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 Dipeptides On Product Quality In Chinese Hamster Ovary Cells, Louise C. Herman

All Theses

Biopharmaceuticals are medicines produced in cells which are used to treat diseases such as cancer. Monoclonal antibodies (mAbs) represent the majority of approved biotherapeutics. MAbs are mostly produce using Chinese hamster ovary (CHO) cells. CHO cells are favored for resiliency, scalability, and established safety, and are considered the “workhorse” organism for biopharmaceuticals.  Amino acids are a key component of cell culture media, however several amino acids have solubility limits, such as cysteine, cystine, and tyrosine.  At neutral pH, these amino acids are poorly soluble.  However, dipeptides, such as N,N'-di-L-Alanyl-L-Cystine (ACCA) have been reported to increase cysteine and cystine availability. Further, …

Fluorescence-Based Quantification Of Hepatic Ldl Uptake And Donor Hepatocyte Engraftment Using Atto 700–Labeled Ldl, Victor Agustin Cortes Flores

All Theses

Familial hypercholesterolemia (FH) is an inherited disorder characterized by elevated low-density lipoprotein cholesterol (LDL-C) that increases the risk of premature atherosclerotic cardiovascular disease. In severe FH, including low-density lipoprotein receptor (LDLR), hepatic LDL clearance is profoundly impaired and many lipid lowering treatment strategies have limited effectiveness. Current advances in ex vivo CRISPR–Cas9 hepatocyte gene editing in vivo and ex vivo approaches for treatment of FH are under development.

However, a central barrier to translation is the ability to quantitatively measure the percentage of healthy gen edited hepatocytes in the liver while simultaneously assessing functional hepatic LDL handling. This thesis establishes …

Quantitative Fluorescence Imaging Using A Paired Agent Approach For Improving Surgical Decision Making, Sanjana Pannem

Dartmouth College Ph.D Dissertations

Surgical resection remains to be the cornerstone of treatment for most solid tumors, where the primary objective is to remove all visible and microscopic disease. However, the extent of tumor resection is inherently limited by the surgeon’s ability to distinguish tumor from normal tissue, intraoperatively. As a result, positive surgical margins (PSM), where residual tumor remains after surgery, are a common occurrence and are associated with increased recurrence, need for adjuvant therapies, dismal patient outcomes, and high healthcare costs.

Fluorescence guided surgery (FGS) has emerged as a promising technique to enhance intraoperative visualization of tumors and may help reduce the …

Advancing Computational Analysis Of Fluorescence Imaging For Head And Neck Cancer Surgical Guidance, Yao Chen

Dartmouth College Ph.D Dissertations

The main goal of surgery for head and neck squamous cell carcinoma (HNSCC) is to remove the tumor completely while preserving as much healthy tissue as possible to enhance patient survival and reduce complications. However, achieving clear surgical margins is difficult due to the complex anatomy of the head and neck and the reliance on a surgeon’s visual and tactile assessments. Recently, near-infrared (NIR) fluorescence molecular imaging has shown potential for better tumor visualization during surgery, using targeted imaging agents like epidermal growth factor receptor (EGFR) probes. This has led to the development of fluorescence-guided surgery (FGS), which helps surgeons …

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 …

The Effects Of Low-Frequency, Low-Intensity Ultrasound On Gene Expression In Macrophage-Endothelial Cell Co-Culture, Evelyn Hergenhan

Honors Theses

Introduction

Low-frequency, low-intensity ultrasound (LFLI US) has been shown to be a promising treatment for noninvasive accelerated chronic wound healing. However, the biological mechanisms of this ultrasound-assisted healing are not yet understood, limiting clinical adoption. The objective of this study is to characterize the effects of LFLI US on gene expression in a simulated chronic wound environment to gain a better understanding of the biological mechanisms underlying this therapeutic.

Methods

A simulated chronic wound environment consisting of endothelial cells and inflammatory M1 macrophages co-cultured in 3D was subject to LFLI US at 50, 100, or 150 mW/cm² Spatial-Peak, Temporal-Peak …

Stentrx, Emma Barna, Minuthi Cooray, Bruce Newlands, Joseph A. Sabetta

Williams Honors College, Honors Research Projects

This project aims to develop a biodegradable, drug-eluting biliary stent for the treatment of biliary cancers. Current metal stents that restore bile flow bit are non-degradable, prone to migration, and unable to deliver drugs locally. The proposed stent will be biocompatible, biodegradable, and capable of controlled drug release to both maintain bile flow and treat tumors. The end goal is to produce a functional prototype that can be deployed via a minimally invasive catheter and maintain bile duct patency for at least three months. The project is expected to be involved in the NEOMED Bench-to-Bedside competition and provisionally patented.

Design And Validation Of A Novel Patterned Microfluidic Culture Platform For Hipsc-Cm Maturation, Anisia Gabriela Țiplea

Dartmouth College Master’s Theses

Adult cardiomyocytes (CMs) are among the most challenging cells to culture in vitro, given their poor adherence to plastic culture wells and poor maturation outside the native extracellular matrix (Narkar et al., 2022). Human induced pluripotent stem cell-derived CMs (hiPSC-CMs) offer a viable alternative to study heart development and disease progression. While not comparable to adult CMs in their maturity (Narkar et al., 2022), they mimic the clinical phenotypes of heart failure and are more feasible to culture. Nevertheless, hiPSC-CMs require specialized culture conditions that cannot be achieved with 2D static culturing, which fail to replicate native tissue environments (X. …

Synthesis And Characterization Of Glass-Ceramic Nanoparticles And Nanobubbles For Integration Into Bone Regenerative Biomaterials, Sun Latt

Honors Undergraduate Theses

Osteoporosis affects over 200 million individuals worldwide and is characterized by an imbalance between osteoclastic bone resorption and osteoblastic bone formation, leading to fragility fractures that significantly diminish quality of life. Current clinical bone cements, such as poly(methyl methacrylate), are limited by poor osseointegration, toxic monomers, and an inability to address the underlying pathophysiology of osteoporosis. This work presents the synthesis, characterization, and systematic in vitro evaluation of a sol-gel-derived, magnesium-incorporated glass-ceramic (GC) nanoparticle within the CaO-MgO-SiO₂ system for incorporation into a novel hydrogel bone cement and bioresorbable magnesium implants. XRD confirmed a biphasic amorphous-crystalline diopside composition, and TEM …

Optimization Of Research Pipeline To Characterize The Effects Of Atp-Sensitive Potassium Channels Mutations On Native Skeletal Muscle Fibers And Evaluate Potential Drug Therapy, Yuezhou Chen

McKelvey School of Engineering Graduate Student Theses & Dissertations

The ATP-sensitive potassium (KATP) channel is a critical metabolic sensor in skeletal muscle, yet its definitive molecular composition and functional roles remain contested. In this study, I conclusively demonstrate that the Kir6.2/SUR2 complex forms sarcolemmal KATP channel in mouse fast-twitch muscle. Genetic knock-out (KO) of Kir6.2 (Kcnj11−/−) or SUR2 (Abcc9−/−) resulted in a similar phenotype with increased fatigue resistance and a pathological rise in unstimulated resting tension at high-frequency stimulation. In contrast, SUR1 (Abcc8−/−) KO was ineffective. Furthermore, a CRISPR/Cas9 knock-in mouse model of a human SUR2A truncation variant (KCGV/KCGV) recapitulated the abnormal force accumulation, demonstrating that even loss-of-function of …

Functional Characterization Of Abcc8 Mutations Potentially Linked To The Transition From Hyperinsulinemic Hypoglycemia To Diabetes, Hao Zhang

McKelvey School of Engineering Graduate Student Theses & Dissertations

ATP-sensitive potassium (KATP) channels, composed of the SUR1 and Kir6.2 subunits encoded by the ABCC8 and KCNJ11 genes, are critical regulators of membrane excitability and insulin secretion in pancreatic β-cells. Gain-of-function (GOF) mutations in these genes cause neonatal diabetes mellitus through impaired insulin secretion and persistent hyperglycemia, whereas loss-of-function (LOF) mutations lead to congenital hyperinsulinism (CHI) with hypoglycemia due to β-cell hyperexcitability. I have addressed a paradoxical form of maturity-onset diabetes of the young (MODY) arising from KATP mutations, in which patients transition from CHI to glucose intolerance later in life. My experiments indicate that many KATP mutations associated with …

2d-Nanosilicate Carriers For Sustained Delivery Of Vaccine Components, Chih Yun Liu

McKelvey School of Engineering Graduate Student Theses & Dissertations

Laponite® 2D Nanosilicates (NS) are disc-shaped platelets (~30 nm diameter, ~1 nm thick) with positively charged edges and negatively charged faces, capable of loading proteins, drugs, and nucleic acids via electrostatic interactions to form a house-of-cards gel that sequesters cargo. Here, we evaluated NS for sustained delivery of antigens and Toll-like receptor (TLR) agonists to extend germinal center duration and enhance antibody quality. The model antigen ovalbumin (OVA) was adsorbed onto NS at acidic pH in a ~1:1 ratio, with partial unfolding that remained stable at physiological pH. NS-OVA complexes were efficiently internalized by dendritic cells, with uptake driven primarily …

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 …

Diabetes-Induced Mitral Valve Degeneration: Tissue-Engineered Solutions And Antioxidant Therapy, Samantha Hahn

All Theses

Mitral valve (MV) fibrosis is a progressive pathological condition characterized by leaflet thickening and stiffening, ultimately compromising valve function. This process is accelerated in diabetes, where chronic hyperglycemia elevates reactive oxygen species (ROS) and alters the mechanics of the extracellular matrix (ECM), collectively promoting the activation of valvular interstitial cells (VICs). Under healthy conditions, VICs maintain a quiescent, fibroblast-like phenotype responsible for sustaining ECM homeostasis. However, pathological biochemical cues or mechanical stress drive VICs toward a myofibroblast phenotype. While this transformation is essential for short-term repair, its persistence contributes to maladaptive MV remodeling and fibrosis. Diabetic stressors may be mitigated …

Mitral Annular Calcification In Diabetes: Valvular Interstitial Cells, Antioxidant Therapy, And Tissue Engineering Approaches, Raleigh J. Keenan

All Theses

Mitral annular calcification (MAC) is a chronic, progressive, and degenerative condition that impairs mitral valve function and increases the risk of cardiovascular disease. The mitral annulus is a saddle-shaped, fibrous structure that is primarily composed of collagen and elastin. The annulus also contains interspersed lipids that are involved in cellular metabolism, as well as inflammatory and osteogenic signaling. These lipids increase the annulus’ susceptibility to calcification, under conditions that increase mechanical stress (hypertension) and promote atherosclerosis (diabetes mellitus). These conditions intensify calcification by promoting the activation of valvular interstitial cells (VICs), increasing fibrosis, and inducing osteogenic differentiation. Although it is …

Strategic Enhancement Of Biofuel Production Using Bacillus Subtilis As A Model Organism: A Dual Approach Of Gene Essentiality Mapping And Optimization, Angela A. Zebede

Electronic Theses and Dissertations

With rising global demand for renewable energy, enhancing microbial platforms for biofuel production is vital. This study engineered Bacillus subtilis to improve resilience to solvent stress by targeting membrane lipid biosynthesis pathways essential for integrity. Growth curve analysis (OD₆₀₀) showed that phosphatidylserine (PS) and phosphatidylethanolamine (PE) knockout strains grew significantly slower than wild type (168), indicating the importance of these lipids under stress. Laurdan fluorescence spectroscopy was used to assess membrane properties. Generalized polarization (GP) analysis revealed a concentration-dependent reduction in membrane order upon 1%–2% 1-butanol exposure. GP values declined progressively from control to 2% treatment, reflecting increased membrane fluidity …

Characterization Of Lactobacilli-Containing Catheter Segments Developed By Using Selective Laser Sintered Mold., Caden Maners

Master of Engineering Theses

Catheter-associated urinary tract infections (CAUTIs) are a major healthcare burden with current prevention strategies offering limited success. This study presents a novel, self-coating biomaterial approach using a silicone-based bioink embedded with thermally sensitive probiotic strains (L. crispatus, L. rhamnosus, and L. reuteri). Catheter segments were fabricated via bio-injection molding using 3D-printed CAD molds. These Lactobacilli, selected for their probiotic and antimicrobial potential, serve as a non- antibiotic strategy to reduce infection risk. These segments were evaluated for mechanical strength (Shore A hardness), thermal stability (TGA/DSC), porosity and bacterial presence (SEM), biofilm formation (crystal-violet assay), as well as mass retention in …

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, …

Examining The Effects Of Quercetin On Phenotypic Characteristics Of Human Mesenchymal Stem Cells, Thomas M. Needy

Biomedical Engineering Undergraduate Honors Theses

Introduction A significant obstacle to mesenchymal stem cell (MSC) potency and therapeutic utility is in vitro senescence, an irreversible cessation of replication associated with age-related complications. Senolytic drugs, such as quercetin, may be helpful in selectively culling senescent cells while leaving non-senescent cells unaffected, thereby increasing potency of high-passage MSCs.

Methods The phenotypic, genotypic, and immunomodulatory effects of quercetin were assessed using in vitro models. Senescent cells, created through repeated subculturing of MSCs in vitro, and non-senescent cells were treated with 10 μM quercetin, differentiated into osteocytes, adipocytes, and chondrocytes, and analyzed to observe the effect of quercetin.

Results Quercetin …

Optimizing The Effects Of Yoda1 And Assessing The Effects Of Mrna Silencing Of Piezo1 Gene In Mesenchymal Stem Cells, Addison R. Mclaughlin

Biomedical Engineering Undergraduate Honors Theses

Mesenchymal stem cells (MSCs) are multipotent adult stem cells that are capable of differentiating into various skeletal tissues, making them promising candidates for regenerative medicine. A critical factor influencing their differentiation potential is mechanosensitivity, the ability to respond to mechanical stimuli. The mechanosensitive calcium ion channel PIEZO1 has emerged as a key regulator in this process, particularly in osteogenic differentiation. This study investigated the role of PIEZO1 in MSC osteogenesis through pharmacological activation using YODA1 and gene silencing via siRNA. Human bone marrow-derived MSCs were cultured and induced for osteogenic differentiation under varying concentrations and durations of YODA1 treatment. Alizarin …

Efficacy Of Mitophagy Inhibition As A Potentiating Agent For Conventional Chemotherapy, Morghan Relich, Shelby Bess

Biomedical Engineering Undergraduate Honors Theses

Cancer is a result of uncontrolled growth and reproduction of cells. Damaged or abnormal cells grow and multiply when they should not and in turn the cells continue to proliferate and metastasize. Malignant cells rely on functional mitochondria for proliferation, metastasis, and survival. These mitochondria maintain a functional mitochondrial network through a quality control mechanism called mitophagy. Mitophagy is a conserved intracellular process to maintain quality control and remove damaged mitochondria.

Previous research has been done to examine the effects of mitophagy in tumor cells, as well as the effects of mitophagy inhibition. One focus of current research has been …

Metabolic Responses Of Adipose-Derived Stem Cells In Collagen I Hydrogels, Caroline R. King

Biological Sciences Undergraduate Honors Theses

Peripheral nerve injury affects approximately 44 in 1 million people in the United States every year and can lead to incomplete nerve recovery and loss of motor and sensory function. A significant amount of energy is required for nerve injury repair. A lack of energy allocation to the injury site hinders repair. Current treatments include hollow nerve guidance conduits, decellularized nerve grafts, and autografts. However, they are limited by factors such as donor site morbidity and sustained immunosuppressive treatments. Using three-dimensional collagen I hydrogels laden with adipose-derived stem cells (ASCs) can assist in neural regeneration. Three-dimensional collagen scaffolds aim to …

In Vitro Investigation Of Macrophage Functional And Metabolic Modulation, Shelby Nicole Bess

Graduate Theses and Dissertations

Macrophages are a subset of immune cells with a variety of functions that are abundant in numerous tumor types such as colorectal and breast cancer. It has been suggested that macrophages may serve an important function as immune mediators in tumor behavior; however, macrophage activity is complex and remains not completely understood. The overall goal of this dissertation was to explore and exploit tumor-associated macrophage phagocytic function and the changes in their interactions with cancer cells in an in vitro mouse model and in 2D and 3D in vitro culture models. First, we wanted to better understand macrophage behavior under …

Exploring Veto Activity: Dnam-1-Cd155 Axis In Overcoming Nk Cell-Mediated Allo-Rejection And Applications For Car-T Cell Therapy, Wei-Hsin Liu

Dissertations and Theses (Open Access)

As Miller et al. defined veto activity, it enables cells to target host cytotoxic T lymphocyte (CTL) precursors specific to veto cells' antigens, selectively eliminating anti- donor T cell clones without inducing rejection. By leveraging this unique immune property, two key research aims were proposed: “Exploration of the Impact of Veto Activity on Natural Killer (NK) Cell-Mediated Allo-rejection” and “Development of Off-the- shelf Chimeric Antigen Receptor (CAR)-T Therapy: Engineering Anti-Viral Veto CD8+ T Cells”. As demonstrated in our previous research, in a murine model with mild conditioning, anti-third-party central memory CD8+ veto T cells (veto Tcm) can prevent T cell- …

The Effects Of Cigarette Smoke Extract On Osteoarthritis In A Primary Joint Tissue Explant Model, Emily Sawvell

All Dissertations

Osteoarthritis (OA) is the most common degenerative joint disease in the world with an economic burden of nearly $140 billion annually. Factors identified to increase the risk of developing OA include age, obesity, sex, race, genetics, and previous joint injury. However, one risk factor that has been scarcely studied in OA etiology is cigarette smoking. While some studies have reported smoking to protect against OA, increasing evidence suggests that cigarette smoking results in more severe and painful OA, increases the rate of developing OA, and reduces the efficacy of pain medications and arthritis-targeting therapies. Despite this, the impact of cigarette …