Angiogenesis Project

 In order for a biomaterial implant to be optimally functional, it must be able to be integrated into the host's body. A key aspect of this successful integration is whether there is vascularization of the tissue surrounding the implant. The more vascular the tissue, the better the integration of the material in the host.

 In this specific study, the focus is to identify how age and biomaterial type affects this vascularization.  In order to test for this 8 week and 18 month old mice were given some biomaterial, polypropylene mesh or urinary bladder matrix (UBM), along with a abdominal defect to simulate the scenario in which the biomaterial would be used.  The mice were given 7 or 14 days to recover before the biomaterial and surrounding tissue were explanted. After the biomaterial explants were taken, the condition medium surrounding the explant was extracted to be grown on its own for 24 hours. This condition medium contains the macrophages as well as its related growth factors.  The condition medium were then used for tests as mentioned below.   

 To test the effects of the macrophage shifts and biomaterial on angiogenesis, two tests have been planned: a tubule formation assay and a migration assay. The tubule formation assay involves placing cells(endothelial cells) on a gel membrane and then the condition medium on top of the cells. The cells are allowed to grow for a certain period of time before being imaged for analysis. The migration assay involves placing endothelial cells on a well above a gold plate with another well full of condition medium below the gold plate. The growth factors within the condition medium will trigger migration of the cells toward the condition medium. To do this, the cells move down onto the gold plate which has a constant voltage running through it. As more cells migrate onto the plate, the resistance of the plate gets should get higher. Thus, a condition medium that promotes more migration should have a plate with lower current reading with a higher resistance. 

 I was personally was tasked with culturing the human umbilical vein endothelial cells (HUVEC) to be used for the tubule formation and migration assay. Additionally, I assisted the conducting of the tubule formation assay as well as the analysis of its data. To collect data from the tubule formation, the amount of nodes(connection of 2+ vessels) and total blood vessel length were collected.
I was personally was tasked with culturing the human umbilical vein endothelial cells (HUVEC) to be used for the tubule formation and migration assay. Additionally, I assisted the conducting of the tubule formation assay as well as the analysis of its data. To collect data from the tubule formation, the amount of nodes(connection of 2+ vessels) and total blood vessel length were collected. 

Below is the preliminary data and pictures relating to the project: