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软骨形态发生蛋白1诱导SD仔鼠脂肪干细胞修复兔膝关节软骨缺损的研究
杨亚军1,2,朱庆生2*
0
(1.宁夏回族自治区人民医院骨科,银川 750001;2.第四军医大学西京医院骨科,西安 710032)
摘要:
目的:应用软骨形态发生蛋白1(CDMP1),以诱导SD仔鼠脂肪干细胞(ADSCs)以修复兔膝关节软骨缺损,探讨异种细胞作为软骨组织工程种子细胞的可行性。 方法:自SD仔鼠腹股沟脂肪组织分离、培养ADSCs,复合于自制牛松质骨支架上,经CDMP1诱导,体外继续培养2周,免疫组化鉴定后备用。建立兔双侧髌骨关节缺损模型。左侧缺损处植入ADSCs-支架复合物,为实验侧;右侧缺损处植入空支架,为对照侧。术后8、16、24和48周各处死9只兔子,缺损处行H-E染色和番红O染色。结果:实验侧8周时可见缺损周围表面充填有薄层白色半透明组织,与周围软骨的界线清楚;16周时缺损表面界限连接较8周时模糊,但仍可辨,24周时,修复良好,修复区新生软骨细胞与周围正常软骨细胞形态相近,呈球形,可见软骨陷窝,H-E染色和番红O染色阳性;48周时,能较容易分清缺损处与修复区的界限,修复效果不及24周时。对照侧8、16、24和48周4个时期标本基本相同,软骨缺损处与周围正常组织边界清楚,缺损处凹陷空洞,被肉芽组织填充,新生细胞呈长梭形,H-E染色和番红O染色阴性。结论:利用CDMP1诱导SD仔鼠ADSCs复合自制牛松质骨支架可较好修复兔膝关节软骨缺损,为异种软骨细胞成为软骨组织工程的种子细胞提供可能。
关键词:  膝关节缺损;脂肪干细胞  异种移植;软骨细胞; 组织工程  软骨形态发生蛋白1
DOI:10.3724/SP.J.1008.2009.01363
投稿时间:2008-11-04修订日期:2009-07-30
基金项目:
Cartilage-derived morphogenetic protein growth factor 1-induced rat adipose-derived stem cells in repairing knee joint defect in rabbits
YANG Ya-jun1,2,ZHU Qing-sheng2*
(1.Department of Orthopedics,People’s Hospital of Ningxia Hui Autonomous Region,Yinchuan 750001,China;2.Department of Orthopedics,Xijing Hospital,Fourth Military Medical University,Xi’an 710032)
Abstract:
Objective:To repair knee joint defects in rabbits with rat adipose-derived stem cells induced by cartilage-derived morphogenetic protein growth factor 1 (CDMP1),so as to assess the feasibility of using heterogeneity cells as the seed cells for cartilage tissue engineering.Methods:The second generation ADSCs were seeded on scaffold,cultured for another two weeks in presence of CDMP1(50 μg/L),and identified by immunohistochemistory method.Bilateral rabbit knee joint defect model was established.The left side defect was embedded with ADSCs-scaffold composite (experimental group); the right side was embedded only with the scaffold(control group).Nine rabbits were killed in each group 8,16,24, and 48 weeks after embedding and the tissues were made into slices for safranine O and haematoxylin eosin staining.Results:In the experimental group the defects were filled with white semi-transparent tissues 8 weeks after embedding,with clear boundary to the surrounding cartilage; 16 weeks after embedding,the boundary of defect was further improved but still could be seen; 24 weeks after embedding,the repair outcomes were satisfactory,with the newly-generated chondrocytes having a nearly normal morphology (sphere shape,cartilage lacuna),and safranine O and haematoxylin eosin staining results were both positive; and 48 weeks after embedding,the boundary of the repair region could be clearly seen,and the repair effects were not as satisfactory as those of after 24 weeks.In the control group the boundary between the repairing area and the normal circumjacent area was visible at all 4 time points,with clear boundary and granulation tissues; the newly generated cells took a spindle shape and were negative for H-E and safranine O staining.Conclusion:The knee joint defects of rabbits can be satisfactorily repaired by using CDMP1-induced ADSCs seeded on spongy bone scaffold of cattle,which provides a theoretical basis for using heterogeneity cells as the seed cells for cartilage tissue engineering.
Key words:  knee joint defect  adipose-derived stem cells  heterogenic transplantation  chondrocyte  tissue engineering  cartilage-derived morphogenetic protein growth factor 1