病理改变的关节软骨吸引强烈的软骨细胞侵入细胞外基质:体外初步研究。

Q2 Medicine
Victoria A Shestakova, Ilya D Klabukov, Ilya V Kolobaev, Longfeng Rao, Dmitry A Atiakshin, Michael A Ignatyuk, Mikhail E Krasheninnikov, Bagavdin G Ahmedov, Sergey A Ivanov, Peter V Shegay, Andrey D Kaprin, Denis S Baranovskii
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引用次数: 0

摘要

背景:由于无血管化和神经结构,关节软骨具有有限的自我修复能力。本研究的目的是探讨人鼻软骨细胞(hNC)对炎症损伤关节软骨基质的恢复作用。材料和方法:软骨基质是通过对术中获得的成人膝关节置换术患者关节软骨标本进行失活制备的。hNC是从天然组织中通过酶消化得到的,并在两个传代上进一步扩增。获得的鼻腔软骨细胞用于植入去细胞支架,然后在软骨培养基中体外培养7、14或21天。用快绿、红素- o、苏木精染色和扫描电镜观察迁移。采用二甲亚甲基蓝和CyQuant细胞增殖分析试剂盒进行生化分析,测定软骨中糖胺聚糖(GAG)和DNA含量。结果:在健康和炎症软骨中植入鼻软骨细胞,发现这些细胞积极侵入病理改变的软骨。生化定量分析结果显示,DNA数量在第7天显著增加,在第14天显著减少,而GAGs的定量值则相反。组织学染色显示,第7天软骨形成,第14天细胞间隙充满全新合成的软骨基质,GAG含量显著降低,第21天观察到新形成的富含GAG的软骨。获得的软骨再生数据经扫描电镜证实。结论:我们的初步结果表明,人鼻软骨细胞能够浸润到关节炎损伤的关节软骨病理改变的细胞外基质中,从而促进其修复到生理相关状态。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
Pathologically altered articular cartilage attracts intense chondrocyte invasion into the extracellular matrix: in vitro pilot study.

Background: Due to non-vascularized and aneural structure, articular cartilage has limited self-repairing capacity. The aim of this study was to investigate the revitalization of inflammatory injured articular cartilage matrices by human nasal chondrocytes (hNC).

Materials and methods: Cartilage matrix was prepared by devitalization of articular cartilage samples obtained intraoperatively from an adult patient undergoing knee joint replacement. hNC were obtained from native tissues by enzymatic digestion with further expansion over two passages. The obtained nasal chondrocytes were used to seed decellularized scaffolds, which were then cultured in vitro for 7, 14, or 21 days in chondrogenic medium. Migration was observed by histologic staining with fast green, safranin-O, and hematoxylin and scanning electron microscopy. Biochemical analysis was performed to determine the glycosaminoglycan (GAG) and DNA content of the cartilage using dimethylmethylene blue and CyQuant Cell Proliferation Assay Kit.

Results: We seeded healthy and inflamed cartilage with nasal chondrocytes and found that the cells actively invade mainly pathologically altered cartilage. The results of biochemical quantitative analysis showed that the amount of DNA significantly increased by day 7 and decreased by day 14, while the quantitative values of GAGs had the opposite trend. Histological staining showed that cartilage formation occurred on day 7, intercellular spaces were filled with de novo synthesized cartilage matrix with significantly low GAG content on day 14, and newly formed GAG-rich cartilage was observed on day 21. The obtained data on cartilage regeneration were confirmed by scanning electron microscopy.

Conclusions: Our preliminary results showed that human nasal chondrocytes are capable of infiltrating the pathologically altered extracellular matrix of articular cartilage damaged by arthritis, thereby promoting its repair to a physiologically relevant state.

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来源期刊
CiteScore
6.50
自引率
0.00%
发文量
42
审稿时长
19 weeks
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