Assessing Effects of Decellularization and Culture Conditions on Small-Diameter Vascular Graft Seeding
Abstract
In tissue engineering, detergent-based decellularization of a tissue can be used to create tissue-specific scaffolds. The process “clears the way” for a desired cell type(s) to then be seeded into or onto the scaffold in place of the displaced cells. Tissue engineered grafts represent an attractive alternative to current graft options for vascular surgeries, which can suffer from complications such as thrombosis, graft-rejection, and reduced patency. In our work to contribute to the production of a small-diameter tissue engineered vascular graft, we are working with decellularized porcine interior thoracic artery (PITA) scaffolds. In the work presented here, we tested two hypotheses: 1) a higher percentage detergent used in decellularization will lead to greater porosity of the medial layer of PITA scaffolds, and 2) extensive rinsing of PITA scaffolds following decellularization will allow endothelial cells to adhere to and grow on the luminal surface. The following results supported our hypotheses: 1) Scanning electron microscopy of the PITA scaffolds showed that the ultrastructure of the medial layer appeared more porous with increasing detergent concentration, and 2) a resazurin reduction assay demonstrated that extensive rinsing of the scaffolds allowed for greater viability of cells in the presence of PITA scaffolds and directly on PITA scaffolds. Fluorescence microscopy was also used to confirm seeding of endothelial cells onto PITA scaffolds. In the future, we plan to simultaneously seed smooth muscle cells and/or adipose-derived mesenchymal stem cells along with endothelial cells into PITA scaffolds.
Assessing Effects of Decellularization and Culture Conditions on Small-Diameter Vascular Graft Seeding
In tissue engineering, detergent-based decellularization of a tissue can be used to create tissue-specific scaffolds. The process “clears the way” for a desired cell type(s) to then be seeded into or onto the scaffold in place of the displaced cells. Tissue engineered grafts represent an attractive alternative to current graft options for vascular surgeries, which can suffer from complications such as thrombosis, graft-rejection, and reduced patency. In our work to contribute to the production of a small-diameter tissue engineered vascular graft, we are working with decellularized porcine interior thoracic artery (PITA) scaffolds. In the work presented here, we tested two hypotheses: 1) a higher percentage detergent used in decellularization will lead to greater porosity of the medial layer of PITA scaffolds, and 2) extensive rinsing of PITA scaffolds following decellularization will allow endothelial cells to adhere to and grow on the luminal surface. The following results supported our hypotheses: 1) Scanning electron microscopy of the PITA scaffolds showed that the ultrastructure of the medial layer appeared more porous with increasing detergent concentration, and 2) a resazurin reduction assay demonstrated that extensive rinsing of the scaffolds allowed for greater viability of cells in the presence of PITA scaffolds and directly on PITA scaffolds. Fluorescence microscopy was also used to confirm seeding of endothelial cells onto PITA scaffolds. In the future, we plan to simultaneously seed smooth muscle cells and/or adipose-derived mesenchymal stem cells along with endothelial cells into PITA scaffolds.
