Available at: http://digitalcommons.calpoly.edu/theses/961
Date of Award
MS in Biomedical Engineering
Biomedical and General Engineering
Kristen O'Halloran Cardinal
Cardiovascular disease is the leading cause of death in developing countries. Because thousands of people are affected by this disease, medical device companies are constantly developing new intravascular devices in attempt to cure or alleviate the symptoms of the disease. However, before these devices can be brought to market or implanted in to patients, they must complete three stages of testing to receive FDA approval. These testing stages include: in vitro testing, in vivo testing, and clinical trials. Currently, there is a large gap between in vitro and in vivo testing because it is difficult to obtain physiologic information about a device during in vitro testing. Therefore, to obtain this information, most devices move on to in vivo testing , increasing the amount of time, money and animal models used during the approval process. In attempt to overcome this limitation of the approval process, Dr. Kristen Cardinal developed a tissue engineered blood vessel mimic (BVM) to bridge the gap between in vitro and in vivo testing to efficiently test intravascular devices. However, before the BVM can be utilized to test intravascular devices, key limitations must be overcome. The limitation addressed in this thesis is the lack of endothelial cell adhesion to the PLGA scaffold used in the BVM. In attempt to overcome this limitation, protein pre-coatings were characterized in 6-well plates and then implemented in to the BVM to determine if endothelial cell adhesion could be increased.
The aim of this thesis was to analyze and compare the effects of different protein pre-coatings coatings on endothelial cell adhesion in 6-well plates and the BVM. The first phase of this thesis sought to characterize the coatings and their effects on cell attachment in a controlled and efficient setting. Different aspects of coating protocols were developed during this phase including the optimization of incubation periods and analysis protocols. The second phase of this thesis included the implementation of the most effective pre-coatings from the 6-well plate studies (Conditioning Media and ProNectin-F) in to the BVM to determine which coating was most effective in increasing cell attachment and reducing cell loss after flow exposure. Using fluorescent staining and image analysis, it was concluded that Conditioning Media was as effective as ProNectin-F in increasing cell attachment and retention in the BVM. While both coatings significantly increased the number of cells adhered compared to a non-coated scaffold, the endothelial lining in the lumen was not 100% confluent. Therefore, other coatings and/or combinations of coatings can be studied in the future to continue to improve cell attachment in the BVM system.