Hemp Seed-Derived Vesicle-like Nanoparticles Modulate Dermal Collagen Homeostasis and Attenuate UVB-Induced...

Plant-derived extracellular vesicle-like nanoparticles (EVNPs) have emerged as bioactive nanomaterials with potential applications in skin biology; however, their isolation from complex plant matrices and their integrated biological effects remain insufficiently characterized. In this study, hemp seed-derived vesicle-like nanoparticles (hs- extracellular VLNPs) were isolated using a multi-step workflow combining pre-clarification, sequential filtration, electrokinetic capture, and tangential flow filtration. The isolated hs-VLNPs exhibited membrane-enclosed vesicle-like morphology, a nanoscale size distribution predominantly within 100-200 nm, and a negative surface charge. hs-VLNPs maintained cellular viability within the concentration range used for most functional assays and exhibited multiple biological activities associated with skin cellular homeostasis. In human dermal fibroblasts, hs-VLNPs increased procollagen type I production and attenuated UVB-induced matrix metalloproteinase-1 (MMP-1) expression following UVB exposure. In HaCaT keratinocytes, hs-VLNPs enhanced scratch closure under non-cytotoxic treatment conditions. Inflammatory mediator responses were cell- and mediator-dependent, with differential effects observed in RAW264.7 macrophages and human dermal fibroblasts rather than uniform suppression of inflammatory mediators. In addition, hs-VLNPs reduced α-melanocyte-stimulating hormone (α-MSH)-induced melanin production while exhibiting little direct inhibition of tyrosinase activity, suggesting that the anti-melanogenic effect may involve cellular regulatory mechanisms rather than direct enzymatic inhibition. Collectively, these findings demonstrate that electrokinetic capture combined with membrane-based filtration enables the enrichment of biologically active vesicle-like nanoparticles from hemp seeds. Rather than exerting a single biological effect, hs-VLNPs modulated multiple interconnected processes involving extracellular matrix homeostasis, cellular migration, immune responses, and melanogenesis, providing a foundation for further investigation of hemp seed-derived EVNPs as plant-derived nanomaterials for skin-related applications.