Protein Biocargo and Anti-Inflammatory Effect of Tomato Fruit-Derived Nanovesicles Separated by Density Gradient Ultracentrifugation and Loaded with Curcumin.
Ramila MammadovaSerena MaggioImmacolata FiumeRamesh BokkaManeea MoubarakGabriella GellenGitta SchlosserGiorgia AdamoAntonella BongiovanniFrancesco TrepiccioneMichele GuesciniGabriella PocsfalviPublished in: Pharmaceutics (2023)
Plant-derived nanovesicles (PDNVs) have become attractive alternatives to mammalian cell-derived extracellular vesicles (EVs) both as therapeutic approaches and drug-delivery vehicles. In this study, we isolated tomato fruit-derived NVs and separated them by the iodixanol density gradient ultracentrifugation (DGUC) into twelve fractions. Three visible bands were observed at densities 1.064 ± 0.007 g/mL, 1.103 ± 0.006 g/mL and 1.122 ± 0.012 g/mL. Crude tomato PDNVs and DGUC fractions were characterized by particle size-distribution, concentration, lipid and protein contents as well as protein composition using mass spectrometry-based proteomics. Cytotoxicity and anti-inflammatory activity of the DGUC fractions associated to these bands were assessed in the lipopolysaccharide (LPS)-stimulated human monocytic THP-1 cell culture. The middle and the low-density visible DGUC fractions of tomato PDNVs showed a significant reduction in LPS-induced inflammatory IL-1β cytokine mRNA production. Functional analysis of proteins identified in these fractions reveals the presence of 14-3-3 proteins, endoplasmic reticulum luminal binding proteins and GTP binding proteins associated to gene ontology (GO) term GO:0050794 and the regulation of several cellular processes including inflammation. The most abundant middle-density DGUC fraction was loaded with curcumin using direct loading, sonication and extrusion methods and anti-inflammatory activity was compared. The highest entrapment efficiency and drug loading capacity was obtained by direct loading. Curcumin loaded by sonication increased the basal anti-inflammatory activity of tomato PDNVs.
Keyphrases
- drug delivery
- lps induced
- inflammatory response
- mass spectrometry
- anti inflammatory
- cancer therapy
- endoplasmic reticulum
- oxidative stress
- binding protein
- protein protein
- endothelial cells
- amino acid
- wound healing
- toll like receptor
- drug release
- genome wide
- liquid chromatography
- high resolution
- gene expression
- emergency department
- dna methylation
- small molecule
- high performance liquid chromatography
- fatty acid
- induced pluripotent stem cells
- cell wall