Novel genipin-crosslinked acellular biogenic conduits for tissue engineering applications García García, Óscar Darío Escalante-Quirós, Sandra Llinares-Monllor, Claudia Ávila-Fernández, Paula Sánchez Porras, David Etayo-Escanilla, Miguel Campos Sánchez, Fernando Chato Astrain, Jesús Carriel Araya, Víctor Biogenic Decellularization Extracellular matrix Genipin Tissue engineering Biogenic matrix Bioconduit Neural engineering Crosslinking Macrophages This research was financed by the Spanish “Plan Nacional de Investigación Científica, Desarrollo e Innovación Tecnológica” (I+D+i) of the Ministry of Science and Innovation of Spain (Instituto de Salud Carlos III) and co-financed by FEDER Funds (European Union) (Grants FIS PI20/00318, PI23/00337 and PI22/00059)". D.S.-P. by PFIS Fellowship Grant FIS PI20/00318 and M.E.-E. by FPU Fellowship Grant FPU21/06183 of the Spanish Ministry of Universities. Tissue Engineering Group, Department of Histology, Faculty of Medicine, University of Granada, Granada, Spain Background: Collagen-based conduits have been generated in-vivo stimulating a fibrotic response through the implantation of a non-resorbable material in animal models, creating biogenic substitutes. However, they often exhibit clinical limitations due to prolonged generation times, exclusive autologous use and insufficient mechanical strength. Consequently, decellularization and cross-linking could solve the aforementioned drawbacks, providing a non-immunogenic and ready-to-use natural substitute with enhanced biomechanical properties. Nevertheless, these processes may alter microarchitecture and biocompatibility. Hence, this is the first study to characterize ex-vivo the biogenic conduits of 1-and 2-months maturation time which were subjected to decellularization and genipin (GP) cross-linking procedures performing histological, structural, biomechanical, biocompatibility, and immunological analyses to identify the most suitable option for peripheral nerve regeneration. Results: Histological examination indicated consistent uniformity of the biogenic conduits at both timepoints post-implantation, maintaining their overall structural integrity and collagen pattern following decellularization and GP crosslinking treatments. Furthermore, no evidence of nuclear debris was observed in the decellularized groups at either stage of maturation, confirming the decellularization protocol’s efficiency. The substitutes with longer maturation time presented a generally higher preservation of ECM key components. In addition, the GP crosslinking significantly increased the resistance values of decellularized biogenic conduits, without drastically affecting the ex-vivo cell biocompatibility nor macrophage polarization rate phenotype. Conclusions: These findings indicate the suitability of our decellularization protocol for biogenic conduits, and subsequent crosslinking with GP improves their biomechanical properties without altering their biocompatibility or immunological profile, suggesting their potential as a ready-to-use tubular substitute for nerve and other tissue engineering applications. 2025-10-22T06:42:43Z 2025-10-22T06:42:43Z 2025-04-21 journal article O.D. García-García et al. Biomedicine & Pharmacotherapy 187 (2025) 118064. https://doi.org/10.1016/j.biopha.2025.118064 https://hdl.handle.net/10481/107266 10.1016/j.biopha.2025.118064 eng http://creativecommons.org/licenses/by-nc-nd/4.0/ open access Attribution-NonCommercial-NoDerivatives 4.0 Internacional Elsevier