应用 miR-29a-Exosome 和多功能支架治疗全厚软骨缺损

Fan Yang , Zewen Wang , Mingjian Wu, Jingyi Xu, Junlei Li, Jiahe Liu, Ting He, Tao Zhang, Baoyi Liu
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引用次数: 0

摘要

背景全厚软骨缺损是骨科常见的难治性疾病。在这项研究中,我们设计了一种新型复合支架,由丝绸纤维素-壳聚糖(用于软骨层)和多孔钽(用于软骨下骨层)组成,并负载了工程骨间充质干细胞外泌体,以评估其修复全厚软骨缺损的功效。方法将多孔钽进行三维打印,并与丝绸纤维素-壳聚糖结合形成复合支架。在支架上培养软骨细胞,并用 CCK-8 法评估其生长情况。甲苯胺蓝染色证实了细胞形态,免疫荧光显示了胶原蛋白Ⅱ型的表达。利用各种技术制作了装载有 miR-29a 的工程外泌体,并对其进行了表征。与软骨细胞的共培养显示,它们的增殖持续了10天。免疫荧光显示了细胞核、II型胶原蛋白和Aggrecan的染色。结果丝纤维素-壳聚糖支架表现出良好的生物相容性,支持软骨细胞的粘附、生长和软骨组织的形成。工程外泌体表现出良好的生物活性,有利于骨和软骨的再生。结论丝纤维素-壳聚糖与多孔钽复合支架搭载的miR-29a工程外泌体具有良好的全厚软骨缺损再生潜力。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
Application of miR-29a-Exosome and multifunctional scaffold for full-thickness cartilage defects

Background

Full-thickness cartilage defect is a common refractory disease in orthopedics. In this study, we designed a novel composite scaffold composed of silk fibroin-chitosan for the cartilage layer and porous tantalum for the subchondral bone layer, loaded with engineered bone mesenchymal stem cell exosomes, to evaluate its efficacy in repairing full-thickness cartilage defect.

Methods

Porous tantalum was 3D printed and combined with silk fibroin-chitosan to form a composite scaffold. Chondrocytes were cultured on the scaffold, and their growth was assessed using the CCK-8 method. Toluidine blue staining confirmed cell morphology, while immunofluorescence revealed collagen type Ⅱ expression. Engineered exosomes loaded with miR-29a were created and characterized using various techniques. Co-culturing with chondrocytes demonstrated their proliferation over 10 days. Immunofluorescence revealed staining for the nucleus, collagen type II, and Aggrecan. In vivo experiments were performed on rats to assess cartilage defect repair, utilizing histological staining and micro-CT scanning at 4 and 8 weeks post-operation.

Results

The silk fibroin-chitosan scaffold demonstrated good biocompatibility, supporting chondrocyte adhesion, growth, and cartilage tissue formation. Engineered exosomes exhibited promising biological activity, conducive to bone and cartilage regeneration. The implantation of the silk fibroin-chitosan/porous tantalum composite scaffold loaded with engineered exosomes promoted integration with the surrounding bone and cartilage tissues, facilitating repair and regeneration.

Conclusions

The silk fibroin-chitosan combined with porous tantalum scaffold carrying engineered exosomes loaded with miR-29a has good potential for full-thickness cartilage defects regeneration.
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来源期刊
Extracellular vesicle
Extracellular vesicle Biochemistry, Genetics and Molecular Biology (General)
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