Anisotropy in Matrix Bioblocks: A Platform for Microvascular Engineering Lung damage has devastating consequences for pulmonary and cardiovascular health, with few treatment alternatives beyond chronic mechanical ventilation and lung transplantation. The study of vascular development is essential to developing 3D engineered grafts for regenerative lung therapies. This proposal addresses two main problems in vascular tissue engineering: 1) the need for a natural, complex scaffold material and 2) the requirement for pre-vascularized tissue constructs. These two issues will be addressed in a model context by developing human lung microvascular networks in natural, tissue-specific matrices produced by human lung fibroblasts.
The first aim i s to examine the assembly of natural, fibroblast matrix in confined, 3D geometries and to identify the molecular mechanisms driving anisotropic cellular growth and matrix development in response to cues from environmental geometry. This will be tested using biochemical and genetic manipulation of cytoskeletal modulators.
This aim will test the hypothesis that actin is the major component influencing anisotropic cellular response to 3D confinement. Biochemical inhibitors and siRNA will be used to target specific cytoskeletal functions. The spatial organization and anisotropy of cytoskeletal assembly and matrix deposition will be evaluated for each treatment group using pairs of 3D shapes that have constant volume and area, but varying anisotropies with confocal laser scanning microscopy and quantitative autocorrelation analysis.
The second aim will analyze the effects of confined, geometric microenvironments on matrix remodeling and vessel formation by human lung microvascular endothelial cells. The hypothesis that 3D microvascular network formation can be controlled using a natural matrix model will be tested. Microvessel formation and alignment in wells of varying geometries will be examined quantitatively. In summary, this predoctoral training proposal will analyze the effects of confined 3D geometries on the alignment of matrix deposition and growth patterns of microvessels. This research provides an avenue for the in vitro control of vasculogenesis orientation and can potentially bridge the gap between the biology of matrix-cell interactions and translational therapy in pulmonary medicine. By bringing together multidisciplinary areas through the integration of engineering and the life sciences, this proposal addresses fundamental questions in tissue engineering to accelerate the development of functional tissue grafts for the lung, while developing a paradigm that may also be used for other vital organs.
This research training project offers a unique opportunity to bridge the gap between developmental cell biology and tissue engineering for regenerative lung therapy. By analyzing effects of 3D, confined geometries on blood vessel growth, this proposal aims to control the orientation of blood vessel formation outside the body and build a foundation for the development of 3D vascular implants. This project paves the way to improve tissue engineering techniques for a wide variety of regenerative treatments by deepening our understanding of blood vessel formation and the interactions between vascular cells and their natural 3D microenvironment.