Bioactivity and multi-omics profiling of purslane-derived nanovesicles with therapeutic implications in diabetic wounds

• Compare fresh vs dried Portulaca oleracea L. (Purslane) and different extraction methods (ultracentrifugation and PEG precipitation) for vesicle isolation. • Vesicles isolated from fresh purslane via ultracentrifugation (F-Po-UC) exhibited superior antioxidative, anti-inflammatory, and pro-proliferative properties. • Proteomic and metabolomic analyses revealed that F-Po-UC vesicles are enriched in bioactive components linked to tissue regeneration pathways. • F-Po-UC-loaded hydrogel dressings significantly accelerated diabetic wound healing in vivo, supporting their potential as a plant-based regenerative therapy. As an emerging natural nanomedicine, traditional Chinese medicine-derived exosome-like nanovesicles have shown unique potential in the treatment of chronic non-healing wounds due to their advantages such as good biocompatibility, high safety and large-scale production. However, at present, there are relatively few studies on different states and extraction processes of traditional Chinese medicine, which restricts its clinical transformation process. This study aims to systematically compare the properties and biological activities of Portulaca oleracea L. (Purslane) derived vesicles (Po-DENs) extracted from fresh and dried materials by using ultracentrifugation and polyethylene glycol extraction methods. The key functional metabolites and proteins were identified through multi-omics analysis, and their therapeutic potential in diabetic wound healing was verified both in vivo and in vitro. Fresh and dried Portulaca oleracea L . samples were treated by ultracentrifugation and polyethylene glycol precipitation methods, and four kinds of plant nanovesicles were separated. Their physical properties such as particle size were characterized. The composition of the vesicles was determined through metabolomics and proteomics. The proliferation, antioxidant, anti-apoptotic and anti-inflammatory effects of four types of vesicles on skin keratinocytes and fibroblasts were investigated, and the mechanism was analyzed through proteomics. And it was finally verified in the wound healing model of diabetic mice to confirm its therapeutic activity and biological safety. The results demonstrated that fresh Purslane-derived nanovesicles isolated by ultracentrifugation (F-Po-UC) exhibited superior bioactivity compared to other extraction methods and dried material preparations. Omics analyses further revealed that F-Po-UC is enriched with a wide range of metabolites and proteins, which act through signaling pathways related to tissue regeneration. Subsequent vivo studies demonstrated that dressings based on Purslane-derived vesicles significantly accelerated wound healing in a diabetic mouse model, confirming their potential as an innovative therapeutic strategy. Fresh Purslane-derived nanovesicles demonstrate exceptional therapeutic potential for diabetic wound healing through their potent proliferative, antioxidant, and anti-inflammatory effects, supported by multi-omics characterization and robust in vivo validation.