| Tissue generation within the airways is a complex process and it is well established that the extracellular matrix (ECM) plays a key role in mediating remodeling, repair and healing of tissues weakened by inflammation, infection, congenital defect or disease. However, the mechanisms governing these processes are not fully understood. Therefore, the goal here is to characterize ECM proteins of the trachea. This knowledge will provide insight into understanding how the airway responds during processes such as inflammation, infection, regeneration and wound healing. Having this information will enhance understanding of how the airway responds to inhaled particles (e.g., agricultural dusts, microbes) and surgical intervention for congenital respiratory diseases such as tracheomalacia (softening of the trachea), traumatic airway injury or tumor removal. For this study, proteins were isolated from native and decellularized porcine tracheas and quantified using the Bradford Assay and the NanoDrop techniques. Gel-based proteomics was used to separate proteins via size and isoelectric point, followed by in-gel or in-solution protein digestion. Proteins were identified using liquid chromatography-mass spectrometry (LC/MS) analysis and UniProt database searching, and were validated via western blot. For statistical analysis, one-way analysis of variance (ANOVA) with post-test corrections was performed. Gel-based proteomic analysis revealed an abundance of high molecular weight proteins with isoelectric points close to neutral. Porcine extracellular matrix proteins including collagen IV, elastin, decorin, aggrecan core protein, and tenascin-C were identified and validated via western blot. Knowledge of airway extracellular matrix proteins will provide insight for understanding wound healing and repair processes within the airways. |
