Investigation of Plant-derived Nanovesicles as Bioactive Agents for Chronic Wound Healing

Abstract

Introduction Chronic wounds can severely affect a patient’s quality of life due to their long healing process, increased likelihood of hospitalisation, and recurrence. They often lead to persistent pain, discomfort, and reduced mobility, which may contribute to social isolation and emotional distress, ultimately impacting overall well-being [1,2]. To address the complexities of chronic wound healing, we investigated the potential of plant-derived nanovesicles (PDNV) as a new therapeutic solution. The unique structure of PDNVs ensures higher bioavailability of bioactive compounds naturally found in plants, protecting them from harsh external factors. However, PDNVs are still relatively new scientific objects, and it is necessary to investigate their biological effects. Aim To investigate the potential of PDNV as bioactive agents for promoting skin wound healing. Methods Isolation of PDNVs by polymeric precipitation method; characterisation of PDNV using Bradford method, TRIzolTM reagent, Nanoparticle Tracking Analysis, Dynamic Light Scattering Analysis, Cryogenic electron microscopy; Secondary metabolites profile analysis by LC-MS using ACQUITY UPLC I-Class system (Waters, Milford, MA) coupled with a SELECT SERIES Cyclic IMS (Waters, Wilmslow, U.K.) and a Q-ToF mass spectrometer equipped with a cyclic ion mobility separation (cIM) device; controlled PDNVs structure destruction; PrestoblueTM Cell Viability Reagent; Fluorescent Microscopy; Wound Test in vitro. Statistical analysis was conducted using ANOVA followed by the LSD post hoc test. Results This study screened various plant sources to identify the most potent PNDV for wound healing, with cucumber-derived nanovesicles demonstrating superior bioactivity. Cucumber fruit and leaf PDNVs were internalised by human keratinocytes (HaCaT) and skin fibroblasts (BJ-5ta). Cucumber PDNVs induced HaCaT and BJ-5ta proliferation and migration. The effect of proliferation was eliminated after destroying the PDNV structure, highlighting the significance of maintaining an intact structure for bioactivity. Additionally, the analysis of secondary metabolites identified 56 compounds in the cucumber fruit and leaf samples of PDNV, some of which, including phenols, may contribute to positive effects on skin cell proliferation and migration. Conclusions In conclusion, cucumber PDNV hold great promise for improving chronic wound healing. Further research is necessary to explore the mechanisms of action to develop innovative therapeutic solutions for chronic wound care.