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EphA2 receptor and its ephrin ligands are involved in virus infection, epithelial permeability, and chemokine secretion. We hypothesized that ephrinA1/ephA2 signaling participates in rhinovirus (RV)-induced antiviral immune response in sinonasal mucosa of patients with chronic rhinosinusitis (CRS). Therefore, we investigated the expression of ephrinA1/ephA2 in normal and inflamed sinonasal mucosa and evaluated whether they regulate chemokine secretion and the production of antiviral immune mediators including interferons (IFNs) in RV-infected human primary sinonasal epithelial cells. For this purpose, the expression and distribution of ephrinA1/ephA2 in sinonasal mucosa were evaluated with RT-qPCR, immunofluorescence, and western blot. Their roles in chemokine secretion and the production of antiviral immune mediators such as type I and III IFNs, and interferon stimulated genes were evaluated by stimulating ephA2 with ephrinA1 and inactivating ephA2 with ephA2 siRNA or inhibitor in cells exposed to RV and poly(I:C). We found that ephrinA1/ephA2 were expressed in normal mucosa and their levels increased in inflamed sinonasal mucosa of CRS patients. RV infection or poly(I:C) treatment induced chemokine secretion which were attenuated by blocking the action of ephA2 with ephA2 siRNA or inhibitor. The production of antiviral immune mediators enhanced by rhinovirus or poly (I:C) is increased by blocking ephA2 compared with that of cells stimulated by either rhinovirus or poly(I:C) alone. In addition, blocking ephA2 attenuated RV replication in cultured cells. Taken together, these results describe a novel role of ephrinA1/ephA2 signaling in antiviral innate immune response in sinonasal epithelium, suggesting their participation in RV-induced development and exacerbations of CRS., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2021 Lee, Kang, Han, Kwak, Kim, Lee, Lee and Kim.)

Background: Little is known about the role of lymphotoxins (LTs) family in the sinonasal mucosa of patients with chronic rhinosinusitis (CRS). This study aims at investigating the expression of LIGHT, LTα, LTβ, and their receptors, LTβR and HVEM in normal and inflammatory sinus mucosa, and the effect of LIGHT and LTalpha1beta2 on chemokine secretion in epithelial cells, epithelial permeability, and leukocyte migration., Material and Methods: The expression of LTs family in sinonasal mucosa was evaluated with real-time PCR, immunohistochemistry, and western blot. In LTβR, HVEM siRNA, or control siRNA-transfected epithelial cells treated with LIGHT or LTalpha1beta2, the expression of chemokines, the epithelial permeability, and the expression of junctional complex proteins were evaluated using real-time PCR, ELISA, western blot, confocal microscopy, and FITC-dextran. In cultured endothelial cells treated with LIGHT or LTalpha1beta2, the expression of ICAM-1 and VCAM-1, and leukocyte migration were elucidated., Results: LTs family was expressed in normal mucosa and their levels were increased in inflammatory mucosa of CRS patients. Recombinant LIGHT and LTalpha1beta2 induced chemokine secretion, increased epithelial permeability, and promoted leukocyte migration. However, the activity of LIGHT and LTalpha1beta2 was attenuated in cells transfected with LTβR and HVEM siRNA., Conclusions: LIGHT and LTs may participate in the ongoing process of chronic inflammation, inducing chemokine secretion, leukocyte migration, and dysregulated epithelial barrier through LTβR and HVEM in sinonasal mucosa., (Copyright © 2021 Elsevier Ltd. All rights reserved.)

The pathogenesis of nasal inflammatory diseases is related to various factors such as anatomical structure, heredity, and environment. The nasal microbiota play a key role in coordinating immune system functions. Dysfunction of the microbiota has a significant impact on the occurrence and development of nasal inflammation. This review will introduce the positive and negative roles of microbiota involved in immunity surrounding nasal mucosal diseases such as chronic sinusitis and allergic rhinitis. In addition, we will also introduce recent developments in DNA sequencing, metabolomics, and proteomics combined with computation-based bioinformatics., Competing Interests: The authors declare no conflicts of interest.

Background: Neutrophil extracellular traps (NETs) participate in innate immunity by trapping microorganisms. Their pathophysiological implications have not been defined in chronic rhinosinusitis (CRS)., Objective: We investigated the presence of NETs in nasal secretion of patients with stable or exacerbated CRS and evaluated whether NETs participate in the secretion of chemokines in sinonasal epithelial cells, the epithelial permeability, and transendothelial leucocyte migration, and elucidate whether NETs are released by macrolides and dexamethasone., Methods: The presence of NETs in nasal secretion and the release of NETs from neutrophils stimulated with macrolides or dexamethasone were evaluated by dsDNA Assay kit and fluorescence microscope. The chemokine secretion, epithelial permeability, and transendothelial leucocyte migration were measured in cultured cells incubated with NETs, the supernatant of unstimulated neutrophils (unstim), NETs inhibitor (DPI), or H3Cit, where the expression of junctional complex proteins and ICAM-1 was evaluated by real-time PCR, Western blots, and confocal microscope., Results: The amount of NETs and NETs-forming neutrophils in nasal secretion increased in exacerbated CRS. Epithelial cells treated with NETs or H3Cit secreted chemokines and showed decreased permeability associated with up-regulated junctional complex proteins. Increased transendothelial leucocyte migration associated with up-regulated ICAM-1 was noted in endothelial cells treated with NETs or H3Cit. These findings were not found in cells treated with unstim, or DPI. NETs were released by macrolides, but not by dexamethasone., Conclusions and Clinical Relevance: NETs formation increased in exacerbated CRS, inducing chemokine secretion, strengthening the epithelial barrier, and promoting the neutrophils infiltration. Therefore, the release of NETs in CRS might be beneficial or detrimental to CRS patients., (© 2019 John Wiley & Sons Ltd.)

Background: In airway epithelium, thymus and activation-regulated chemokine (CCL17) and macrophage-derived chemokine (CCL22) are induced by defective epithelial barriers such as E-cadherin and attract the effector cells of Th2 immunity. However, the association between the epithelial barrier and CCL17 expression has not been studied in chronic rhinosinusitis with nasal polyp (CRSwNP). Thus, we aimed to evaluate the expression of CCL17 and its regulation by Th cytokines in nasal polyp (NP) epithelial cells., Methods: The expression and distribution of CCL17, CCL22, E-cadherin and/or epidermal growth factor receptor (EGFR) were measured using real-time PCR, western blot, and immunohistochemistry and compared between normal ethmoid sinus epithelium and NP epithelium. In addition, the expression level of CCL17 was determined in cultured epithelial cells treated with IL-4, IL-5, IL-13, TNF-α, and IFN-γ., Results: The expression of CCL17 was decreased in the NP epithelium compared to the epithelium of normal ethmoid sinus, whereas the expression of CCL22 was not decreased. E-cadherin was differentially distributed between the epithelium of normal ethmoid sinus and NP epithelium. EGFR was also decreased in NPs. Interestingly, the stimulation of cultured epithelial cells with Th2 cytokines, IL-4 and IL-5, resulted in an upregulation of CCL17 expression only in NP epithelial cells whereas the expression of CCL17 was increased in both normal epithelial cells and NP epithelial cells by Th1 cytokines., Conclusion: Our results suggest that the decreased expression of CCL17 in defective NP epithelium may be closely connected to NP pathogenesis and can be differentially regulated by cytokines in the NP epithelium of patients with CRSwNP.

Numerous peripheral tissues possess self-sustaining daily biologic rhythms that are regulated at the molecular level by clock genes such as PER1, PER2, CLOCK, and BMAL1. Physiological function of nasal mucosa exhibits rhythmic variability to a day-night environmental cycle. Nevertheless, little is known of the expression and distribution pattern of clock genes in nasal mucosa. The present study investigates the expression level and distribution pattern of PER1, PER2, CLOCK, and BMAL1 genes in nasal mucosa of healthy controls, allergic rhinitis patients, and normal rats. In human and rat nasal mucosa, the levels of these genes are asymmetrically expressed in nasal mucosa derived from right and left cavities in normal controls, allergic patients, and rat. In human nasal mucosa, the expression levels of these genes were higher in the decongested side than the congested mucosa. In rat nasal mucosa, these clock genes are expressed in a rhythmic circadian manner under the regular light/dark cycles. The expression levels of MUC5AC, a key mucin genes produced in superficial epithelium, are higher in decongested side than that congested side in human nasal mucosa. In rat nasal mucosa, MUC5AC levels showed a circadian rhythm which was associated with different expression levels in nasal mucosa derived from the right and left nasal cavities. Taken together with these results, the present study shows that the clock genes such as PER1, PER2, CLOCK, and BMAL1 are present in human and rat nasal mucosa, and suggest that these clock genes may control the pathophysiological function of nasal mucosa as circadian oscillators and affect the maintenance of the nasal cycle.

Objective: Laryngopharyngeal reflux disease (LPRD) is one of potential factors in recalcitrant chronic rhinosinusitis with or without polyps. An increase in junctional permeability in the nasal mucosa in LPRD may be due to disrupted protein bridge formation with cell-to-cell adhesion molecules such as E-cadherin. Despite the relationship between nasal mucosal inflammation and LPRD, the clear mechanism by which acid reflux affects the nasal epithelium remains unclear., Methods: The expression levels and distribution patterns of E-cadherin in primary culture of nasal epithelial cells after acid exposure with or without dexamethasone and matrix metalloproteinase (MMP) inhibitor were determined using Western blot and immunocytochemistry. The functional roles of MMP inhibitor in maintaining junctional permeability in the nasal epithelium were elucidated by transepithelial permeability test., Results: By acid exposure to nasal epithelial cells, mature E-cadherin was decreased and cleaved E-cadherin was increased. This was thought to be caused by cleavage of mature E-cadherin between cells and was confirmed by the increment of E-cadherin inside a cell in immunocytochemical evaluation. Whereas disruption of E-cadherin was not recovered by steroid medication with various treatments of dexamethasone, disrupted E-cadherin was restored to normal by inhibition of MMPs with actinonin, a broad MMP inhibitor. This recovery was functionally demonstrated by transepithelial permeability test., Conclusion: Our results suggest that altered expression of E-cadherin in the nasal epithelium by acid exposure may be a possible mechanism for nasal tissue injury in chronic nasal inflammation with LPRD, and that MMP inhibition is a potential treatment., Level of Evidence: NA. Laryngoscope, 128:E1-E7, 2018., (© 2017 The American Laryngological, Rhinological and Otological Society, Inc.)

Background: Pollutants produced by industrial and traffic-related activities have been linked to allergic responses. These noxious agents induce their effects through the aryl hydrocarbon receptor (AhR)., Objective: We analyzed the expression and distribution pattern of AhR in normal and allergic nasal mucosa, and cytokine-driven regulation of its expression. The production levels of chemokine in cultured nasal epithelial cells were evaluated after stimulation with AhR ligand., Methods: The expression levels and distribution pattern of AhR in normal, mild, and moderate-severe persistent allergic nasal mucosa were assessed by using real-time polymerase chain reaction, Western blot, and immunohistochemistry. The expression levels of AhR were determined in cultured nasal epithelial cells treated with T-helper 2 cytokines. In cultured epithelial cells stimulated with 2-(10H-indole-30-carbonyl)-thiazole-4-carboxylic acid methyl ester, the expression levels of granulocyte macrophage colony-stimulating factor, thymus and activation regulated chemokine, macrophage inflammatory protein 1 α, monocyte chemotactic protein 1, regulated on activation normal T-cell expressed and secreted, eotaxin, and interleukin 8 were measured with real-time polymerase chain reaction and enzyme-linked immunosorbent assay., Results: Expression of AhR was observed in normal and allergic nasal mucosa where it is distributed in the epithelial layer, submucosal glands, endothelial cells, and inflammatory cells. Its expression levels are increased in allergic nasal mucosa and upregulated after stimulation with T-helper 2 cytokines. The stimulation with 2-(10H-indole-30-carbonyl)-thiazole-4-carboxylic acid methyl ester resulted in increased production of chemokines in cultured epithelial cells., Conclusion: Analysis of the study results indicated that increased expression levels of AhR may play a role in the pathogenesis of allergic rhinitis, which contributes to chemokine production in nasal mucosa.

Background: Allergic rhinitis is a chronic nasal inflammatory disease mediated by an immunoglobulin E mediated process to environmental allergens. Although atopy is a potent predisposing risk factor for allergic rhinitis, local tissue susceptibilities are inevitable for disease expression. The nasal epithelium maintains tissue homeostasis by providing a physical barrier controlled by epithelial junctional proteins. However, the expression of epithelial junctional proteins has not been studied in patients with allergic rhinitis. We sought to elucidate the expression and the regulation of epithelial junctional proteins in the nasal epithelium of patients with allergic rhinitis., Methods: The expression of E-cadherin and zonula occludens (ZO) 1 in epithelium of turbinate was measured by using real-time polymerase chain reaction, Western blot, and immunohistochemical assays, and was compared between control subjects and patients with allergic rhinitis. In addition, the expression levels of E-cadherin and ZO-1 were determined in cultured epithelial cell treated with interleukin (IL) 4, IL-5, tumor necrosis factor (TNF) alpha, and interferon gamma., Results: The expression and the immunoreactivity of E-cadherin and ZO-1 were decreased in the nasal epithelium of patients with allergic rhinitis. Interestingly, the stimulation of cultured epithelial cells with IL-4, IL-5, and TNF-alpha resulted in downregulation of E-cadherin expression only in cultured epithelial cells of patients with allergic rhinitis, whereas E-cadherin expression in cultured epithelial cells of controls was not affected by stimulation with the same panel of cytokines., Conclusion: Decreased expression of epithelial junctional proteins was found in patients with allergic rhinitis. The disruption of epithelial integrity by IL-4, IL-5, and TNF-alpha in vitro indicated a possible role for these cytokines in the pathogenesis of patients with allergic rhinitis.

Background and Purpose: The anti-inflammatory and immunomodulatory effects of macrolides include the ability to decrease mucus secretion and inhibit inflammatory mediators in chronic rhinosinusitis. Nevertheless, their mechanisms of action remain to be determined. Here we have investigated the effects of macrolide antibiotics (clarithromycin, azithromycin and josamycin; representating the 14-, 15- and 16-membered macrolides) on endogenous steroids in human sinonasal epithelial cells and mouse nasal mucosa., Experimental Approach: The effects of macrolides on the expression of steroid-converting enzymes [11β-hydroxysteroid dehydrogenase (11β-HSD1 and 11β-HSD2)], steroid-synthesizing enzymes (3β-HSD, CYP21, CYP11B1 and CYP11A1) and cortisol levels were assessed in cultured human epithelial cells. In control and adrenalectomized mice , these enzymes and corticosterone levels were evaluated in nasal mucosa and serum after administration of macrolides., Key Results: The expression levels of 3β-HSD, CYP21, 11β-HSD1 and CYP11B1 increased in human epithelial cells treated with clarithromycin and azithromycin, whereas the expression levels of 11β-HSD2 and CYP11A1 were not affected. Josamycin had no effects on the expression of these enzymes. Cortisol levels increased in epithelial cells treated with clarithromycin or azithromycin. The expression of 3β-HSD, CYP11A1, CYP21, CYP11B1 and 11β-HSD1 was upregulated in nasal mucosa of mice treated with clarithromycin or azithromycin, but not in adrenalectomized mice., Conclusions and Implications: This study provides evidence that 14- and 15-membered macrolide antibiotics may affect the expression of steroid-synthesizing and steroid-converting enzymes in human sinonasal epithelial cells and mouse nasal mucosa, increasing the endogenous cortisol levels in sinonasal mucosa., (© 2015 The British Pharmacological Society.)

Purpose: The nasal mucosa is the first site to encounter pathogens, and it forms continuous barriers to various stimuli. This barrier function is very important in the innate defense mechanism. Additionally, inflammation of the nasal sinus is known to be a hypoxic condition. Here, we studied the effect of hypoxia on barrier function in normal human nasal epithelial (NHNE) cells., Materials and Methods: The expression levels of various junction complex proteins were assessed in hypoxia-stimulated NHNE cells and human nasal mucosal tissues. We performed real-time polymerase chain reaction analysis, western blotting, and immunofluorescence assays to examine differences in the mRNA and protein expression of ZO-1, a tight junction protein, and E-cadherin in NHNE cells. Moreover, we evaluated the trans-epithelial resistance (TER) of NHNE cells under hypoxic conditions to check for changes in permeability. The expression of ZO-1 and E-cadherin was measured in human nasal mucosa samples by western blotting., Results: Hypoxia time-dependently decreased the expression of ZO-1 and E-cadherin at the gene and protein levels. In addition, hypoxia decreased the TER of NHNE cells, which indicates increased permeability. Human nasal mucosa samples, which are supposed to be hypoxic, showed significantly decreased levels of ZO-1 and E-cadherin expression compared with control., Conclusion: Our results demonstrate that hypoxia altered the expression of junction complex molecules and increased epithelial permeability in human nasal epithelia. This suggests that hypoxia causes barrier dysfunction. Furthermore, it may be associated with innate immune dysfunction after encountering pathogens.

Background: Psoriasin is known to be expressed in diverse organs, where it exerts antimicrobial activity. Psoriasin is also involved in the local host defense mechanism against pathogens. We hypothesized that allergy-related T-helper cell type 2 (Th2) cytokines may regulate the expression of psoriasin., Methods: We treated normal human nasal epithelial (NHNE) cells with IL-4 or IL-13. Using human nasal tissues, we compared the expression level of psoriasin. We performed real-time polymerase chain reaction and Western blot assays using NHNE cells. Immunohistochemical staining and Western blot assays were performed with human nasal tissues. Furthermore, we studied the antimicrobial activity of nasal secretions from normal and allergic rhinitis patients., Results: IL-13 markedly down-regulated psoriasin expression at the gene and protein levels in NHNE cells, and it also decreased the amount of psoriasin protein that was secreted into the extracellular compartment in NHNE cells. IL-4 had no statistically significant effect. Results from immunohistochemical staining and Western blot assays showed that psoriasin expression was decreased in allergic rhinitis patients compared with control subjects. Nasal secretions of allergic rhinitis patients exhibited decreased antimicrobial activity compared with control subjects., Conclusion: We found that Th2 cytokines regulated psoriasin expression in NHNE cells, and psoriasin expression was decreased in allergic rhinitis patients compared with control subjects. The decreased expression of psoriasin may be related to the reduction in antimicrobial capacity of nasal secretions under allergic conditions, resulting in an increase in susceptibility to viruses or bacterial infections.

Background: It has been suggested that glucocorticoids might act in target tissues to increase their own intracellular availability in response to inflammatory stimuli. These mechanisms depend on the local metabolism of glucocorticoids catalyzed by 11β-hydroxysteroid dehydrogenase 1 (11β-HSD1) and 11β-hydroxysteroid dehydrogenase 2 (11β-HSD2)., Objective: This study is to investigate the effect of chronic rhinosinusitis (CRS) on expression of 11β-HSD1, 11β-HSD2, steroidogenic enzymes (cytochrome P450, family 11, subfamily B, polypeptide 1 [CYP11B1] and cytochrome P450, family 11, subfamily A, polypeptide 1 [CYP11A1]), and endogenous cortisol levels in human sinus mucosa. Expression levels were compared with those of healthy control subjects., Methods: The expression levels of 11β-HSD1, 11β-HSD2, CYP11B1, CYP11A1, and cortisol were measured in healthy control subjects, patients with CRS with nasal polyps, and patients with CRS without nasal polyps by using real-time PCR, Western blotting, immunohistochemistry, and ELISA. Expression levels of 11β-HSD1, 11β-HSD2, CYP11B1, CYP11A1, and cortisol were determined in cultured epithelial cells treated with CRS-relevant cytokines. The conversion ratio of cortisone to cortisol was evaluated by using the small interfering RNA technique, 11β-HSD1 inhibitor, and measurement of 11β-HSD1 activity., Results: 11β-HSD1, CYP11B1, and cortisol levels increased in patients with CRS with nasal polyps and those with CRS without nasal polyps, but 11β-HSD2 expression decreased. In cultured epithelial cells treated with IL-4, IL-5, IL-13, IL-1β, TNF-α, and TGF-β1, 11β-HSD1 expression and activity increased in parallel with expression levels of CYP11B1 and cortisol, but the production of 11β-HSD2 decreased. The small interfering RNA technique or the measurement of 11β-HSD1 activity showed that the sinus epithelium activates cortisone to cortisol in an 11β-HSD-dependent manner., Conclusion: These results indicate that CRS-relevant cytokines can modulate the expression of 11β-HSD1, 11β-HSD2, and CYP11B1 in the sinus mucosa, resulting in increasing intracellular concentrations of bioactive glucocorticoids., (Copyright © 2014. Published by Elsevier Inc.)

Background: Chronic sinusitis with nasal polyps (CRSwNPs) or CRS without NPs (CRSsNPs) is associated with expression of various cytokines. Lysophosphatidic acid (LPA) generated by autotaxin (ATX), LPA-producing enzyme, initiates signaling cascade involved in the inflammatory responses and participates in diverse biological processes through LPA receptors, including cytokine production. We analyzed the expression and distribution patterns of LPA-related molecules in nasal secretion and sinus mucosa of normal controls and patients with CRSwNPs and CRSsNPs, to evaluate the possible effects of the ATX-LPA receptor axis on the pathogenesis of CRS., Methods: LPA levels in nasal secretion and the expression and distribution patterns of ATX and LPA receptors 1-3 (LPA1-3) in sinus mucosa were investigated using ELISA, real-time polymerase chain reaction, Western blot, and immunohistochemistry. We elucidated the effect of CRS-relevant cytokines on the expression of ATX and LPA receptors, using cultured sinus epithelial cells, and investigated the effect of LPA on the expression of CRS-relevant cytokines, using sinus mucosa explant culture., Results: LPA, ATX, and LPA1-3 levels are increased in CRSwNPs and CRSsNPs. ATX and LPA1-3 were localized to superficial epithelium, submucosal glands in normal and inflammatory mucosa, but in inflammatory mucosa, they were found in inflammatory cells. LPA1-3 were noted in endothelium. Sinus mucosa explant stimulated with LPA increasingly produced IL-4, IL-5, interferon gamma, and TNF-alpha, and in cultured epithelial cells stimulated with CRS-relevant cytokines, ATX, and LPA1-3 were differentially induced., Conclusion: LPA in human sinus mucosa may play important roles in the pathogenesis of CRS, contributing to produce CRS-related cytokines. LPA-related molecules were increased in CRS, which may attribute to CRS-related cytokines.

Background: Chronic rhinosinusitis with nasal polyps (CRSwNPs) or CRS without nasal polyps (CRSsNPs) is characterized by persistent inflammation of sinonasal mucosa. No one causative factor fully explains for the pathological manifestations of CRS. Endogenous hydrogen sulfide (H2S) has been shown to participate in inflammatory diseases, functioning as an inflammatory mediator in various organs. We analyzed the contents and synthesis activity of H2S, the expression and distribution pattern of H2S-generating enzymes, cystathione β-synthase (CBS), and cystathione γ-lyase (CSE) in CRSwNPs and CRSsNPs. The effects of H2S on the expression of CRS-relevant cytokines and the effects of cytokines on the expression of CBS and CSE were assessed in an in vitro experiment., Methods: The contents and synthesis activity of H2S and the expression and distribution pattern of CBS and CSE in sinus mucosa were evaluated using spectrophotometry, real-time polymerase chain reaction, Western blot, and immunohistochemistry. Cultured epithelial cells were used to elucidate the effects of H2S donor, sodium hydrosulfide (NaHS), on the expression of CRS-relevant cytokines and the effects of cytokines on H2S-generating enzymes expression., Results: The contents and synthesis activity of H2S were increased in CRSwNPs and CRSsNPs. CBS and CSE were localized to the superficial epithelium and submucosal glands, but CSE was also found in vascular endothelium. NaHS induced increased expression of IL-4, IL-5, interferon γ, and TNF-α. CBS and CSE expression in cultured cells was up-regulated by CRS-relevant cytokines., Conclusion: H2S levels are increased in CRS, contributing to increased production of cytokines. These results suggest that H2S may function as inflammatory mediator in CRS.

Background: Prominent expression of YKL-40 has been associated with pathological conditions characterized by tissue remodeling. We determined the expression level and distribution pattern of YKL-40 in allergic nasal mucosa and evaluated the effect of YKL-40 on the proliferation and migration of fibroblasts, the production of the mediators related to tissue remodeling, and collagen production. Additionally, the cytokine-driven regulation of YKL-40 expression was evaluated in cultured epithelial cells., Methods: The expression of YKL-40 in normal, mild, and moderate/severe allergic nasal mucosa was evaluated using real-time polymerase chain reaction (PCR), Western blot, and immunohistochemistry. Fibroblast migration was observed using a scratch wound method, and proliferation was determined by the MTT methods. Expression of proteoglycans, transforming growth factor (TGF) beta1, MMP2, MMP9, TIMP1, TIMP2, and collagen concentration were analyzed in fibroblasts treated with YKL-40. The expression levels of YKL-40 in cultured epithelial cells were examined after stimulation with mediators including Th2 cytokines, interferon (IFN)gamma, and TNF-alpha with real-time PCR and ELISA., Results: The expression of YKL-40 was up-regulated in allergic rhinitis and distributed in superficial epithelium, submucosal glands, and vascular endothelium, in addition to infiltrating cells. TGF-beta1, TIMP1, MMP9, and biglycan were up-regulated in fibroblasts on stimulation with YKL-40, accompanying increased proliferation and migration, and collagen production. IL-13, IFN-gamma, and TNF-alpha induced the increased production of YKL-40 in cultured epithelial cells., Conclusion: YKL-40 is up-regulated in mild and moderate/severe persistent allergic rhinitis, and its expression can be regulated differentially by different cytokines, possibly contributing to the remodeling of nasal mucosa in allergic rhinitis.

Objectives/hypothesis: Hydrogen sulfide (H(2) S), the third endogenous gaseous transmitter, may be a crucial mediator in airway hyper-responsiveness and airway inflammation, including asthma. To elucidate the role of H(2) S in allergic rhinitis, the present study was undertaken to determine the level of expression of H(2) S in healthy nasal mucosa and mild and moderate/severe persistent allergic nasal mucosa as well as peripheral blood obtained from each patient. The expression and distribution pattern of the H(2) S-synthesizing enzymes cystathione γ-lyase (CSE) and cystathione β-synthase (CBS) were investigated in healthy and allergic nasal mucosa., Study Design: Controlled, prospective study., Methods: The concentration of H(2) S in nasal mucosa and plasma was determined by zinc trap spectrophotometry. The expression levels and patterns of distribution of CSE and CBS mRNA and proteins were evaluated using real time polymerase chain reaction, Western blot, and immunohistochemistry., Results: The levels of expression of H(2) S in nasal mucosa and plasma were increased in patients with mild and moderate/severe persistent allergic rhinitis compared with healthy controls. CSE was localized in vascular endothelium and surrounding muscles, and submucosal glands, whereas CBS was exclusively distributed in the superficial epithelium and submucosal glands. Their expression levels were increased in mild and moderate/severe persistent allergic rhinitis., Conclusions: The current findings indicate that, in parallel with increased expression levels of CSE and CBS, H(2) S is upregulated in nasal mucosa and plasma of allergic patients. Based on localization of CSE and CBS, H(2) S may play multiple functions in human nasal mucosa, contributing to the development of allergic symptoms such as rhinorrhea, sneezing, and nasal stuffiness., (Copyright © 2013 The American Laryngological, Rhinological, and Otological Society, Inc.)

Background: The aim of this study was to evaluate the role of suppressor of cytokine signaling (SOCS) molecules, SOCS1 and SOCS3, which act as negative regulators of cytokine signaling in various allergic diseases, in patients with mild and moderate/severe persistent allergic rhinitis., Methods: The expression and distribution pattern of SOCS1 and SOCS3 were analyzed in nasal mucosa and peripheral blood mononuclear cells (PBMC) of healthy controls, and patients with mild and moderate/severe persistent allergic rhinitis using RT-PCR, immunohistochemistry and Western blotting. IL-4, IL-13, IL-15 and IFN-γ expression was also analyzed in the nasal mucosa of each individual using RT-PCR and Western blotting., Results: SOCS1 and SOCS3 mRNA and protein expression was significantly increased in the nasal mucosa and PBMC of patients with mild and moderate/severe persistent allergic rhinitis compared with healthy controls. In healthy and allergic nasal mucosa, they were commonly localized to the epithelium, submucosal glands and endothelium, showing stronger staining intensity in mild and moderate/severe persistent allergic nasal mucosa than in healthy nasal mucosa. Tissue levels of IL-4 and IL-13 were increased in moderate/severe persistent allergic nasal mucosa whereas IL-15 and IFN-γ were decreased in moderate/severe persistent allergic nasal mucosa., Conclusions: Upregulation of SOCS1 and SOCS3 in mild and moderate/severe persistent allergic rhinitis suggests that SOCS proteins may be important regulators in the pathogenesis of allergic rhinitis and play a role as molecular determinants of allergic rhinitis persistence., (Copyright © 2012 S. Karger AG, Basel.)

Background: Adiponectin, one of the adipokines, has been implicated in the inflammatory process in patients with allergic rhinitis. The level of adiponectin is affected by immunotherapy. Considering the fact that adiponectin receptors (AdipoRs) mediate intracellular signaling events in response to the binding of adiponectin, the role of AdipoRs in healthy and allergic nasal mucosa should be determined. This study investigates the level of expression and distribution pattern of AdipoR1 and AdipoR2 in healthy, mild, and moderate/severe persistent allergic nasal mucosa to understand the role of adiponectin in allergic rhinitis., Methods: The level of expression and distribution pattern of AdipoR1 and AdipoR2 were evaluated in healthy, mild, and moderate/severe persistent allergic nasal mucosa, using semiquantitative reverse-transcriptase-polymerase chain reaction (RT-PCR), immunohistochemistry, and Western blot analysis., Results: AdipoR1 was expressed in healthy, mild, and moderate/severe persistent allergic nasal mucosa where it was commonly localized to the vascular endothelium. However, AdipoR2 was not expressed in any samples of nasal mucosa tested in the present study. Semiquantitative RT-PCR and Western blot analysis showed that the level of expression of AdipoR1 mRNA and protein was decreased in mild and moderate/severe persistent allergic nasal mucosa in comparison with healthy nasal mucosa, but not significantly different between mild and moderate/severe persistent allergic nasal mucosa., Conclusion: These results indicate that AdipoR1 may play an important role in the pathogenesis of allergic nasal mucosa, suggesting a role for AdipoR1 in vascular dysfunction in mild and moderate/severe persistent allergic nasal mucosa.

Background: A disintegrin and metalloproteases (ADAMs) constitute a family of cell surface proteins containing disintegrin and metalloprotease domain, which associate features of adhesion molecules and proteases. ADAM8 is a member of the ADAM family and has recently been implicated to have a role in allergic lung inflammation. Therefore, our aims were to determine the expression of ADAM8 mRNA and protein and to localize ADAM8 protein in the nasal mucosa of patients with allergic rhinitis and control subjects., Methods: Inferior turbinate mucosa samples were obtained from 20 patients with allergic rhinitis and 20 matched healthy normal controls. ADAM8 mRNA was extracted from the inferior turbinate mucosa, and then reverse transcription-polymerase chain reaction was performed. Western blot testing was used to analyze differences in the level of ADAM8 protein expression between patients with allergic rhinitis and normal controls, and the ADAM8 protein was localized with immunohistochemical staining., Results: The level of expression of ADAM8 mRNA and protein in the nasal mucosa was significantly increased in patients with allergic rhinitis compared with normal controls. ADAM8 protein was expressed in the epithelium, infiltrating inflammatory cells, and submucosal glands., Conclusion: ADAM8 is expressed in human nasal mucosa and is increased in patients with allergic rhinitis. These results suggest a possible contribution for ADAM8 in chronic inflammation of the nasal mucosa in patients with allergic rhinitis.

Conclusions: These results suggest that nestin and BMI-1 are candidates for stem cell markers and renewal factors in human nasal mucosa, may contribute to tissue homeostasis and differentiation in the epithelium and submucosal glands of normal nasal mucosa, and may play a role in proliferation of nasal polyps., Objectives: The stem cell marker, nestin, and the stem cell renewal factor, BMI-1, have been identified in a variety of inflammatory and normal tissues, implicating these markers in tissue regeneration., Materials and Methods: We investigated the expression and distribution of nestin and BMI-1 in normal nasal mucosa and nasal polyps, using RT-PCR, immunohistochemistry, and Western blotting., Results: Nestin and BMI-1 were localized to the epithelium and submucosal glands of normal nasal mucosa and nasal polyps. The expression of nestin was confined to the plasma membrane and cytoplasm, whereas BMI-1 showed a nuclear staining pattern. In normal nasal mucosa and nasal polyps, nestin and BMI-1 expression was strongest in the basal portion of the epithelial layer, and decreased toward the upper portion. In the submucosal glands, weak to strong expression was commonly detected in the glandular acini. There was no significant difference in the level of expression of nestin and BMI-1 between normal nasal mucosa and nasal polyps.

Objectives: To investigate the expression levels and distribution patterns of elafin and cystatin C in normal and inflammatory human sinus mucosa and to evaluate their roles in chronic sinusitis., Design: A controlled, prospective study., Setting: A tertiary academic institution., Patients: Normal sinus mucosa was obtained from the ethmoid sinus during surgery in 30 patients with blowout fractures. Inflammatory sinus mucosa was obtained from 30 patients undergoing endoscopic sinus surgery for chronic polypoid sinusitis., Interventions: Reverse transcription-polymerase chain reaction, immunohistochemical analysis, and Western blotting., Main Outcome Measures: Expression levels and distribution patterns of elafin and cystatin C in normal and inflammatory mucosa., Results: Expression of elafin and cystatin C messenger RNAs and proteins analyzed by means of reverse transcription-polymerase chain reaction and Western blot was detected in all normal and inflammatory sinus mucosa tested. Their expression levels were increased in inflammatory vs normal mucosa. Elafin in normal and inflammatory sinus mucosa was distinctly expressed in goblet cells, which are increased in inflammatory sinus mucosa. Elafin in submucosal glands was usually weak in staining intensity, except for a few scattered submucosal glands showing moderate intensity in inflammatory sinus mucosa. Cystatin C was also localized in goblet cells and submucosal glands in normal and inflammatory mucosa. Staining intensity was increased more in inflammatory vs normal sinus mucosa., Conclusion: Elafin and cystatin C may play an important role in the protection of normal sinus mucosa and further in regulation of the inflammatory condition in chronic sinusitis.

Background: The purpose of this study was to investigate the expression and distribution of superoxide anion, NADPH oxidase (NOX)1, and NOX4 in healthy, allergic nasal mucosa and nasal polyps to evaluate the possible influence of oxidative stress on the development of allergic rhinitis and nasal polyps., Methods: The expression and distribution of superoxide anion, NOX1 and NOX4 were evaluated in healthy and allergic nasal mucosa and nasal polyps, using dihydroethidium fluorescence, semiquantitative reverse transcriptase-polymerase chain reaction, immunohistochemistry, and Western blot., Results: NOX1 and NOX4 were localized mainly in the epithelial layer, submucosal glands, vascular endothelium, and inflammatory cells in healthy and allergic nasal mucosa and nasal polyps. The cellular source that generated superoxide anion is also localized in the epithelial cells, submucosal glands, vascular endothelium, and inflammatory cells, demonstrating the similar sites of expression of NOX1 and NOX4 in healthy and allergic nasal mucosa and nasal polyps. NOX1 and NOX4 mRNA and proteins and superoxide anions had increased levels of expression in allergic nasal mucosa and nasal polyps compared with healthy nasal mucosa., Conclusions: These results indicate that NOX1 and NOX4 may play an important role in reactive oxygen species production, contributing to the oxidative stress in allergic rhinitis and nasal polyp tissues.

Background: Heparanase (HPA) is known to be involved in tissue remodeling of various organs with inflammatory diseases. The aim of this study was to investigate the level of expression and the pattern of distribution of HPA in normal human sinus mucosa, inflammatory sinus mucosa, and nasal polyps to evaluate the possible effect of HPA on the tissue remodeling of chronic inflammatory sinus mucosa and nasal polyps., Methods: Normal sinus mucosa was obtained from the ethmoid sinus during endoscopic reduction in 25 patients with blowout fractures. Inflammatory sinus mucosa and nasal polyps were obtained from 25 patients undergoing endoscopic sinus surgery for chronic sinusitis with nasal polyps. The levels of expression and the pattern of distribution of HPA were evaluated in normal human sinus mucosa, inflammatory sinus mucosa, and nasal polyps, using reverse transcriptase-polymerase chain reaction, immunohistochemical analysis, and Western blot analysis., Results: HPA mRNA and protein were detected in inflammatory sinus mucosa and nasal polyps but not in normal sinus mucosa. HPA was mainly localized in the vascular endothelium, epithelium, submucosal glands, and inflammatory cells of inflammatory sinus mucosa. In nasal polyps, inflammatory cells and vascular endothelium showed immunopositivity in the entire portion, whereas glands and epithelial cells did not show positivity., Conclusion: Our results indicate that HPA is not constitutively expressed in normal sinus mucosa and is upregulated in chronic inflammatory sinus mucosa and nasal polyps, suggesting that HPA may play an important role in the tissue remodeling in chronic sinusitis.

Background: Although the complement anaphylatoxin peptides, complement anaphylatoxin 3a (C3a) and complement anaphylatoxin 5a (C5a), are implicated in the inflammatory process in allergic rhinitis, a direct interaction between allergic mucosa and complement receptors has not been demonstrated., Objective: We investigated the expressional levels and distributional patterns of the C3a receptor (C3aR) and C5a receptor (C5aR) in normal, mild, and severe persistent allergic nasal mucosa., Methods: Immunohistochemistry and Western blotting using C3aR and C5aR antibodies were applied to normal nasal, mild, and severe persistent allergic nasal mucosa., Results: Complement anaphylatoxin 5a receptor was detected in the inflammatory cells of normal and allergic nasal mucosa, and its expression level was significantly higher in allergic nasal mucosa than normal nasal mucosa. C3aR in normal and allergic nasal mucosa was commonly expressed in nonmyeloid cells such as epithelial cells, submucosal glands, and nerve fibers. In addition, C3aR was expressed in the endothelium of cavernous sinuses and the surrounding perivascular muscle layer in severe persistent allergic nasal mucosa, but not in normal and mild allergic nasal mucosa. Western blotting demonstrated that the expression level of C3aR was significantly increased in severe persistent allergic nasal mucosa compared with normal and mild nasal mucosa., Conclusion: On the basis of the location of C3aR and C5aR, C5aR may play a role in activation of inflammatory cells, whereas C3aR may mediate mucus secretion and mucosal swelling in allergic nasal mucosa, especially severe persistent allergic nasal mucosa.

Objectives: To investigate the expression and distribution of glycoprotein 340 (gp340), a secretory glycoprotein, in normal human sinus mucosa and inflammatory sinus mucosa and evaluate the possible effects of gp340 on the development of chronic sinusitis. Glycoprotein 340 was identified as a key element in the innate host defense mechanism on many mucosal surfaces and is directly involved in defense functions aimed at clearing gram-positive and gram-negative bacteria., Design: Prospective study., Setting: Tertiary academic institution., Patients: Normal sinus mucosa was obtained from the ethmoid sinus mucosa of 8 patients with blowout fractures undergoing endoscopic reduction. Inflammatory sinus mucosa was taken from 25 patients with chronic polypoid sinusitis during endoscopic sinus surgery., Intervention: Semiquantitative reverse transcription-polymerase chain reaction (RT-PCR), immunohistochemical analysis, and Western blotting were performed., Main Outcome Measures: The expression level and distributional pattern of gp340 in normal and inflammatory sinus mucosa were analyzed., Results: Transcripts of the gp340 gene were detected in all human sinus tissues analyzed by RT-PCR. Immunohistochemical analysis revealed that gp340 is mainly localized in submucosal gland of both normal and inflammatory sinus mucosa. Semiquantitative RT-PCR and Western blot analysis showed the increased expression levels of gp340 in the inflammatory sinus mucosa compared with the normal sinus mucosa., Conclusion: These results suggest that gp340 may play a constitutive role in nasal defense and may be up-regulated in response to inflammation, participating in antimicrobial defense in chronic sinusitis.

Objectives: Urocortin (UCN) is a member of the corticotropin releasing factor (CRF) neuropeptide family. UCN act as locally expressed proinflammatory factor and induce mast cell degranulation, cytokine secretion, and trigger vascular permeability, which are mediated by CRF receptors in peripheral tissues. Considering its functional roles, UCN and its receptors may play a role in the pathogenesis of allergic nasal mucosa. Therefore, we investigated the expression profile and distribution of UCN and CRF receptors in normal and allergic nasal mucosa., Methods: Reverse transcriptase-polymerase chain reaction, immunohistochemistry, and Western blotting were applied to the normal and allergic nasal mucosa., Results: The expression levels of UCN and CRF receptors were increased in allergic nasal mucosa in comparison with normal nasal mucosa. In normal nasal mucosa, UCN and CRF receptors were restricted to the vascular endothelium of submucosal cavernous sinusoids where faint staining was found. However, in allergic nasal mucosa, UCN was expressed in small vessels distributed in lamina propria and the vascular endothelium of cavernous sinusoid located in submucosa. Many scattered positive cells were also found in allergic nasal mucosa, probably UCN-positive leukocytes. CRF receptors were also localized in the vascular endothelium of small vessels and cavernous sinusoid., Conclusions: These results indicate that UCN may play a role in the regulation of vascular swelling in normal nasal mucosa. Moreover, in allergic nasal mucosa, increased expression levels of UCN and its receptors may contribute to increased mucosal swelling and vascular permeability, playing an important role in the pathogenesis of allergic rhinitis.

An immunohistochemical staining technique with the D2-40 antibody was undertaken to examine the functional and morphological features of lymphatic networks in tissue sections and whole-mount preparations of normal nasal mucosa and ethmoid sinus mucosa. In normal nasal mucosa, most lymphatic vessels were found in the superficial mucosa beneath the epithelial layer. Some of these vessels were dilated, whereas others were compressed and had a slit-like lumen. Whole-mount preparations revealed the extent of lymphatic vessels in normal ethmoid sinus mucosa. A network of lymphatic vessels was mainly found in the subepithelial layer, where lymphatic vessels represented rich networks, possessing antler-like branches and typical blind ends. However, these lymphatic networks were not arranged in the form of lymphangion chains, with each lymphangion consisting of a contractile compartment and valve. Thus, recognition of the distinctive features of the lymphatic network in normal nasal and sinus mucosa might aid investigations of lymphatic involvement in sinonasal diseases, such as rhinitis, sinusitis, and malignancy.

Background: Nasal polyps and inverted papillomas often look similar. Clinically, it is difficult to distinguish the masses by endoscopic examination. Therefore, in this study, we aimed to develop a deep learning algorithm for computer-aided diagnosis of nasal endoscopic images, which may provide a more accurate clinical diagnosis before pathologic confirmation of the nasal masses. Methods: By performing deep learning of nasal endoscope images, we evaluated our computer-aided diagnosis system's assessment ability for nasal polyps and inverted papilloma and the feasibility of their clinical application. We used curriculum learning pre-trained with patches of nasal endoscopic images and full-sized images. The proposed model's performance for classifying nasal polyps, inverted papilloma, and normal tissue was analyzed using five-fold cross-validation. Results: The normal scores for our best-performing network were 0.9520 for recall, 0.7900 for precision, 0.8648 for F1-score, 0.97 for the area under the curve, and 0.8273 for accuracy. For nasal polyps, the best performance was 0.8162, 0.8496, 0.8409, 0.89, and 0.8273, respectively, for recall, precision, F1-score, area under the curve, and accuracy. Finally, for inverted papilloma, the best performance was obtained for recall, precision, F1-score, area under the curve, and accuracy values of 0.5172, 0.8125, 0.6122, 0.83, and 0.8273, respectively. Conclusion: Although there were some misclassifications, the results of gradient-weighted class activation mapping were generally consistent with the areas under the curve determined by otolaryngologists. These results suggest that the convolutional neural network is highly reliable in resolving lesion locations in nasal endoscopic images. [ABSTRACT FROM AUTHOR]

This study aimed to identify and evaluate drug candidates targeting the kinase inhibitory region of suppressor of cytokine signaling (SOCS) 3 for the treatment of allergic rhinitis (AR). Utilizing an artificial intelligence (AI)-based new drug development platform, virtual screening was conducted to identify compounds inhibiting the SH2 domain binding of SOCS3. Luminescence assays assessed the ability of these compounds to restore JAK-2 activity diminished by SOCS3. Jurkat T and BEAS-2B cells were utilized to investigate changes in SOCS3 and STAT3 expression, along with STAT3 phosphorylation in response to the identified compounds. In an OVA-induced allergic rhinitis mouse model, we measured serum levels of total IgE and OVA-specific IgE, performed real-time PCR on nasal mucosa samples to quantify Th2 cytokines and IFN-γ expression, and conducted immunohistochemistry to analyze eosinophil levels. Screening identified 20 hit compounds with robust binding affinities. As the concentration of SOCS3 increased, a corresponding decrease in JAK2 activity was observed. Compounds 5 and 8 exhibited significant efficacy in restoring JAK2 activity without toxicity. Treatment with these compounds resulted in reduced SOCS3 expression and the reinstatement of STAT3 phosphorylation in Jurkat T and BEAS-2B cells. In the OVA-induced allergic rhinitis mouse model, compounds 5 and 8 effectively alleviated nasal symptoms and demonstrated lower levels of immune markers compared to the allergy group. This study underscores the promising nonclinical efficacy of compounds identified through the AI-based drug development platform. These findings introduce innovative strategies for the treatment of AR and highlight the potential therapeutic value of targeting SOCS3 in managing AR. [ABSTRACT FROM AUTHOR]

Chronic rhinosinusitis (CRS) is an inflammation of the nasal and paranasal sinus mucosa, and eosinophilic CRS (eCRS) is a subtype characterized by significant eosinophil infiltration and immune response by T-helper-2 cells. The pathogenesis of eCRS is heterogeneous and involves various environmental and host factors. Proteases from external sources, such as mites, fungi, and bacteria, have been implicated in inducing type 2 inflammatory reactions. The balance between these proteases and endogenous protease inhibitors (EPIs) is considered important, and their imbalance can potentially lead to type 2 inflammatory reactions, such as eCRS. In this review, we discuss various mechanisms by which exogenous proteases influence eCRS and highlight the emerging role of endogenous protease inhibitors in eCRS pathogenesis. [ABSTRACT FROM AUTHOR]

Dendritic cells (DCs) are antigen-presenting cells derived from the bone marrow that play an important role in the association between the innate and adaptive immune responses. The onset and development of chronic rhinosinusitis (CRS) involve a serious imbalance in immune regulation and mechanical dysfunction caused by an abnormal remodeling process. Recent studies have shown that an increase in DCs in CRS and their function of shaping the nasal mucosal immune response may play an important role in the pathogenesis of CRS. In this review, we discuss DC subsets in mice and humans, as well as the function of DCs in the nasal sinus mucosa. In addition, the mechanism by which DCs can be used as targets for therapeutic intervention for CRS and potential future research directions are also discussed. [ABSTRACT FROM AUTHOR]

Nasal drug delivery is advantageous when compared with other routes of drug delivery as it avoids the hepatic first-pass effect, blood–brain barrier penetration, and compliance issues with parenteral administration. However, nasal administration also has some limitations, such as its low bioavailability due to metabolism on the mucosal surface, and irreversible damage to the nasal mucosa due to the ingredients added into the formula. Moreover, the method of nasal administration is not applicable to all drugs. The current review presents the nasal anatomy and mucosal environment for the nasal delivery of vaccines and drugs, as well as presents various methods for enhancing nasal absorption, and different drug carriers and delivery devices to improve nasal drug delivery. It also presents future prospects on the nasal drug delivery of vaccines and drugs. [ABSTRACT FROM AUTHOR]

Objectives/Hypothesis: Carbonic anhydrase (CA) is a zinc metalloenzyme that participate in the biological processes of various fluid transporting epithelia, including ion and water transport. CA may thus play a role in the pathophysiology of normal nasal mucosa and nasal polyp. We evaluated the expression and pattern of distribution of mRNAs and proteins for CA isoenzymes in normal nasal mucosa and polyps. Study Design: This was a controlled, prospective study. Methods: The expression levels of 11 isoenzyme genes (I, II, III, IV, VA, VB, VI, VII, IX, XII, XIV) were evaluated in normal mucosa and polyps using semiquantitative reverse-transcriptase polymerase chain reaction. The expression and pattern of distribution of CA I, II, and IX were also investigated using immunohistochemistry and Western blot. Results: mRNAs for the 11 CA isoenzymes were detected in all normal nasal mucosa and polyps tested. Their expression levels were decreased in nasal polyp in comparison with normal nasal mucosa. CA I was detected in the epithelial cells, endothelial cells, and submucosal gland of normal nasal mucosa, but only in the endothelial cells in nasal polyp. CA II in normal mucosa was intensely expressed in the submucosal gland cells and CA IX was noted in the cytoplasm and plasma membrane of epithelial cells and submucosal glands. However, these findings were not found in nasal polyp. Western blot also showed differences in their expression levels. Conclusions: These results indicate that CA may participate in the physiology of normal nasal mucosa, but also suggest that altered their expression in nasal polyps may cause impaired electrolyte and water transport across the epithelial cells. [ABSTRACT FROM AUTHOR]

Chronic rhinosinusitis (CRS) is a chronic inflammatory condition of the nasal and paranasal sinus mucosa that affects up to 10% of the population worldwide. CRS is the most representative disease of the upper respiratory tract where airway remodeling occurs, including epithelial damage, thickening of the basement membrane, fibrosis, goblet cell hyperplasia, subepithelial edema, and osteitis. CRS is divided into two phenotypes according to the presence or absence of nasal polyps: CRS with nasal polyp (CRSwNP) and CRS without nasal polyps (CRSsNP). Based on the underlying pathophysiologic mechanism, CRS is also classified as eosinophilic CRS and non-eosinophilic CRS, owing to Type 2 T helper (Th2)-based inflammation and Type 1 T helper (Th1)/Type 17 T helper (Th17) skewed immune response, respectively. Differences in tissue remodeling in CRS are suggested to be based on the clinical phenotype and endotypes; this is because fibrosis is prominent in CRSsNP, whereas edematous changes occur in CRSwNP, especially in the eosinophilic type. This review aims to summarize the latest information on the different mechanisms of airway remodeling in CRS according to distinct endotypes. [ABSTRACT FROM AUTHOR]

Background: Exposure to air particulate matter (PM) is associated with various diseases in the human respiratory system. To date, most in vitro studies showing cellular responses to PM have been performed in cell culture using a single cell type. There are few studies considering how multicellular networks communicate in a tissue microenvironment when responding to the presence of PM. Here, an in vitro three-dimensional (3D) respiratory mucosa-on-a-chip, composed of human nasal epithelial cells, fibroblasts, and endothelial cells, is used to recapitulate and better understand the effects of urban particulate matter (UPM) on human respiratory mucosa. Results: We hypothesized that the first cells to contact with UPM, the nasal epithelial cells, would respond similar to the tissue microenvironment, and the 3D respiratory mucosa model would be a suitable platform to capture these events. First, whole transcriptome analysis revealed that UPM induced gene expression alterations in inflammatory and adhesion-related genes in human nasal epithelial cells. Next, we developed an in vitro 3D respiratory mucosa model composed of human nasal epithelial cells, fibroblasts, and endothelial cells and demonstrated that the model is structurally and functionally compatible with the respiratory mucosa. Finally, we used our model to expose human nasal epithelial cells to UPM, which led to a disruption in the integrity of the respiratory mucosa by decreasing the expression of zonula occludens-1 in both the epithelium and endothelium, while also reducing vascular endothelial cadherin expression in the endothelium. Conclusions: We demonstrate the potential of the 3D respiratory mucosa model as a valuable tool for the simultaneous evaluation of multicellular responses caused by external stimuli in the human respiratory mucosa. We believe that the evaluation strategy proposed in the study will move us toward a better understanding of the detailed molecular mechanisms associated with pathological changes in the human respiratory system. [ABSTRACT FROM AUTHOR]

Background: Chronic rhinosinusitis (CRS) is an inflammatory disease of the sinonasal mucosa. Thymic stromal lymphopoietin (TSLP) is associated with T‐helper 2 (Th2) response and induced by pathogen, allergen, toll‐like receptor (TLR) ligands, and cytokines. Fibroblasts are known to be modulators of wound‐healing, from inflammation to tissue remodeling. We examined effect of lipopolysaccharide (LPS) on TSLP production and the underlying mechanisms. We aimed to determine whether the effects of commonly used medications in CRS, namely corticosteroids, and macrolides, are related to LPS‐induced TSLP in nasal fibroblasts. Methods: Fibroblasts were isolated from inferior turbinate tissues of CRS patients. TSLP and TLR4 expressions were determined by reverse transcript‒polymerase chain reaction (RT‐PCR), Western blot, enzyme‐linked immunoassay, and immunofluorescence staining. Mitogen‐activated protein kinase (MAPK), protein kinase B (Akt), and nuclear factor‐kappaB (NF‐κB) phosphorylation was determined by Western blot and/or luciferase assay. Results: LPS increased TSLP expression in a dose‐ and time‐dependent manner. LPS antagonist and corticosteroids inhibited TLR4 expression in LPS‐stimulated fibroblasts. LPS‐RS, macrolides, corticosteroids, and specific inhibitors suppressed LPS‐induced alterations. Ex vivo culture showed similar results. Conclusion: LPS induces TSLP production via the TLR4, MAPK, Akt, and NF‐κB pathways. The effects of corticosteroids and macrolides are related to LPS‐induced TSLP expression. We explored new treatment modalities targeting LPS‐induced TSLP production that could replace the currently used corticosteroid and macrolides for treatment of CRS. [ABSTRACT FROM AUTHOR]