&lt;em&gt;In Vivo&lt;/em&gt; Imaging of Reactive Oxygen Species in a Murine Wound Model

Rabbani, Piul S.; Abdou, Salma A.; Sultan, Darren; Kwong, Jennifer Q.; Duckworth, April; Ceradini, Daniel J.

doi:10.3791/58450

Cited by 9 publications

(5 citation statements)

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“…Alternative dyes such as probes specific to detect mitochondria-generated superoxide anions could be used 10 . A repertoire of chemiluminescent probes was also developed to detect specific ROS species with high sensitivity, such as luciferin-based probes 15,16 . These have the advantage of being compatible with in vivo imaging but can't be used to map ROS production with specific cell types.…”

Section: Discussionmentioning

confidence: 99%

Analyzing Oxidative Stress in Murine Intestinal Organoids using Reactive Oxygen Species-Sensitive Fluorogenic Probe

Stedman

Lévy

Sansonetti

et al. 2021

JoVE

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Section: Discussionmentioning

confidence: 99%

Analyzing Oxidative Stress in Murine Intestinal Organoids using Reactive Oxygen Species-Sensitive Fluorogenic Probe

Stedman

Lévy

Sansonetti

et al. 2021

JoVE

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“…To measure in vivo ROS response from a pressure wound, a ROS-sensing probe, L-012 luminol (10 mg/mL, Labchem Wako) stock solution, was prepared in normal saline and intraperitoneally injected with 10 mL/kg of stock solution to achieve a final dosage of 100 mg/kg ( 56 ). After injection, animals were imaged using the Ami HTX (Spectral Instruments Imaging, Tucson, AZ, USA).…”

Section: Methodsmentioning

confidence: 99%

L-arginine supplementation abrogates hypoxia-induced virulence of Staphylococcus aureus in a murine diabetic pressure wound model

Baker,

Seo,

Park

et al. 2024

mSphere

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Diabetic foot ulcers (DFUs) are the most common complications of diabetes resulting from hyperglycemia leading to ischemic hypoxic tissue and nerve damage. Staphylococcus aureus is the most frequently isolated bacteria from DFUs and causes severe necrotic infections leading to amputations with a poor 5-year survival rate. However, very little is known about the mechanisms by which S. aureus dominantly colonizes and causes severe disease in DFUs. Herein, we utilized a pressure wound model in diabetic TALLYHO/JngJ mice to reproduce ischemic hypoxic tissue damage seen in DFUs and demonstrated that anaerobic fermentative growth of S. aureus significantly increased the virulence and the severity of disease by activating two-component regulatory systems leading to expression of virulence factors. Our in vitro studies showed that supplementation of nitrate as a terminal electron acceptor promotes anaerobic respiration and suppresses the expression of S. aureus virulence factors through inactivation of two-component regulatory systems, suggesting potential therapeutic benefits by promoting anaerobic nitrate respiration. Our in vivo studies revealed that dietary supplementation of L-arginine (L-Arg) significantly attenuated the severity of disease caused by S. aureus in the pressure wound model by providing nitrate. Collectively, these findings highlight the importance of anaerobic fermentative growth in S. aureus pathogenesis and the potential of dietary L-Arg supplementation as a therapeutic to prevent severe S. aureus infection in DFUs. IMPORTANCE S. aureus is the most common cause of infection in DFUs, often resulting in lower-extremity amputation with a distressingly poor 5-year survival rate. Treatment for S. aureus infections has largely remained unchanged for decades and involves tissue debridement with antibiotic therapy. With high levels of conservative treatment failure, recurrence of ulcers, and antibiotic resistance, a new approach is necessary to prevent lower-extremity amputations. Nutritional aspects of DFU treatment have largely been overlooked as there has been contradictory clinical trial evidence, but very few in vitro and in vivo modelings of nutritional treatment studies have been performed. Here we demonstrate that dietary supplementation of L-Arg in a diabetic mouse model significantly reduced duration and severity of disease caused by S. aureus . These findings suggest that L-Arg supplementation could be useful as a potential preventive measure against severe S. aureus infections in DFUs.

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“…For select studies, L012 (25 mg/kg) was administered intraperitoneally 5 minutes prior to euthanasia. Extracted lungs were imaged immediately using the IVIS Spectrum running Living Image 4.5.5 software (Perkin Elmer, MA) 9,[17][18][19] . Platelet counts were analysed by flow cytometry and calculated relative to time 0, prior to IgG and heparin injection.…”

Section: Microfluidics Studiesmentioning

confidence: 99%

Inhibition of NADPH oxidase blocks NETosis and reduces thrombosis in heparin-induced thrombocytopenia

et al. 2021

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Heparin-induced thrombocytopenia (HIT) is associated with severe and potentially lethal thrombotic complications. NETosis was recently shown to be an important driver of thrombosis in HIT. We investigated the role of reactive oxygen species (ROS) and NADPH oxidase 2 (NOX2) and their contributions to thrombus development in HIT. We showed that neutrophil activation by HIT immune complexes induced ROS-dependent NETosis. Analysis of thrombi formed in a microfluidics system showed ROS production in both platelets and neutrophils, and abundant NETs and ROS distributed throughout the clot. Neutrophil-targeted ROS inhibition was sufficient to block HIT-induced NETosis and thrombosis using human blood. Inhibition of NOX2 with diphenyleneiodonium chloride or GSK2795039 abrogated HIT-induced thrombi in vivo using FcγRIIa+/hPF4+ transgenic mice. Thrombocytopenia in mice remained unaffected by ROS inhibition. Increased ROS production in activated neutrophils were also confirmed using fresh blood from patients with active HIT. Our findings show that ROS and NOX2 play a crucial role in NETosis and thrombosis in HIT. This enhances our understanding of the processes driving thrombosis in HIT and identifies NOX2 as a potential new therapeutic target for antithrombotic treatment for HIT.

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<em>In Vivo</em> Imaging of Reactive Oxygen Species in a Murine Wound Model

Cited by 9 publications

References 0 publications

Analyzing Oxidative Stress in Murine Intestinal Organoids using Reactive Oxygen Species-Sensitive Fluorogenic Probe

Analyzing Oxidative Stress in Murine Intestinal Organoids using Reactive Oxygen Species-Sensitive Fluorogenic Probe

L-arginine supplementation abrogates hypoxia-induced virulence of Staphylococcus aureus in a murine diabetic pressure wound model

Inhibition of NADPH oxidase blocks NETosis and reduces thrombosis in heparin-induced thrombocytopenia

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