Fabrication of multi-layered 3D bioprinted lower respiratory model for in vitro disease development and drug screening
Research Highlights: Novel Facile Decellularization Method, Affordable Lung ECM based Hydrogel, 3D BioPrinting, Innovative and Multifunctional 3D Lung Model, Drug Screening Applications
Chronic respiratory diseases (CRDs) such as asthma, COPD, and lower respiratory tract infections (LRTIs) impose significant socioeconomic burdens in developing countries. Pre-existing progressive lung disease is a significant comorbidity associated with Covid-19 related deaths. By 2027, the market for in vitro lung models is expected to reach around USD 800 million. Commercially available bronchial mucociliary models (e.g., MucilAir and EpiAirway-FT®) that lack complex submucosal layers are unable to reproduce lung pathophysiology and persistent inflammatory states accurately. Various techniques have been attempted to create biomimetic in vitro lung models that replicate the organotypic environment at the tissue level. However, the challenge is in developing structurally and functionally relevant human lung tissue models that are reliably manufactured and customizable for the study of disease pathogenesis and drug efficacy. Herein, we propose to fabricate an immunocompetent 3D in vitro biomimetic model with complex microarchitecture with potential innervation to mimic the chronic inflammatory condition in the bronchial microenvironment and structural remodeling of the airway wall during pathophysiological events. Decellularized extracellular matrix (dECM) contains major biochemical and biophysical cues that provide biocompatibility, porosity, and elasticity requirements supporting cell attachment, cell-cell interactions, and underlying ECM-mediated differentiation, proliferation, and migration of cells. dECM gelates at physiological temperature and pH, resulting in hydrogel formation due to thermally induced in vitro self-assembly of collagen molecules. Therefore, it is ideal biomaterial for engineering 3D printed lung tissue designed to mimic complex lung microenvironment, which represents a better strategy than artificial matrices. We plant to fabricate a physiologically relevant bronchial wall model with 3D bioprinting using cell-laden dECM bioink followed by culturing in the appropriate environment for in vitro biological validation. Furthermore, the designed models can be utilized for investigating infection mechanisms of respiratory viruses, screening targeted gene-editing technology, and personalized medicine applications. We expect this model to help reduce the use of preclinical animal models and provide a precise, low-cost, and industry-relevant human lung-tissue biomimicking platform for disease modeling, drug screening, and pharmaceutics, which is an urgent need of the hour.
Objectives
Research Highlights: Novel Facile Decellularization Method, Affordable Lung ECM based Hydrogel, 3D BioPrinting, Innovative and Multifunctional 3D Lung Model, Drug Screening Applications
Chronic respiratory diseases (CRDs) such as asthma, COPD, and lower respiratory tract infections (LRTIs) impose significant socioeconomic burdens in developing countries. Pre-existing progressive lung disease is a significant comorbidity associated with Covid-19 related deaths. By 2027, the market for in vitro lung models is expected to reach around USD 800 million. Commercially available bronchial mucociliary models (e.g., MucilAir and EpiAirway-FT®) that lack complex submucosal layers are unable to reproduce lung pathophysiology and persistent inflammatory states accurately. Various techniques have been attempted to create biomimetic in vitro lung models that replicate the organotypic environment at the tissue level. However, the challenge is in developing structurally and functionally relevant human lung tissue models that are reliably manufactured and customizable for the study of disease pathogenesis and drug efficacy. Herein, we propose to fabricate an immunocompetent 3D in vitro biomimetic model with complex microarchitecture with potential innervation to mimic the chronic inflammatory condition in the bronchial microenvironment and structural remodeling of the airway wall during pathophysiological events. Decellularized extracellular matrix (dECM) contains major biochemical and biophysical cues that provide biocompatibility, porosity, and elasticity requirements supporting cell attachment, cell-cell interactions, and underlying ECM-mediated differentiation, proliferation, and migration of cells. dECM gelates at physiological temperature and pH, resulting in hydrogel formation due to thermally induced in vitro self-assembly of collagen molecules. Therefore, it is ideal biomaterial for engineering 3D printed lung tissue designed to mimic complex lung microenvironment, which represents a better strategy than artificial matrices. We plant to fabricate a physiologically relevant bronchial wall model with 3D bioprinting using cell-laden dECM bioink followed by culturing in the appropriate environment for in vitro biological validation. Furthermore, the designed models can be utilized for investigating infection mechanisms of respiratory viruses, screening targeted gene-editing technology, and personalized medicine applications. We expect this model to help reduce the use of preclinical animal models and provide a precise, low-cost, and industry-relevant human lung-tissue biomimicking platform for disease modeling, drug screening, and pharmaceutics, which is an urgent need of the hour.
Objectives
- Comparative characterization of detergent-based and detergent-free method for efficient decellularization of caprine lung tissue matrix (dLTM) and hydrogel formulation.
- Evaluation of multifunctional decellularized lung matrix hydrogel bioink in terms of growth of fibroblasts and smooth muscle cells and differentiation of epithelial cells.
- Development of a physiologically relevant 3D bioprinted biomimetic multicellular lower respiratory model and in vitro biological validation thereof.
- Development of respiratory pathogenesis in immunocompetent 3D lung model in a dynamic cell culture environment to validate the efficacy for disease modeling and in vitro drug evaluation.
- Recapitulating the neural regulation of airway smooth muscle layer functionality in the normal and diseased conditions through 3D culturing