摘要
背景:随着组织工程技术的发展,使用天然的生物材料作为支架可以加快新生软骨组织的生成,有利于软骨缺损的修复。目的:探究软骨细胞外基质源性微粒修复负重区软骨大面积缺损的能力。方法:刮取山羊膝关节软骨,通过脱细胞技术制备细胞外基质源性微粒。取12只中国山羊(解放军总医院动物实验中心提供),制作右股骨内外髁负重区直径8 mm、深2 mm的全层骨软骨缺损模型,随机分为2组,实验组(n=6)于缺损处植入同种异体软骨细胞外基质源性微粒,并用纤维蛋白胶固定;对照组(n=6)仅填入纤维蛋白胶。植入3,6个月取右膝关节股骨远端样本,进行组织学及生物力学评价。结果与结论:(1)苏木精-伊红染色:对照组植入3个月时缺损基本无修复,有少量纤维组织填充,缺损区凹陷,基质无异染,两侧交界面整合差;植入后6个月缺损区仍主要为纤维组织,少量纤维软骨填充,新生组织与周围正常组织仍有明显界限。实验组植入3个月时缺损处面积较对照组小,新生组织为纤维软骨与透明软骨复合体,内部可见典型软骨陷窝,细胞排列较为有序;植入6个月时缺损区组织透明软骨比例增大,表面较为平滑,与周围正常软骨组织类似,并整合良好,放大可见典型软骨陷窝,细胞排列有序;(2)番红O-固绿染色:植入3个月时,对照组缺损区基本无蛋白多糖红染,实验组缺损区显示明显蛋白多糖着色;植入6个月时,实验组新生组织中蛋白多糖着色明显多于对照组,且实验组新生组织透明软骨比例大,与周围正常软骨类似;(3)生物力学:植入后6个月,实验组平均杨氏模量明显高于对照组(P <0.05);(4)结果表明:使用软骨细胞外基质源性微粒可促进软骨缺损的修复。
BACKGROUND:With the development of tissue engineering technology,the use of natural biological materials as scaffolds can accelerate the formation of new cartilage tissue and facilitate the repair of cartilage defects.OBJECTIVE:To explore the ability of cartilage extracellular matrix-derived particles to repair cartilage defects in the weight-bearing area.METHODS:Extracellular matrix-derived particles were prepared by decellularization of goat knee cartilage.Twelve Chinese goats(provided by the Experimental Animal Center of PLA General Hospital in China)were selected to make a full-thickness osteochondral defect model with a diameter of8mm and a depth of2mm in the weight-bearing area of the inner and outer condyles of the right femur.These goat models were randomly divided into two groups.In the experimental group(n=6),the cartilage extracellular matrix-derived particles were implanted to the defect and fixed with fibrin glue.The control group(n=6)was only filled with fibrin glue.General,histological and biomechanical evaluations were conducted with distal right femur samples taken at3and6months after implantation.RESULTS AND CONCLUSION:(1)Hematoxylin-eosin staining:The defect was basically not repaired at3months after implantation in the control group,with a small amount of fibrous tissue filling,sag in the defect area,matrix staining and poor integration of the bilateral interface;at6months after implantation,the defect area in the control group was still mainly filled with fibrous tissues and contained a small amount of fibrous cartilage,and there was still a clear boundary between the new tissue and surrounding normal tissue.At3months after implantation,the defect area of the experimental group was smaller than that of the control group,and the new tissue was a complex of fibrocartilage and hyaline cartilage.At6months after implantation,the proportion of tissue hyaline cartilage in the defect area of the experimental group increased,and the surface was relatively smooth,similar to the surrounding normal cartilage tissue,and well integrated.Enlarged cartilage lacunae were visible and the cells are arranged in order.(2)Safranine O-Fast Green staining:At3months after implantation,the defect area of the control group was basically free of proteoglycan red stain,while the defect area of the experimental group showed obvious proteoglycan coloring.At6months after implantation,the amount of proteoglycan-stained new tissue in the experimental group was significantly higher than that in the control group,and the proportion of hyaline cartilage in the new tissue was higher in the experimental group,similar to that of surrounding normal cartilage.(3)Biomechanical analysis:Six months after implantation,the average Young’s modulus of the experimental group was significantly higher than that of the control group(P<0.05).All the findings reveal that the use of cartilage extracellular matrix-derived microparticles can promote the repair of cartilage defects.
作者
侯昂扬
陈鹏
汤贺
韩公海
刘平
陈凯鹏
彭江
张宇明
Hou Angyang;Chen Peng;Tang He;Han Gonghai;Liu Ping;Chen Kaipeng;Peng Jiang;Zhang Yuming(Shanxi Medical University, Taiyuan 030001, Shanxi Province, China;People’s Hospital of Shanxi Medical University, Taiyuan 030012, Shanxi Province, China;PLA Orthopedics Institute of PLA General Hospital, Beijing 100853, China)
出处
《中国组织工程研究》
CAS
北大核心
2019年第6期849-854,共6页
Chinese Journal of Tissue Engineering Research
基金
解放军总医院转化医学项目(2016TM-020)
项目负责人:彭江
山西省重点研发计划项目(201703D321027-4)
项目负责人:张宇明~~
