Novel exosome-like vesicles from Dendrobium officinale: Unraveling a pioneering extraction protocol and their skin anti-aging potentials

Numerous studies suggest that exosome-like nanovesicles (EVs) derived from medicinal plants are crucial for their therapeutic and cosmetic benefits. Nevertheless, the precise role of EVs derived from Dendrobium officinale Kimura et Migo ( D. officinale ), a treasured traditional Chinese medicine, in promoting skin health has not been extensively studied. Thus, the objective of this study was to establish an effective isolation method to isolate these EVs and to investigate their potential anti-aging benefits for skin. We developed a sequential filtration-based isolation process to extract D . officinale EVs (DO-EVs). We compared their physical properties and phytochemical profiles with those of EVs obtained via the standard ultracentrifugation method (UC-EVs) using Nanoparticle Tracking Analysis (NTA), Transmission Electron Microscopy (TEM), and LC-MS/MS. At the cellular level, we assessed the anti-aging efficacy of DO-EVs against UC-EVs and ethanol-extracted polysaccharides from D. officinale (EE-PS) by analyzing pro-COL1A2 levels, cytoskeletal organization, and senescence markers. We further examined DO-EVs’ anti-photodamage effects on UV-treated 3D skin models through histological and immunological staining, and elasticity measurements. Finally, we observed DO-EVs’ impact on human skin using a two-photon microscope. Both isolation methods yielded EVs of similar size and structure but distinct metabonomic profiles. DO-EVs uniquely contained compounds vital for skin health. Compared to UC-EVs and EE-PS, DO-EVs more effectively enhanced collagen I production, restored cytoskeletal structures, and reduced senescence markers in UV-exposed fibroblasts. In 3D skin models, DO-EVs significantly improved epidermal thickness and skin elasticity post-UV exposure, upregulated hyaluronic acid, collagen I & IV, integrin α6β4, plectin, laminin 5, and nidogen expression, and decreased DNA damage. Human trials confirmed DO-EVs’ effectiveness in enhancing skin structures within two weeks. Our isolation method successfully extracted D. officinale EVs with superior anti-aging bioactivity compared to UC-EVs or ethanol-extracted polysaccharides. These findings suggest DO-EVs’ great potential as an anti-aging cosmetic ingredient.