Chemical constituents of exosome-like nanoparticles-enriched vesicle fraction derived from Sparassis crispa and its biological and molecular docking studies.

Exosome-like nanovesicles (ELNs) are extracellular vesicles (50-200 nm) with a bilayer membrane derived from endosomes. They contain nucleic acids, proteins, lipids and secondary metabolites, and function as mediators of intercellular communication. ELNs isolated from plants and mushrooms have been reported to exhibit diverse biological activities. ELN-enriched vesicle fraction derived from Sparassis crispa was extracted using low-temperature pressure squeeze pre-treatment and tangential flow filtration system. The ethyl acetate portion was purified using multiple chromatography techniques to yield three compounds. Their structures were determined by spectroscopic analysis and comparison with reported data. The effects of the compounds on type I collagen gene (COL1A1) expression were evaluated in CCD-1064Sk human fibroblasts. Molecular docking was conducted to assess their interactions with collagenases (MMP-1, MMP-8 and MMP-13). Three compounds were isolated from ELN-enriched vesicle fraction derived from S. crispa and identified as sparalide C (1), 5-hydroxy-7-methoxyphthalide (2) and methyl orsellinate (3). All compounds increased COL1A1 gene expression in a dose-dependent manner. These compounds exhibited binding energies in the range of -5.8 to -7.0 kcal/mol against three collagenases. This study identified three bioactive compounds, sparalide C (1), 5-hydroxy-7-methoxyphthalide (2) and methyl orsellinate (3), from ELN-enriched vesicle fraction derived from S. crispa. All isolated compounds enhanced COL1A1 gene expression in CCD-1064Sk human fibroblasts, suggesting their potential to promote collagen biosynthesis. Molecular docking analysis further supported their relevance by indicating moderate binding affinities to collagenase enzymes (MMP-1, MMP-8 and MMP-13), which are known to degrade type I collagen. These findings suggest that ELN-enriched vesicle fraction derived from S. crispa may serve as a promising natural source for skin health applications, particularly in maintaining extracellular matrix homeostasis.