Highlights • We studied the mechanical consequences of airway remodeling in mice. • We examined the relationship between maximal tidal expiratory flow and airflow resistance following ...methacholine-induced bronchoconstriction in mice subjected to acute and chronic antigen challenge protocols. • The results we obtained show that the technique employed has promise for providing novel insight on the role of airway remodeling in airway hyperresponsiveness.
ABSTRACT
Asthma is a chronic disorder of the airways associated in many instances with structural changes of the airways, termed airway remodelling. Irritant and allergen‐induced murine models have ...been used to further understand the mechanisms of airway remodelling. The infiltration of the airways by inflammatory cells, such as T lymphocytes, mast cells, eosinophils, neutrophils and macrophages after repeated allergen challenges may be important effectors in the initiation and perpetuation of airway remodelling through the release of inflammatory mediators and growth factors. Interleukins‐4 and ‐13 have been widely studied in experimental models, and have been shown to play a significant role in airway remodelling. Recently, a role for Th17 cells has been established. Other mediators involved in this process are ligands of the epidermal growth factor receptor, matrix metalloproteases and cysteinyl leukotrienes. A better understanding of the mechanisms leading to airway remodelling in allergic diseases may lead to the identification of novel therapeutic strategies but validation in human subjects is required for potential targets.
Abstract There is a need for particles which exhibit controlled release of therapeutic agents delivered via the inhalational route, for tissue specific applications such as anti-cancer, ...bronchodilators and antiviral agents as well as drugs for systemic action. The aim of this study was to assess the acute toxicity, distribution and capacity of the microspheres to exhibit controlled release properties in an in vivo model of airway inflammation. Calcium pyrophosphate nanofibrous microspheres were loaded with dexamethasone phosphate (Dex-P); the profile of drug release was studied in vitro and validated in vivo . Unloaded microspheres were administered intra-tracheally (i.t.) to rats to assess the tissue reaction. The anti-inflammatory properties of the Dex-P loaded microspheres against an inflammatory agent (compound 48/80), were evaluated in vivo . Unloaded microspheres did not cause an inflammatory response when given at doses below 3 mg, and appeared to be eliminated through mucus clearance mechanisms. Microspheres loaded with Dex-P but not Dex-P alone, were capable of inhibiting eosinophil and total inflammatory cell increases in bronchoalveolar lavage fluid for 42 h following a single application. These observations demonstrated that calcium pyrophosphate nanofibrous microspheres displayed in vivo controlled release properties, were well tolerated and did not accumulate in the lung.
The covalent chemical functionalization of exfoliated hexagonal boron–nitride nanosheets (BNNSs) is achieved by the solution-phase oxygen radical functionalization of boron atoms in the h-BN lattice. ...This involves a two-step procedure to initially covalently graft alkoxy groups to boron atoms and the subsequent hydrolytic defunctionalization of the groups to yield hydroxyl-functionalized BNNSs (OH-BNNSs). Characterization of the functionalized-BNNSs using HR-TEM, Raman, UV–vis, FTIR, NMR, and TGA was performed to investigate both the structure of the BNNSs and the covalent functionalization methodology. OH-BNNSs were used to prepare polymer nanocomposites and their mechanical properties analyzed. The influence of the functional groups grafted to the surface of the BNNSs is investigated by demonstrating the impact on mechanical properties of both noncovalent and covalent bonding at the interface between the nanofiller and polymer matrixes.
We report on the modification of graphene oxide (GO) with poly(vinyl alcohol) (PVA) leading to the mechanical improvement of GO based materials. First, GO was covalently functionalised with PVA by ...esterification of carboxylic groups on GO with hydroxyl groups of PVA resulting in functionalised f-(PVA)GO. This was carried out for PVA of six different molecular weights. This functionalised graphene oxide could be formed into a paper-like material by vacuum filtration. Papers prepared from f-(PVA)GO showed significant increases in mechanical properties compared to those prepared with GO or with simple mixtures of GO and PVA. The best performance was achieved for PVA functional groups with molecular weights between 50 and 150kg/mol. Improvements in Young’s moduli of 60% and tensile strength of 400% were observed relative to GO-only paper. The improved mechanical properties are attributed to enhanced inter-flake stress transfer due to the covalently bonded PVA. Second, functionalised f-(PVA)GO was used as filler in PVA-based composites. The application of a pre-selection method allowed the use of only the largest functionalised f-(PVA)GO flakes. This resulted in substantially reinforced PVA–f-(PVA)GO composites. Both modulus and strength increased by 40% relative to the pure polymer for f-(PVA)GO loadings below 0.3 vol.%.
The bronchoconstrictive and proinflammatory properties of cysteinyl leukotrienes (cysLTs) in allergic asthma mediate their effects predominantly through the cysLT1 receptor (cysLT1R). However, the ...role of cysLTs and cysLT1R in innate immune-triggered asthma is largely unexplored. We explored the synthesis of cysLTs and cysLT1R as determinants of airway responses in an oxidative stress-induced model of irritant asthma. Wild-type (WT) mice exposed to 100 ppm Cl2 for 5 min had airway neutrophilia, increased cysLT production, and pulmonary expression of cysLT-related biosynthetic genes. CysLT1R-deficient (CysLTr1(-/-)) mice that were exposed to Cl2 demonstrated airway hyperresponsiveness to inhaled methacholine significantly greater than in WT BALB/c mice. Compared to WT mice, airway neutrophilia and keratinocyte chemoattractant production levels were higher in CysLTr1(-/-) mice and airway hyperresponsiveness was ameliorated using a granulocyte depletion Ab. CysLTr1(-/-) mice also demonstrated prolonged bronchial epithelial cell apoptosis following Cl2 WT mice showed increased antioxidant and NF erythroid 2-related factor 2 (Nrf2) gene expression, Nrf2 nuclear translocation in bronchial epithelial cells, and increased reduced glutathione/oxidized glutathione following Cl2 exposure whereas CysLTr1(-/-) mice did not. Furthermore, CysLTr1(-/-) mice demonstrated increased pulmonary E-cadherin expression and soluble E-cadherin shedding compared with WT mice. Loss of a functional cysLT1R results in aberrant antioxidant response and increased susceptibility to oxidative injury, apparently via a cysLT1R-dependent impairment of Nrf2 function.
Inhalation of organic dust (OD) from swine confinement facilities leads to pulmonary inflammation, airway hyperresponsiveness, and oxidative stress. In mice, pretreatment with a hydroxyl radical ...scavenger prevents airway inflammation and airway hyperresponsiveness (AHR) induced by OD exposure. We sought to determine a mechanism by which OD could induce oxidative stress in bronchial epithelial cells. Human bronchial epithelial cells (BEAS-2B or NHBE) were treated with various concentrations of OD, followed by evaluation of intracellular oxidative stress using 2',7'-dichlorofluorescein diacetate (DCFDA). After stimulation with OD, gene expression of antioxidant genes was assessed by real-time quantitative PCR followed by quantification of Nrf2 nuclear translocation using a luciferase reporter assay. Phagocytic markers (CD36 and CD68) were analyzed by FACS. Cells were treated with an actin inhibitor, cytochalasin D, before OD exposure and evaluated for Nrf2 nuclear translocation and DCFDA. Mice were pretreated with sulforaphane, the Nrf2 activator, before OD exposure and evaluated for pulmonary inflammation and airway reactivity. OD induced a time- and concentration-dependent increase in DCFDA. mRNA expression levels of Nrf2-dependent genes and Nrf2 nuclear translocation were increased after OD exposure. OD exposure increased the expression of CD68 and CD36. Cytochalasin D prevented oxidative stress and Nrf2 nuclear translocation after OD. Pretreatment with sulforaphane prevented OD-induced inflammation and AHR while increasing the uptake of OD in bronchial epithelial cells. Bronchial epithelial cells can phagocytose OD, resulting in an increase in endogenous oxidative stress. Nrf2-dependent mechanisms mediate the antioxidant response to OD.
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