Cell and tissue culture model of airway mucosa and its applications

Epithelial cells and fibroblast are two important cells involved in the inflammation and remodeling process. In our study, we established the cell culture model of epithelial cells and fibroblast cells and the co-culture model to study the airway inflammation and remodeling processes. We studied the modulatory effect of anti-inflammatory steroids on cytokines in airway inflammations in cultured epithelial cells and the dual effects of soluble chitosan in proinflammatory cytokine and wound healing process. Co-culture modulated their effects. We identified the underlying mechanisms by which steroids and soluble chitosans modulate the effect on proinflammatory/inhibitory cytokines in lung cell(s). We studied the differential response of proteins to anti-inflammatory steroids and soluble chitosan use a novel proteomic analysis.; Corticosteroids are widely used in suppressing airway inflammation. However a number of serious systemic side effects limit the clinical use of corticosteroids. An intensive effort has been conducted to modify the structure of corticosteroids by reducing the systemic side effects while still keep their anti-inflammatory function. In the present study, we hypothesize that anti-inflammatory steroidal antedrug may enhance/restore the secretion of inhibitory cytokines or cytokine receptors or mimic their effects on target cells in asthma. To test the hypotheses, an newly modified anti-inflammatory steroids 21-acetyloxy-9alpha-fluoro-11beta-hydroxy-3,20-dioxo-1,4-pregnadieno-[16(,17(-d] isoxazoline FP-ISO-21AC, which is designed with 'antedrug' concept to exert desirable effects at the application site, but is rapidly transformed to an inactive metabolite by enzymatic reaction upon entry into the circulation and prednisolone (PRED) were evaluated. Pro-inflammatory cytokine-induced nitric oxide (NO) production was significantly reduced when human alveolar epithelial cells (A549) were pre-treated with anti-inflammatory steroidal antedrugs, FP-ISO-21AC and PRED. When we added exogenous IL-22 to the media of A549 which express IL-22 cell surface receptors (IL-22R), further reduction of NO production was induced only in A549 pretreated with anti-inflammatory steroidal antedrugs but not in human lung fibroblasts (HFL1) which do not express IL-22R. This suggests that anti-inflammatory steroidal antedrugs FP-ISO-21AC and PDS may enhance the expression of cell surface receptors (IL-22R) to increase the cell's ability to respond to exogenous inhibitory cytokines (e.g. IL-22), which inhibit nuclear factor kappaB (NF-kappaB) activation.; Airway inflammation and remodeling are two key factors in chronic airway disease such as asthma. Inflammation in the airways is the major effect leading fixed narrowing of small airways and alveolar wall destruction in chronic obstructive pulmonary disease such as asthma. Airway remodeling is another important factor in airway disease. It is apparent that well-defined airway changes occur within the airway wall in disease condition. The inflammatory proteins expressed in the inflammation process and the matrix metalloproteinase (MMPs) released in the remodeling process may be affected by soluble chitosan. To investigate the effect of soluble chitosan, we examined the production of inflammatory proteins IL-8 and nitric oxide (NO) on human alveolar epithelial cells A549 stimulated with cytomix IL-1beta, TNF-alpha, and IFN-gamma. We also examined the MMPs release from human alveolar epithelial cells A549 and normal fibroblast HFL1 cells in epithelial/fibroblast co-culture systems in the presence of soluble chitosan. Our results indicate that the expression of IL-8 and NO induced by cytomix were patially inhibited by pretreatment of soluble chitosan. We also demonstrate the regulatory effect of soluble chitosan on the release of MMPs.; Airway consists of epithelial cells, fibroblasts, and smooth muscle cells. These cells play an important role in the airway. Especially, the interaction between...