CHARACTERIZATION AND IMPLEMENTATION OF A DECELLULARIZED PORCINE VESSEL AS A BIOLOGIC SCAFFOLD FOR A BLOOD VESSEL MIMIC

Every 34 seconds, someone in the United States suffers from a heart attack. Most heart attacks are caused by atherosclerotic build up in the coronary arteries, occluding normal blood flow. Balloon angioplasty procedures in combination with a metal stent often result in successful restoration of normal blood flow. However, bare metal stents often lead to restenosis and other complications. To compensate for this problem, industry has created drug-eluting stents to promote healing of the artery wall post stenting. These stents are continually advancing toward better drug-eluting designs and methods, resulting in a need for fast and reliable pre-clinical testing modalities. Dr. Kristen Cardinal recently developed a tissue engineered blood vessel mimic, with the goal of testing intravascular devices. However, the scaffold component of this model exhibits several physiological limitations that must be addressed to create a truly biomemtic BVM. The current model uses expanded poly(terafluorethylene) [ePTFE] or poly(lactic-go-glycolide) [PLGA] as the choice material for the scaffold. EPTFE has several advantages as it is a widely recognized biomaterial. However, ePTFE is very expensive and lacks native mechanical properties. PLGA is another polymer that is created in-house to produce a uniquely tailored scaffold for use in the BVM; resulting in a cheaper alternative scaffold material. However, PLGA again lacks the necessary native mechanical properties to properly mimic an in-vivo artery. The creation of a biological scaffold will provide a unique biomimetic material to most accurately recapitulate the artery in-vitro. Decellularization is the process of removing all cellular components from a tissue, leaving an acellular structure of extracellular matrix. Understanding the clinical problem and the potential of the BVM, the aim of this thesis is to develop the decellularization process for the creation of a biologic scaffold as a replacement to the non-physiologic polymer scaffolds for the BVM. The first phase of this thesis was to develop and optimize an acceptable protocol for the decellularization of porcine arteries. The use of a 0.075% sodium dodecyl sulfate detergent was sufficient for complete removal of all vascular cell types, without significant degradation to the scaffold wall. In the second phase of this thesis, the decellularized scaffolds were mechanically tested to ensure retention of their native properties. The longitudinal and radial properties of the scaffold were found to be similar to the native artery, indicating the decellularized scaffold improves several physiologically aspects when compared to a polymer scaffold. These mechanical attributes improve the testing environment when evaluating sent deployment or new balloon angioplasty devices; as the decellularized scaffold has an phsyiolgical compliance. The final phase of this thesis examined the cellular adhesion capacities of the scaffold through recellularization with human umbilical vein endothelial cells (hUVECS). Fluorescent microscopy analysis suggests uniform attachment of cells along the length of the scaffold creating a monolayer. These results indicate this new scaffold type can develop an endothelium to complete the ideal, most physiologically relevant BVM system. Further optimization of the decellularization procedures could enhance the ability of the scaffold to be cultured for long-term interaction with intravascular devices

Removal of cardiovascular obstructions by spark erosion

Coronary atherosclerosis, leading to coronary artery stenosis, is the main cause for ischemic health disease in the Westem countries. Stenoses manifest themselves by limiting blood supply to the myocardium thus causing complaints. A long history of degenerative atherosclerotic disease of the intimal wall of the coronary vessels has usually preceded these events. Probably because of this long term process the composition of the accumulated obstructive tissue is quite heterogeneous and consists of a variety of cells and extra cellular material like lipid containing macrophages, smooth muscle cells, Illonocytes, collagen. cholesterol crystals and calcium. In addition, fresh or organized thrombi may have been deposited on these plaques. Regression of these lesions may be obtained by lifestyle changes or lipid lowering therapy. The acute invasive removal of such complex lesions, however, cannot be achieved by applying simple mechanical or chemical means

Aerospace medicine and biology: A continuing bibliography with indexes (supplement 401)

This bibliography lists 140 reports, articles and other documents introduced into the NASA Scientific and Technical Information System during May 1995. Subject coverage includes: aerospace medicine, behavioral sciences, man/system technology and life support, and space biology

Primary Angioplasty

Medicine; Cardiolog

Aerospace medicine and biology: A continuing bibliography with indexes (supplement 403)

This bibliography lists 217 reports, articles and other documents introduced into the NASA Scientific and Technical Information System during July 1995. Subject coverage includes: aerospace medicine and physiology, life support systems and man/system technology, protective clothing, exobiology and extraterrestrial life, planetary biology, and flight crew behavior and performance

Certified Registered Nurse Anesthetist Performance and Perceptions: Use of a Handheld, Computerized, Decision Making Aid During Critical Events in a High-fidelity Human Simulation Environment

With the increasing focus on patient safety and human error, understanding how practitioners make decisions during critical incidents is important. Despite the move towards evidence-based practice, research shows that much decision making is based on intuition and heuristics (“rules of thumb”). The purpose of this study was to examine and evaluate the methodologic feasibility of a strategy for comparing traditional cognition versus the use of algorithms programmed on a personal digital assistant (FDA) in the management of unanticipated critical events by certified registered nurse anesthetists (CRNAs). A combined qualitative-quantitative methodology was utilized. The quantitative element consists of a pilot study using a cross-over trial design. Two case scenarios were carried out in a full-scale, high fidelity, simulated anesthesia care delivery environment. Four subjects participated in both scenarios, one without and one with a PDA containing a catalog of approximately 30 events with diagnostic and treatment related information in second scenario. Audio—videotaping of the scenarios allowed for definitive descriptive analysis of items of interest, including time to correct diagnosis and definitive intervention. The qualitative approach consisted of a phenomenological investigation of problem solving and perceptions of FDA use and the simulation experience by the participants using “think aloud” and retrospective verbal reports, semi-structured group interviews, and written evaluations. Qualitative results revealed that participants found the PDA algorithms useful despite some minor technical difficulties and the simulated environment and case scenarios realistic, but also described feelings of expectation, anxiety, and pressure. Problem solving occurred in a hypothetico-deductive manner. More hypotheses were considered when using the PDA. Time to correct diagnosis and treatment varied by scenario, taking less time with the PDA for one but taking longer with the PDA for the other, likely due to differences in pace and intensity of the two scenarios. The methodologic investigation revealed several areas for improvement including more precise control of case scenarios. All participants agreed with the value of using high fidelity simulation, particularly for problem solving of critical events, and provided useful information for more effective utilization of this tool for education and research

Primary Angioplasty

Medicine; Cardiolog

Development of an In-Vitro Tissue Engineered Blood Vessel Mimic Using Human Large Vessel Cell Sources

Tissue engineering is an emerging field that offers novel and unmatched potential medical therapies and treatments. While the vast aim of tissue engineering endeavors is to provide clinically implantable constructs, secondary applications have been developed to utilize tissue-engineered constructs for in-vitro evaluation of devices and therapies. Specifically, in-vitro blood vessel mimics (BVM) have been developed to create a bench-top blood vessel model using human cells that can be used to test and evaluate vascular disease treatments and intravascular devices. Previous BVM work has used fat derived human microvascular endothelial cells (EC) sodded on an ePTFE scaffold. To create a more physiologically accurate model, a dual layer of large vessel endothelial and smooth muscle cells (SMC) on an ePTFE tube is investigated throughout this thesis. Human umbilical vein endothelial cells (HUVEC) and human umbilical vein smooth muscle cells (HUVSMC) were chosen as the large vessel cell types and cultivated according to standard procedures. Before dual sodding, sodding density experiments with HUVSMC were performed to determine the number of cells required to create a confluent cell layer. HUVSMC sodded by trans-luminal pressure at densities ranging from 3.5x10^5 cells/cm^2 to 1.0x10^6 cells/cm^2 were run for one day to observe luminal coverage. After determining the desirable range for HUVSMC sodding, HUVSMC experiments with 5.0x10^5 cells/cm^2 and 7.5x10^5 cells/cm^2 were run over seven days to evaluate progression of the graft over time. Histology and SEM methods were used for analysis. A HUVEC study was next conducted over 7 days to confirm that the large vessel endothelial cell could be sodded and sustained on ePTFE in-vitro. Next, dual sodding was performed by pressure sodding HUVSMC at 7.5x10^5 cells/cm^2 followed by trans-luminal flow for 30 minutes. HUVECs were subsequently trans-luminally pressure sodded at 5.0x10^5 cells/cm^2 followed by an additional 30 minutes of trans-luminal flow; perfusion flow began following the final 30 minutes of trans-luminal flow. Experiments for the dual layered grafts were run for both one and seven days to evaluate and develop the dual sodding protocol as well as observe the co-culture over time. Analysis of the dual layered grafts was performed by SEM, histology, and fluorescence microscopy. HUVECs were incubated with Cell Tracker™ prior to dual sodding and both cell types with bisbenzimide after graft harvest to attempt to distinguish between cell types. Results from the thesis illustrate that large vessel smooth muscle and endothelial cells can be sodded onto ePTFE scaffolds and sustained within the in-vitro BVM system for up to 7 days. Furthermore, cost analysis demonstrates that the addition of a smooth muscle cell layer adds minimal costs to the BVM system. In conclusion, the studies contained within this thesis culminate in a protocol for the dual sodding of smooth muscle and endothelial cells with the aim of creating a physiologically representative co-culture blood vessel mimic