Customized decellularization protocol of rabbit dermal matrix for tissue engineering applications

Decellularization is a challenging process that converts biological tissues into acellular matrices which can then be used in tissue engineering (TE). The goal is to remove cellular content while preserving extracellular matrix (ECM) components. Diverse chemical, physical, and biological reagents or combinations can be employed to produce specific scaZolding materials for TE and biomedical applications. A thorough understanding of the interactions between selected reagents and matrix components is crucial when customizing a decellularization protocol. This study proposes a new decellularization protocol of rabbit dermis in collaboration with Cortadoria Nacional de Pêlo, S.A., a supplier of a valuable agro-food by-product exceeding 1 ton/day. The selected chemical decellularization agents, including SDS, SDC, calcium hydroxide, and EDTA, were chosen for their proven eZicacy and according to previous work [3]. The impact of these decellularization reagents on matrix preservation was examined in this study and the impact of parameters such as exposure time, reagent concentration, and pH eZects on matrix components and mechanical properties were analyzed and optimized for an eZicient decellularization process. Our results indicate that reagent plays an important role in the ECM molecular conformation and consequently in expanding the matrix structure and allowing for optimal detergent interaction. The evaluation of the mechanical properties confirmed the matrix integrity, highlighting the protocol's impact on elastin and collagen content. DNA quantification demonstrated varied decellularization eZicacies depending on the applied reagents, methods, and decellularization steps. This study marks the first demonstration of successfully clean chemical methods for rabbit dermal decellularization, preserving ECM components and yielding high-quality polymeric matrices with superior biological performance for TE applications.