Wild-Type and SOD1-G93A SH-SY5Y under oxidative stress: EVs characterization and topographical distribution of budding vesicles

Extracellular vesicles (EVs) are important mediators of intercellular communication in several physiopathological conditions. Oxidative stress alters EVs release and cargo composition depending on the cell type and stimulus. Recently, most of the EVs studies have focused on the characterization of their cargo, rather than on the morphological features (i.e., size distribution, shape, and localization on the cell surface). Due to their high heterogeneity, to fully characterize EVs both the functional and morphological characterization are required. Atomic force microscopy (AFM), introduced for cell morphological studies at the nanoscale, represents a promising method to characterize in detail EVs morphology, dynamics along the cell surface, and its variations reflecting the cell physiological status. In the present study, untreated or H2O2-treated wild-type and SOD1-G93A SH-SY5Y cells have been compared performing a transmission electron microscopy (TEM) and AFM morpho-quantitative analysis of budding and released vesicles. Intriguingly, our analysis revealed a differential EVs profiling, with an opposite behavior and implying different cell areas between WT and SOD1-G93A cells, on both physiological conditions and after H2O2 exposure. Our results empower the relationship between the morphological features and functional role, further proving the efficacy of EM/AFM in giving an overview of the cell physiology related to EVs trafficking

Therapeutic Effect of Polymeric Nanomicelles Formulation of LY2157299-Galunisertib on CCl4-Induced Liver Fibrosis in Rats

Hepatic fibrosis (HF) is a major cause of liver-related disorders and together with cancer-associated fibroblasts can favor liver cancer development by modulating the tumor microenvironment. Advanced HF, characterized by an excess of extracellular matrix (ECM), is mediated by TGF- β1, that activates hepatic stellate cells (HSCs) and fibroblasts. A TGF-β1 receptor inhibitor, LY2157299 or Galunisertib (GLY), has shown promising results against chronic liver progression in animal models, and we show that it can be further improved by enhancing GLYs bioavailability through encapsulation in polymeric polygalacturonic-polyacrylic acid nanomicelles (GLY-NMs). GLY-NMs reduced HF in an in vivo rat model of liver fibrosis induced by intraperitoneal injection of CCl4 as shown by the morphological, biochemical, and molecular biology parameters of normal and fibrotic livers. Moreover, GLY-NM was able to induce recovery from HF better than free GLY. Indeed, the encapsulated drug reduces collagen deposition, hepatic stellate cells (HSCs) activation, prevents fatty degeneration and restores the correct lobular architecture of the liver as well as normalizes the serum parameters and expression of the genes involved in the onset of HF. In summary, GLY-NM improved the pharmacological activity of the free TGF- β1 inhibitor in the in vivo HF treatment and thus is a candidate as a novel therapeutic strategy