Endothelial Barrier Dysfunction Induced by Zinc Oxide Nanoparticles In Vivo and In Vitro and Their Mechanism of Crossing the Endothelial Barrier

Paxillin, a multi-domain scaffold-adapter focal adhesion (FA) protein, plays an important role in facilitating protein networking and efficient signaling transduction. Paxillin is phosphorylated at multiple serine/threonine and tyrosine residues; however, the role of tyrosine phosphorylation of paxillin in endothelial barrier dysfunction and the acute respiratory distress syndrome (ARDS) remains unclear. In this study, we used paxillin-specific siRNA and site-specific non-phosphorylatable mutants of paxillin to abrogate the function of paxillin, both in vitro and in vivo, to determine its role in the regulation of lung endothelial permeability and ARDS. In vitro, lipopolysaccharide (LPS) challenge of human lung microvascular endothelial cells (ECs) resulted in paxillin accumulation at focal adhesions, enhanced tyrosine phosphorylation of paxillin at Y31 and Y118, and significant endothelial barrier dysfunction. However no significant changes in Y181 phosphorylation by LPS challenge was observed. Paxillin silencing (siRNA) attenuated LPS-induced endothelial barrier dysfunction and dissociation of VE-cadherin from adherens junctions. LPS-induced paxillin phosphorylation at Y31 and Y118 was mediated by c-Abl tyrosine kinase and not by Src or focal adhesion kinase (FAK) in human lung microvascular ECs. Furthermore, down-regulation of c-Abl (siRNA) significantly reduced LPS-mediated endothelial barrier dysfunction. Transfection of human lung microvascular ECs with paxillin Y31, Y118 and Y31/Y118 mutants mitigated LPS-induced barrier dysfunction and VE-cadherin destabilization at adherens junctions. In vivo, knockdown of paxillin with siRNA in mouse lungs ameliorated LPS-induced pulmonary protein leak and lung inflammation. Together, these results suggest that c-Abl-mediated tyrosine phosphorylation of paxillin at Y31 and Y118 regulates LPS-mediated pulmonary vascular permeability and injury.

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Effects of Zinc Oxide Nanoparticles Synthesized Using Aspergillus niger on Carbapenem-Resistant Klebsiella pneumonia In Vitro and In Vivo

Frontiers in Cellular and Infection Microbiology ◽

10.3389/fcimb.2021.748739 ◽

2021 ◽

Vol 11 ◽

Author(s):

Elsayim Rasha ◽

Manal M. Alkhulaifi ◽

Monerah AlOthman ◽

Ibrahim Khalid ◽

Elnagar Doaa ◽

...

Keyword(s):

Zinc Oxide ◽

Aspergillus Niger ◽

Zinc Oxide Nanoparticles ◽

Diffusion Method ◽

Oxide Nanoparticles ◽

Zno Nps ◽

X Ray ◽

Carbapenem Resistant

Currently, the mortality rate in Saudi Arabia’s ICUs is increasing due to the spread of Klebsiella pneumoniae carbapenemase (KPC)-producing bacteria. This study was carried out to evaluate the ability of biologically synthesized zinc oxide nanoparticles (ZnO-NPs) using Aspergillus niger to overcome carbapenem-resistant K. pneumoniae (KPC) in vitro and in vivo. ZnO-NPs were synthesized via a biological method and characterized using UV–Vis spectroscopy, Zetasizer and zeta potential analyses, x-ray diffraction spectroscopy, Fourier transform infrared spectroscopy, scanning electron microscopy (SEM), and energy-dispersive x-ray spectroscopy (EDX). In vitro sensitivity of KPC to ZnO-NPs was identified using the well diffusion method. The minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC) were determined by a macro-dilution method. The morphological alteration of KPC cells after ZnO-NPs treatment was observed by SEM. The in vivo susceptibility of KPC cells to ZnO-NPs ointment was evaluated using wound healing in experimental rats. The chemical characterization findings showed the formation, stability, shape, and size of the synthesized nanoparticles. The MIC and MBC were 0.7 and 1.8 mg/ml, respectively. The in vivo results displayed reduced inflammation and wound re-epithelialization of KPC-infected rats. These findings demonstrated that ZnO-NPs have great potential to be developed as antibacterial agents.

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Atrial natriuretic peptide protects against Staphylococcus aureus-induced lung injury and endothelial barrier dysfunction

Journal of Applied Physiology ◽

10.1152/japplphysiol.00284.2010 ◽

2011 ◽

Vol 110 (1) ◽

pp. 213-224 ◽

Cited By ~ 24

Author(s):

Junjie Xing ◽

Nurgul Moldobaeva ◽

Anna A. Birukova

Keyword(s):

Staphylococcus Aureus ◽

Lung Inflammation ◽

Endothelial Barrier ◽

Protective Effects ◽

Barrier Dysfunction ◽

Gram Positive ◽

Gram Negative ◽

Endothelial Barrier Dysfunction

Lung inflammation and alterations in endothelial cell (EC) permeability are key events to development of acute lung injury (ALI). Protective effects of atrial natriuretic peptide (ANP) have been shown against inflammatory signaling and endothelial barrier dysfunction induced by gram-negative bacterial wall liposaccharide. We hypothesized that ANP may possess more general protective effects and attenuate lung inflammation and EC barrier dysfunction by suppressing inflammatory cascades and barrier-disruptive mechanisms shared by gram-negative and gram-positive pathogens. C57BL/6J wild-type or ANP knockout mice (Nppa−/−) were treated with gram-positive bacterial cell wall compounds, Staphylococcus aureus-derived peptidoglycan (PepG) and/or lipoteichoic acid (LTA) (intratracheal, 2.5 mg/kg each), with or without ANP (intravenous, 2 μg/kg). In vitro, human pulmonary EC barrier properties were assessed by morphological analysis of gap formation and measurements of transendothelial electrical resistance. LTA and PepG markedly increased pulmonary EC permeability and activated p38 and ERK1/2 MAP kinases, NF-κB, and Rho/Rho kinase signaling. EC barrier dysfunction was further elevated upon combined LTA and PepG treatment, but abolished by ANP pretreatment. In vivo, LTA and PepG-induced accumulation of protein and cells in the bronchoalveolar lavage fluid, tissue neutrophil infiltration, and increased Evans blue extravasation in the lungs was significantly attenuated by intravenous injection of ANP. Accumulation of bronchoalveolar lavage markers of LTA/PepG-induced lung inflammation and barrier dysfunction was further augmented in ANP−/− mice and attenuated by exogenous ANP injection. These results strongly suggest a protective role of ANP in the in vitro and in vivo models of ALI associated with gram-positive infection. Thus ANP may have important implications in therapeutic strategies aimed at the treatment of sepsis and ALI-induced gram-positive bacterial pathogens.

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Alteration of gene expression by zinc oxide nanoparticles or zinc sulfate in vivo and comparison with in vitro data: A harmonious case

Theriogenology ◽

10.1016/j.theriogenology.2016.03.006 ◽

2016 ◽

Vol 86 (3) ◽

pp. 850-861.e1 ◽

Cited By ~ 11

Author(s):

Wei-Dong Zhang ◽

Yong Zhao ◽

Hong-Fu Zhang ◽

Shu-Kun Wang ◽

Zhi-Hui Hao ◽

...

Keyword(s):

Gene Expression ◽

Zinc Oxide ◽

Zinc Oxide Nanoparticles ◽

Zinc Sulfate ◽

Oxide Nanoparticles ◽

Vitro Data ◽

In Vitro Data

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Zinc Oxide Nanoparticles Prime a Protective Immune Response in Galleria mellonella to Defend Against Candida albicans

Frontiers in Microbiology ◽

10.3389/fmicb.2021.766138 ◽

2021 ◽

Vol 12 ◽

Author(s):

Mei-nian Xu ◽

Li Li ◽

Wen Pan ◽

Huan-xin Zheng ◽

Meng-lei Wang ◽

...

Keyword(s):

Immune Response ◽

Zinc Oxide ◽

Candida Albicans ◽

Zinc Oxide Nanoparticles ◽

Galleria Mellonella ◽

Oxide Nanoparticles ◽

Protective Effects ◽

Zno Nps

Purpose: Zinc oxide nanoparticles (ZnO-NPs) have exerted antimicrobial properties. However, there is insufficient evaluation regarding the in vivo antifungal activity of ZnO-NPs. This study aimed to investigate the efficacy and mechanism of ZnO-NPs in controlling Candida albicans in the invertebrate Galleria mellonella.Methods:Galleria mellonella larvae were injected with different doses of ZnO-NPs to determine their in vivo toxicity. Non-toxic doses of ZnO-NPs were chosen for prophylactic injection in G. mellonella followed by C. albicans infection. Then the direct in vitro antifungal effect of ZnO-NPs against C. albicans was evaluated. In addition, the mode of action of ZnO-NPs was assessed in larvae through different assays: quantification of hemocyte density, morphology observation of hemocytes, characterization of hemocyte aggregation and phagocytosis, and measurement of hemolymph phenoloxidase (PO) activity.Results: Zinc oxide nanoparticles were non-toxic to the larvae at relatively low concentrations (≤20 mg/kg). ZnO-NP pretreatment significantly prolonged the survival of C. albicans-infected larvae and decreased the fungal dissemination and burden in the C. albicans-infected larvae. This observation was more related to the activation of host defense rather than their fungicidal capacities. Specifically, ZnO-NP treatment increased hemocyte density, promoted hemocyte aggregation, enhanced hemocyte phagocytosis, and activated PO activity in larvae.Conclusion: Prophylactic treatment with lower concentrations of ZnO-NPs protects G. mellonella from C. albicans infection. The innate immune response primed by ZnO-NPs may be part of the reason for the protective effects. This study provides new evidence of the capacity of ZnO-NPs in enhancing host immunity and predicts that ZnO-NPs will be attractive for further anti-infection applications.

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Using Biosynthesized Zinc Oxide Nanoparticles to Alleviate the Toxicity on Banana Parasitic-Nematode

10.21203/rs.3.rs-186764/v1 ◽

2021 ◽

Author(s):

Mostafa elansary ◽

Ragaa Hamouda ◽

Maha Elshamy

Keyword(s):

Zinc Oxide ◽

Zinc Oxide Nanoparticles ◽

Oxide Nanoparticles ◽

Root Knot Nematode ◽

Zno Nps ◽

Second Stage ◽

Highest Weight ◽

Direct Exposure

Abstract We appraised the use of zinc oxide nanoparticles, (ZnO-NPs) and zinc oxide bulk (ZnO-bulk) or zinc acetate, as a natural nematocide, alone or in combination with oxamyl in vitro and in vivo trials in order to improve systems for root-knot nematode (RKNs) control in banana plants. Especially, ZnO-NPs were biosynthesized from the alga, Ulva fasciata. In general, all applications of ZnO-NPs were more effective to control RKNs than ZnO-bulk as well oxamyl alone (chemical control). In in vitro conditions, ZnO-NPs with oxamyl showed 98.91% second stage juveniles2 (J2s) mortality of Meloidogyne incognita after 72 hrs, while 72.86% mortality was observed at the same NPs treatment without oxamyl at the same exposure time. The same treatment was the most effective in diminution of J2s community (82.77%) in soil and galls number (81.87%) in roots under in vivo conditions. In contrast, the highest weight and height of the shoot was observed in Zn-bulk treatment in combination with oxamyl as well oxamyl only (nematocides check). Scanning electron microscopy (SEM) reports displayed the distributions and accumulations of ZnO-NPs on the nematode (J2s) body under direct exposure, which might be the reason of NP-mediated toxicity and disruption for M. incognita.

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