Date of Award

7-2019

Document Type

Dissertation

Degree Name

Doctor of Philosophy (PhD)

Department

Biomedical and Chemical Engineering and Sciences

First Advisor

Chris Bashur

Second Advisor

James Brenner

Third Advisor

Vipuil Kishore

Fourth Advisor

Yi Liao

Abstract

Cardiovascular disease (CVD) accounts for one in every five deaths in the United States alone due to occlusion in small diameter (< 6 mm) vessels. The current treatment options includes bypass grafting; however, more than 30 percent of patients do not have viable saphenous veins for autologous grafting [1]. Therefore, tissue engineering is being considered as an alternative approach. The overall goals of this project were to develop a tissue engineered vascular graft with the incorporation of carbon monoxide releasing molecules (CORMs) and to determine the impacts of carbon monoxide (CO) on endothelial cells (ECs) with the goal of promoting a functional endothelium within the graft. This was accomplished through two complementary studies: investigating the impacts of CO-loaded electrospun scaffolds on ECs for cardiovascular applications and diffusion-based modeling of drug delivery of gasotransmitters from tissue engineered scaffolds. In the first study, we extended the maximum in vitro incubation time to permit better cellular attachment and proliferation with a newly-synthesized, more hydrophobic CORM (DK3) and established the impact of CORMs on EC viability and function (e.g. reactive oxygen species (ROS) products, and ROS levels). We further investigated toxicity and biocompatibility of a newly synthesized CORM (DK4) loaded within PCL thin films and nanoparticles. We concluded that the DK4 and other compounds are not toxic at doses ranging from 0 – 50 μg/mL to the ECs both within nanoparticles that can be internalized within cells. We also conducted an in vivo pilot study to determine graft biocompatibility and preliminary results showed that CORM implants maintain mechanical integrity, support blood flow, and do not show toxicity for up to six weeks. In the second study, we validated a computational model and analyzed the output of CO delivery to better understand and control local dose. For CO release, this model is necessary because of the limitations with real-time experimental analysis and the need to better understand the dose available to the cells. We demonstrated that the validated model can be used to predict drug availability to cells for a variety of scaffolds and drug molecules. Our simulated results suggest that only a fraction of the initial concentration of gasotransmitters released from fibers that enters the interstitial fluid in vivo, or culture media in vitro, will be available to cells. We also demonstrated that fiber orientation and fiber diameter are important for drug delivery, but fiber density provides even more important information. The more contact area within the fiber scaffolds is equivalent to experimental conditions with more cell attachment and spreading. These parameters are not only important for traditional tissue engineering, but also for drug delivery. Overall, these results demonstrate the feasibility of making a tissue engineered vascular graft with the incorporation of CORMs and validating he importance of computational modeling of diffusion-based transport of CO. Future work will involve performing surface modifications to enhance cell attachment and proliferation, using the developed computational model to better predict dose available to cells, and experimentally determining impact of CO dose on ECs.

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