Human Monocyte Response to S-Nitrosoglutathione-Loaded Nanoparticles: Uptake, Viability, and Transcriptome

Abstract

S-Nitrosoglutathione (GSNO) is a good candidate for nitric oxide (NO(•)) delivery, and its nanoformulation improves NO(•) stability and bioavailability. We have compared the effect of empty Eudragit nanoparticles (eENP), GSNO-loaded ENP (gENP), and free GSNO on THP-1 human monocytic cell line. We investigated cellular viability and growth by WST-1 and trypan blue tests. ENP uptake was studied using transmission electron microscopy, confocal microscopy, and flow cytometry. Transcriptomic profiles were obtained using microarray. ENP entered cells by clathrin- and caveolae-mediated endocytosis. Exposure to either free GSNO or gENP induced an activation of genes from the same clusters, in favor of intracellular delivery of GSNO by ENP. GSNO nanoformulation might be a therapeutic option for NO(•) delivery.

Synthesis of S-nitrosoglutathione-alginate for prolonged delivery of nitric oxide in intestines

“…Therefore, based on this MTT assay, our polymer was not toxic at 10 mM NO and exhibits 100% metabolic activity in the cells after 24 h of incubation. Similar results were obtained in a study performed by Ronzani et al (2014) and Safar et al (2015), where empty nanoparticles consisting of Eudragit RL were seeded into the human THP-1 monocytic cell line, and a significant increase in the viability of the cells up to 40% was observed compared with the control. The cell viability increased due to an increase in the metabolic activity and growth of the cells after contact with the nanoparticles, and this phenomenon was related to an augmentation of the mitochondrial volume and activity.…”

“…The mitochondrial activity may be related to cell proliferation, which leads to an increase in the mitochondrion number. The upregulation of genes involved in metabolism and proliferation may be related to this increased mitochondrial activity, which belongs to ''transcription'', ''nucleus'', ''chromosomal part'', ''metabolic process'', and ''proliferation'' (Safar et al, 2015).…”

Drug Release and Targeting: the Versatility of Polymethacrylate Nanoparticles for Peroral Administration Revealed by Using an Optimized In Vitro-Toolbox

“…By assessing cytotoxicity of the TMP-001-loaded drug delivery system, an effect of reduced cell viability on further experiments was excluded. Consistent with some earlier findings concentrations in a range between 0.05 to 3 mg/mL of the TMP-001-loaded nanoparticles were tested on Caco-2 and HCA-7 cells (4,32,33).…”

“…The selected concentrations corresponded to previous observations focusing on the safety of polymethacrylic-based formulations in monocytes and cancer cell lines (4,32,33). Even with the highest concentration no significant reduction of cell viability was observed.…”

“…The medium was removed and 100 μL of the nanoparticle suspensions diluted in white DMEM were pipetted into each well. The applied concentrations ranged from 0.05 to 3 mg/mL loaded Eudragit® RS 100 nanoparticles (4,32,33). After 3 h of incubation the medium was removed and replaced with fresh white DMEM.…”