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Journal of Zhejiang University SCIENCE B

ISSN 1673-1581(Print), 1862-1783(Online), Monthly

Repair of articular cartilage defects in rabbits through tissue-engineered cartilage constructed with chitosan hydrogel and chondrocytes

Abstract: Objective: In our previous work, we prepared a type of chitosan hydrogel with excellent biocompatibility. In this study, tissue-engineered cartilage constructed with this chitosan hydrogel and costal chondrocytes was used to repair the articular cartilage defects. Methods: Chitosan hydrogels were prepared with a crosslinker formed by combining 1,6-diisocyanatohexane and polyethylene glycol. Chitosan hydrogel scaffold was seeded with rabbit chondrocytes that had been cultured for one week in vitro to form the preliminary tissue-engineered cartilage. This preliminary tissue-engineered cartilage was then transplanted into the defective rabbit articular cartilage. There were three treatment groups: the experimental group received preliminary tissue-engineered cartilage; the blank group received pure chitosan hydrogels; and, the control group had received no implantation. The knee joints were harvested at predetermined time. The repaired cartilage was analyzed through gross morphology, histologically and immunohistochemically. The repairs were scored according to the international cartilage repair society (ICRS) standard. Results: The gross morphology results suggested that the defects were repaired completely in the experimental group after twelve weeks. The regenerated tissue connected closely with subchondral bone and the boundary with normal tissue was fuzzy. The cartilage lacuna in the regenerated tissue was similar to normal cartilage lacuna. The results of ICRS gross and histological grading showed that there were significant differences among the three groups (P<0.05). Conclusions: Chondrocytes implanted in the scaffold can adhere, proliferate, and secrete extracellular matrix. The novel tissue-engineered cartilage constructed in our research can completely repair the structure of damaged articular cartilage.

Key words: Articular cartilage, Chitosan hydrogel, Repair, Tissue engineering

Chinese Summary  <23> 新型壳聚糖水凝胶结合软骨细胞修复兔关节软骨缺损的实验研究

目的:评估壳聚糖水凝胶结合肋软骨细胞构建的新型组织工程软骨对兔关节软骨缺损的修复效果。
创新点:利用自主研发的具有良好生物相容性和稳定性的壳聚糖水凝胶与软骨细胞,在体外初步构建组织工程软骨,并尝试利用其修复缺损的关节软骨,从而为关节软骨缺损的修复提供了一种新的治疗方法。
方法:取兔肋软骨体外培养扩增,获得P2代软骨细胞,将其种植到冻干的壳聚糖水凝胶上,体外培养一周,获得初步构建的组织工程软骨。构建兔膝关节软骨缺损模型,并分为3组:实验组植入组织工程软骨;对照组植入壳聚糖水凝胶;空白组不做任何处理。分别于术后4、8和12周取材,通过大体观察、苏木精-伊红染色、番红-O染色及II型胶原免疫组化染色等方法观察缺损关节软骨的修复情况,并用国际关节软骨修复协会(ICRS)制定的评分法进行大体及组织学评分。
结论:兔肋软骨细胞能在实验室自主构建的壳聚糖水凝胶上增殖并分泌细胞外基质(图2),植入到兔关节软骨缺损处后,对缺损关节软骨具有良好的修复作用(图3和5),且修复是一种完全的结构性的修复(图5~7)。

关键词组:关节软骨修复;组织工程软骨;壳聚糖水凝胶


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DOI:

10.1631/jzus.B1500036

CLC number:

R318.08

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On-line Access:

2015-11-04

Received:

2015-02-04

Revision Accepted:

2015-06-09

Crosschecked:

2015-10-20

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