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胶原酶治疗似乎能改善软骨组织整合,但对胶原网络的损伤可能是永久性的。

Collagenase treatment appears to improve cartilage tissue integration but damage to collagen networks is likely permanent.

作者信息

Shajib Md Shafiullah, Futrega Kathryn, Jacob Klein Travis, Crawford Ross W, Doran Michael Robert

机构信息

School of Biomedical Science, Faculty of Health, Queensland University of Technology, Brisbane, QLD, Australia.

Centre for Biomedical Technologies, School of Mechanical, Medical, and Process Engineering, Faculty of Engineering, Queensland University of Technology, Brisbane, QLD, Australia.

出版信息

J Tissue Eng. 2022 Jan 23;13:20417314221074207. doi: 10.1177/20417314221074207. eCollection 2022 Jan-Dec.

Abstract

When repairing cartilage defects a major challenge is achieving high-quality integration between the repair tissue and adjacent native cartilage. Matrix-rich cartilage is not easily remodeled, motivating several studies to trial enzyme treatment of the tissue interface to facilitate remodeling and integration. Studying and optimizing such processes is tedious, as well as potentially expensive, and thus simpler models are needed to evaluate the merits of enzyme treatment on cartilage tissue integration. Herein, we used engineered cartilage microtissues formed from bone marrow-derived stromal cells (BMSC) or expanded articular chondrocytes (ACh) to study the impact of enzyme treatment on cartilage tissue integration and matrix remodeling. A 5-min treatment with collagenase appeared to improve cartilage microtissue integration, while up to 48 h treatment with hyaluronidase did not. Alcian blue and anti-collagen II staining suggested that collagenase treatment did facilitate near seamless integration of cartilage microtissues. Microtissue sections were stained with Picrosirius red and characterized using polarized light microscopy, revealing that individual microtissues contained a collagen network organized in concentric shells. While collagenase treatment appeared to improve tissue integration, assessment of the collagen fibers with polarized light indicated that enzymatically damaged networks were not remodeled nor restored during subsequent culture. This model and these data paradoxically suggest that collagen network disruption is required to improve cartilage tissue integration, but that the disrupted collagen networks are unlikely to subsequently be restored. Future studies should attempt to limit collagen network disruption to the surface of the cartilage, and we recommend using Picrosirius red staining and polarized light to assess the quality of matrix remodeling and integration.

摘要

在修复软骨缺损时,一个主要挑战是实现修复组织与相邻天然软骨之间的高质量整合。富含基质的软骨不易重塑,这促使多项研究尝试对组织界面进行酶处理,以促进重塑和整合。研究和优化此类过程既繁琐又可能成本高昂,因此需要更简单的模型来评估酶处理对软骨组织整合的优点。在此,我们使用由骨髓来源的基质细胞(BMSC)或扩增的关节软骨细胞(ACh)形成的工程化软骨微组织,来研究酶处理对软骨组织整合和基质重塑的影响。用胶原酶处理5分钟似乎可改善软骨微组织的整合,而用透明质酸酶处理长达48小时则没有效果。阿尔新蓝和抗胶原蛋白II染色表明,胶原酶处理确实促进了软骨微组织的近乎无缝整合。用天狼星红对微组织切片进行染色,并使用偏光显微镜进行表征,结果显示单个微组织包含以同心壳形式组织的胶原网络。虽然胶原酶处理似乎改善了组织整合,但用偏光对胶原纤维进行评估表明,在随后的培养过程中,酶损伤的网络并未重塑或恢复。这个模型和这些数据自相矛盾地表明,需要破坏胶原网络来改善软骨组织整合,但被破坏的胶原网络随后不太可能恢复。未来的研究应尝试将胶原网络破坏限制在软骨表面,并且我们建议使用天狼星红染色和偏光来评估基质重塑和整合的质量。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/825c/8793122/bc89897a3425/10.1177_20417314221074207-fig1.jpg

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