Cytotoxicity In Epithelial Cells Research Articles

Nitric oxide-generating microparticles: An in vitro evaluation of anti-biofilm efficacy and sinonasal epithelial cell cytotoxicity.

International Forum of Allergy & RhinologyAccepted Articles RESEARCH NOTE Nitric Oxide Generating Microparticles: An In Vitro Evaluation of Anti-Biofilm Efficacy and Sinonasal Epithelial Cell Cytotoxicity Kevin L. Li MD, Kevin L. Li MD Department of Otorhinolaryngology – Head and Neck Surgery, Albert Einstein College of Medicine, Bronx, NY, USASearch for more papers by this authorDaniel Z. Miranda PhD, Daniel Z. Miranda PhD Department of Medicine (Division of Infectious Diseases) and Department of Microbiology and Immunology, Albert Einstein College of Medicine, Bronx, NY, USASearch for more papers by this authorLevi G. Cleare BA, Levi G. Cleare BA Department of Medicine (Division of Infectious Diseases) and Department of Microbiology and Immunology, Albert Einstein College of Medicine, Bronx, NY, USASearch for more papers by this authorNadeem A. Akbar MD, Nadeem A. Akbar MD Department of Otorhinolaryngology – Head and Neck Surgery, Albert Einstein College of Medicine, Bronx, NY, USASearch for more papers by this authorJoel M. Friedman MDPhD, Joel M. Friedman MDPhD Department of Physiology and Biophysics, Albert Einstein College of Medicine, Bronx, NY, USASearch for more papers by this authorAndrew Draganski PhD, Andrew Draganski PhD Zylö Therapeutics, Greenville, SC, USASearch for more papers by this authorJoshua D. Nosanchuk MD, Joshua D. Nosanchuk MD Department of Medicine (Division of Infectious Diseases) and Department of Microbiology and Immunology, Albert Einstein College of Medicine, Bronx, NY, USASearch for more papers by this authorWaleed M. Abuzeid MD, Corresponding Author Waleed M. Abuzeid MD wabuzeid@uw.edu Department of Otolaryngology: Head and Neck Surgery, University of Washington, Seattle, WA, USA Correspondence Waleed M. Abuzeid, MD, Associate Professor, Director of Rhinology Research, Director of Equity, Diversity, and Inclusion, Department of Otolaryngology: Head and Neck Surgery, University of Washington. Email: wabuzeid@uw.eduSearch for more papers by this author Kevin L. Li MD, Kevin L. Li MD Department of Otorhinolaryngology – Head and Neck Surgery, Albert Einstein College of Medicine, Bronx, NY, USASearch for more papers by this authorDaniel Z. Miranda PhD, Daniel Z. Miranda PhD Department of Medicine (Division of Infectious Diseases) and Department of Microbiology and Immunology, Albert Einstein College of Medicine, Bronx, NY, USASearch for more papers by this authorLevi G. Cleare BA, Levi G. Cleare BA Department of Medicine (Division of Infectious Diseases) and Department of Microbiology and Immunology, Albert Einstein College of Medicine, Bronx, NY, USASearch for more papers by this authorNadeem A. Akbar MD, Nadeem A. Akbar MD Department of Otorhinolaryngology – Head and Neck Surgery, Albert Einstein College of Medicine, Bronx, NY, USASearch for more papers by this authorJoel M. Friedman MDPhD, Joel M. Friedman MDPhD Department of Physiology and Biophysics, Albert Einstein College of Medicine, Bronx, NY, USASearch for more papers by this authorAndrew Draganski PhD, Andrew Draganski PhD Zylö Therapeutics, Greenville, SC, USASearch for more papers by this authorJoshua D. Nosanchuk MD, Joshua D. Nosanchuk MD Department of Medicine (Division of Infectious Diseases) and Department of Microbiology and Immunology, Albert Einstein College of Medicine, Bronx, NY, USASearch for more papers by this authorWaleed M. Abuzeid MD, Corresponding Author Waleed M. Abuzeid MD wabuzeid@uw.edu Department of Otolaryngology: Head and Neck Surgery, University of Washington, Seattle, WA, USA Correspondence Waleed M. Abuzeid, MD, Associate Professor, Director of Rhinology Research, Director of Equity, Diversity, and Inclusion, Department of Otolaryngology: Head and Neck Surgery, University of Washington. Email: wabuzeid@uw.eduSearch for more papers by this author First published: 08 October 2022 https://doi.org/10.1002/alr.23096 This article has been accepted for publication and undergone full peer review but has not been through the copyediting, typesetting, pagination and proofreading process, which may lead to differences between this version and the Version of Record. Please cite this article as https://doi.org/10.1002/alr.23096 AboutPDF ToolsExport citationAdd to favoritesTrack citation ShareShare Give accessShare full text accessShare full-text accessPlease review our Terms and Conditions of Use and check box below to share full-text version of article.I have read and accept the Wiley Online Library Terms and Conditions of UseShareable LinkUse the link below to share a full-text version of this article with your friends and colleagues. Learn more.Copy URL Share a linkShare onFacebookTwitterLinked InRedditWechat Accepted ArticlesAccepted, unedited articles published online and citable. The final edited and typeset version of record will appear in the future. RelatedInformation

Protective Effects of Theaflavins and Epigallocatechin Gallate against ZnO-NP-Induced Cell Apoptosis In Vitro

Zinc oxide nanoparticles (ZnO-NPs) are commonly used in various commercial applications, causing toxic effects on organisms and destroying biodiversity, but information about their protective approaches remains unknown. This study aims to evaluate the protective effects of theaflavins (TFs) and epigallocatechin gallate (EGCG) against ZnO-NP-induced cytotoxicity in rat tracheal epithelial (RTE) cells. Herein, RTE cells were exposed to 100 μg/L ZnO-NPs for 12 h, then treated with 0, 10, 100, and 1000 μg/L TFs or EGCG for another 12 h; subsequently, oxidative stress, inflammation, and apoptosis analyses were conducted. Relative to the control groups, TFs and EGCG treatment significantly inhibited the levels of reactive oxygen species and malondialdehyde content. Exposure to 1000 μg/L TFs or EGCG treatment downregulated cytochrome C gene expression levels by 59.10% and 77.27%; Caspase 3 gene expression by 50.03% and 60.01%; Caspase 8 gene expression by 45.11% and 55.57%; and Caspase 9 gene expression by 51.33% and 66.67%, respectively. Meanwhile, interleukin 1β and interleukin 6, tumor necrosis factor-α, and the other inflammatory chemokines such as C-C motif chemokine 2 and C-X-C motif chemokine 8 expression were all gradually rescued after the addition of TFs or EGCG. These results imply that TFs or EGCG possibly ameliorated ZnO-NPs-induced toxicity through antiapoptotic, antioxidant, and anti-inflammatory effects. This study provides novel approaches which mitigate the emerging nanoparticle pollutant toxicity in organisms, which may potentially slow down the destruction of biodiversity.

Clinical features of organizing pneumonia in anti-aquaporin-4 antibody-positive neuromyelitis optica spectrum disorders

Anti-aquaporin-4 (AQP4) antibody is an autoantibody marker often observed in patients with neuromyelitis optica spectrum disorder (NMOSD). The pathological relevance of complicated pulmonary disorders in anti-AQP4 antibody-positive NMOSD remains unclear. We aimed to assess the clinical and histological relevance of complicated pulmonary disorders in anti-AQP4 antibody-positive NMOSD. We retrospectively reviewed the medical records of 52 patients with anti-AQP4 antibody-positive NMOSD and conducted immunohistochemical evaluations of the lung biopsy specimens. Among 52 patients with anti-AQP4 antibody-positive NMOSD, 4 patients showed pulmonary involvement with a diagnosis of organizing pneumonia (OP). The proportion of males was larger (75% vs. 12.5%; p=0.013) and creatine kinase levels were higher (458.3 U/L vs. 83.9 U/L; p=0.003) in patients with OP than in those without OP. OP development preceded or coincided with the NMOSD symptoms. Chest computed tomography findings were consistent with OP in all four patients. Bronchoalveolar lavage fluid predominantly contained lymphocytes. Transbronchial lung biopsy revealed intraluminal plugs of inflammatory debris within the alveoli. Alveolar epithelial cells covering the OP lesions exhibited AQP4 loss, immunoglobulin G deposition, and complement activation. Corticosteroid treatment resulted in clinical improvement of OP. OP may be considered a pulmonary manifestation of anti-AQP4 antibody-positive NMOSD beyond the central nervous system. Complement-dependent cytotoxicity of the lung epithelial cells caused by anti-AQP4 antibody is at least partly involved in OP development. When diagnosing NMOSD, the possibility of OP should be carefully evaluated based on the detailed history and chest imaging findings.

The Impact of Heat Treatment of Quercetin and Myricetin on their Activities to Alleviate the Acrylamide-Induced Cytotoxicity and Barrier Loss in IEC-6 Cells.

Two flavonols quercetin and myricetin were assessed for their in vitro activities to attenuate the acrylamide-induced cytotoxicity and barrier loss in rat intestinal epithelial (IEC-6) cells and to identify whether heat treatment of the flavonols might cause activity changes. The results showed that the flavonols could alleviate the acrylamide-caused cell injury, resulting in higher cell viability, lower lactate dehydrogenase release, and less formation of reactive oxygen species. Meanwhile, the flavonols could antagonize the acrylamide-induced barrier dysfunction via decreasing the paracellular permeability, increasing the transepithelial resistance of cell monolayer, and enhancing the expression of three tight junction proteins namely occludin, claudin-1, and zonula occludens-1. The flavonols also could down-regulate the expression of JNK/Src proteins and thus cause lower relative protein ratios of p-JNK/JNK and p-Src/Src, resulting in a suppressed JNK/Src activation. Totally, quercetin was more potent than myricetin to exert these assessed activities, while the heated flavonols obtained lower activity than the unheated ones. It is thus concluded that the flavonols had beneficial activities towards the intestinal epithelial cells with acrylamide exposure by alleviating the acrylamide-induced cytotoxicity and barrier disruption, while heat treatment of the flavonols was unfavorable because it led to a reduced flavonol activity to the cells.

Digested Human Colostrum Reduces Interleukin-8 Production in Induced Human Intestinal Epithelial Cells.

Little is known about the impact of human colostrum on infant intestinal health following digestion. The aim of this study was to compare the effect of digested versus undigested human colostrum on inflammation and cytotoxicity in human intestinal epithelial cells (Caco2BBe) stimulated with lipopolysaccharides (LPS) or tumor necrosis factor (TNF). Colostrum samples (days 2–8 postpartum) from ten mothers of preterm infant were applied. Caco2BBe cells were pretreated by digested or undigested colostrum before stimulation with LPS or TNF. The inflammatory response was determined by measuring the production of interleukin-8 (IL-8) from cells using enzyme linked immunosorbent assay (ELISA). Cytotoxicity was examined by measuring the release of lactate dehydrogenase (LDH) from the cells. Digested colostrum significantly reduced IL-8 production under LPS and TNF stimulation compared with undigested colostrum. Individual colostrum samples exhibited wide variance in the ability to suppress IL-8 production and cytotoxicity in Caco2BBe cells. In vitro-digested human colostrum suppressed an inflammatory response more than undigested human colostrum in an induced intestinal cell culture model.

Evaluating electronic cigarette cytotoxicity and inflammatory responses in vitro.

INTRODUCTIONCigarette smoking poses many health risks and can cause chronic obstructive pulmonary disease (COPD), cardiovascular disease, cancer of the lung and other organs. Smokers can substantially reduce their risks of these diseases by quitting, but nicotine addiction makes this difficult. Alternatives, such as electronic cigarettes (e-cigarettes), may provide a similar dose of nicotine, but expose users to fewer toxic chemicals than traditional cigarettes and may still be harmful especially for dual users, therefore, we sought to develop bioassays that can assess the potential toxicity and inflammatory response induced by e-cigarette liquids (e-liquids) with and without flavors.METHODSE-liquids with varying nicotine content and flavors were aerosolized through growth media and exposed to human bronchial epithelial cell line (BEAS-2B) and human monocyte-macrophage cell line (THP-1) in vitro. Cytotoxicity in response to e-cigarette aerosols was measured by MTT assay in BEAS-2B cells and inflammatory response was measured by TNF-α, IL-6, IL-8, and MCP-1 released from THP-1 cells. In addition, the oxidative stress marker, REDD1, and impact on phagocytosis, was assessed following exposure of BEAS-2B and THP-1 derived macrophages, respectively. Cigarette smoke extract was used as a positive control with known cytotoxicity and impairment of inflammatory response.RESULTSE-cigarette aerosols induced moderate cellular toxicity in bronchial epithelial cells. Our data also show that low nicotine levels are less damaging to the bronchial epithelial cells, and flavors in e-liquids influence the combined inflammatory response markers, phagocytosis, and REDD1 when examined in vitro.CONCLUSIONSOur in vitro bioassays can be utilized to effectively measure flavor and nicotine-induced effects of e-cigarettes on combined inflammatory response and cytotoxicity in human macrophages and human bronchial epithelial cells, respectively.

N-acetylcysteine functionalized chitosan oligosaccharide-palmitic acid conjugate enhances ophthalmic delivery of flurbiprofen and its mechanisms

An N-acetylcysteine functionalized chitosan oligosaccharide-palmitic acid conjugate (NAC-COS-PA) with bioadhesive and permeation promoting properties was synthesized to enhance transocular drug delivery. Flurbiprofen (FB) loaded self-assembled NAC-COS-PA nanomicelles (NAC-COS-PA-FB) were prepared and the drug loading was 7.35 ± 0.32%. Human immortalized corneal epithelial (HCE-T) cell cytotoxicity and hen's egg test-chorioallantoic membrane assays confirmed that the conjugate had good biocompatibility. The transportation efficiency of coumarin-6 (C6) loaded nanomicelles in the HCE-T cell monolayer was approximately 1.97 times higher than that of free C6. Decreased intracellular Ca2+ concentration and cell membrane potential, increased cell membrane fluidity, and reversible changes in the F-actin cytoskeleton are presumed to be responsible for the enhanced drug permeation. NAC-COS-PA exhibited strong binding capacity with mucin and rabbit eyeball. In vivo pharmacokinetics indicated that the area under the curve (AUC0–6 h) and the maximum concentration (Cmax) of NAC-COS-PA-FB were approximately 1.92 and 2.44 times that of the FB solution, respectively. NAC-COS-PA-FB demonstrated the best in vivo anti-inflammatory efficacy compared to unfunctionalized nanomicelles (COS-PA-FB) and FB solution. Consequently, NAC-COS-PA appears to be a promising bioadhesive carrier for ophthalmic delivery.

Influence and Implications of the Molecular Paradigm of Nitric Oxide Underlying Inflammatory Reactions of the Gastrointestinal Tract of Dog: A Major Hallmark of Inflammatory Bowel Disease.

Nitric oxide (NO), a pleiotropic free radical messenger molecule, is responsible for the various cellular function of the gastrointestinal mucosa. It plays a major role in the maintenance of perfusion, regulation of microvascular, epithelial permeability, and immune functions. Nitric oxide exerts its beneficial effect on the initiation and maintenance of inflammation in human inflammatory bowel disease (IBD). But the accelerated production of NO triggers activation of the inducible form of the NO synthase enzyme (iNOS) that leads to damages of the intestinal membrane. Nitric oxide synthase enzyme is responsible for the higher production of NO from l-arginine and causes an inflammatory condition in the intestinal epithelium. Nitric oxide induces nitrative DNA damage and oxidative DNA damage in the cellular system. Accelerated production of NO enhances iNOS activity that is associated with cytotoxicity and apoptosis of gastrointestinal epithelial cells in the dog. Chronic inflammation leads to angiogenesis that is modulated by the immune system in IBD. Chronic inflammation is a major risk factor for the development of gastrointestinal malignancies. Nitric oxide participates in mucosal inflammation in the intestine through invigoration of NO synthase enzyme. The intrinsic complex mechanism is correlated with the inflammation in the gastrointestinal tract and is also correlated with the expression of iNOS, enzymatic activity and NO production. Nitric oxide employs a significant role in modulating epithelial permeability with accelerated immune response in acute colitis. But the enormous generation of NO causes adverse effects on the mucosal cell during the inflammatory process in IBD. In this review, a complex episode of NO generation with altered biochemical pathways was assessed for the regulation of mucosal inflammation in inflammatory bowel disease of dogs. This review is a unique compilation of the role of NO in the pathogenesis of inflammatory bowel disease of dogs. Nitric oxide plays a key role in modulating cancer in the gastrointestinal tract. This review seeks to explore the characteristics of NO as a major hallmark of canine inflammatory bowel diseases.

Anticancer effects of Pimaric acid is mediated via endoplasmic reticulum stress, caspase-dependent apoptosis, cell cycle arrest, and inhibition of cell migration in human ovarian cancer cells.

Pimaric acid is a naturally occurring resin and has been found to perform many pharmacological activities including, anticancer activity. However, the role of Pimaric acid in ovarian cancer is still not known. This investigation aimed to evaluate the anticancer effects of Pimaric acid and its molecular mechanism in human ovarian cancer cells. MTT assay was used to examine cell viability. Cell morphology was determined through phase contrast microscopy. DAPI staining and TUNEL assay were performed for apoptotic study. Examination of cell cycle phase distribution was carried out through flow cytometry. In vitro wound healing assay was used for cell migration determination. Pimaric acid induced cytotoxicity in human ovarian cancer cells (PA-1) in a dose-dependent manner without causing too much cytotoxicity in human ovarian epithelial cells (T1074). Cell morphology in treated cancer cells showed significant changes compared to untreated controls. Furthermore, it was observed that the cytotoxic effects of Pimaric acid were apoptosis-mediated and caspase-dependent cascade. Western blotting analysis showed that the expression of apoptosis-associated proteins like BAX, p-53 and caspase-3 was enhanced and BCL-2 expression was diminished. The induction of cytotoxicity was mediated via endoplasmic reticulum stress through expressions of related proteins which showed a tremendous increase in p-PERK, PERK, AT-4, CHOP and IRE-1 levels after treatment. Cell cycle analysis through cytometry showed significant results as it revealed G2/M phase cell cycle arrest. Furthermore, the in vitro wound healing assay showed specific anti-migratory effects of Pimaric acid on PA-1 cells. In conclusion it can be assumed that Pimaric acid may act as a potential anticancer agent against ovarian carcinoma, however further investigations are required to validate this initial claim.

Ex Vivo and In Vitro Analysis Identify a Detrimental Impact of Neutrophil Extracellular Traps on Eye Structures in Equine Recurrent Uveitis

Equine recurrent uveitis (ERU) is a common ocular disease of horses and described as a model for human autoimmune uveitis. This immune-mediated, inflammatory condition progressively destroys the eye, ultimately leading to blindness. Genetic and autoimmune factors, next to infections with Leptospira, are discussed as key factors in the pathogenesis. Furthermore, a release of neutrophil extracellular traps (NETs) by activated neutrophils is involved. NETs are composed of decondensed chromatin and proteins that can immobilize invading pathogens. However, if NETs accumulate, they can contribute to detrimental autoimmune processes. Thus, we aimed to investigate the impact of NETs in ERU patients. Therefore, we quantified several NET-markers (cell-free DNA, nucleosomes, citrullinated histone H3, histone-myeloperoxidase complexes, interleukin-17, equine cathelicidin 1 and DNase I activity) and NET-autoantibodies in sera and vitreous body fluids (VBF) of ERU-diseased horses and correlated the data with the disease status (signalment, ERU scores and Leptospira infection status). NET markers were detected to varying degrees in VBF of diseased horses, and partially correlated to disease severity and the presence of Leptospira spp. Cell-free DNA and nucleosomes as NET markers correlate with ERU severity in total and VBF scores, despite the presence of active DNases. Additionally, a significant correlation between fundus affection in the eye and NET autoantibodies was detectable. Therefore, we further investigated the influence of VBF samples from equine patients and isolated NETs on the blood-retina barrier in a cell culture model. VBF of diseased horses significantly induced cytotoxicity in retinal pigment epithelial cells. Moreover, partially digested NETs also resulted in cytotoxic effects. In the presence of lipopolysaccharide (LPS), the main component of the leptospiral surface, both undigested and completely digested NETs were cytotoxic. Correlations between the ERU-scores and Leptospira were also calculated. Detection of leptospiral DNA, and antibody titers of the serovar Grippotyphosa correlated with disease severity. In addition, a correlation between Leptospira and several NET markers was observed in VBF. Altogether, our findings suggest a positive correlation between NET markers with disease severity and involvement of Leptospira in the VBF of ERU-diseased horses, as well as a cytotoxic effect of NETs in eyes.

Cadmium and Cadmium/BDE (47 or 209) Exposure Affect Mitochondrial Function, DNA Damage/Repair Mechanisms and Barrier Integrity in Airway Epithelial Cells

Heavy metals and Brominated diphenyl ether flame-retardants (BDEs) often coexist in the environment and are capable of inducing injury, cytotoxicity or genotoxicity in human epithelial cells of the lung. We studied the effects of single Cadmium chloride (CdCl2) or CdCl2/BDE (47 or 209) mixtures in airway epithelial cells, using A549 cell line cultured at submerged conditions and air–liquid interface (ALI) (an in vitro model described as physiologically relevant in vivo-like). We evaluated cell viability, oxidative stress, apoptosis, DNA damage/repair (Comet assay, γH2AX phosphorylation ser139), mitochondrial redox balance (NOX-4, Nrf2 and TFAM) and cell barrier integrity (TEER, ZO-1, Claudin-1, E-cadherin-1) in A549 cells exposed to CdCl2 (1 nM to 10 µM), or to CdCl2 (100 nM)/BDEs (47 or 209) (100 nM). CdCl2 (10 μM) reduced cell viability and increased apoptosis. CdCl2 (100 nM) significantly affected DNA-damage/repair (Olive Tail length production), γH2AX phosphorylation and oxidative stress (ROS/JC-1 production) in submerged cell cultures. CdCl2 (100 nM) decreased viability, TEER, ZO-1, Claudin-1 and E-cadherin-1 mRNA expression, and Nrf2 and TFAM while increased NOX-4, in ALI culture of cells. In both cell culture approaches, the cells stimulated with Cadmium/BDEs mixtures did not show a significant increase in the effects observed in the cells treated with CdCl2 alone. CdCl2 inhalation might exert cytotoxicity and genotoxicity, playing a pivotal role in the uncontrolled oxidative stress, damaging DNA and gene expression in airway epithelial cells. No additional or synergistic adverse effects of CdCl2/BDEs mixture were observed in comparison to CdCl2 alone in lung epithelium.

Protective Effects of Gamma-mangostin on Hydrogen Peroxideinduced Cytotoxicity in Human Retinal Pigment Epithelial Cells.

The role of epigenetic alterations in the pathogenesis of retinal degenerative diseases, such as macular degeneration is not well established. This study aimed to evaluate whether treatments with gamma-mangostin can rescue the hydrogen peroxide (H2O2)-induced cytotoxicity in human retinal pigment epithelial (ARPE-19) cells. ARPE-19 cells were treated with H2O2 alone or with gamma-mangostin plus H2O2 to investigate changes relating to cell viability, appearance of sub-G1 cells, antioxidant enzymes, and apoptotic-related proteins. The data showed that under H2O2 treatment of 400 μM, there was a significant decrease in cell viability and enhanced apoptosis, together with an increased expression of Bax, Bad, cleaved-caspase-3, -8, and -9 at the protein level. On the contrary, the protein expression levels of Bcl2 and Bcl-xl were decreased. Gamma-mangostin pre-treatments (2-16 μM) could effectively prevent all alterations. Gamma-mangostin may conduct its eye-protective effects against H2O2-induced oxidative damage via anti- apoptotic and antioxidant mechanisms in ARPE-19 cells.

PTX3 alleviates hard metal-induced acute lung injury through potentiating efferocytosis

Prolonged exposure to hard metal dust results in hard metal lung disease (HMLD) characterized by respiratory symptoms. Understanding the pathogenesis and pathological process of HMLD would be helpful for its early diagnosis and treatment. In this study, we established a mouse model of hard metal-induced acute lung injury through one-time intratracheal instillation of WC-Co dust suspension. We found that WC-Co treatment damaged the lungs of mice, leading to increased production of IL-1β, TNF-α, IL-6 and IL-18, inflammatory cells infiltration and apoptosis. In vitro, WC-Co induced cytotoxicity, inflammatory response and apoptosis in macrophages (PMA-treated THP-1) and epithelial cells (A549) in a dose-dependent manner. Moreover, RNA-sequence and validation experiments verified that Pentraxin 3 (PTX3), an important mediator in the regulation of inflammation, was elevated both in vivo and in vitro induced by WC-Co. Functional experiments confirmed the PTX3, which was located on the membrane of apoptotic cells, promoted macrophage efferocytosis efficiently. This progress could help block the lung inflammation and contribute to the rapid recovery of WC-Co-induced acute lung injury. These observations provide a further understanding of the molecular mechanism of WC-Co-induced pulmonary injury and disclose PTX3 as a new potential therapeutic approach to relieve WC-Co-induced acute lung injury via efferocytosis.

Combined ataxia telangiectasia mutated and DNA-dependent protein kinase inhibition radiosensitizes Madin-Darby canine kidney cells.

Uncovering radiation toxicity is critical for the adaptation and expansion of advanced radiation therapies and for the development of novel cancer radiotherapy. In the near future, advanced radiotherapies, including heavy ion beam treatment, are expected to be applied in the treatment of dogs, but further basic research on the effects of radiation using canine normal and cancer cells is necessary to actually apply these techniques and achieve high therapeutic efficacy. The radiation sensitivity is varied by the activities of DNA damage response (DDR) and DNA repair. The development of radiosensitizers that target DDR- and DNA repair-kinases, like ataxia telangiectasia mutated (ATM) and DNA-dependent protein kinase (DNA-PK), is progressing and is expected to be introduced into canine radiotherapy. However, there are no cytotoxicity reports on using the combination of radiation and these sensitizers as treatment in canine cells. In this study, we examined the cytotoxic effects of X-rays and/or radiosensitizers on the Madin-Darby canine kidney (MDCK) cell line. Our results show that X-rays suppress MDCK cell colony formation and proliferation in a dose-dependent manner. Additionally, our observations imply that the combination treatment with ATM inhibitor KU-55933 and DNA-PK inhibitor NU7441 significantly increased X-ray cytotoxicity in MDCK cells compared with the drugs alone. Furthermore, our findings further suggest that MDCK cells might be useful in clarifying the cytotoxicity in canine epithelial cells due to radiation and/or radiosensitizers, such as molecule-targeted drugs.

WSG, a glucose-enriched polysaccharide from Ganoderma lucidum, suppresses tongue cancer cells via inhibition of EGFR-mediated signaling and potentiates cisplatin-induced apoptosis

Tongue cancer, a kind of oral cancer, is common in Southeast Asian countries because of dietary habits. However, there is no specific targeted drug that could effectively inhibit oral cancer. WSG, as a water soluble glucose-enriched polysaccharide from Ganoderma lucidum, exerts excellent pharmacological efficacy of anti-lung cancer. However, its anticancer functions and mechanisms in human tongue cancer need to be further explored. Herein, we showed that WSG dramatically reduced cell viability and colony formation of tongue cancer cells. WSG increased subG1 and G2/M populations as well as induced apoptotic responses. In parallel, WSG enhanced apoptosis-related Bax/Bcl2 ratio. Mechanistic studies showed that WSG reduced phosphorylation of EGFR and AKT. In addition, we found a synergistic effect of WSG with cisplatin in inhibition of cell viability and induction of apoptosis. WSG significantly reduced the inhibition concentration 50% (IC50) of cisplatin. More importantly, WSG ameliorated cisplatin-induced cytotoxicity in normal human oral epithelial SG cells. In conclusion, our findings provided important insights into the anti-tongue cancer effects of WSG via inhibition of EGFR/AKT axis and induction of apoptosis, which indicated that WSG could be a promising supplement for tongue cancer treatment.

Spray-freeze-dried inhalable composite microparticles containing nanoparticles of combinational drugs for potential treatment of lung infections caused by Pseudomonas aeruginosa

The multi-drug resistance of Pseudomonas aeruginosa is an overwhelming cause of terminal and persistent lung infections in cystic fibrosis (CF) patients. Antimicrobial synergy has been shown for colistin and ivacaftor, and our study designed a relatively high drug-loading dry powder inhaler formulation containing nanoparticles of ivacaftor and colistin. The ivacaftor-colistin nanosuspensions (Iva-Col-NPs) were prepared by the anti-solvent method with different stabilizers. Based on the aggregation data, the formulation 7 (F7) with DSPG-PEG-OMe as the stabilizer was selected for further studies. The F7 consisted of ivacaftor, colistin and DSPG-PEG-OMe with a mass ratio of 1:1:1. The F7 powder formulation was developed using the ultrasonic spray-freeze-drying method and exhibited a rough surface with relatively high fine particle fraction values of 61.4 ± 3.4% for ivacaftor and 63.3 ± 3.3% for colistin, as well as superior emitted dose of 97.8 ± 0.3% for ivacaftor and 97.6 ± 0.5% for colistin. The F7 showed very significant dissolution improvement for poorly water soluble ivacaftor than the physical mixture. Incorporating two drugs in a single microparticle with synchronized dissolution and superior aerosol performance will maximize the synergy and bioactivity of those two drugs. Minimal cytotoxicity in Calu-3 human lung epithelial cells and enhanced antimicrobial activity against colistin-resistant P. aeruginosa suggested that our formulation has potential to improve the treatment of CF patients with lung infections.

Synthesis, Bioactivity, Pharmacokinetic and Biomimetic Properties of Multi-Substituted Coumarin Derivatives.

A series of novel multi-substituted coumarin derivatives were synthesized, spectroscopically characterized, and evaluated for their antioxidant activity, soybean lipoxygenase (LOX) inhibitory ability, their influence on cell viability in immortalized human keratinocytes (HaCaT), and cytotoxicity in adenocarcinomic human alveolar basal epithelial cells (A549) and human melanoma (A375) cells, in vitro. Coumarin analogues 4a–4f, bearing a hydroxyl group at position 5 of the coumarin scaffold and halogen substituents at the 3-phenyl ring, were the most promising ABTS•+ scavengers. 6,8-Dibromo-3-(4-hydroxyphenyl)-4-methyl-chromen-2-one (4k) and 6-bromo-3-(4,5-diacetyloxyphenyl)-4-methyl-chromen-2-one (3m) exhibited significant lipid peroxidation inhibitory activity (IC50 36.9 and 37.1 μM). In the DCF-DA assay, the 4′-fluoro-substituted compound 3f (100%), and the 6-bromo substituted compounds 3i (80.9%) and 4i (100%) presented the highest activity. The 3′-fluoro-substituted coumarins 3e and 4e, along with 3-(4-acetyloxyphenyl)-6,8-dibromo-4-methyl-chromen-2-one (3k), were the most potent lipoxygenase (LOX) inhibitors (IC50 11.4, 4.1, and 8.7 μM, respectively) while displaying remarkable hydroxyl radical scavenging ability, 85.2%, 100%, and 92.9%, respectively. In silico docking studies of compounds 4e and 3k, revealed that they present allosteric interactions with the enzyme. The majority of the analogues (100 μΜ) did not affect the cell viability of HaCaT cells, though several compounds presented over 60% cytotoxicity in A549 or A375 cells. Finally, the human oral absorption (%HOA) and plasma protein binding (%PPB) properties of the synthesized coumarins were also estimated using biomimetic chromatography, and all compounds presented high %HOA (>99%) and %PPB (60–97%) values.

Protective Action of Hydrogen Sulfide-Releasing Compounds against Oxidative Stress-Induced Cataract Formation in Cultured Bovine Lenses

ABSTRACT Purpose The gaseous signalling molecule, hydrogen sulfide (H2S) has antioxidant, anti-inflammatory and anti-apoptotic properties. Since oxidative stress has been implicated in the pathogenesis of cataracts and lenticular hydrogen peroxide (H2O2) is elevated in some cataract patients, the present study investigated the ability of H2S-releasing compounds to prevent H2O2-induced cataract formation in cultured bovine lenses. Methods Lenses were cultured in either Dulbecco's Modified Eagle Medium (DMEM; control); H2O2 (50 mM); ascorbic acid (AA; 3 mM) (positive control); and the H2S-releasing compounds (diallyl trisulfide [DATS] or GYY4137) in the presence of H2O2 (50 mM). Lens opacity was determined using a plate reader to measure transmittance. Lens glutathione content (GSH), superoxide dismutase (SOD) activity and lactate dehydrogenase (LDH) cytotoxicity were assessed before and after treatment with the H2S-releasing compounds. Results Both DATS (10−7M – 10−4M) and GYY4137 (10−7M – 10−4M) significantly (p < .001) attenuated H2O2 (50 mM)-induced loss in transmittance, with DATS (10−4M) and GYY4137 (10−7M) achieving a maximal reversal of opacity by 56.86 ± 0.01% (n = 6) and 8.39 ± 0.11% (n = 6) after 120 hours, respectively. These observations were corroborated by photographic evaluation, where DATS (10−5M – 10−4M) and GYY4137 (10−7M – 10−5M)-treated lenses had relatively clear grids after 120 hours, compared to H2O2 (50 mM)-treated lenses. The H2O2 (50 mM)-induced decline in total GSH content and total SOD activity were significantly (p < .001; n = 5) reversed by DATS (10−4M) and GYY4137 (10−7M). After 24 hours, DATS (10−4M) and GYY4137 (10−7M) significantly (p < .001; n = 4) reduced cytotoxicity of primary bovine lens epithelial cells by 33.88 ± 4.59% and 36.19 ± 10.53%, respectively. Conclusion Both H2S-releasing compounds protected cultured bovine lenses against oxidative stress-induced cataract formation. The slow-releasing H2S compound, GYY4137 was more potent than DATS in restoring lenticular total GSH content and total SOD activity along with reducing H2O2 (50 mM)-induced cytotoxicity

Potential cytotoxicity of trace elements and polycyclic aromatic hydrocarbons bounded to particulate matter: a review on in vitro studies on human lung epithelial cells.

A large number of studies have been conducted for clarifying toxicological mechanisms of particulate matter (PM) aimed to investigate the physicochemical properties of PM and providing biological endpoints such as inflammation, perturbation of cell cycle, oxidative stress, or DNA damage. However, although several studies have presented some effects, there is still no consensus on the determinants of biological responses. This review attempts to summarize all past research conducted in recent years on the physicochemical properties of environmental PM in different places and the relationship between different PM components and PM potential cytotoxicity on the human lung epithelial cells. Among 447 papers with our initial principles, a total of 50 articles were selected from 1986 to April 2020 based on the chosen criteria for review. According to the results of selected studies, it is obvious that cytotoxicity in human lung epithelial cells is created both directly or indirectly by transition metals (such as Cu, Cr, Fe, Zn), polycyclic aromatic hydrocarbons (PAH), and ions that formed on the surface of particles. In the selected studies, the findings of the correlation analysis indicate that there is a significant relationship between cell viability reduction and secretion of inflammatory mediators. As a result, it seems that the observed biological responses are related to the composition and the physicochemical properties of the PMs. Therefore, the physicochemical properties of PM should be considered when explaining PM cytotoxicity, and long-term research data will lead to improved strategies to reduce air pollution.

In vitro evaluation of a novel oxygen-generating biomaterial for chronic rhinosinusitis therapy.

Hypoxia due to closure at the ostiomeatal complex is widely considered one of the major pathogenic mechanisms leading to chronic inflammation in chronic rhinosinusitis (CRS). The objective of this study was to develop and characterize an oxygen-generating biomaterial (OGB) as an innovative treatment strategy for CRS. An OGB was fabricated by coating hydrophobic beeswax (BW, 15mg or 30mg) on the surface of calcium peroxide - catalase complex (CPO-CA, 30mg) and characterized using scanning electron microscopy (SEM). In vitro releases of both oxygen and hydrogen peroxide (H2 O2 ) were spectrophotometrically quantified, and cytotoxicity in human sinonasal epithelial cells (HSNECs) was evaluated. The influence of OGB on transepithelial Cl- secretion was also determined by pharmacologically manipulating HSNECs, cultured under hypoxic conditions, in Ussing chambers. Three groups of OGBs: (1) CPO only; (2) CPO coated with CA and BW (1:1 ratio, CPO-CA(1)-BW(1)); and (3) CPO coated with CA and BW (1:0.5 ratio, CPO-CA(1)-BW(0.5) were analyzed for accumulated oxygen release over 7 days: highest release (mmol/mg) was observed in CPO-CA(1)-BW(1)=0.11±0.003, followed by CPO-CA(1)-BW(0.5)=0.08 ± 0.010, and CPO=0.05 ± 0.004 (p<0.0001). H2 O2 production (mM) was significantly higher in CPO (1.87±0.50) compared to CPO-CA (1)-BW(1) (0.00±0.00) (p<0.001) after 24h. CPO-CA(1)-BW(1) showed significantly reduced cytotoxicity and increased Cl- transport compared to the CPO group. A novel OGB (CPO-CA-BW complex) exhibited sustained oxygen release over 7 days without significant cytotoxicity after 24h in vitro. Preclinical studies evaluating the efficacy of OGB in CRS are warranted, especially for potential therapy in an obstruction-based CRS model.