chondrogenesis by co-culture of bone marrow stromal cells （BMSC） and chondrocytes. Methods Rabbit MSCs and chondrocytes were in vitro isolated, cultured and expanded respectively, then mixed up with the ratio of 7：3 （MSCs： chondrocytes）. The mixed cells were seeded onto PGA （polyglycolic acid） scaffold at the ultimate concentration of 5.0x107/ml. One week later, the cell-scaffold constructs were randomly divided into four groups according to the different imposed stresses： experimental group A in which a centrifugal stress was imposed. Experimental group B with rotative stress; experimental group C with fluid shear stress, and the control group in which the constructs were statically cultured. All specimens were collected for relative detections after 6 weeks. Results The constructs in three experimental groups were basically maintained in their original sizes and shapes with optimal elasticity. Massive lacuna -like structures in most areas of the construct and extra cellular matrix were evenly deposited on HE stain. Safranin-O and toluidine blue staining showed massive GAG formation. IHC （immunohistochemistry） staining of collagen type II was observed with strong positive result. In three experiment groups, weights of wet tissue, sizes and GAG contents were higher respectively than the control group. Conclusions Mechanical stimulation promotes in vitro chondrogenesis by co-culture of BMSCs and a small amount of chondrocytes, and constructs tissue engineered cartilages on 3-dimensional scaffolds.

关 键 词：软骨细胞 骨髓基质干细胞 共培养 力学 组织工程 

分 类 号：R318[医药卫生—生物医学工程]