Effects of Mucosal Decongestion on Nasal Aerodynamics: A Pilot Study.

OBJECTIVE: Mucosal decongestion with nasal sprays is a common treatment for nasal airway obstruction. However, the impact of mucosal decongestion on nasal aerodynamics and the physiological mechanism of nasal airflow sensation are incompletely understood. The objective of this study is to compare nasal airflow patterns in nasal airway obstruction (NAO) patients with and without mucosal decongestion and nondecongested healthy subjects. STUDY DESIGN: Cross-sectional study of a convenience sample. SETTING: Academic tertiary medical center. METHODS: Forty-five subjects were studied (15 nondecongested healthy subjects, 15 nondecongested NAO patients, and 15 decongested NAO patients). Three-dimensional models of the nasal anatomy were created from computed tomography scans. Steady-state simulations of airflow and heat transfer were conducted at 15 L/min inhalation rate using computational fluid dynamics. RESULTS: In the narrow side of the nose, unilateral nasal resistance was similar in decongested NAO patients and nondecongested healthy subjects, but substantially higher in nondecongested NAO patients. The vertical airflow distribution within the nasal cavity (inferior vs middle vs superior) was also similar in decongested NAO patients and nondecongested healthy subjects, but nondecongested NAO patients had substantially less middle airflow. Mucosal cooling, quantified by the surface area where heat flux exceeds 50 W/m2, was significantly higher in decongested NAO patients than in nondecongested NAO patients. CONCLUSION: This pilot study suggests that mucosal decongestion improves objective measures of nasal airflow, which is consistent with improved subjective sensation of nasal patency after decongestion.

[Correlation between nasal mucosal temperature change and nasal airflow perception].

The mechanism of nasal airflow perception remains little known. It is currently believed that the main mechanism for perceiving nasal patency is to activate transient receptor potential melastatin subtype 8. Computer fluent dynamics show that increased airflow and heat flux are associated with higher subjective scores. Similarly, physical measurements of the nasal cavity using a temperature probe show a correlation between the lower nasal mucosa temperature and better results. Trigeminal function detection also indirectly confirms this. This literature review aimed to explore the role of nasal mucosal temperature change in the subjective perception of nasal patency and the secondary aim was to appraise the relevant evidence about the mechanism.

The role of nasal congestion as a defence against respiratory viruses.

INTRODUCTION: This review discusses how nasal congestion may have benefits as a mechanism of defence against respiratory viruses. METHODS: A literature research was conducted on respiratory viruses and nasal congestion, following a recently published review on how temperature sensitivity is important for the success of common respiratory viruses. RESULTS: The literature reported that common respiratory viruses are temperature sensitive and replicate well at the cooler temperatures of the upper airways (32°C), but replication is restricted at body temperature (37°C). The amplitude of the phases of congestion and decongestion associated with the nasal cycle was increased on infection with respiratory viruses and this caused unilateral nasal congestion and obstruction. Nasal congestion and obstruction increase nasal mucosal temperature towards 37°C and therefore restricted the replication of respiratory viruses. CONCLUSION: Nasal congestion associated with the nasal cycle may act as a mechanism of respiratory defence against infection with respiratory viruses.

Effects of Hangeshashinto on the nasal physiological function: An in vitro study.

OBJECTIVE: Hangeshashinto is a Japanese Kampo medicine applied for the treatment of oral mucositis and gastroenteritis. Hangeshashinto exhibits broad-spectrum antibacterial activity and suppresses prostaglandin (PG)E2 production in the mucosa and has the ability to improve the inflammatory condition. In addition to these effects, because cAMP, a composition of Hangeshashinto, facilitates ciliary beat, Hangeshashinto could also improve the physiological function of the nasal mucosa, consist of ciliated epithelium, but details were unknown. METHODS: This study was aimed to investigate the effects of Hangeshashinto on the nasal mucosa. Healthy nasal mucosal sections were collected from the nasal septum of ten Japanese white rabbits, placed in a collagen dish for tissue culture, and rinsed with two different concentrations of Hangeshashinto solution (1.0%, n = 10 and 2.5%, n = 10) and cAMP solution (50µM, n=10 and 100 µM, n=10) or saline (control, n = 10). Ciliary beat frequency (CBF) as a physiological function of the nasal mucosa was recorded at 1, 3 and 7 days after rinsing, and histological evaluation of epithelial damage was performed at 7 days after rinsing. RESULTS: CBF in the 1.0% but not in the 2.5% Hangeshashinto group, increased at 3 and 7 days compared with that in the control group (p < 0.05). This trend was also observed in the CBF in the 100 µM cAMP group, significant difference was not observed between the CBF of the 1.0% Hangeshashinto group and the 100 µM cAMP group at 1, 3 and 7 days after rinsing (p > 0.05). Histological score only in the 2.5% Hangeshashinto group was lower than that in the control group (p < 0.05), while a significant decline was not observed in the other groups compared to that in the control group (p > 0.05). CONCLUSION: Our results suggest that 1.0% Hangeshashinto solution facilitates the physiological function of the nasal mucosa by promoting ciliary functions without histological damage of cilia epithelium. When applied with the appropriate concentration, Hangeshashinto could have ability to improve the physiological functions of the nasal mucosal epithelium.

Rapid activation of brainstem nuclei following TLR stimulation of the nasal mucosa.

We recently identified a novel neuroimmune mechanism in the nasal mucosa, in which activation of neuronal Toll­like receptor (TLR) 7 results in upregulation of epithelial TLRs, via release of substance P. In the present study, we assessed whether intranasal challenge with the TLR7 agonist R­837 additionally activated neurons in the central nervous system. Within one hour, R­837 induced activation of the nucleus of the solitary tract, as well as a small increase in nasal IL­6, but otherwise in the absence of an overt inflammatory response. It is tempting to speculate that it might be a direct interaction of R­837 with trigeminal neurons in order to alert the central nervous system of invading pathogens.

Ciliary beat frequency of in vitro human nasal epithelium measured with the simple high-speed microscopy is applicable for safety studies of nasal drug formulations.

Nasal drug formulations can be effective for local delivery of therapeutic drugs to the sinonasal mucosa or for systemic drug delivery by absorption directly into the bloodstream. The growing field of potential nasal therapies includes nasal vaccination and even treatment of neurodegenerative diseases. However, it is important that nasal drug formulations don't have a disruptive effect on the cilia and mucosa of nasal epithelium. Mucociliary clearance represents the first host defence of the respiratory tract that requires the coordinated beating of cilia. A key parameter to determine mucociliary clearance is ciliary beat frequency (CBF). The objective of this study was to validate the high-speed digital imaging for CBF measurements in nasal MucilAir™ in vitro model and to test its potential for ciliotoxicity studies to evaluate the safety of investigational nasal drug formulations. Our CBF measuring setup was first validated by benzalkonium chloride, a common-practice preservative with cilio-inhibiting effect. Next, MucilAir™ model was treated with mometasone nasal spray (Mommox®/Mometasone Sandoz®). Short term cilio-stimulatory effect and dose dependent effect of mometasone nasal spray were demonstrated. Post-treatment analysis showed un-altered ultrastructure of MucilAir™ model. In conclusion, characterization of the ciliary activity of nasal MucilAir™ in vitro model and its response to relevant agents with herein developed efficient and reproducible set up for CBF analysis show great potential of this model for airway ciliotoxicity studies.

Spaceflight-Associated Changes in the Opacification of the Paranasal Sinuses and Mastoid Air Cells in Astronauts.

Importance: Head congestion is one of the most common somatic symptoms experienced by astronauts during spaceflight; however, changes in the opacification of the paranasal sinuses or mastoid air cells in astronauts have not been adequately studied. Objectives: To quantify preflight to postflight changes in the opacification of the paranasal sinuses and mastoid air cells in Space Shuttle astronauts and International Space Station (ISS) astronauts and to assess whether there are differences between the 2 groups of astronauts. Design, Setting, and Participants: This cohort study examined preflight and postflight head magnetic resonance images (MRIs) of 35 astronauts who had participated in either a short-duration (≤30 days) Space Shuttle mission or a long-duration (>30 days) ISS mission and had undergone both preflight and postflight MRI. Images were obtained before and after spaceflight. Images were evaluated by 2 neuroradiologists blinded to which mission each astronaut had flown and to which images were preflight or postflight images. Exposure: Spaceflight on the Space Shuttle or the ISS. Main Outcomes and Measures: Measured outcomes included preflight to postflight changes in Lund-Mackay scores for the paranasal sinuses and in scores grading mastoid effusions. Results: Most astronauts in both the Space Shuttle group (n = 17; 15 men; mean [SD] age at launch, 47.7 [3.1] years) and the ISS group (n = 18; 14 men; mean [SD] age at launch, 48.6 [4.7] years) exhibited either no change or a reduction in paranasal sinus opacification as seen on postflight MRI scans (Space Shuttle group: 6 [35.3%] had no sinus opacification before or after spaceflight, 5 [29.4%] had less sinus opacification after spaceflight, 3 [17.6%] had the same amount of sinus opacification before and after spaceflight, and 3 [17.6%] had increased paranasal sinus opacification after spaceflight; ISS group: 8 [44.4%] had no sinus opacification before or after spaceflight, 4 [22.2%] had less sinus opacification after spaceflight, 1 (5.6%) had the same amount of sinus opacification before and after spaceflight, and 5 [27.8%] had scores consistent with increased paranasal sinus opacification after spaceflight). Long-duration spaceflight (ISS group) was associated with an increased risk of mastoid effusion relative to short-duration spaceflight (relative risk, 4.72; 95% CI, 1.2-18.5). Images were obtained a mean (SD) 287.5 (208.6) days (range, 18-627 days) prior to and 6.8 (5.8) days (range, 1-20 days) after spaceflight. Astronauts had undergone either a mean (SD) of 13.6 (1.6) days of spaceflight on the Space Shuttle (17 astronauts) or 164.8 (18.9) days on the ISS (18 astronauts). Conclusions and Relevance: This study found that exposure to spaceflight conditions on the ISS is associated with an increased likelihood for the formation of mastoid effusions. There was no association between exposure to spaceflight conditions and changes in paranasal sinus opacification. The limitations of this study include lack of information concerning medical history and mission-specific operational experience for individual astronauts. Further studies are indicated to determine the cause and composition of the mastoid effusions.

Measurement of mucociliary clearance in the patients with multiple sclerosis.

OBJECTIVES: The aim of the present study was to measure nasal mucociliary clearance (NMC) time in the patients with MS and to compare the findings with healthy population. METHODS: Totally 97 individuals including 47 patients with relapsing-remitting multiple sclerosis and 50 healthy volunteers were enrolled into the study. Saccharin clearance test was performed on both groups and NMC time was measured. Data analysis was performed by SPSS version 24.0 statistics program (SPSS Inc., Chicago, Illinois, USA). Statistical tests were interpreted at p < 0.05 significance level. RESULTS: The NMC time averages in MS patients and healthy control group were 12.43 ± 4.05 min and 8.14 ± 2.87 min, respectively; the difference between the groups was significant (p < 0.001). There was a statistically strong association between NMC time values and Expanded Disability Status Scale (EDSS) values in MS patients (r = 0.817, p < 0.001). CONCLUSION: We found nasal mucociliary transport time longer in MS patients than healthy population in the present study. To the best of our knowledge, there is not any study conducted about this topic in the literature. We believe that our findings would shed a light on further studies.

Integration of PEG 400 into a self-nanoemulsifying drug delivery system improves drug loading capacity and nasal mucosa permeability and prolongs the survival of rats with malignant brain tumors.

Introduction: Kolliphor® EL (K-EL) is among the most useful surfactants in the preparation of emulsions. However, it is associated with low hydrophobic drug loading in the resulting emulsified formulation. Methods: In this study, a formulation for intranasal administration of butylidenephthalide (Bdph), a candidate drug against glioblastoma (GBM), was prepared. Physical characteristics of the formulation such as particle size, zeta potential, conductivity, and viscosity were assessed, as well as its cytotoxicity and permeability, in order to optimize the formulation and improve its drug loading capacity. Results: The optimized formulation involved the integration of polyethylene glycol 400 (PEG 400) in K-EL to encapsulate Bdph dissolved in dimethyl sulfoxide (DMSO), and it exhibited higher drug loading capacity and drug solubility in water than the old formulation, which did not contain PEG 400. Incorporation of PEG 400 as a co-surfactant increased Bdph loading capacity to up to 50% (v/v), even in formulations using Kolliphor® HS 15 (K-HS15) as a surfactant, which is less compatible with Bdph than K-EL. The optimized Bdph formulation presented 5- and 2.5-fold higher permeability and cytotoxicity, respectively, in human GBM than stock Bdph. This could be attributed to the high drug loading capacity and the high polarity index due to DMSO, which increases the compatibility between the drug and the cell. Rats bearing a brain glioma treated with 160 mg/kg intranasal emulsified Bdph had a mean survival of 37 days, which is the same survival time achieved by treatment with 320 mg/kg stock Bdph. This implies that the optimized emulsified formulation required only half the Bdph dose to achieve an efficacy similar to that of stock Bdph in the treatment of animals with malignant brain tumor.

Experimental Acute Exposure to Thirdhand Smoke and Changes in the Human Nasal Epithelial Transcriptome: A Randomized Clinical Trial.

Importance: No previous studies have shown that acute inhalation of thirdhand smoke (THS) activates stress and survival pathways in the human nasal epithelium. Objective: To evaluate gene expression in the nasal epithelium of nonsmoking women following acute inhalation of clean air and THS. Design, Setting, and Participants: Nasal epithelium samples were obtained from participants in a randomized clinical trial (2011-2015) on the health effects of inhaled THS. In a crossover design, participants were exposed, head only, to THS and to conditioned, filtered air in a laboratory setting. The order of exposures was randomized and exposures were separated by at least 21 days. Ribonucleic acid was obtained from a subset of 4 healthy, nonsmoking women. Exposures: By chance, women in the subset were randomized to receive clean air exposure first and THS exposure second. Exposures lasted 3 hours. Main Outcomes and Measures: Differentially expressed genes were identified using RNA sequencing with a false-discovery rate less than 0.1. Results: Participants were 4 healthy, nonsmoking women aged 27 to 49 years (mean [SD] age, 42 [10.2] years) with no chronic diseases. A total of 389 differentially expressed genes were identified in nasal epithelium exposed to THS, while only 2 genes, which were not studied further, were affected by clean air. Enriched gene ontology terms associated with stress-induced mitochondrial hyperfusion were identified, such as respiratory electron transport chain (q = 2.84 × 10-3) and mitochondrial inner membrane (q = 7.21 × 10-6). Reactome pathway analysis identified terms associated with upregulation of DNA repair mechanisms, such as nucleotide excision repair (q = 1.05 × 10-2). Enrichment analyses using ingenuity pathway analysis identified canonical pathways related to stress-induced mitochondrial hyperfusion (eg, increased oxidative phosphorylation) (P = .001), oxidative stress (eg, glutathione depletion phase II reactions) (P = .04), and cell survival (z score = 5.026). Conclusions and Relevance: This study found that acute inhalation of THS caused cell stress that led to the activation of survival pathways. Some responses were consistent with stress-induced mitochondrial hyperfusion and similar to those demonstrated previously in vitro. These data may be valuable to physicians treating patients exposed to THS and may aid in formulating regulations for the remediation of THS-contaminated environments.

Real-time In Vitro Monitoring of Odorant Receptor Activation by an Odorant in the Vapor Phase.

Olfactory perception begins with the interaction of odorants with odorant receptors (OR) expressed by olfactory sensory neurons (OSN). Odor recognition follows a combinatorial coding scheme, where one OR can be activated by a set of odorants and one odorant can activate a combination of ORs. Through such combinatorial coding, organisms can detect and discriminate between a myriad of volatile odor molecules. Thus, an odor at a given concentration can be described by an activation pattern of ORs, which is specific to each odor. In that sense, cracking the mechanisms that the brain uses to perceive odor requires the understanding odorant-OR interactions. This is why the olfaction community is committed to "de-orphanize" these receptors. Conventional in vitro systems used to identify odorant-OR interactions have utilized incubating cell media with odorant, which is distinct from the natural detection of odors via vapor odorants dissolution into nasal mucosa before interacting with ORs. Here, we describe a new method that allows for real-time monitoring of OR activation via vapor-phase odorants. Our method relies on measuring cAMP release by luminescence using the Glosensor assay. It bridges current gaps between in vivo and in vitro approaches and provides a basis for a biomimetic volatile chemical sensor.

Comparison of telomere lengths in leukocytes and in nasal and vaginal epithelial cells from Water Buffaloes (Bubalus bubalis) of different ages.

Telomeres are short and repetitive sequences at the ends of linear chromosomes which shorten with every cell-division in vitro. Telomere length (TL) is reported to decrease with age and stress. The domesticated water buffalo (Bubalus bubalis) is the second most important milk producing animal worldwide. The productive lifespan of water buffalo cows is reported to be longer than that of dairy cows (Bos taurus). With this background, we aimed to compare TL in leukocytes obtained from blood samples from water buffaloes across different ages. In addition, we tested the suitability of assessing TL in DNA derived from nasal and vaginal epithelial cells via swabs as potential non-invasive alternatives to blood sampling Samples were collected from 20 calves (3 months of age), 20 heifers (2 years old), 20 cows (1st lactation, 3 years old), and 13 cows (3rd lactation, about 5 years old). We found that TL in leukocytes from water buffalo calves, heifers, and from cows in their first lactation was not different, but shorter telomeres were observed in cows in their third lactation. The results thus support an age-dependent decrease of TL in water buffaloes. Leukocyte TL was weakly correlated with TL measured in DNA from nasal epithelial cells (r = 0.327; P = .025), but not with TL from vaginal epithelial cells. Due to the poor correlation between epithelial cell and leukocyte TL and to the difficulties with collecting nasal swabs, we conclude that they are no suitable alternatives to blood samples for telomere studies in water buffaloes.

TGF-ß1 Induces Epithelial-Mesenchymal Transition of Chronic Sinusitis with Nasal Polyps through MicroRNA-21.

OBJECTIVES: To characterize the epithelial-mesenchymal transition (EMT) in chronic rhinosinusitis with nasal polyps (CRSwNP) and to investigate the mechanism by which microRNA-21 (miR-21) regulates EMT in CRSwNP. METHOD: (1) Tissue experiments: Mucosa tissues were collected from 13 patients with CRSwNP and 12 patients with CRS without nasal polyps (CRSsNP), as well as 11 patients without CRS (controls). Protein localization and quantification were achieved by immunofluorescence staining and Western blotting, involving the epithelial marker protein E-cadherin and the mesenchymal marker proteins α-smooth muscle actin (α-SMA), fibronectin, and vimentin. Quantitative RT-PCR was used to detect the relative expression levels of miR-21 and TGF-ß1 mRNAs. (2) Cellular experiments: Primary human nasal epithelial cells (PHNECs) treated with TGF-ß1, or TGF-ß1 with miR-21 inhibitor, or miR-21 mimics alone were observed for morphology changes under a phase-contrast microscope. The expression levels of epithelial/mesenchymal marker proteins were determined as aforementioned. PTEN and phosphorylated Akt were detected by Western blotting. RESULTS: (1) Tissue experiments: Compared with the CRSsNP and control groups, the expression of E-cadherin was downregulated in the CRSwNP group, whereas the expression of TGF-ß1, α-SMA, fibronectin, and vimentin was upregulated. The expression levels of miR-21 and TGF-ß1 mRNAs in CRSwNP were significantly higher than those in CRSsNP and controls. (2) Cellular experiments: TGF-ß1 induced EMT-like transformation in PHNECs, featured by changes in cell morphology and upregulation of mesenchymal proteins and miR-21. The miR-21 inhibitor, as well as the Akt-specific -inhibitor, suppressed TGF-ß1-induced EMT. Mechanically, downregulation of miR-21 resulted in increased PTEN and decreased Akt phosphorylation. Furthermore, overexpression of miR-21 had the opposite effects. CONCLUSIONS: Our findings suggest that the TGF-ß1-miR-21-PTEN-Akt axis may contribute to the pathogenesis of CRSwNP. miR-21 might be a reliable target for treating nasal polyp genesis through EMT suppression. Moreover, miR-21 inhibitors could be a novel class of antipolyp drug that modulates PTEN expression and Akt activation. In addition, further investigation regarding the reason underlying miR-21 overexpression in CRSwNP could provide a molecular target for novel treatment strategies for nasal polyposis.

Dendritic cells modified with Der p1 antigen as a therapeutic potential for allergic rhinitis in a murine model via regulatory effects on IL-4, IL-10 and IL-13.

OBJECTIVES: House dust mites, including Der p1, are common allergens. The current study was designed to explore the allergen-specific immune tolerance effects of Der p1-modified dendritic cells (DCs) through IL-4, IL-10 and IL-13 on an allergic rhinitis (AR) mouse model. METHODS: A lentivirus was modified to express Derp1. Then, immature DCs from mice were infected with this modified lentivirus to generate a lenti-Derp1-GFP DCs. 24 mice were random divided into four groups (n = 6 each), AR mouse were sensitized by Derp1 allergens and treated with lenti-GFP DCs (GFP-DC/AR group), or lenti-Derp1-GFP DCs (Der p1-DC/AR group) and dexamethasone (Dex/AR group), mice in the control group were treated with PBS instead of Der p1 then also intraperitoneally injected with 5 × 106 lenti-GFP DCs/mouse. AR symptoms expressed by each mouse were recorded. The proportions of CD4+CD25+Foxp3+ regulatory T cells among CD4+ T cells in the peripheral blood, and mRNA and protein expression levels of IL-4, IL-10, and IL-13 were measured. RESULTS: DCs infected with lenti-Derp1-GFP stimulated the maturation of DCs. Compared with the GFP-DC/AR group, mice in the Der p1-DC/AR group showed an ameliorated allergic response, a significant decrease in the levels of serum IgE, IgG1, and histamine, and a decrease in the expression of IL-4 and IL-13 mRNA and protein in the nasal mucosa. The expression of IL-10 increased in the Der p1-DC/AR group to a level similar to that observed in the Dex/AR group. CONCLUSIONS: These results indicate that Der p1-modified DCs have therapeutic potential for AR via downregulation of IL-4 and IL-13, and upregulation of IL-10.

The role of autophagy in the overexpression of MUC5AC in patients with chronic rhinosinusitis.

BACKGROUND: Autophagy is a lysosomal degradation pathway that protects the body and is essential for cell survival and differentiation. Mucins (MUCs) are important components of secreted mucus, mucin (MUC)5 AC is the major MUC secreted in the normal airway. OBJECTIVE: Investigated the role of autophagy in pathogenic mucin (MUC)5 AC production during chronic rhinosinusitis (CRS). METHODS: The expression of human neutrophil elastase (HNE) and the autophagic proteins microtubule-associated protein 1 light chain (LC)3B-II, c-Jun N-terminal kinase (JNK), c-Jun, and MUC5AC were analyzed in the sinonasal mucosa and human nasal epithelial cells (HNECs) using immunohistochemistry, enzyme-linked immunosorbent assay (ELISA), and quantitative real-time polymerase chain reaction (qRT-PCR). Autophagic vacuoles were studied using transmission electron microscopy (TEM). Primary HNECs were treated with HNE, bafilomycin A1, and SP600125. In some experiments, cultured primary HNECs were transfected with small interfering RNAs (siRNAs) to target Beclin-1 (BECN1; BECN1-siRNA), autophagy-related gene 5 (Atg5; Atg5-siRNA), and c-Jun (c-Jun-siRNA). Cultured cells were analyzed using western blotting, qRT-PCR, and ELISA. RESULTS: In CRS patients, both with and without nasal polyps, the expression levels of HNE, LC3B, JNK, c-Jun, and MUC5AC were upregulated. Bafilomycin A1 upregulated LC3B-II expression and inhibited MUC secretion in HNE-treated normal primary HNECs. Autophagosomes were observed in HNE-treated primary HNECs using TEM. HNE-induced secretion of MUC5AC was suppressed in normal primary HNECs by BECN1-siRNA, Atg5-siRNA, c-Jun-siRNA, and SP600125. CONCLUSIONS: In HNE-induced CRS, autophagy increases the secretion of MUC5AC by promoting the phosphorylation of JNK and c-Jun.

Evaluation of the Osteogenic Potential of Different Scaffolds Embedded with Human Stem Cells Originated from Schneiderian Membrane: An In Vitro Study.

BACKGROUND: Novel treatments for bone defects, particularly in patients with poor regenerative capacity, are based on bone tissue engineering strategies which include mesenchymal stem cells (MSCs), bioactive factors, and convenient scaffold supports. OBJECTIVE: In this study, we aimed at comparing the potential for different scaffolds to induce osteogenic differentiation of human maxillary Schneiderian sinus membrane- (hMSSM-) derived cells. Methods. hMSSM-derived cells were seeded on gelatin, collagen, or Hydroxyapatite ß-Tricalcium phosphate-Fibrin (Haß-TCP-Fibrin) scaffolds. Cell viability was determined using an MTT assay. Alizarin red staining method, Alkaline phosphatase (ALP) activity assay, and quantitative real-time PCR analysis were performed to assess hMSSM-derived cells osteogenic differentiation. RESULTS: Cell viability, calcium deposition, ALP activity, and osteoblastic markers transcription levels were most striking in gelatin scaffold-embedded hMSSM-derived cells. CONCLUSION: Our findings suggest a promising potential for gelatin-hMSSM-derived cell construct for treating bone defects.

Acellular dermal matrix contributes to epithelialization in patients with chronic sinusitis.

BACKGROUND: Nasal endoscopic surgery is widely used for nasal diseases, including sinusitis and tumors. However, scar hyperplasia, nasal irritation, scab, and nasal obstruction delay nasal mucosal recovery, with prolonged cleaning exacerbating the patient's financial burden. Here, we presented a novel approach for the treatment of nasal mucosal defects, termed acellular dermal matrix. METHODS: A total of 31 patients with bilateral chronic sinusitis (maxillary sinusitis and ethmoid sinusitis) underwent nasal surgery and nasal mucosal repair in September-October 2016. We divided the nasal cavities of each patient into control and acellular dermal matrix groups, randomly selected one side for nasal mucosal repair by surgery. A suitable acellular dermal matrix size was selected according to the defect in each patient. After pruning, the acellular dermal matrix was placed on the wound surface and filled with gelatin sponge. All patients were followed up for 14 weeks to compare nasal mucosal epithelialization between the control and acellular dermal matrix groups. Results：No obvious complications and adverse reactions were observed after nasal surgery. Lund-Kennedy scores in the acellular dermal matrix group were significantly decreased compared with the control group at 8 (0 (0, 1) vs. 2 (2, 4); P<0.05) weeks. Epithelialization time of eight weeks in the acellular dermal matrix groups was significantly decreased than the control group of 14 weeks. CONCLUSION: Acellular dermal matrix provides a growth framework for the healthy mucosa on the wounded surface and reduces postoperative epithelialization time.

Broncho-Vaxom® (OM-85 BV) soluble components stimulate sinonasal innate immunity.

BACKGROUND: Broncho-Vaxom® (OM-85 BV) is an extract of infectious respiratory bacteria that is used as an immunostimulant outside of the United States for the prevention and treatment of bronchitis and rhinosinusitis. Prior studies have shown that use of OM-85 BV is associated with reduction in frequency of respiratory infection and decreased duration of antibiotic usage. However, the effects of OM-85 BV on respiratory mucosal innate immunity are unknown. METHODS: Human sinonasal epithelial cells were grown at an air-liquid interface (ALI). Ciliary beat frequency (CBF) and nitric oxide (NO) production in response to stimulation with OM-85 BV was measured in vitro. Pharmacologic inhibitors of bitter taste receptor (T2R) signaling were used to determine if this pathway was taste-receptor-mediated. RESULTS: Apical application of OM-85 BV resulted in an NO-mediated increase in CBF (p < 0.05) and increased NO production (p < 0.0001) when compared to saline-stimulated control cultures. ALI pretreatment with taste receptor pathway inhibitors blocked OM-85 BV-induced increases in NO. CONCLUSION: OM-85 BV has ciliostimulatory and immunogenic properties that may be partially responsible for its observed efficacy as a respiratory therapeutic. These responses were NO-dependent and consistent with T2R activation. Further work is necessary to elucidate specific component-receptor signaling relationships.

Resveratrol and ivacaftor are additive G551D CFTR-channel potentiators: therapeutic implications for cystic fibrosis sinus disease.

BACKGROUND: Mutations in the cystic fibrosis transmembrane conductance regulator (CFTR) gene result in defective Cl- transport and cause chronic bacterial infections in the upper and lower airways of cystic fibrosis (CF) patients. Ivacaftor is a CFTR potentiator that improves Cl- transport in CF patients with at least 1 copy of the G551D mutation. Resveratrol is also a potent CFTR potentiator that increases determinants of mucociliary transport. The objective of this study is to determine whether resveratrol and ivacaftor improve Cl- secretion in G551D CFTR over either agent alone. METHODS: Fisher rat thyroid cells (FRT) transfected with G551D CFTR and human sinonasal epithelial cells (HSNE) containing the CFTR G551D mutation were subjected to pharmacologic manipulation of transepithelial ion transport in Ussing chambers. Activity was further evaluated using whole-cell patch clamp methods in G551D FRT cells. RESULTS: In G551D FRT cells, resveratrol (100 µM) and ivacaftor (10 µM) significantly increased Cl- transport (change in short-circuit current, δISC = µA/cm2 ) compared with single-agent and dimethylsulfoxide vehicle controls (resveratrol + ivacaftor 4.97 ± 0.57 vs ivacaftor 0.74 ± 0.12 vs resveratrol 2.96 ± 0.52 vs control 0.74 ± 0.12; p < 0.001). Maximal Cl- secretion (20 µM forskolin) was also significantly enhanced (p < 0.0001). Activity was confirmed in G551D HSNE (resveratrol + ivacaftor 4.48 ± 0.39 vs ivacaftor 1.05 ± 0.11 vs. resveratrol 0.84 ± 0.3 vs control, 0.0 ± 0.02; p < 0.001), and whole-cell patch clamp analysis in G551D FRT cells (resveratrol + ivacaftor -2535 ± 179.3 pA vs ivacaftor -1408.9 ± 101.3 pA vs resveratrol; -766.2 ± 71.2 pA; p < 0.0001). CONCLUSION: Additive improvement in G551D CFTR-mediated Cl- secretion suggests that resveratrol could enhance ivacaftor therapy in these patients and improve CF-related rhinosinusitis.