Abstract
Angiogenesis is critical for effective wound healing and relies on the successful coordination of various cell types, including endothelial cells, macrophages, and fibroblasts. Adipose-derived stem cell extracellular vesicles (ADSC-EVs) have demonstrated proangiogenic properties and have been posited as a novel therapeutic to aid wound healing; however, their functional impact within human-derived multicellular models remains largely uncharacterized. This study explores the development and application of a 3D multicellular in vitro model to assess the effects of ADSC-EVs on vascularization in the context of wound healing. 3D multicellular in vitro models were developed by coculturing human umbilical vein endothelial cells (HUVECs), monocyte-derived macrophages, and fibroblasts within Matrigel to recapitulate the in vivo wound healing microenvironment. A five-color confocal microscopy panel was developed to visualize each cell type and EVs within the models. The optimized models were then treated with ADSC-EVs or control to determine their impact on angiogenesis and cell colocalization. We determined that vessel formation was significantly enhanced when HUVECs were cocultured in multicellular models compared with monocultures, with the greatest effect observed in the full three-cell-type model. This effect was even more pronounced with the addition of ADSC-EVs. ADSC-EV treatment also enhanced macrophage colocalization within endothelial structures. This study developed a multicellular model that can be used for future work assessing wound healing in vitro and will be additive to currently used single-cell and in vivo models. We have applied these models to demonstrate that ADSC-EVs significantly enhance tube formation in HUVECs and the development of tissue-like structures in multicell systems, highlighting their potential as a promising therapeutic approach for improving wound healing.
Impact Statement
This study presents the development of a 3D in vitro multicellular wound healing model, designed to serve as a platform for assessing angiogenesis and cellular interactions. Using this model, we demonstrate that extracellular vesicles derived from adipose-derived stem cells (ADSC-EVs) significantly enhance wound healing by promoting angiogenic activity. The model established in this study provides a valuable tool for future investigations into the mechanisms of wound healing and the evaluation of novel therapeutic strategies.
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