1 research outputs found
Exosomes for Wound Healing: Purification Optimization and Identification of Bioactive Components
Human mesenchymal stem cell exosomes have been shown to promote cutaneous wound healing. Their bioactivity is most often attributed to their protein and nucleic acid components, while the function of exosomal lipids remains comparatively unexplored. This work specifically assesses the involvement of lipids and the transmembrane enzyme CD73 in the exosomesâ biological activity in stimulating the cutaneous wound healing process. Since exosome preparation processes are not harmonized yet, certain production and purification parameters are first systematically investigated, enabling the optimization of a standardized protocol delivering high exosome integrity, yield, and purity. An in situ enzymatic assay is introduced to specifically assess the vesicle functionality, and quantitative proteomics is employed to establish the cell culture conditions yielding a stable exosome protein profile. Using a combination of in vitro approaches, CD73 and constitutional lipids of HPVâ16 E6/E7 transformed human bone marrow stromal cellâderived exosomes are identified as key bioactive components promoting the exosomeâdriven acceleration of processes required for wound repair. A pilot wound healing study in mice indeed suggests a role of lipids in the exosomesâ biological activity. Strikingly, the extent of the bioactivity of these exosomal components is found to be dependent on the target cell type.ISSN:2198-384