Inducing a Mucosal Barrier–Sparing Inflammatory Response in Laboratory‐Grown Primary Human Nasal Epithelial Cells

Hayes

et al. 2019

JCM

Background: Chronic rhinosinusitis (CRS) is defined as a chronic inflammation of the nose and paranasal sinus mucosa associated with relapsing infections—particularly with S. aureus. Long-term treatments with protein synthesis inhibitor antibiotics have been proposed to reduce inflammation in the context chronic severe inflammatory airway pathologies, including CRS. This study assessed the effect of subinhibitory clindamycin and azithromycin on S. aureus exoprotein induced inflammation, toxicity and invasiveness. Methods: S. aureus ATCC51650 and two clinical isolates grown in planktonic and biofilm form were treated with subinhibitory clindamycin and azithromycin. Exoproteins were collected and applied to primary human nasal epithelial cells (HNECs) in monolayers and at air-liquid interface. This was followed by lactate dehydrogenase (LDH), enzyme-linked immunosorbent assay (ELISA), Transepithelial Electrical Resistance (TEER) and paracellular permeability assays to assess the effect on cell toxicity, inflammatory cytokine production and mucosal barrier structure and function, respectively. The effect of these treatments was tested as well on the S. aureus invasiveness of HNECs. Results: Subinhibitory clindamycin reduced S. aureus exoprotein production in planktonic and biofilm form, thereby blocking exoprotein-induced toxicity, reversing its detrimental effects on mucosal barrier structure and function and modulating its inflammatory properties. Sub-inhibitory azithromycin had similar effects—albeit to a lesser extent. Furthermore, clindamycin—but not azithromycin—treated S. aureus lost its invasive capacity of HNECs. Conclusion: Subinhibitory clindamycin and azithromycin reduce S. aureus exoprotein production, thereby modulating the inflammatory cascade by reducing exoprotein-induced toxicity, inflammation, mucosal barrier disruption and invasiveness.

Section: Methodsmentioning

confidence: 99%

Sub-Inhibitory Clindamycin and Azithromycin reduce S. aureus Exoprotein Induced Toxicity, Inflammation, Barrier Disruption and Invasion

Hayes

et al. 2019

JCM

“…The extent of adhesion formation is multifactorial, and depends significantly on the level of inflammation, ROS production and fibroblast migration during that time (Eming et al, 2007). Fibroblasts are responsible for collagen synthesis, and a decrease in collagen synthesis and fibroblast cell migration results in slower, more measured wound repair that is less likely to result in an adhesion (Rajan and Murray, 2008;Munireddy et al, 2010). Our data implies that some, but not all, antibiotics might be beneficial in preventing adhesions after nasal surgery as they restrict fibroblast cell migration without negatively affecting the reepithelialization.…”

Section: Discussionmentioning

confidence: 99%

“…In an in vitro wound model of hypertrophic scar fibroblasts, a microarray analysis indicated the interleukin 6 (IL-6) signaling pathway to be the main pathway involved in the early response to injury in those cells (Tosa et al, 2005). Moreover IL-6, along with other pro-inflammatory factors such as interleukin (IL)-1α, IL-1β and tumor necrosis factor-α are up regulated in hypertrophic scar tissues (Dong et al, 2013;Ramezanpour et al, 2019b). Certain limitations must be considered for the present study.…”

Section: Discussionmentioning

confidence: 99%

“…On day 3 after seeding, the apical media was removed and the basal media replaced with B-ALI TM differentiation media, exposing the apical cell surface to the atmosphere. Human nasal epithelial cultures at air liquid interface (HNEC-ALI) were maintained for a minimum of 21 days prior to experimentation for development of tight junctions (Ramezanpour et al, 2019b).…”

Section: Air Liquid Interface Culturementioning

confidence: 99%

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Antibiotics Affect ROS Production and Fibroblast Migration in an In-vitro Model of Sinonasal Wound Healing

Gouzos

Front. Cell. Infect. Microbiol.

Bassiouni

et al. 2020

Self Cite

Introduction: Antibiotics are often administered to patients perioperatively and have been shown to affect ROS production of nasal cells in vitro, but their effect in the setting of active wound healing remains unclear. Reactive oxygen species (ROS) are known to play a significant role in wound healing. This study analyzed a broad array of antibiotics used after sinus surgery to assess their effect on wound healing and ROS production in vitro. It was hypothesized that ROS production would be affected by these antibiotics and there would be a negative relationship between ROS activity and cell migration speed. Methods: Monolayers of primary human nasal epithelial cells (HNEC) and primary fibroblasts were disrupted with a linear wound, treated with 10 different antibiotics or a ROS inhibitor and observed over 36 h in a controlled environment using confocal microscopy. ROS activity and migration speed of the wound edge were measured at regular intervals. The relationship between the two parameters was analyzed using mixed linear modeling. Results: Performing a linear scratch over the cell monolayers produced an immediate increase in ROS production of ∼35% compared to unscratched controls in both cell types. Incubation with mitoquinone and the oxazolidinone antibiotic linezolid inhibited ROS activity in both fibroblasts and HNEC in association with slowed fibroblast cell migration (p < 0.05). Fibroblast cell migration was also reduced in the presence of clarithromycin and mupirocin (p < 0.05). A significant correlation was seen between ROS suppression and cell migration rate in fibroblasts for mitoquinone and all antibiotics except for azithromycin and doxycycline, where no clear relationship was seen. Treatments that slowed fibroblast cell migration compared to untreated controls showed a significant correlation with ROS suppression (p < 0.05). Conclusion: Increased ROS production in freshly wounded HNEC and fibroblast cell monolayers was suppressed in the presence of antibiotics, in correlation with reduced fibroblast cell migration. In contrast, HNEC cell migration was not significantly affected by any of the antibiotics tested. This differential effect of antibiotics on fibroblast and HNEC migration might have clinical relevance by reducing adhesion formation without affecting epithelial healing in the postoperative setting.

“…The apical media was then removed and the basal media replaced with B‐ALI™ differentiation media. HNEC‐ALI were maintained for a minimum of 14 days for development of tight junctions and 28 days for cilia generation 13 …”

Section: Methodsmentioning

confidence: 99%

In vitro safety evaluation of a povidone‐iodine solution applied to human nasal epithelial cells

Int Forum Allergy Rhinol

Smith

Psaltis

et al. 2020

Self Cite

Background: Nasal topical treatments can provide an effective method of disease control for patients suffering from chronic rhinosinusitis (CRS). However, some frequently used formulations lack adequate evaluation on their safety. This study investigated the effect of 0.5% povidone-iodine (Nasodine) on the sinonasal epithelial barrier and ciliated human nasal epithelial cells (HNECs) in vitro. Methods: Nasodine was applied to air-liquid interface (ALI) cultures of primary HNECs from CRS patients. Epithelial barrier structure was assessed by measuring the transepithelial electrical resistance (TEER), paracellular permeability, and immunolocalization of the zona occludens-1 (ZO-1) tight junction protein. Toxicity and ciliary beat frequency (CBF) were also studied. Results: Nasodine was not toxic and did not have detrimental effects on the paracellular permeability or CBF. Na-sodine did not show a significant reduction in TEER with a 5-minute exposure; however, with a 30-minute exposure there was a significant reduction in TEER at 1 hour and at 4 hours a er exposure. Conclusion: Application of Nasodine to HNEC-ALI cultures in vitro for up to 30 minutes was not toxic and did not affect the paracellular permeability or CBF.