Page 268 - IJB-10-6

International

Journal of Bioprinting

RESEARCH ARTICLE
Bioprinted keratinocyte and stem cell-laden

constructs for skin tissue engineering

Eva Bettendorf , Rafael Schmid 1† id , Raymund E. Horch 1 id ,
1†
Annika Kengelbach-Weigand 1 id , Yvonne Kulicke , Stefan Schrüfer 2 id ,
1
Dirk W. Schubert 2 id , Zan Lamberger 3 id , Philipp Stahlhut 3 id , Gregor Lang 3 id ,
and Celena A. Sörgel *
1
1 Department of Plastic and Hand Surgery and Laboratory for Tissue Engineering and Regenerative
Medicine, University Hospital of Erlangen, Friedrich-Alexander University Erlangen-Nürnberg,
Erlangen, Germany
2 Department of Materials Science and Engineering, Institute of Polymer Materials, Friedrich-
Alexander University Erlangen-Nürnberg, Erlangen, Germany
3 Department for Functional Materials in Medicine and Dentistry, University Hospital of Würzburg,
Julius-Maximilians University Würzburg, Würzburg, Germany

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
† These authors contributed equally production. Herein, this study aims to evaluate the feasibility of biofabrication and
to this work. develop a customized three-dimensional (3D) bioprinted skin construct containing
*Corresponding author: immortalized HaCaT keratinocytes and adipose-derived stem cells (ADSCs).
Celena A. Sörgel Keratinocytes were cultured in various hydrogels (e.g., containing alginate [Alg],
(celena.soergel@uk-erlangen.de) fibrin [Fib], collagen, gelatin [Gel], gelatin methacryloyl [GelMA], hyaluronic acid
Citation: Bettendorf E, Schmid R, [HA], and a pre-fabricated collagen-elastin-matrix) for 7 days. The metabolic activity

Horch RE, et al. Bioprinted of cultured keratinocytes was then evaluated during the co-cultivation of HaCaT

keratinocyte and stem
cell-laden constructs for and ADSCs in a transwell model. The metabolic activity in all groups increased over
skin tissue engineering. the experimental period. Alg/HA/Gel and GelMA hydrogels demonstrated good
Int J Bioprint. 2024;10(6):3925. printability and high diffusion rates. There was no significant difference in pore size
doi: 10.36922/ijb.3925
between all hydrogels. Based on the results of printability and diffusion assays, as well
Received: June 12, 2024 as scanning electron microscopy (SEM) and rheological measurements, Alg/HA/Gel
Revised: July 25, 2024
Accepted: August 6, 2024 and GelMA hydrogels were selected for the bioprinted 3D model. Fib hydrogel was
Published Online: August 7, 2024 integrated into the biofabricated constructs for its excellent metabolic activity in the
transwell model. Hydrogel stability, cell survival, and metabolic activity in bioprinted
Copyright: © 2024 Author(s).
This is an Open Access article 3D models containing keratinocytes and ADSCs were evaluated over 14 days. On
distributed under the terms of the day 14, metabolic activity and live cell count within the bioprinted constructs of the
Creative Commons Attribution co-cultured groups were significantly higher compared to day 1. The biofabricated
License, permitting distribution,
and reproduction in any medium, GelMA constructs displayed higher cell viability than Alg/HA/Gel constructs.
provided the original work is Additionally, to evaluate cell migration out of the constructs, the metabolic activity
properly cited. and viability of the cells on the well bottom were examined. After 14 days, an average
Publisher’s Note: AccScience of 50% of the well bottom was covered by HaCaT cells, which were initially printed
Publishing remains neutral with in co-culture into the constructs. These findings indicate that GelMA constructs
regard to jurisdictional claims in
published maps and institutional containing keratinocytes and ADSCs may offer a promising therapeutic option in the
affiliations. treatment of large chronic wounds.

Volume 10 Issue 6 (2024) 260 doi: 10.36922/ijb.3925