Posted on January 24, 2022
Our hypothesis was that among the inflammatory cytokines involved in NPs, IL-6 could alter epithelial repair mechanisms and mucociliary clearance
Our hypothesis was that among the inflammatory cytokines involved in NPs, IL-6 could alter epithelial repair mechanisms and mucociliary clearance. ciliated edges of primary cultures were analyzed on IL-6 modulation by digital high-speed video-microscopy to measure: ciliary beating frequency (CBF), ciliary length, relative ciliary density, metachronal wavelength and the ciliary beating efficiency index. Results Our results showed that: (i) IL-6 accelerated airway wound repair in vitro, with a doseCresponse effect whereas no effect was observed after other ILs-stimulation. After 24?h, 79% of wounded wells with IL6-100 were fully repaired, vs 46% in the IL6-10 group, 28% in the IL6-1 group and 15% in the control group; (ii) specific migration analyses of closed wound at late repair stage (Day 12) showed IL-6 had the highest migration compared with other ILs (iii) The study of the IL-6 effect on ciliary function showed that CBF and metachronal wave increased but without significant modifications of ciliary density, length of cilia and efficiency index. Conclusion The up-regulated epithelial cell proliferation observed in polyps could be induced by IL-6 in the case of prior epithelial damage. IL-6 could be a major cytokine in NP physiopathology. repair of the nasal airway epithelium has been described in NPs [3, 11, 12]. In addition to epithelial cell dysfunction, a type 2 inflammatory pattern involving expression of interleukins (IL) Pyridoxine HCl IL-4, -5, and -13 and increased concentrations of IgE, has been reported in the NPs of 85% of patients with CRSwNP in western countries . Evidence of high levels of IL-6 expression has already been reported in NPs [14, 15]. IL-6 plays an Pyridoxine HCl important role in the development and progression of inflammatory responses, autoimmune diseases, and cancers. IL-6 can induce tissue damage, inflammation and cell proliferation [16C18]. To date, no study has precisely described the role of IL-6 in CRSwNP, and particularly its effect on mucociliary clearance, although one study does describe the effect of IL-6 on the regeneration of airway ciliated cells from basal stem cells . More recently, high concentrations of IL-9 and IL-10 have been described in NPs but their influence on nasal airway epithelial cell dysfunction are unknown [16, 20]. Our hypothesis was that inflammatory cytokines in NPs, particularly IL-6, could be responsible for alteration of sinonasal epithelial cell functions (i.e. dysfunction of repair mechanisms and mucociliary clearance) thus creating favorable conditions for chronic inflammation and polyp growth. We thus set out to investigate in vitro the relationship between nasal epithelial cell functions and ILs. We developed air-liquid interface (ALI) cultures of primary differentiated human nasal epithelial cells (HNEC) that can be used as an in vitro wound repair and ciliary beating evaluation model. Our results suggest new mechanisms of epithelial cell-IL relationships and may lead to the identification of novel therapeutic pathways that could improve treatment for patients with CRSwNP . Methods In healthy conditions, after a mechanical wound, epithelium repair mechanisms involve cell migration, followed by Mouse monoclonal to Fibulin 5 a cell proliferation phase, epithelial junction and finally a differentiation phase of basal cells in ciliated cells . The restoration of barrier integrity and mucociliary clearance after epithelial injury represent a key step in the defense capacity of the airway epithelium . We aimed to evaluate these mechanisms of epithelial repair with and without IL modulation in cultures of HNEC from NPs. Primary Cultures of Human Nasal Epithelial Cells (HNEC) NPs were obtained from 11 patients with CRSwNP undergoing ethmoidectomy. CRSwNP is a heterogeneous inflammatory disease with various underlying pathophysiologic mechanisms which correspond to?different endotypes and clinical manifestations of the disease . In this study, our samples were obtained from the most severe patients, i.e. those with medically uncontrolled CRSwNP and needing surgery. However, to ensure the homogeneity of the samples, all patients were required to stop oral corticosteroids treatment 1?month before surgery, and in all cases, surgery was decided after at least 3?months of well-conducted Pyridoxine HCl medical treatment with daily intranasal corticosteroids. All the patients had given informed consent and the study was approved by the local ethics committee (CPP IDF X 2016-01-01). HNECs were isolated from NPs as previously described . Briefly, the NPs were immediately placed in DMEM/F-12 supplemented with antibiotics (100 U/ml penicillin, 100?mg/ml streptomycin, 2.5?g/ml amphotericin B, and 100?mg/ml gentamicin) and sent to the laboratory for.