Biomedical Engineering and Bioengineering Commons^™

Engineering An Injectable Hydrogel With Self-Assembling 3d Vasculature, Kendyl Cohn

Master's Theses

This research developed methods for culturing self-assembling capillaries in an injectable gel as a potential method for vascularizing tissue-on-a-chip models to mimic physiological drug delivery. Additionally, a mathematical model was developed as a tool for understanding nutrient delivery and comparison of potential delivery systems. Organs-on-a-chip provide novel platforms for studying biology and physiology in 3D, allow exploration of tissue engineering on a manageable scale, and serve as models for drug screening and drug-delivery testing.

Methods were first developed for co-culture of endothelial cells and fibroblasts (3T3s or HDFs) in 2D, evaluating culture time, seeding density and ratio of HUVECs and …

Manufacturing Silicone In-House For The Creation Of Customized Neurovascular Blood Vessel Mimics, Jacob Wilbert Perisho

Master's Theses

The Tissue Engineering Lab at California Polytechnic State University San Luis Obispo focuses on creating tissue-engineered Blood Vessel Mimics (BVMs) designed for the preclinical testing of neurovascular devices. These BVMs are composed of silicone models, representing anatomically accurate neurovasculatures, that are sodded with vascular cell types and then cultivated in bioreactors (which maintain physiologic conditions). These silicone models are currently sourced externally from industry partners, so the primary goal of this thesis was to develop the means and methods for the Tissue Engineering Lab to manufacture silicone models in-house.

The first aim of this thesis was to develop and explore …

Development Of An In Vitro 3-Dimensional Co-Culture Human Colorectal Cancer Model In Microfluidic Devices, Abby Jens

Master's Theses

Colorectal cancer is the second most common cause of cancer-related deaths in the United States, with the relative 5-year survival rate for distant stage cancer being only 14%. The most common treatment for colorectal cancer is with chemotherapeutic drugs; however, the discovery of these drugs is costly, time-consuming, and often requires the use of animal models that do not yield results that translate to clinical trials. Due to these shortcomings, researchers seek to develop physiologically relevant in vitro tumor models that more accurately mimic the tumor microenvironment for cheaper and faster high-throughput drug screening. The aim of this research was …

Development Of An Angiogenic Tissue-On-A-Chip Microenvironment, Eric Stuehr

Master's Theses

Preclinical testing is necessary to investigate the safety and efficacy of novel therapeutics before moving to clinical trials, yet approximately 90% of these therapies fail once tested in humans. This has led to increased interest in developing robust preclinical models that accurately mimic the complex human in vivo physiology. Microfluidic devices that can introduce dynamic conditions to 3D cell/organoid cultures, also known as tissue-on-a-chip, have emerged as physiologically relevant in vitro preclinical models that can achieve high throughput screening of therapeutics. The research presented here aimed to develop an angiogenic environment within a novel microfluidic device to stimulate formation of …

Implementation Of Methods For Cell Therapy Research And Assessment Of The Impact Of Cryopreservation, Samantha Lynes

Master's Theses

Cell therapy is the administration of living cells into a patient to prevent or treat a variety of diseases and illnesses. The cell therapy industry is rapidly expanding, and continued research is necessary for manufacturing safe and effective therapies. Although cell therapy manufacturing generally involves cryopreservation processes for storage, there are limited standards for cryopreservation processes and assays required to evaluate cell therapies post-thaw, and limited understanding exists about how a recovery period post-thaw could impact cell health. The overall goal of this thesis was to evaluate the effect of cryopreservation and potential subsequent recovery time on cell viability.

Because …

Hypoxic Incubation Chamber, Simone Lisette Helfrich, Makenzie Nicole Jones

Master's Theses

This paper describes the design, manufacturing, and testing of a novel controllable hypoxic incubator with fully functional oxygen gas control and temperature control in a humid environment. On the current market, a majority of the few hypoxic incubators use pre-mixed gas that does not offer precise control over gas concentration. The objective for this project was to create a chamber that allows the user to set the O2 concentration to varying set points of % O2 while maintaining the chamber at a constant body temperature, CO2 level, humidity, and sterility. To start the project, multiple concepts were developed for the …

Optimization Of A Novel Nipam-Based Thermoresponsive Copolymer For Intramuscular Injection As A Myoblast Delivery Vehicle To Combat Peripheral Artery Occlusive Disease, Quentin R. Klueter

Master's Theses

There is a need for a minimally invasive delivery method to enable cell therapies to combat peripheral artery occlusive disease (PAOD) in end stage patients. Myoblasts show promise as a cell mediated therapy but warrant an improved delivery method to increase cell retention in the region of interest because of their adherent nature, relative to previously used BM-MNC’s that are non-adherent. Contemporary issues with achieving successful cell therapies of vasculature can be mainly characterized by the lack of clinical translation from promising animal studies and absence of cell delivery scaffolding. Naturally, polymers have been widely experimented with as grafts to …

Assessing The Effects Of Oxaliplatin On An In Vitro Three-Dimensional Human Colorectal Cancer Model, Sabrina Nelson

Master's Theses

Colorectal cancer is the third most common cancer in the United States with a 5-year late-stage survival rate of only 14%. Due to the lack of translation between animal models and clinical trials as well as the inefficacy of many chemotherapeutics in initial clinical trials, researchers are turning to in vitro drug screening models in an effort to mimic the conditions in vivo. This research project aimed to validate an in vitro tumor culture model within a microfluidic device using a clinically relevant chemotherapy drug. The first experiment consisted of a cell density and drug concentration study to determine …

Development Of A Robust Methodology To Obtain And Assess Myogenic Precursor Cells For Their Use In Regenerative Therapies, Ricardo Lasa

Master's Theses

Peripheral arterial occlusive disease (PAOD) is characterized by buildup of atherosclerotic plaque in peripheral arteries that leads to an occlusion that can interrupt the supply of blood to the peripheral tissue, causing downstream tissue ischemia/hypoxia. PAOD is estimated to affect over 200 million patients worldwide. Current surgical revascularization treatments can be effective in about half of the patient population, leading to a significant number of patients with no treatment options beyond pharmacological intervention and lifestyle modification. The decrease in blood flow downstream of the occlusion leads to increased blood pressure gradient in the microvasculature, specifically in vessels that connect arterial …

Studying The Efficacy Of An Injectable 3-Dimensional Fibrin Extracellular Matrix To Characterize The Effects Of Antitumor Agents On Sw620 Cells In A Microfluidic Device, Thèo Anastos

Master's Theses

Colorectal cancer is the third most common cancer in the United States and there is currently a lot of research going into new antitumor agents to kill the cancer. One method for replicating the tumor response to a drug in vivo is by creating an in vitro drug testing model to replicate the in vivo condition. This research project was conducted to determine the efficacy of testing tumor cultures in a microfluidic device as a way to provide accurate drug responses in vitro instead of using in vivo subjects in clinical trials. A total of four experiments were conducted with …

Development And Optimization Of Imaging And Image Quantification Techniques For Tissue-Engineered Blood Vessel Mimics, Ashley Turcott

Master's Theses

Blood vessels mimics (BVMs) are tissue-engineered blood vessels used to test vascular devices in an environment that mimics some simple anatomical factors of native blood vessels. It is important to accurately and consistently assess tissue-engineered blood vessels, although there is currently a lack of standardization in Cal Poly’s Tissue Engineering Lab and in the entirety of the field. The goal of this thesis was to develop and optimize imaging and image quantification techniques for tissue-engineered blood vessels.

The first aim of this thesis optimized and compared imaging and assessment techniques for electrospun scaffolds. Images from different SEMs were compared to …

Investigation Of Experimental Variation Of Bovine Sphingomyelin As A Novel Ingredient For Ultraviolet Protection, Esther Chen

Master's Theses

Skin cancer is a prevalent disease that globally affects 2-3 million people per year [1]. This number is expected to grow tenfold as depletion of the ozone layer contributes to harsher rays reaching Earth’s surface [2]. A common way to protect against those ultraviolet waves is to apply sunscreen, however, recent reports call into question the safety of some active ingredients as they can enter through the skin into the bloodstream [3]. This thesis aims to investigate an alternative solution that uses bovine sphingomyelin (BSM) as photoprotective solution against UV irradiation.

In order to evaluate the effectiveness of BSM against …

Evaluation Of Human Umbilical Vein Endothelial Cells In Blood Vessel Mimics Through Changes In Gene Expression And Caspase Activity, Conor Charles Hedigan

Master's Theses

Blood vessel mimics (BVMs) are simple tissue engineered blood vessel constructs intended for preclinical testing of vascular devices. This thesis developed and implemented methods to characterize two of these components. The first aim of this thesis investigated the effect of cell culture duration and flow conditions on endothelial cell gene expression, especially regarding endothelial-to-mesenchymal transition (EndMT). A trend of decreased endothelial marker gene expression and increased mesenchymal marker gene expression would indicate EndMT. qPCR analysis revealed that increased cell culture duration did not result in EndMT, and in fact increased endothelial marker expression as cell culture duration increased. Disturbed flow …

In Vitro Growth Of Osteoblasts On Poly Lactic-Co-Glycolic Acid Scaffolds Created Via Gas Foaming, Matthew James Thomas

Master's Theses

This study analyzed the feasibility of using gas foaming to create Poly Lactic-co-Glycolic Acid (PLGA) scaffolds for use as a substrate in bone tissue engineering and set out to determine whether the presence of osteoblasts on these scaffolds enhanced their material stiffness. The process of bone formation involves osteoblasts depositing extracellular matrix and calcifying this matrix with calcium phosphate crystals (Hasegawa et al., 2017) and pits between 30-40μm in diameter on tissue engineering scaffold surfaces have been shown to best promote osteogenic activity in the presence of bone-forming cells (Halai et al., 2014).The scaffolds were determined to contain pits within …

Development And Characterization Of Tissue Engineered Blood Vessel Mimics Under "Diabetic" Conditions, Shelby Gabrielle Kunz

Master's Theses

The development of tissue engineered blood vessel mimics for the testing of intravascular devices in vitro has been established in the Cal Poly tissue engineering lab. Due to the prevalence of cardiovascular disease in diabetic patients and minimal accessible studies regarding the interactions between diabetes and intravascular devices used to treat vascular disease, there is a need for the development of diabetic models that more accurately represents diabetic processes occurring in the blood vessels, primarily endothelial dysfunction. This thesis aimed to create a diabetic blood vessel mimic by implementing a high glucose environment for culturing human endothelial cells from healthy …

Matrix Remodeling Accompanies In Vitro Articular Cartilage Spherical Shaping, Nathan Thomas Balcom

Master's Theses

Introduction: Articular cartilage (AC) is a low friction load bearing material found in synovial joints. The natural repair of damaged tissue is difficult and often requires surgical intervention. With large defects it becomes necessary to match the original tissue geometry. We hypothesized that localized collagen (COL) and/or proteoglycan (PG) remodeling occurs during AC spherical reshaping. The objective of this study was to determine the presence, magnitude and depth dependence of COL and PG remodeling that accompanies AC reshaping. Methods: Full thickness AC blocks (7x7 mm² surface area) were harvested from the ridges of the patellofemoral groove of immature (1-3 …

Development Of In Vitro Tissue Engineered Blood Vessel Mimics In Complex Geometries For Coronary Stent Testing, Robert Dalton Chavez

Master's Theses

Coronary heart disease is the leading cause of death in the United States and occurs when plaque occludes coronary arteries. Coronary stents, which may be used to treat coronary occlusions, are small metal tubes that are implanted in coronary arteries to restore blood flow. After stent implantation, endothelial cells grow over the stent so that blood contacts the endothelial cells instead of the stent surface; this event is known as re-endothelialization. Re-endothelialization prevents blood from clotting on the stent surface and is a good predictor of stent success. Blood vessel mimics (BVMs) are in vitro tissue engineered models of human …

Characterization Of Tight Junction Formation In An In-Vitro Model Of The Blood-Brain Barrier, Michael Robert Machado

Master's Theses

Active and passive transport of substances between the microcirculation in the brain and the central nervous system is regulated by the Blood-Brain Barrier (BBB). This barrier allows for chronic and acute modulation of the CNS microenvironment, and protects the brain from potentially noxious compounds carried in the circulatory system. In-vitro modeling of the BBB has become the target of much research over the past decade, as there are many unanswered questions regarding modulations in the permeability of this barrier. Additionally, the development of a practical and inexpensive model of the BBB would facilitate a much more efficient drug development process. …

Tissue Engineering A Blood Vessel Mimic While Monitoring Contamination Through Sterility Assurance Testing, Navid Djassemi

Master's Theses

Tissue Engineering A Blood Vessel Mimic While Monitoring Contamination Through Sterility Assurance Testing

Navid Djassemi

Tissue engineering blood vessel mimics has been proposed as a method to analyze the endothelial cell response to intravascular devices that are used in today’s clinical settings for the treatment of cardiovascular disease. Thus, the development of in vitro blood vessel mimics (BVMs) in Cal Poly’s Tissue Engineering Lab has introduced the possibility of assessing the characteristics of cellular response to past, present, and future intravascular devices that aim at treating coronary artery disease.

This thesis aimed at improving the methods and procedures utilized in …

Development And Characterization Of An In-House Custom Bioreactor For The Cultivation Of A Tissue Engineered Blood-Brain Barrier, Amin Hadi Mirzaaghaeian

Master's Theses

The development of treatments for neurological disorders such as Alzheimer’s and Parkinson’s disease begins by understanding what these diseases affect and the consequences of further manifestation. One particular region where these diseases can produce substantial problems is the blood-brain barrier (BBB). The BBB is the selective diffusion barrier between the circulating blood and the brain. The barrier’s main function is to maintain CNS homeostasis and protect the brain from the extracellular environment. The progression of BBB research has advanced to the point where many have modeled the BBB in vitro with aims of further characterizing and testing the barrier. Particularly, …

Viscoelastic Anisotropic Finite Element Mixture Model Of Articular Cartilage Using Viscoelastic Collagen Fibers And Validation With Stress Relaxation Data, Matthew Alexander Griebel

Master's Theses

Experimental results show that collagen fibers exhibit stress relaxation under tension and a highly anisotropic distribution. To further develop the earlier model of Stender [1], the collagen constituent was updated to reflect its intrinsic viscoelasticity and anisotropic distribution, and integrated with an existing mixture model with glycosaminoglycans and ground substance matrix. A two-term Prony series expansion of the quasi-linear viscoelastic model was chosen to model the viscoelastic properties of the collagen fibers. Material parameters were determined by using the simplex method to minimize the sum of squared errors between model results and experimental stress relaxation data of tissue in tension. …

Design Of Experimentation To Systematically Determine The Interaction Between Electrospinning Variables And To Optimize The Fiber Diameter Of Electrospun Poly (D, L-Lactide-Co-Glycolide) Scaffolds For Tissue Engineered Constructs, Yvette S. Castillo

Master's Theses

Cardiac disease causes approximately a third of the deaths in the United States. Furthermore, most of these deaths are due to a condition termed atherosclerosis, which is a buildup of plaque in the coronary arteries, leading to occlusion of normal blood flow to the cardiac muscle. Among the methods to treat the condition, stents are devices that are used to restore normal blood flow in the atherosclerotic arteries. Before advancement can be made to these devices and changes can be tested in live models, a reliable testing method that mimics the environment of the native blood vessel is needed. Dr. …

Protective Effects Of Milk Phospholipids Against Uv Photodamage In Human Skin Equivalents, Zyra Achay

Master's Theses

The ultraviolet (UV) spectrum has been known to cause damage to skin in varying degrees. UVB radiation (290-320 nm) in particular, has been proven to be highly mutagenic and carcinogenic in many animal experiments compared to either UVA or UVC. The alarming rate of increase in skin cancer incidence has prompted many investigators to pursue other alternatives to sunscreens including changes in lifestyle habits and dietary consumption in order to boost our efforts in tackling this widespread disease. Previous studies employing confocal reflectance, MTT assay and histology suggest that milk phospholipids may possess protective properties against UVB-mediated damage but the …

Implementation And Assessment Of Hyperglycemic Conditions For The Creation Of A Diabetic Blood Vessel Mimic, Vikramaditya Mediratta

Master's Theses

Introduction: Diabetes Mellitus is a metabolic disorder that affects a person’s ability to either produce insulin (Type I diabetes mellitus) or properly use insulin (Type II diabetes mellitus) in order to maintain adequate blood glucose levels. The most severe diabetic complications arise due to hyperglycemia – a state of extremely high blood glucose levels – such as, coronary artery disease (CAD), in which coronary stent therapy is a popular method of treatment. However, research has shown a high rate of in-stent restenosis in diabetic patients with CAD, most likely due to activation of cellular adhesion molecules on endothelial cells exposed …

Ischemia Impairs Vasodilation In Skeletal Muscle Resistance Artery, Kyle Remington Struthers

Master's Theses

Functional vasodilation in arterioles is impaired with chronic ischemia. We sought to examine the impact of chronic ischemia and age on skeletal muscle resistance artery function. To examine the impact of chronic ischemia, the femoral artery was resected from young (2-3mo) and adult (6-7mo) mice and the profunda femoris artery diameter was measured at rest and following gracilis muscle contraction 14 days later using intravital microscopy. Functional vasodilation was significantly impaired in ischemic mice (14.4±4.6% vs. 137.8±14.3%, p<0.0001 n=8) and non-ischemic adult mice (103.0±9.4% vs. 137.8±14.3%, p=0.05 n=10). In order to analyze the cellular mechanisms of the impairment, a protocol was developed to apply pharmacological agents to the experimental preparation while maintaining tissue homeostasis. Endothelial and smooth muscle dependent vasodilation were impaired with ischemia, 39.6 ± 13.6% vs. 80.5 ± 11.4% and 43.0 ± 11.7% vs. 85.1 ± 10.5%, respectively. From this data, it can be supported that smooth muscle dysfunction is the reason for the observed impairment in arterial vasodilation.

The Electrical Properties Of Human Tissue For The Diagnosis And Treatment Of Melanoma Skin Cancer, Glenn Cameron Stante

Master's Theses

This thesis discusses the research, experimental methods, and data gathered for the investigation of a novel method for the diagnosis of melanoma skin cancer. First, a background about human skin tissue is presented. Then, a detailed description of melanoma along with current diagnosis techniques and treatment options are presented. In the experimental methods, the electrical properties of several types of tissue were analyzed, the purpose of which was to discover if a tissue type can be distinguished by its electrical properties alone. This would allow for the diagnosis of melanoma to be done by examining the electrical properties of the …

Regression Analysis Of Fracture Toughness For Secondary Osteons Located In Human Cortical Bone, Chase A. Fetzer

Master's Theses

An experiment was carried out in order to locate and quantify osteon types within a sample of cortical bone taken from a human tibia. This was done using a microscope-camera assembly and the BioQuant computer software. The results of this were correlated with a previous experiment’s results on fracture toughness so that an analysis could be run on the data in order to determine the factors that most affect the value of fracture toughness of this cortical bone. Results were examined closely and the analysis repeated until the author was satisfied that the best possible model for fracture toughness had …

Assessment Of Electrospinning As An In-House Fabrication Technique For Blood Vessel Mimic Cellular Scaffolding, Colby M. James

Master's Theses

Intravascular devices, such as stents, must be rigorously tested before they can be approved by the FDA. This includes bench top in vitro testing to determine biocompatibility, and animal model testing to ensure safety and efficacy. As an intermediate step, a blood vessel mimic (BVM) testing method has been developed that mimics the three dimensional structure of blood vessels using a perfusion bioreactor system, human derived endothelial cells, and a biocompatible polymer scaffold used to support growth of the blood vessel cells. The focus of this thesis was to find an in-house fabrication method capable of making cellular scaffolding for …

Preparation And Characterization Of Electrospun Poly(D, L-Lactide-Co-Glycolide) Scaffolds For Vascular Tissue Engineering And The Advancement Of An In Vitro Blood Vessel Mimic, Tiffany Richelle Pena

Master's Theses

PREPARATION AND CHARACTERIZATION OF ELECTROSPUN POLY(D,L-LACTIDE-CO-GLYCOLIDE) SCAFFFOLDS FOR VASCULAR TISSUE ENGINEERING AND THE ADVANCEMENT OF AN IN VITRO BLOOD VESSEL MIMIC

Tiffany Richelle Peña

Currently, an estimated 1 in every 3 adult Americans are affected by one or more cardiovascular complications. The most common complication is coronary artery disease, specifically atherosclerosis. Outcomes of balloon angioplasty treatments have been significantly improved with the addition of drug eluting stents to the process. Although both bare metal and drug eluting stents have greatly increased the effectiveness of angioplasty and decreased the occurrence of restenosis, several complications still exist. For this reason, the stent …

Development Of An In-Vitro Tissue Engineered Blood Vessel Mimic Using Human Large Vessel Cell Sources, Dimitri E. Delagrammaticas

Master's Theses

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 …