Abstract
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.
Highlights
Effective engineering approaches for cartilage regeneration involve a combination of cells and biomaterial scaffolds
It is well known that the regenerative capacity of articular cartilage after damage or disease is limited by low chondrocyte density with an extremely slow metabolic rate, abundant extracellular matrix (ECM), and no vascularity[3]
We evaluated differences in human mesenchymal stem cell (MSC) (hMSCs) chondrogenic potential according to diverse compositions of fibrin, thrombin, and atelocollagen
Summary
Effective engineering approaches for cartilage regeneration involve a combination of cells and biomaterial scaffolds. Histological analysis revealed lacunae-like structures, while staining demonstrated glycosaminoglycan accumulation These results indicate that atelocollagen scaffolds improve hMSC chondrogenic differentiation and are a potential approach for cartilage regeneration. Many therapeutic approaches have been developed for cartilage regeneration, including osteochondral transfer[4], bone marrow stimulation techniques such as microfracture, and cell-based therapies including autologous chondrocyte[5,6] and mesenchymal stem cell (MSC) implantation. These approaches do not yield complete cartilage regeneration as newly formed tissues have fibro-cartilaginous tissue characteristics and are mechanically weak[7]. Growth factors and cytokines such as TGF-β1, TGF-β2, TGF-β3, BMP-2, BMP-6, and BMP-7 can be used to enhance chondrogenic d ifferentiation[12]
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