Protein Biocargo and Anti-Inflammatory Effect of Tomato Fruit-Derived Nanovesicles Separated by Density Gradient Ultracentrifugation and Loaded with Curcumin
Ramila Mammadova,
Serena Maggio,
Immacolata Fiume,
Ramesh Bokka,
Maneea Moubarak,
Gabriella Gellén,
Gitta Schlosser,
Giorgia Adamo,
Antonella Bongiovanni,
Francesco Trepiccione,
Michele Guescini,
Gabriella Pocsfalvi
Affiliations
Ramila Mammadova
Extracellular Vesicles and Mass Spectrometry Laboratory, Institute of Biosciences and BioResources, National Research Council of Italy, 80131 Naples, Italy
Serena Maggio
Department of Biomolecular Sciences, University of Urbino Carlo Bo, 61029 Urbino, Italy
Immacolata Fiume
Extracellular Vesicles and Mass Spectrometry Laboratory, Institute of Biosciences and BioResources, National Research Council of Italy, 80131 Naples, Italy
Ramesh Bokka
Extracellular Vesicles and Mass Spectrometry Laboratory, Institute of Biosciences and BioResources, National Research Council of Italy, 80131 Naples, Italy
Maneea Moubarak
Extracellular Vesicles and Mass Spectrometry Laboratory, Institute of Biosciences and BioResources, National Research Council of Italy, 80131 Naples, Italy
Gabriella Gellén
MTA-ELTE Lendület Ion Mobility Mass Spectrometry Research Group, Institute of Chemistry, ELTE Eötvös Loránd University, H-1117 Budapest, Hungary
Gitta Schlosser
MTA-ELTE Lendület Ion Mobility Mass Spectrometry Research Group, Institute of Chemistry, ELTE Eötvös Loránd University, H-1117 Budapest, Hungary
Giorgia Adamo
Institute for Biomedical Research and Innovation, National Research Council of Italy, 90146 Palermo, Italy
Antonella Bongiovanni
Institute for Biomedical Research and Innovation, National Research Council of Italy, 90146 Palermo, Italy
Francesco Trepiccione
Department of Translational Medical Sciences, University of Campania Luigi Vanvitelli, 80138 Naples, Italy
Michele Guescini
Department of Biomolecular Sciences, University of Urbino Carlo Bo, 61029 Urbino, Italy
Gabriella Pocsfalvi
Extracellular Vesicles and Mass Spectrometry Laboratory, Institute of Biosciences and BioResources, National Research Council of Italy, 80131 Naples, Italy
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.
