Molecular, Cellular, and Tissue Engineering Commons^™

Isolating Human Monocytes And Characterizing Human Macrophage Polarization Through Flow Cytometry And Morphological Analysis, Arthur Peter Klujian

Biomedical Engineering

Peripheral Artery Occlusive Disease (PAOD) is a cardiovascular disease that affects more than 200 million people. It is characterized by atherosclerosis, a biologic process where peripheral arteries are restricted due to a buildup in cholesterol and fats. This eventually leads to ischemia, developing complications such as chronic inflammation and possible limb amputation. The current standard of care for patients includes surgical revascularization and exercise therapy. These treatment strategies have had limited success with alleviating symptoms and restoring perfusion, however, many patients still experience ischemia and do not fully restore blood flow. Further exploration in pursuit of treating these shortcomings led …

Evaluation Of Alternative Solvents For Electrospinning Occluded Scaffolds To Model Intracranial Atherosclerotic Disease In Blood Vessel Mimics, Olivia D. Rankin

Master's Theses

Intracranial atherosclerotic disease (ICAD) is characterized by a buildup of fatty plaque within the walls of arteries supplying the brain, resulting in vessel narrowing, restricted blood flow, and increased risk of ischemic stroke. While various treatments exist for ICAD, most are accompanied by high recurrent stroke rates and other adverse events, highlighting the need for further investigation and models for pre-clinical testing. Tissue engineered Blood Vessel Mimics (BVMs), consisting of a polymer scaffold sodded with human vascular cells, serve as physiologically relevant in vitro models for vascular device testing. This thesis focused on the application of alternative solvents to a …

Cardiac Twitch Force Measurement System, Tara Hoffman, Sebastian Carrascal Gómez, Julian Macdonald

Biomedical Engineering

The goal of this project is to develop a low-cost and accurate force measurement system and method of measurement to be used in the testing of grown cardiac tissue samples. The primary stakeholder in this project is Dr. Luke Perreault, a biomedical engineering professor at California Polytechnic State University San Luis Obispo who is studying grown cardiac tissue rings, and any of his student lab personnel. Other stakeholders include other researchers in the tissue engineering field who require a similar form of testing to the methods developed in this project.

This document will introduce the identified problem and background as …

The Effects Of Adipose Tissue Trauma On Vascular Function After Collateral Arteriogenesis, Nathan Tran

Biomedical Engineering

Peripheral artery disease (PAD) affects an estimated 200 million people worldwide and is characterized by arterial narrowing that reduces perfusion to the limbs. Collateral arterioles can naturally restore blood flow through a remodeling process called arteriogenesis, making it a promising therapeutic target. However, the femoral artery ligation (FAL) model used to study arteriogenesis requires surgical access deep to the inguinal fat pad, and approaches for handling the fat pad vary widely across surgeons and protocols. Since adipose tissue harbors a large population of macrophages, the immune cells that govern arteriogenesis, fat pad manipulation may introduce confounding inflammatory stimuli that disrupt …

The Impact Of Human Myoblasts On Macrophage Polarization In A Coculture Model, Elisabeth Ann Stodola

Biomedical Engineering

Peripheral Artery Disease (PAD) is characterized by the buildup of atherosclerotic plaque in the lower extremities that occludes blood flow, and affects over 200 million people worldwide. Without effective treatment, PAD can lead to claudication, ischemia, ulcers, and gangrene. PAD causes more amputations in the United States than any other condition, and for severe cases that require revascularization, procedures benefit fewer than 50% of patients. This demonstrates a clear need for alternative therapies. Potential therapies for PAD target expansion of collateral blood vessels around the area of occlusion – a process called arteriogenesis to compensate for loss of blood flow …

A Novel Microfluidic Device For Preclinical Testing Of Chemotherapeutics, Kaija Saleh Ogan

Master's Theses

This work demonstrates the feasibility of a novel microfluidic tumor model that reproduces physiologically relevant nutrient gradients and endothelium–tumor interactions, validating its potential as a preclinical test platform. Finite element simulations of glutamine diffusion and fluid dynamics aligned with experimental tracer studies, and co-cultures supported directed endothelial migration toward tumor spheroids, confirming the device’s ability to mimic in vivo–like behavior.

Current preclinical models often fail to capture the complexity of the tumor microenvironment (TME), limiting their predictive value for therapeutic testing. Nutrient availability and angiogenic signaling are critical drivers of tumor progression, yet they are poorly represented in traditional …

Computational Investigation Of The Role Of 3d Genome Architecture In The Lifecycle Of The Malaria Parasite, Srish Maulik

College of Engineering Summer Undergraduate Research Program

Malaria, a mosquito-borne infectious disease caused by the Plasmodium parasite, is responsible for more than a half a million deaths per year, the vast majority of which occur in central Africa. The parasite undergoes an incredibly complex cell molecular transformation as it transitions from living in mosquitoes to living in humans with different sets of genes being activated or silenced in order to evade the immune system of the host. Understanding how its genome guides this transition is critical for developing adequate treatments. In this project, we aim to develop a computational framework for investigating the role of the three …

Novel Electrospinning Techniques For Fabricating Scaffolds To Model Intracranial Atherosclerotic Disease In Tissue Engineered Blood Vessel Mimics, Isaac "Squeaky" Buentipo

Master's Theses

Novel Electrospinning Techniques for Fabricating Scaffolds to Model Intracranial

Atherosclerotic Disease in Tissue Engineered Blood Vessel Mimics

Isaac “Squeaky” Buentipo

Dr. Kristen Cardinal’s Tissue Engineering Lab at California Polytechnic State University specializes in fabricating customized vascular models, known as Blood Vessel Mimics (BVMs), for medical device testing. BVMs are simplified in vitro vascular models designed to bridge the gap between bench testing and in vivo evaluation of intravascular medical devices. BVMs are produced by combining a polymer scaffold with human vascular cells. Through novel scaffold fabrication techniques, such as silicone injection molding, silicone dip casting and electrospinning, the capabilities of …

Electroporation Protocol Development For Cal Poly Cell Therapy Laboratory Courses, Gabrielle Melamed

Biomedical Engineering

Cancer is a heterogeneous disease caused by unchecked cell proliferation. The disease contributed to 9.7 million deaths worldwide in 2022, with hematological malignancies (leukemias, lymphomas, and myelomas) accounting for over 1.4 million deaths in the United States from 1999 to 2020. This thesis focuses on the treatment of these cancers, with an emphasis on non-Hodgkin lymphomas (NHLs). R-CHOP (Rituximab-Cyclophosphamide, Doxorubicin hydrochloride, Oncovin/Vincristine, and Prednisone) is the standard of care first-line treatment for diffuse large B-cell lymphoma (DLBCL), the most common subtype of NHL, and encompasses a mixture of chemotherapy, immunotherapy, and corticosteroids. The R-CHOP regimen cures 50% to 70% patients, …

Hypoxic Incubator: Improving Robustness/Reliability And Demonstrating Physiological Efficacy, Damon Dennis Tan

Master's Theses

The Microphysiological Systems Laboratory aims to develop colorectal cancer tumor models under a hypoxic environment to assess model response to pharmaceutical compounds in vitro. To perform relevant studies, researchers have attempted to use different hypoxic inducing strategies such as a nitrogen pod and hypoxic incubator to recreate in vivo physiological responses to hypoxia. However, studies would be interrupted due to incubator functionality failure. To ensure successful and physiologically relevant studies, I improved and verified the robustness and reliability of a hypoxic incubator previously designed and manufactured in the lab. Through the testing and iterating design processes, I engineered and implemented …

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

Biomedical Engineering: Graduate Reports and Projects

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 …

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 …

Bone And Cartilage Compression Simulator, Karoline M. Wucherer, Benjamin A. Parmentier, Thomasina E. Hinkle

Biomedical Engineering

A device was developed that delivers mechanical loads to bone and soft tissue samples under physiological conditions to aid in the research of tissue engineering bone and cartilage. To begin the design process, a Network Diagram and Gantt Chart were produced to create a general timeline for the project to follow. This allowed us to measure our progress and determine what effects (if any) delays could have on our project.

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 …

The Impact Of Myoblast Transplantation On Collateral Capillary Arteriogenesis And Macrophage Phenotype, Christine Hue Do

Biomedical Engineering

Gangrene, pain, loss of limb function, amputation, and death are only few of the grievous consequences associated with peripheral arterial disease (PAD), a vascular disease caused by an obstruction that narrows the blood vessels. Since some patients have collateral vessels that can re-route blood to its downstream destination, much focus has been spotlighted upon discovering the mechanism of this process, termed arteriogenesis, as well as cell therapies to increase arterial diameter of collateral vessels. Since some patients do not have native pre-existing collateral vessels, another method to re-route blood is through arterialized collateral capillaries (ACC), which is the conversion of …

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 …

Mechanisms Of Reduced Vascular Tone Following Arteriogenesis Induced By Femoral Artery Ligation, Christopher Hatch

Biomedical Engineering

The presence of a developed, native collateral network can decrease the severity of ischemic injury proceeding arterial occlusion. The collateral network must under arteriogenesis to enlarge and increase blood flow to the ischemic region. Although there has been tremendous effort attempting to understand the mechanisms of arteriogenesis, no therapies have been successful in improving patient outcome. To better understand the mechanisms involved in arteriogenesis, the effect of nitric oxide production, myogenic tone, and a-adrenergic receptors were evaluated as these have been identified as playing an important role in vascular injury. Arteriogenesis was induced by ligating the femoral artery between the …

Nano Fab Lab 63, Brian K. Deemer, Josh Clemons, Nick Brodine, Delaney Fitzsimmons

Mechanical Engineering

Lawrence Livermore National Laboratories (LLNL) has several steps in the production process for ceramic nanofiber tubes that they would like to improve - electrospinning, cutting, rolling, sealing and heat treating. We undertook the challenge to deliver LLNL with a semi-automated process that efficiently integrates the steps of cutting, rolling, and sealing to save time and improve control over end dimensions. In this document, we discuss the technical background of the manufacturing steps currently followed to create nanofiber tubes, identify which steps are incorporated in our prototype and explain how they will interface with one another, define the design requirements, present …

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 …

Plga Scaffold Device, Tyler D. Tackabery, Bryce Powada, Tate Morell

Biomedical Engineering

No abstract provided.

Fluid Delivery System For A Cell Culture On A Microfluidic Chip, Austin J. Roeder, Colleen A. Richards, Emily A. Matteson

Biomedical Engineering

This project report provides a description of the progress made in the development of a fluid delivery system for a microfluidic cell culture on a chip. The system is intended to be used in a humidified incubator in a university laboratory and the fluid delivery system is required to exist and operate within that incubator for extended periods of time. Therefore, the system will be gravity-driven and contain no electronic components. The key specification of the system is to provide fluid flow at a constant velocity.

After manufacturing and testing the device, all specifications were met except for the fluid …