Characterization of anti-proliferative and anti-oxidant effects of nano-sized vesicles from Brassica oleracea L. (Broccoli)

Abstract

In this in vitro study, we test our hypothesis that Broccoli-derived vesicles (BDVs), combining the anti-oxidant properties of their components and the advantages of their structure, can influence the metabolic activity of different cancer cell lines. BDVs were isolated from homogenized fresh broccoli (Brassica oleracea L.) using a sucrose gradient ultracentrifugation method and were characterized in terms of physical properties, such as particle size, morphology, and surface charge by transmission electron microscopy (TEM) and laser doppler electrophoresis (LDE). Glucosinolates content was assessed by RPLC–ESI–MS analysis. Three different human cancer cell lines (colorectal adenocarcinoma Caco-2, lung adenocarcinoma NCI-H441 and neuroblastoma SHSY5Y) were evaluated for metabolic activity by the MTT assay, uptake by fluorescence and confocal microscopy, and anti-oxidant activity by a fluorimetric assay detecting intracellular reactive oxygen species (ROS). Three bands were obtained with average size measured by TEM based size distribution analysis of 52 nm (Band 1), 70 nm (Band 2), and 82 nm (Band 3). Glucobrassicin, glucoraphanin and neoglucobrassicin were found mostly concentrated in Band 1. BDVs affected the metabolic activity of different cancer cell lines in a dose dependent manner compared with untreated cells. Overall, Band 2 and 3 were more toxic than Band 1 irrespective of the cell lines. BDVs were taken up by cells in a dose- and time-dependent manner. Pre-incubation of cells with BDVs resulted in a significant decrease in ROS production in Caco-2 and NCI-H441 stimulated with hydrogen peroxide and SHSY5Y treated with 6-hydroxydopamine, with all three Bands. Our findings open to the possibility to find a novel “green” approach for cancer treatment, focused on using vesicles from broccoli, although a more in-depth characterization of bioactive molecules is warranted.

Current Knowledge on the Preparation and Benefits of Cruciferous Vegetables as Relates to In Vitro, In Vivo, and Clinical Models of Inflammatory Bowel Disease

“…Broccoli has been widely studied for its anti-inflammatory and antioxidant properties against in vitro IBD models ( Table 1 [ [110] , [111] , [112] , [113] , [116] , [117] , [118] , [119] , [120] , [121] , [122] , [123] , [124] , [125] ] and Figure 1 ). One study showed that broccoli-derived nanoparticles (BDN), made of broccoli juice and lipids, prevented dendritic cell (DC) activation by upregulating AMPK anti-inflammatory pathways, thereby reducing chemoattractant molecules like chemokine (C-C) receptors (CCR), chemokine (CXC) ligand (CXCL), and CCLs in the DCs and monocytes [ 116 ].…”

“…One study showed that broccoli-derived nanoparticles (BDN), made of broccoli juice and lipids, prevented dendritic cell (DC) activation by upregulating AMPK anti-inflammatory pathways, thereby reducing chemoattractant molecules like chemokine (C-C) receptors (CCR), chemokine (CXC) ligand (CXCL), and CCLs in the DCs and monocytes [ 116 ]. Similarly, broccoli-derived vesicles rich in GLR, glucobrassicin, and neoglucobrassicin decreased oxidative stress [ 117 ] in Caco-2 cells (colon cells derived from a 72-y-old man with colorectal adenocarcinoma). The SFN treatment transformed the white blood cell monocytes (M1) to anti-inflammatory macrophages (M2) [ 118 , 119 ] through IL10 and signal-transducer and activator of transcription-3 pathways to ameliorate inflammation in bone marrow-derived macrophages [ 118 ], and decreased COX-2 and iNOS activities and increased IL10 and CD36 against chronic inflammation in human cells [ 119 , 126 ].…”

“… Cruciferous vegetables Image Bioactive compounds Preparation Dose Treatmentperiod Pathways Benefits Model References purified SFN — SFN purified SFN 10,1 and 0.1 μmol/L N.R. IL10 activation of STAT3 Anti-inflammatory Bone marrow-derived macrophages [ 118 ] Broccoli SFN, lipids Edible nanoparticles N.R., mean diameter of 32.4 nm 2 and 3 h CCRs, CXCLs, and CCLs Anti-inflammatory DCs and monocytes [ 116 ] Glucoraphanin, Glucobrassicin, Neoglucobrassicin Nano-vesicles 5 and 22 μg/mL 24 h Hydrogen peroxide Antioxidative Caco-2 and NCI-H441 cells [ 117 ] SFN N.R. 100, 50, 20,10 and 1 μmol 24 h COX-2 and iNOS, IL10 and CD36 Anti-inflammatory Immortalized human blood monocytes [ 119 ] Glucoraphanin, Glucoerucin, Quercetin-3-glucoside Cold-pressed seed 0.1% (vol:vol) 0, 2, 4, 5, and 6 h time point Bacteroidota/Bacillota ratio; ORAC, HOSC, and ABTS scavenging activities; COX-2 and IL1β cytokines Antioxidant, Prebiotic, Anti-inflammatory J774A.1 macrophages [ 110 ] Green cabbage and kimchi Polyphenols Freeze-dried 0.2, 0.5, 1 and 2 mg/mL 24 h MCP-1, IL1β and TNFɑ, E. coli, S. aureus Probiotic (kimchi), Anti-inflammatory ...…”

Solanum lycopersicum (Tomato)-Derived Nanovesicles Accelerate Wound Healing by Eliciting the Migration of Keratinocytes and Fibroblasts

“…The presence of nanoparticulate structures in the samples, their morphology, integrity, and size were analyzed by transmission electron microscopy (TEM), while vesicles Zeta Potential was measured by laser Doppler electrophoresis (LDE) using a Zeta sizer Nano ZS (Malvern Panalytical Ltd., Malvern, UK) and in diffusion barrier mode with water [ 63 ]. The TEM analysis was done by a method previously described [ 63 ].…”

Anti-oxidative effects of broccoli (Brassica oleracea var. italica) sprout extract in RAW 264.7 cell and cisplatin-induced testicular damage

“…4, December 2023 190 (Obrenovich et al, 2011). Members of the Brassica oleracea family include cabbage, broccoli, cauliflower, kale, Brussels sprouts, collard greens, Savoy cabbage, kohlrabi, and gai lan, which are antioxidant-rich foods and are natural sources of antioxidants (Al Talebi et al, 2023;Hossain et al, 2022). Broccoli (Brassica oleracea var.…”