Bioprinted keratinocyte and stem cell-laden constructs for skin tissue engineering

Bettendorf E, Schmid R, Horch RE, Kengelbach-Weigand A, Kulicke Y, Schrüfer S, Schubert DW, Lamberger Z, Stahlhut P, Lang G, Sörgel C (2024)

Publication Language: English

Publication Type: Journal article

Publication year: 2024

Journal

Book Volume: 10

Pages Range: 260-281

Article Number: 3925

Journal Issue: 6

DOI: 10.36922/ijb.3925

Abstract

Treating large-scale skin wounds remains a significant therapeutic challenge, often due to insufficient autologous material for complete coverage. Recent advances in biofabrication offer a solution with reproducible and precise large-scale production. Herein, this study aims to evaluate the feasibility of biofabrication and develop a customized three-dimensional (3D) bioprinted skin construct containing immortalized HaCaT keratinocytes and adipose-derived stem cells (ADSCs). Keratinocytes were cultured in various hydrogels (e.g., containing alginate [Alg], fibrin [Fib], collagen, gelatin [Gel], gelatin methacryloyl [GelMA], hyaluronic acid [HA], and a pre-fabricated collagen-elastin-matrix) for 7 days. The metabolic activity of cultured keratinocytes was then evaluated during the co-cultivation of HaCaT and ADSCs in a transwell model. The metabolic activity in all groups increased over the experimental period. Alg/HA/Gel and GelMA hydrogels demonstrated good printability and high diffusion rates. There was no significant difference in pore size between all hydrogels. Based on the results of printability and diffusion assays, as well as scanning electron microscopy (SEM) and rheological measurements, Alg/HA/Gel and GelMA hydrogels were selected for the bioprinted 3D model. Fib hydrogel was integrated into the biofabricated constructs for its excellent metabolic activity in the transwell model. Hydrogel stability, cell survival, and metabolic activity in bioprinted 3D models containing keratinocytes and ADSCs were evaluated over 14 days. On day 14, metabolic activity and live cell count within the bioprinted constructs of the co-cultured groups were significantly higher compared to day 1. The biofabricated GelMA constructs displayed higher cell viability than Alg/HA/Gel constructs. Additionally, to evaluate cell migration out of the constructs, the metabolic activity and viability of the cells on the well bottom were examined. After 14 days, an average of 50% of the well bottom was covered by HaCaT cells, which were initially printed in co-culture into the constructs. These findings indicate that GelMA constructs containing keratinocytes and ADSCs may offer a promising therapeutic option in the treatment of large chronic wounds.

Authors with CRIS profile

Involved external institutions

Universitätsklinikum Würzburg Germany (DE)

How to cite

APA:

Bettendorf, E., Schmid, R., Horch, R.E., Kengelbach-Weigand, A., Kulicke, Y., Schrüfer, S.,... Sörgel, C. (2024). Bioprinted keratinocyte and stem cell-laden constructs for skin tissue engineering. International Journal of Bioprinting, 10(6), 260-281. https://doi.org/10.36922/ijb.3925

MLA:

Bettendorf, Eva, et al. "Bioprinted keratinocyte and stem cell-laden constructs for skin tissue engineering." International Journal of Bioprinting 10.6 (2024): 260-281.

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