3D-Printed Perfusable Lab-on-a-Chip-Based Engineering of MSC-Derived Exosome-Enriched Vascularized Grafts for Skin Regeneration

Perfusable and functional vascularization is a critical yet unresolved challenge in regenerative medicine, particularly for large tissue constructs where passive diffusion of oxygen and nutrients is insufficient to sustain cellular viability. Without stable microvascular networks, engineered tissues often fail to survive or integrate following implantation, making vascularization a key determinant of clinical success. Conventional strategiesincluding growth factor supplementation, gene therapy, and endothelial coculturehave demonstrated limited efficacy due to inadequate vasculogenic signaling and compromised cell viability in the engineered niche. In this study, we report the development of an exosome-enriched biocomposite bioink composed of chitosan, collagen, and fibrinogen, incorporating vitamin D3 and periodontal ligament stem cell-derived exosomes (CCF-D3-exo). This formulation was used to fabricate perfusable three-dimensional (3D) vascular channel scaffolds via extrusion-based bioprinting, followed by the coculture of human adult dermal fibroblasts (HADF) and dental pulp stem cells (DPSC) within a dynamic lab-on-a-chip system. CCF-D3-exo scaffolds demonstrated significantly (<i>p</i> < 0.05) enhanced endothelial transdifferentiation and vasculogenic activity compared to exosome-free controls, with transcriptomic analysis showing upregulation of key angiogenic and vasculogenic markers, followed by proteomic validation via immunofluorescence, revealing robust expression and organized localization of α-SMA, vimentin, VEGF-A, VEGF-R1 CD34, and F-actin at different time points, indicative of early vascular morphogenesis and lumen formation. Together, these results establish a functional, perfusable, and biomimetic 3D vasculogenic niche, offering a promising strategy for fabricating vascularized skin grafts and advancing the translational potential of regenerative tissue constructs.