Recellularization of bronchial extracellular matrix with primary bronchial smooth muscle cells

Abstract

Severe asthma is associated with an increased airway smooth muscle (ASM) mass and an altered composition of the extracellular matrix (ECM). Studies have suggested that ECM-ASM cell interactions contribute to this remodeling and its limited reversibility with current therapy.Three-dimensional matrices allow the study of complex cellular responses to different stimuli in an almost natural environment. Our goal was to obtain acellular bronchial matrices and then develop a recellularization protocol with ASM cells. We studied equine bronchi as horses spontaneously develop a human asthma-like disease. The bronchi were decellularized using Triton/Sodium Deoxycholate. The obtained scaffolds retained their anatomical and histological properties. Using immunohistochemistry and a semi-quantitative score to compare native bronchi to scaffolds revealed no significant variation for matrixial proteins. A DNA quantification and electrophoresis revealed that DNA was < 10 ng/μl ± 2, with remaining fragments of less than 100 bp. Primary ASM cells were then seeded on the scaffolds. Histological analysis of the recellularizations showed that ASM cells migrated and proliferated within the scaffolds primarily in the decellularized smooth muscle matrix, suggesting a chemotactic effect. This is the first report of primary ASM cells preferentially repopulating the smooth muscle matrix layer in bronchial matrices. This protocol is now being used to study the molecular interactions occurring between the asthmatic ECMs and ASM to identify effectors of asthmatic bronchial remodeling.