In human dystrophies, progressive muscle wasting is exacerbated by ectopic deposition of fat and fibrous tissue originating from fibro/adipogenic progenitors (FAPs). In degenerating muscles, the ability of these cells to promote successful healing is attenuated, and FAPs aberrantly expand and differentiate into adipocytes and fibroblasts. Thus, arresting the fibro/adipogenic fate of FAPs, without affecting their physiological role, represents a valuable therapeutic strategy for patients affected by muscle diseases. Here, using a panel of adipose progenitor cells, including human-derived FAPs, coupled with pharmacological perturbations and proteome profiling, we report that LY2090314 interferes with a genuine adipogenic program acting as WNT surrogate for the stabilization of a competent β-catenin transcriptional complex. To predict the beneficial impact of LY2090314 in limiting ectopic deposition of fat in human muscles, we combined a poly-ethylene-glycol-fibrinogen biomimetic matrix with these progenitor cells to create a miniaturized 3D model of adipogenesis. Using this scalable system, we demonstrated that a two-digit nanomolar dose of this compound effectively represses adipogenesis at higher 3D scale, thus indicating the potential for LY2090314 to limit FAP-derived fat infiltrates in dystrophic muscles.

Background/Objectives: Skin regeneration is a rapidly advancing field with significant implications for regenerative medicine, particularly in treating wounds and burns. This study explores the potential of hydrogels functionalized with fibroblast-derived extracellular vesicles (EVs) to enhance skin regeneration in vivo. Being immunoprivileged, EVs minimize immune rejection, offering an attractive alternative to whole-cell therapies by replicating fibroblasts’ key roles in tissue repair. Methods: To promote EVs’ versatility and effective application across different conditions, a lyophilization method with lyoprotectants was optimized. Then, EVs were used to functionalize a hydrogel to perform experiments on murine cutaneous wound models. Results: Gelatin methacrylate (GelMA) was selected as the polymeric hydrogel due to its biocompatibility, tunable mechanical properties, and ability to support wound healing. Mechanical tests confirmed GelMA’s strength and elasticity for this application. Fibroblast-derived EVs were characterized using Western blot, Transmission Electron Microscopy, and NanoSight analysis, proving their integrity, size distribution, and stability. miRNome profiling identified enriched biological pathways related to cell migration, differentiation, and angiogenesis, emphasizing the critical role of EV cargo in promoting wound repair. In a murine model, hydrogels loaded with fibroblast-derived EVs significantly accelerated wound healing compared to controls (mean wound area 0.62 mm² and 4.4 mm², respectively), with faster closure, enhanced epithelialization, increased vascularization, and reduced fibrosis. Notably, the lyoprotectants successfully preserved the EVs’ structure and bioactivity during freeze-drying, reducing EVs loss by 35% compared to the control group and underscoring the feasibility of this approach for long-term storage and clinical application. Conclusions: This study introduces a novel scalable and adaptable strategy for regenerative medicine by combining fibroblast-derived EVs with GelMA, optimizing EVs’ stability and functionality for enhanced wound healing in clinical settings, even in challenging contexts such as combat zones or large-scale natural disasters.

The development of advanced in vitro systems to replicate vascularized tissue environments is critical for studying cancer progression, immune interactions, and therapeutic responses. Traditional models often lack physiological perfusion, scale flexibility, and compatibility with complex microenvironments, limiting their translational impact. The small Vessel Environment Bioreactor (sVEB) represents a promising advancement in microfluidic and organ-on-chip technologies, enabling the replication of dynamic environments that mimic key features of vascular and tumor biology. Validated through in vitro experiments and computational flow simulations, the sVEB supports vascular network formation, dynamic cell cultures, and tumor-immune interactions. iPSC-derived endothelial cells in the sVEB formed stable perfusable microvessels with secondary branching into the surrounding matrix, while fluidic simulations confirmed laminar flow and shear stress conditions compatible with physiological parameters. In parallel, breast cancer organoids were assembled within the hydrogel compartment surrounding the sVEB and cultured under dynamic flow conditions. Moreover, CD8⁺ T lymphocytes were delivered using a magnetic nanoparticle-based approach, enabling immune-tumor contact within the model. Advancing this technology will require continued efforts on biomaterial development, integration of patient-derived cells, and standardized protocols to ensure scalability and reproducibility, ultimately establishing the sVEB as a versatile platform for precision medicine capable of modeling patient-specific microenvironments to support the discovery of innovative therapeutic approaches.