Biofabrication of a bioactive three-layered skin hydrogel based on ECM components effectively promotes wound healing and regeneration

Biofabrication of a bioactive three-layered skin hydrogel based on ECM components effectively promotes wound healing and regeneration Paula Pleguezuelos-Beltrán1,2,3,†, Carlos Chocarro-Wrona1,2,3,†, Julia López de Andrés1,2,3, Cristina Antich1,3,4, Juan de Vicente5, Patricia Gálvez-Martín6,*, Elena López-Ruiz2,3,7,*, Juan Antonio Marchal1,2,3,* 1BioFab i3D - Biofabrication and 3D (bio)printing laboratory, Department of Human Anatomy and Embryology, University of Granada, Spain. 2Biopathology and Regenerative Medicine Institute (IBIMER), Center for Biomedical Research (CIBM), University of Granada, Spain. 3 Instituto de Investigación Biosanitaria ibs.GRANADA, University Hospitals of Granada, University of Granada, Spain. 4National Center for Advancing Translational Sciences, National Institute of Health, United States. 5F2N2Lab, Magnetic Soft Matter Group, Department of Applied Physics, University of Granada, Spain. 6Bioibérica S.A.U., Barcelona, Spain. 7Department of Health Sciences, University of Jaén, Spain. † Equal contribution; * Corresponding authors paulapb[email protected]gr.es INTRODUCTION Skin wounds Chocarro-Wrona et al. (2019) •May be due to diverse causes. •Classified depending on the depth of the injury: Savoji et al. Front Bioeng Biotechnol. (2018); Papini. Br Med J. (2004) •Epidermal •Superficial partial-thickness •Deep partial-thickness •Full thickness Can be regenerated naturally Require therapeutic intervention to heal INTRODUCTION •Market skin substitutes: dermal and epidermal layers (subcutaneous layer is often neglected) •Increased interest in three-layered skin substitutes •Most of them only use one-component hydrogels ECM of each layer was customized to biofabricate a bioactive and biologically-inspired skin substitute: BT Skin. •Supplementation of the hydrogels with: •Specific components of each skin layer: keratin, sphingolipids, dermatan sulfate, HA, or elastin •Provides the BT Skin with enhanced ECM complexity. BT SKIN BT SKIN AB C 2 mm 3 - 4 mm MECHANICAL CHARACTERIZATION MECHANICAL CHARACTERIZATION BIOCOMPATIBILITY 500 µm BIOCOMPATIBILITY CellTracker Green: hEKs CellTracker Red: hDFs CellTracker Green: hMSCs 16 This research was supported by the Ministry of Economy and Competitiveness, Instituto de Salud Carlos III (FEDER funds, DTS19/00143, DTS21/00098, PID2019-104883 GB-I00 and TED2021129384B-C22), by the Consejería de Economía, Conocimiento, Empresas y Universidad de la Junta de Andalucía (P18-FR-2470, B-CTS-230-UGR18,PYC20.RE.015.UGR and A-CTS-180UGR20, FEDER Funds), by the European Union's Horizon Europe research and innovation program (grant agreement n°101092269, NABIHEAL), and by Modeling Nature (MNat), Project number QUAL21-11. Conflict of interest disclosure: nothing to disclose. I B I MER ACKNOWLEDGEMENTS REFERENCE doi.org/10.1016/j.mtbio.2025.101592 18