Biogenic nanoparticles from liquid and solid matrices: biochemical and biophysical properties of Extracellular Vesicles-enriched samples from human plasma and skeletal muscle tissue.

Aim Studies on extracellular vesicles (EVs) focused on samples enriched from liquid matrices, such as cell culture media and blood. Recent research highlights the roles of EVs derived from the extracellular matrix of solid tissues, and how investigating these specific EVs offers insights into their microenvironment and potential biological influences on surrounding cells. This study presents a shared method to separate and compare EV enriched from solid (human skeletal muscle biopsy) and liquid (human plasma) matrices, addressing technical challenges and minimizing biases in separation techniques.

Plasma and skeletal muscle-EVs were obtained combining serial centrifugation steps and discontinuous sucrose density gradient. EVs characterization employed advanced analytical techniques such as Western Blot, Colorimetric Nanoplasmonic assay, Atomic Force Miscoscopy, Nanoparticle Tracking Analysis and Dynamic Light Scattering to focus on biomolecular composition, nanomechanical properties, particle yield, size distribution, and colloidal stability.

The analysis revealed distinct differences between skeletal muscle-EVs and plasma-EVs including: molecular composition, physical and nanomechanical properties, as well as particle size distribution. Skeletal muscle-EVs exhibited unique colloidal behavior compared to their plasma counterparts, suggesting tissue-specific features that may influence their biological activity and stability.

The findings demonstrate that EVs from skeletal muscle tissue possess unique biochemical and biophysical characteristics when compared to those derived from plasma. These differences reflect their diverse biological origins and microenvironments. Understanding these distinctions could advance the development of EV-based diagnostic tools, particularly for muscular disorders, and broaden our knowledge of EV roles across various tissue contexts. HIGHLIGHTS Shared protocol to enrich and compare extracellular vesicles (EVs) from both solid (skeletal muscle biopsy) and liquid (plasma) human samples, reducing methodological bias across matrices. Skeletal muscle–derived EVs differ significantly from plasma EVs in biochemical and biophysical properties such as molecular composition, size distribution, nanomechanical properties, and colloidal stability. EV heterogeneity across microenvironments emerged as a key biological feature, highlighting their potential for specific diagnostic applications. Competing Interest Statement The authors have declared no competing interest. Footnotes Title and abstract modified. Increased number of samples analyzed and new data described.