Human Fibroblasts Release Reactive Oxygen Species in Response to Treatment with Synovial Fluids from Patients Suffering from Arthritis

Adhesions frequently occur postoperatively, causing morbidity. In this noninterventional observational cohort study, we enrolled patients who presented for repeat abdominal surgery, after a history of previous abdominal myomectomy, from March 1998 to June 20210 at St. Vincent’s Catholic Medical Centers. The primary outcome of this pilot study was to compare adhesion rates, extent, and severity in patients who were treated with intraperitoneal triamcinolone acetonide during the initial abdominal myomectomy (n = 31) with those who did not receive any antiadhesion interventions (n = 21), as documented on retrospective chart review. Adhesions were blindly scored using a standard scoring system. About 32% of patients were found to have adhesions in the triamcinolone group compared to 71% in the untreated group (p < 0.01). Compared to controls, adhesions were significantly less in number (0.71 vs. 2.09, p < 0.005), severity (0.54 vs. 1.38, p < 0.004), and extent (0.45 vs. 1.28, p < 0.003). To understand the molecular mechanisms, human fibroblasts were incubated in hypoxic conditions and treated with triamcinolone or vehicle. In vitro studies showed that triamcinolone directly prevents the surge of reactive oxygen species triggered by 2% hypoxia and prevents the increase in TGF-β1 that leads to the irreversible conversion of fibroblasts to an adhesion phenotype. Triamcinolone prevents the increase in reactive oxygen species through alterations in mitochondrial function that are HIF-1α-independent. Controlling mitochondrial function may thus allow for adhesion-free surgery and reduced postoperative complications.

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Mechanisms of Mitochondrial Damage in Keratinocytes by Pemphigus Vulgaris Antibodies

Journal of Biological Chemistry ◽

10.1074/jbc.m113.472100 ◽

2013 ◽

Vol 288 (23) ◽

pp. 16916-16925 ◽

Cited By ~ 22

Author(s):

Mina Kalantari-Dehaghi ◽

Yumay Chen ◽

Wu Deng ◽

Alex Chernyavsky ◽

Steve Marchenko ◽

...

Keyword(s):

Reactive Oxygen Species ◽

Mouse Skin ◽

Reactive Oxygen ◽

Pemphigus Vulgaris ◽

Mitochondrial Damage ◽

Response To Treatment ◽

Transfer Model ◽

Clear Understanding ◽

Oxygen Species ◽

Mitochondrial Functions

The development of nonhormonal treatment of pemphigus vulgaris (PV) has been hampered by a lack of clear understanding of the mechanisms leading to keratinocyte (KC) detachment and death in pemphigus. In this study, we sought to identify changes in the vital mitochondrial functions in KCs treated with the sera from PV patients and healthy donors. PV sera significantly increased proton leakage from KCs, suggesting that PV IgGs increase production of reactive oxygen species. Indeed, measurement of intracellular reactive oxygen species production showed a drastic increase of cell staining in response to treatment by PV sera, which was confirmed by FACS analysis. Exposure of KCs to PV sera also caused dramatic changes in the mitochondrial membrane potential detected with the JC-1 dye. These changes can trigger the mitochondria-mediated intrinsic apoptosis. Although sera from different PV patients elicited unique patterns of mitochondrial damage, the mitochondria-protecting drugs nicotinamide (also called niacinamide), minocycline, and cyclosporine A exhibited a uniform protective effect. Their therapeutic activity was validated in the passive transfer model of PV in neonatal BALB/c mice. The highest efficacy of mitochondrial protection of the combination of these drugs found in mitochondrial assay was consistent with the ability of the same drug combination to abolish acantholysis in mouse skin. These findings provide a theoretical background for clinical reports of the efficacy of mitochondria-protecting drugs in PV patients. Pharmacological protection of mitochondria and/or compensation of an altered mitochondrial function may therefore become a novel approach to development of personalized nonhormonal therapies of patients with this potentially lethal autoimmune blistering disease.

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Electronic device generated light increases reactive oxygen species in human fibroblasts

Lasers in Surgery and Medicine ◽

10.1002/lsm.22794 ◽

2018 ◽

Vol 50 (6) ◽

pp. 689-695 ◽

Cited By ~ 5

Author(s):

Evan Austin ◽

Amy Huang ◽

Tony Adar ◽

Erica Wang ◽

Jared Jagdeo

Keyword(s):

Reactive Oxygen Species ◽

Electronic Device ◽

Human Fibroblasts ◽

Reactive Oxygen ◽

Oxygen Species

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4-Nitroquinoline 1-Oxide Forms 8-Hydroxydeoxyguanosine in Human Fibroblasts through Reactive Oxygen Species

Toxicological Sciences ◽

10.1093/toxsci/kfj161 ◽

2006 ◽

Vol 91 (2) ◽

pp. 382-392 ◽

Cited By ~ 71

Author(s):

Yaeno Arima ◽

Chikako Nishigori ◽

Toru Takeuchi ◽

Shigenori Oka ◽

Kanehisa Morimoto ◽

...

Keyword(s):

Reactive Oxygen Species ◽

Human Fibroblasts ◽

Reactive Oxygen ◽

Oxygen Species

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Stimulatory Auto-Antibodies to the PDGF Receptor in Patients with Chronic GVHD.

Blood ◽

10.1182/blood.v106.11.454.454 ◽

2005 ◽

Vol 106 (11) ◽

pp. 454-454

Author(s):

Attilio Olivieri ◽

Silvia Svegliati ◽

Nadia Campelli ◽

Michele Maria Luchetti ◽

Silvia Trappolini ◽

...

Keyword(s):

Gene Expression ◽

Reactive Oxygen Species ◽

Type I Collagen ◽

Human Fibroblasts ◽

Reactive Oxygen ◽

Type I ◽

Collagen Gene ◽

Pdgf Receptor ◽

Oxygen Species ◽

Normal Human

Abstract Background Experimental data are consistent with the hypothesis that activation of the PDGF receptor (PDGFR) is characteristic of scleroderma (SSc) fibroblasts and may contribute to their activation. We have recently demonstrated that fibroblasts from SSc patients contain high Ha Ras and ROS (Reactive Oxygen Species) levels and constitutive activation of ERK1/2 (Svegliati et al: JBC in press). Furthermore, SSc patients have circulating auto-antibodies against the PDGFR which induce type I collagen gene expression in normal human fibroblasts through the Ha Ras-ERK1/2- ROS pathway (Svegliati et al: Submitted). These findings suggest that anti PDGFR auto-antibodies play a pivotal role in the pathogenesis of scleroderma. Clinical chronic graft-versus-host disease (cGVHD) can show manifestations that are very similar to those of SSc. Although it is conceivable that the two diseases can share a similar pathophysiological mechanism there are no data supporting this assumption. In view of these considerations we tested the hypothesis that patients with cGVHD have serum auto-antibodies that stimulate PDGFR and activate collagen gene expression in fibroblasts. Methods Serum from 7 patients with extensive cGVHD showing scleroderma-like features either in the skin or in the lung was analyzed for the presence of stimulatory autoantibodies to PDGFR. Patients receiving allogeneic transplantation, but without any signs of cGVHD were used as controls. The median F-U after transplant was 23 months (range 16–36) in patients with cGVH and 42 (range 9–51) in the control group. The assay was carried by incubating purified IgG of the patients with mouse embryo fibroblasts carrying inactive copies of PDGFR α or β chains (PDGFR −/−) or the same cells expressing PDGFR α or β, respectively. Production of reactive oxygen species was assayed in the presence or absence of specific PDGFR inhibitors. The antibodies were characterized by immunoprecipitation, immunoblotting and absorption experiments in primary human fibroblasts and endothelial cells. Result Stimulatory antibodies to the PDGFR were selectively found in all patients with cGVHD and fibrotic lesions. The antibodies specifically recognized PDGFR, induced tyrosine phosphorylation and ROS accumulation. Their activity was completely and selectively abolished by pre-incubation with cells expressing PDGFR α or β chains or by PDGF receptor tyrosine kinase inhibitor. Anti-PDGFR antibodies induced selectively Ha-Ras-ERK1/2 and ROS cascade and stimulated the expression of type I collagen gene and myofibroblast phenotype conversion in normal human primary fibroblasts. Antibodies were absent in all controls. Conclusions Stimulatory auto-antibodies against PDGFR represent a specific hallmark of patients with cGVHD. Their biological activity on fibroblasts strongly argues for a causal role in the pathogenesis of the disease.

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Superoxide Release in Human Fibroblasts upon Treatment with Culture Supernatants of the Arthritogenic Bacteria Erysipelothrix rhusiopathiae and Mycoplasma arthritidis

Zeitschrift für Naturforschung C ◽

10.1515/znc-1998-3-416 ◽

1998 ◽

Vol 53 (3-4) ◽

pp. 254-263

Author(s):

Beate Meier

Keyword(s):

Reactive Oxygen Species ◽

Lipid A ◽

Human Fibroblasts ◽

Primary Cultures ◽

Reactive Oxygen ◽

Oxygen Species ◽

Erysipelothrix Rhusiopathiae ◽

Mycoplasma Arthritidis ◽

Culture Supernatants ◽

Irreversible Injury

Abstract Culture supernatants of the arthritogenic bacteria Mycoplasma pneum onia, Mycoplasma arthritidis, Borrelia burgdorferi and Erysipelothrix rhusiopathiae stimulated primary cultures of human fibroblasts to release reactive oxygen species into the environment, whereas cell walls and membranes of these bacteria had no effects. Lipopolysaccharides of various gramnegative bacteria and lipid A , the lipid moiety of endotoxines, also failed to stimulate the release of reactive oxygen species by fibroblasts. The stimulatory fractions of the culture supernatants of Mycoplasma arthritidis and Erysipelothrix rhusiopathiae exhibited a molecular weight of about 9.5 kDa. After an induction period of 5 min the presence of the stimulant was not necessary any more. The primary radical released by the fibroblasts was the superoxide anion O2-. Radical formation took place continuously over some hours. Additionally, low-level chemiluminescence of fibroblasts was increased upon stimulation with culture supernatants of Mycoplasma arthritidis and Erysipelothrix rhusiopathiae. No irreversible injury of the fibroblast was caused upon stimulation and the cells exhibited normal proliferation pattern after replacing them to the culture medium.

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Sildenafil Reduces Expression and Release of IL-6 and IL-8 Induced by Reactive Oxygen Species in Systemic Sclerosis Fibroblasts

International Journal of Molecular Sciences ◽

10.3390/ijms21093161 ◽

2020 ◽

Vol 21 (9) ◽

pp. 3161 ◽

Cited By ~ 4

Author(s):

Luigi Di Luigi ◽

Paolo Sgrò ◽

Guglielmo Duranti ◽

Stefania Sabatini ◽

Daniela Caporossi ◽

...

Keyword(s):

Oxidative Stress ◽

Reactive Oxygen Species ◽

Systemic Sclerosis ◽

Tissue Damage ◽

Human Fibroblasts ◽

Reactive Oxygen ◽

Dermal Fibroblasts ◽

Vascular Damage ◽

Oxygen Species ◽

Phosphodiesterase Type 5 Inhibitor

Oxidative stress linked to vascular damage plays an important role in the pathogenesis of systemic sclerosis (SSc). Indeed, vascular damage at nailfold capillaroscopy in patients with Raynaud’s Phenomenon (RP) is a major risk factor for the development of SSc together with the presence of specific autoantiobodies. Here, we investigated the effects of the phosphodiesterase type 5 inhibitor (PDE5i) sildenafil, currently used in the management of RP, in modulating the proinflammatory response of dermal fibroblasts to oxidative stress in vitro. Human fibroblasts isolated from SSc patients and healthy controls were exposed to exogenous reactive oxygen species (ROS) (100 µM H2O2), in the presence or absence of sildenafil (1 µM). Treatment with sildenafil significantly reduced dermal fibroblast gene expression and cellular release of IL-6, known to play a central role in the pathogenesis of tissue damage in SSc and IL-8, directly induced by ROS. This reduction was associated with suppression of STAT3-, ERK-, NF-κB-, and PKB/AKT-dependent pathways. Our findings support the notion that the employment of PDE5i in the management of RP may be explored for its efficacy in modulating the oxidative stress-induced proinflammatory activation of dermal fibroblasts in vivo and may ultimately aid in the prevention of tissue damage caused by SSc.

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Protective Activity and Underlying Mechanism of Ginseng Seeds against UVB-Induced Damage in Human Fibroblasts

Antioxidants ◽

10.3390/antiox10030403 ◽

2021 ◽

Vol 10 (3) ◽

pp. 403

Author(s):

Huijin Heo ◽

Hana Lee ◽

Jinwoo Yang ◽

Jeehye Sung ◽

Younghwa Kim ◽

...

Keyword(s):

Reactive Oxygen Species ◽

Fatty Acids ◽

Bioactive Compounds ◽

Transforming Growth Factor ◽

Human Fibroblasts ◽

Reactive Oxygen ◽

Underlying Mechanism ◽

Oxygen Species ◽

Protective Activity ◽

Skin Photoaging

Ginseng seeds are rich in phytosterols, ginsenosides, and fatty acids, and can therefore be used in skincare to delay the aging process. Ginseng seed embryo (GSE) and ginseng seed coat (GSC) were separated from ginseng seeds (Panax ginseng Meyer). This study evaluated the protective activity and underlying mechanism of GSE and GSC on UVB irradiation-induced skin photoaging using Hs68 cells. Their bioactive compounds, including phytosterols, ginsenosides, tocopherols, tocotrienols, and fatty acids were determined by HPLC and GC. The levels of reactive oxygen species, matrix metalloproteinases (MMPs), and collagen levels were measured in human dermal fibroblast cell line, Hs68 cells. The antioxidant capacity and contents of total polyphenols and flavonoids were higher in GSC than those in GSE. Linoleic acid was the major fatty acid in both GSE and GSC. GSE and GSC treatment alleviated UVB-induced increase of reactive oxygen species (ROS), matrix metalloproteinase (MMP)-1, and MMP-3, resulting in reduced collagen degradation. Increased UVB-mediated phosphorylation of mitogen activated protein kinase (MAPK) and activator protein-1 (AP-1) was inhibited by GSE and GSC treatment. Moreover, GSE and GSC effectively upregulated transforming growth factor-β (TGF-β) 1 levels. It was found that ginseng seeds regulate the expression of TGF-β/Smad and MAPK/AP-1 pathways. Ginseng seeds contain various bioactive compounds and have protective activity against UVB-induced skin photoaging. Therefore, ginseng seeds have the potential for use in cosmeceutical preparations.

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