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Nasoseptal chondroprogenitors isolated through fibronectin-adherence confer no biological advantage for cartilage tissue engineering compared to nasoseptal chondrocytes.

Thomas Jovic, Emman Thomson, Nick Jones Orcid Logo, Cathy Thornton Orcid Logo, Shareen Doak Orcid Logo, Iain Whitaker

Frontiers in Bioengineering and Biotechnology, Volume: 12

Swansea University Authors: Thomas Jovic, Emman Thomson, Nick Jones Orcid Logo, Cathy Thornton Orcid Logo, Shareen Doak Orcid Logo, Iain Whitaker

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DOI (Published version): 10.3389/fbioe.2024.1421111

Abstract

The ability to bioprint facial cartilages could revolutionise reconstructive surgery, but identifying the optimum cell source remains one of the great challenges of tissue engineering. Tissue specific stem cells: chondroprogenitors, have been extracted previously using preferential adhesion to fibro...

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Published in: Frontiers in Bioengineering and Biotechnology
ISSN: 2296-4185
Published: Frontiers Media S.A. 2024
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URI: https://cronfa.swan.ac.uk/Record/cronfa68056
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Abstract: The ability to bioprint facial cartilages could revolutionise reconstructive surgery, but identifying the optimum cell source remains one of the great challenges of tissue engineering. Tissue specific stem cells: chondroprogenitors, have been extracted previously using preferential adhesion to fibronectin based on the expression of CD49e: a perceived chondroprogenitor stem cell marker present on <1% of cartilage cells. This study sought to determine whether these fibronectin-adherent chondroprogenitor cells could be exploited for cartilage tissue engineering applications in isolation, or combined with differentiated chondrocytes. Nasoseptal cartilage samples from 20 patients (10 male, 10 female) were digested to liberate cartilage-derived cells (CDCs) from extracellular matrix. Total cell number was counted using the Trypan Blue exclusion assay and added to fibronectin coated plates for 20 min, to determine the proportion of fibronectin-adherent (FAC) and non-adherent cells (NFACs). All populations underwent flow cytometry to detect mesenchymal stem/progenitor cell markers and were cultured in osteogenic, chondrogenic and adipogenic media to determine trilineage differentiation potential. Cell adherence and growth kinetics of the different populations were compared using iCELLigence growth assays. Chondrogenic gene expression was assessed using RT-qPCR for Type 2 collagen, aggrecan and SOX9 genes. Varying proportions of NFAC and FACs were cultured in alginate beads to assess tissue engineering potential. 52.6% of cells were fibronectin adherent in males and 57.7% in females, yet on flow cytometrical analysis, only 0.19% of cells expressed CD49e. Moreover, all cells (CDC, FAC and NFACs) demonstrated an affinity for trilineage differentiation by first passage and the expression of stem/progenitor cell markers increased significantly from digest to first passage (CD29, 44, 49e, 73 and 90, p < 0.0001). No significant differences were seen in adhesion or growth rates. Collagen and aggrecan gene expression was higher in FACs than CDCs (2-fold higher, p = 0.008 and 0.012 respectively), but no differences in chondrogenic potential were seen in any cell mixtures in 3D culture models. The fibronectin adhesion assay does not appear to reliably isolate a chondroprogenitor cell population from nasoseptal cartilage, and these cells confer no advantageous properties for cartilage tissue engineering. Refinement of cell isolation methods and chondroprogenitor markers is warranted for future nasoseptal cartilage tissue engineering efforts. [Abstract copyright: Copyright © 2024 Jovic, Thomson, Jones, Thornton, Doak and Whitaker.]
Keywords: Chondroprogenitor, cartilage, tissue engineering, fibronectin, stem cell
College: Faculty of Medicine, Health and Life Sciences
Funders: The author(s) declare that financial support was received for the research, authorship, and/or publication of this article. This study was supported by The Scar Free Foundation and Health and Care Research Wales Programme of research in Reconstructive Surgery and Regenerative Medicine, which has been established in the ReconRegen Research Centre at Swansea University in partnership with Swansea Bay University Health Board. Additionally, TJ would like to acknowledge funding from Action Medical Research and the VTCT Foundation (GN2782) and Microtia United Kingdom. TJ and ET would like to acknowledge funding from the Welsh Clinical Academic Training Programme.

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