中国组织工程研究

中国组织工程研究 ›› 2019, Vol. 23 ›› Issue (6): 849-854.doi: 10.3969/j.issn.2095-4344.1567

• 组织工程骨及软骨材料 tissue-engineered bone and cartilage materials • 上一篇    下一篇

软骨细胞外基质源性微粒修复山羊负重区大面积软骨缺损

侯昂扬1，2，3，陈 鹏3，汤 贺3，韩公海3，刘 平1，2，3，陈凯鹏1，2，彭 江3，张宇明2   

  1. 1山西医科大学，山西省太原市 030001；2山西医科大学附属人民医院，山西省太原市 030012；3解放军总医院全军骨科研究所，北京市 100853
  • 收稿日期:2018-10-25 出版日期:2019-02-28 发布日期:2019-02-28
  • 通讯作者: 张宇明，硕士，主任医师，山西省人民医院骨科，山西省太原市 030012
  • 作者简介:侯昂扬，男，1993年生，山西省运城市人，汉族，山西医科大学在读硕士，主要从事关节软骨再生研究。
  • 基金资助:

    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

Cartilage extracellular matrix-derived particles for repairing large-area cartilage defects in goat weight-bearing area

Hou Angyang1, 2, 3, Chen Peng3, Tang He3, Han Gonghai3, Liu Ping1, 2, 3, Chen Kaipeng1, 2, Peng Jiang3,  Zhang Yuming2   

  1. 1Shanxi Medical University, Taiyuan 030001, Shanxi Province, China; 2People’s Hospital of Shanxi Medical University, Taiyuan 030012, Shanxi Province, China; 3PLA Orthopedics Institute of PLA General Hospital, Beijing 100853, China
  • Received:2018-10-25 Online:2019-02-28 Published:2019-02-28
  • Contact: Zhang Yuming, Master, Chief physician, People’s Hospital of Shanxi Medical University, Taiyuan 030012, Shanxi Province, China
  • About author:Hou Angyang, Master candidate, 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
  • Supported by:

    the Translational Medicine Program of PLA General Hospital, No. 2016TM-020 (to PJ); the Major Research and Development Plan of Shanxi Province, No. 201703D321027-4 (to ZYM)

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摘要：

文章快速阅读：

 

文题释义：
软骨细胞外基质源性微粒：将新鲜软骨通过粉碎、过筛、脱细胞等一系列处理，得到的富含软骨细胞外基质，具有良好的生物相容性，可快速扩增种子细胞并具有促进干细胞成软骨分化能力的软骨微粒。
细胞外基质：是由细胞合成并分泌到胞外、分布在细胞表面或细胞之间的大分子，主要是一些多糖和蛋白或蛋白聚糖，这些物质构成复杂的网架结构，支持并连接组织结构、调节组织的发生和细胞的生理活动。


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。

  ORCID: 0000-0001-9854-9381(侯昂扬)

关键词: 软骨修复, 软骨细胞外基质源性微粒, 杨氏模量, 负重区软骨缺损, 大面积缺损, 蛋白聚糖

Abstract:

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 of 8 mm and a depth of 2 mm 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 at 3 and 6 months after implantation.

RESULTS AND CONCLUSION: (1) Hematoxylin-eosin staining: The defect was basically not repaired at 3 months 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; at 6 months 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. At 3 months 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. At 6 months 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: At 3 months 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. At 6 months 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.

Key words: Cartilage, Articular, Extracellular Matrix, Tissue Engineering

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