Department of Orthopedic Surgery, College of Medicine, The Catholic University of Korea, Seoul, Republic of Korea.
The Rheumatism Research Center, Catholic Research Institute of Medical Science, College of Medicine, The Catholic University of Korea, Seoul, Republic of Korea.
Sci Rep. 2020 Jun 30;10(1):10678. doi: 10.1038/s41598-020-67836-3.
Effective engineering approaches for cartilage regeneration involve a combination of cells and biomaterial scaffolds. Multipotent mesenchymal stem cells (MSCs) are important sources for cartilage regeneration. Atelocollagen provides a suitable substrate for MSC attachment and enhancing chondrogenic differentiation. Here, we assessed the chondrogenic potential of adipose tissue derived human MSCs (hMSCs) mixed with atelocollagen gel. We observed cell attachment, viability, and microstructures by electron microscopy over 21 days. The levels of Sox9, type II collagen, aggrecan, type I collagen, Runx2, type X collagen, ALP, Osterix, and MMP13 were measured by RT-qPCR. Cartilage matrix-related proteins were assessed by enzyme-linked immunosorbent assay (ELISA), histology, and immunohistochemistry. hMSCs of all groups exhibited well-maintained cell survival, distribution and morphology. Abundant type II collagen fibers developed on day 21; while Sox9, type II collagen, and aggrecan expression increased over time in the atelocollagen group. However, type I collagen, RUNX2, type X collagen (CoL10A1), Osterix, and ALP were not expressed. These results corroborated the protein expression detected by ELISA. Further, histological analysis revealed lacunae-like structures, while staining demonstrated glycosaminoglycan accumulation. Cumulatively, these results indicate that atelocollagen scaffolds improve hMSC chondrogenic differentiation and are a potential approach for cartilage regeneration.
有效的软骨再生工程方法涉及细胞和生物材料支架的结合。多能间充质干细胞（MSCs）是软骨再生的重要来源。Atelocollagen 为 MSC 附着和增强软骨分化提供了合适的基质。在这里，我们评估了脂肪组织来源的人 MSCs（hMSCs）与 Atelocollagen 凝胶混合的软骨生成潜力。我们通过电子显微镜观察了 21 天内的细胞附着、活力和微观结构。通过 RT-qPCR 测量 Sox9、II 型胶原、聚集蛋白聚糖、I 型胶原、Runx2、X 型胶原、ALP、Osterix 和 MMP13 的水平。通过酶联免疫吸附试验（ELISA）、组织学和免疫组织化学评估软骨基质相关蛋白。所有组的 hMSCs 均表现出良好的细胞存活、分布和形态。第 21 天大量 II 型胶原纤维形成；而 Sox9、II 型胶原和聚集蛋白聚糖的表达随时间增加。然而，I 型胶原、RUNX2、X 型胶原（CoL10A1）、Osterix 和 ALP 未表达。这些结果与 ELISA 检测到的蛋白表达相符。此外，组织学分析显示出腔隙样结构，而染色显示出糖胺聚糖的积累。综上所述，这些结果表明 Atelocollagen 支架可改善 hMSC 软骨分化，是软骨再生的一种潜在方法。
