Molecular, Cellular, and Tissue Engineering Commons^™

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 …

Evaluating Tissue Morphology In The Context Of Varied Initial Fabrication Conditions, Cassidy Caffin

Biomedical Engineering Undergraduate Honors Theses

Cardiovascular diseases have been the leading cause of death for years. This includes myocardial infarctions (MI) where blood flow to the myocardium is restricted. This causes damage to cardiac muscle due to insufficient oxygen. There are multiple ways to treat patients following an MI with the most common involving assorted medication. However, there are limited medications that can be used for treating patients following MIs, and the FDA’s decreasing approval rate for new cardiac drugs will not dramatically improve the range of options. The lead-up to drug candidate rejection by the FDA can involve drugs exhibiting promising preliminary research that …

Utilizing Spatial Transcriptomics To Compare Gene Expression In Volumetric Muscle Loss Injury Recovery, Colton Gattis

Biomedical Engineering Undergraduate Honors Theses

Volumetric Muscle Loss (VML) injuries are known to disrupt the normal regenerative process via fibrosis leading to an extensive permanent loss of muscle function. The specific causation of this disruption remains to be defined; however, with new developments in transcriptomics, genetic trends can be identified across regions of tissue over time. This study follows VML injuries within the tibialis anterior of a mouse model for fifteen days after initial injury to attempt to identify expected transitions in healing. Genetic presence began to become more diversified as recovery progressed from four to fifteen days post injury with spatial clusters becoming globalized …

Investigating The Presence Of Patented In Vitro Models For Cancer-Nerve Crosstalk Methods, Christoff Hauptmann

Biomedical Engineering Undergraduate Honors Theses

The purpose of this research project is to investigate the presence of patented in vitro models for perineural invasion (PNI) and tumor innervation, the two most prominent modes of cancer-nerve crosstalk that play an active role in cancer initiation and progression. PNI occurs through a positive feedback loop in which cancer cells invade passive nerves to metastasize. Tumor innervation is the process where tumor cells attract nearby nerves to the tumor microenvironment (TME) for tumor growth and metastasis. These crosstalk paths have been established for many types of cancer and result in poor patient prognosis. Recently, in vitro models have …

Evaluation Of The Efficiency Of Muscle-Specific Promoters To Express Non-Endogenous Proteins In Skeletal Muscle, Gabrielle Bulliard, Made Harumi Padmaswari, Christopher E. Nelson

Biomedical Engineering Undergraduate Honors Theses

Gene therapies are emerging as powerful tools for treating genetic diseases and cancers, offering the potential for a permanent cure. Hemophilia B, affecting approximately 6,000 men in the U.S., results from a deficiency in coagulation factor IX protein (FIX), which is crucial for blood clotting. A substantial portion of patients, over 40%, suffer from severe hemophilia B, experiencing spontaneous, prolonged bleeding. Current prophylactic treatments involve frequent coagulation factor infusions, yet a curative approach would alleviate this burden. Gene editing technologies like CRISPR-Cas systems show promise for correcting genetic mutations, but the diverse nature of factor IX gene (F9) …

Transcriptomic Profiling Of Engineered Human Gene Integration In The Mouse Genome Following In Vitro Gene Editing, Ethan Potts, Made Harumi Padmaswari, Christopher Nelson

Biomedical Engineering Undergraduate Honors Theses

Gene replacement is a promising method of therapy for genetic diseases. However, safety and efficacy are areas that need more research. This experiment aims to use RNA sequencing and bioinformatic techniques to provide answers to these questions and provide direction to future studies to develop a gene replacement therapeutic. C2C12 mouse myoblast cells were transfected with a vector containing a CRISPR-Cas9 system and the Human Factor IX (hF9) gene in order to hijack target genes and integrate the hF9 gene. The two target genes, myoglobin (Mb) and creatine kinase (Ckm), were chosen for their high rate of expression and low …

The Determination Of Macrophage Localization In Muscle Injuries Using Spatial Transcriptomics, Julliana Renales

Biomedical Engineering Undergraduate Honors Theses

The repair of traumatic muscle injury is often a long process which can in many cases be insufficient and incomplete. In such cases, scar tissue may form, leading to high risk of re-injury and the prevention of muscle regeneration. An important part of decreasing this risk is gaining a better understanding of the cellular complexities of injury repair. More specifically, macrophage cells are an important piece in the regulation of inflammation and repair. In this project, computational analysis spatial transcriptomic data was performed to examine mouse muscle injury and its development over time. This analysis was used to examine the …

Investigating The Effect Of Pancreatic Cancer Tumor-Derived Extracellular Vesicles On Schwann Cell Activation, Isabel Powers

Biomedical Engineering Undergraduate Honors Theses

Pancreatic cancer is one of the deadliest diseases because it is hard to identify in its early stages, has high chemoresistance, and early metastasis. Because of these traits, it is often associated with perineural invasion (PNI), where the cancer travels to the nervous system. Schwann cells (SC) are associated with both metastasis and PNI when activated. The purpose of this study was to determine if and how pancreatic tumor-derived extracellular vesicles (TEVs) can activate SC in PNI. This experiment was performed by embedding Schwann cells in a hydrogel to model the tumor microenvironment and treating them with isolated extracellular vesicles …

Optimizing Three-Dimensional Bioengineered Co-Cultures For The Study Of Fibrotic Spinal Cord Injury, Sarah Aly

Biomedical Engineering Undergraduate Honors Theses

Spinal cord injuries (SCIs) have major physical implications on those affected^[1]. There are two main types of scarring that occur after injury; they are known as the glial and fibrotic scar. The injured site causes an altered micro-environment which transforms astrocytes from their normal to reactive state, thus inhibiting axonal regeneration. The effect of reactive astrocytes on fibrotic scarring is not fully understood due to limited study^[2,3]. Using Three-Dimensional (3D) bioengineered co-cultures of astrocytes and neuroblasts the pathological environment can be replicated and manipulated to further the study of fibrotic scarring. Temperature casted co-cultures were successfully …

Exploring The Effects Of Microarchitecture On Astrocyte Behavior And Vascular Remodeling In Spinal Cord Injury, Devin Gerber

Biomedical Engineering Undergraduate Honors Theses

Spinal cord injury (SCI) is a common condition and is characterized by traumatic and progressive damage that results in limited neuron regeneration and thus a restricted recovery. In SCI, the vasculature is also severely damaged, which exacerbates inflammation at the injury site. Specifically, insufficient angiogenesis occurs at the site of injury due to the fibrotic pathological microenvironment1 . SCI also leads to the formation of a fluid-filled cavity and thus gliosis and fibrosis, forming a dense glial and fibrotic scar around the cavity. The microenvironment at the injury site plays a significant role in astrocyte behavior, promoting the transformation of …

Comprehensive Long- And Short- Read Sequencing Approaches To Screen The Mutational Landscape After Therapeutic Crispr-Editing For Duchenne Muscular Dystrophy, Mary Jia, Made Harumi Padmaswari, Landon Burcham, Shilpi Agrawal, Alexis Ivy, Abbey Bryan, Christopher E. Nelson

Biomedical Engineering Undergraduate Honors Theses

Duchenne muscular dystrophy (DMD) is an X-linked muscle wasting disease caused by mutations in the Dmd gene resulting in non-functional dystrophin. Adeno-associated virus (AAV)-mediated delivery of classical CRISPR systems resulting in double-stranded breaks (DSBs) has yielded promise as a curative DMD therapeutic. However, the consequences of AAV-CRISPR require more thorough characterization. PCR-enriched short-read amplicon sequencing at cut sites is the most common method of sequencing CRISPR editing, but PCR bias results in unreliable quantification of on-target edits. We analyze orthogonal long- and short-read sequencing approaches to screen the outcomes of CRISPR technologies at the DNA and RNA-level for DMD. We …

Assessing The Potential Effectiveness Of Diffuse Reflectance Spectroscopy For Quantifying Neonatal Skin Properties Using Tissue Phantoms, Jack Murphy

Biomedical Engineering Undergraduate Honors Theses

Diffuse reflectance spectroscopy (DRS) has previously been shown to be effective at analyzing the optical characteristics of a wide variety of biological tissues, including the skin. In addition to being sensitive to the amount of blood and its oxygenation, DRS can be used to extract bilirubin levels and assess jaundice, a common complication in newborn infants. To assess its potential for being utilized in applications involving neonatal skin, we evaluated the utility of DRS for extracting hemoglobin and bilirubin concentrations from tissue phantoms. Phantom sets were created by including a known amount of scattering and by varying known concentrations of …

The Interaction Of Angiotensin I And Ii On Calcified Aortic Valve Cells, Fah Sysavanh

Biomedical Engineering Undergraduate Honors Theses

Cardiovascular diseases affect the older population with limited treatments available. Aortic valve stenosis is a cardiovascular disease in which the aortic valve narrows, causing blood flow constriction and calcification.¹ The only treatment is to replace the valve, which can be costly and lead to other health complications.

The renin-angiotensin system (RAS) is a pathway that helps with maintaining cardiovascular homeostasis. Angiotensin I (ANGI) and II (ANGII) are hormones in the RAS that cause an increase in inflammation, stress, and blood pressure by binding to the Angiotensin Type 1 Receptor (AT1R).¹ Gelatin-based valve thin film (vTF) platforms were fabricated to investigate …

Sequential Illumination In A Tomographic Microendoscopic Probe For Imaging Tumor Microvasculature, Zachariah Neumeier

Biomedical Engineering Undergraduate Honors Theses

Knowledge of colorectal cancer biology is improving how we approach cancer treatment. Specifically, the tumor microenvironment and abnormal angiogenesis are of particular interest. Optical methods are a prime candidate for research of the tumor microenvironment due to their ability to quantitively assess tissue structure and perfusion in real time. Particularly, the “transport scattering regime” has been identified as a method of obtaining high-resolution images and reflectance spectroscopy data; this light scattering regime has been demonstrated compatible with endoscopic imaging systems. In this study, a proof-of-concept optical imaging system is presented, capable of resolving absorbers within scattering turbid media using a …

The Effects Of Microgravity On Human Endothelial Cells In Relation To Deep Vein Thrombosis, Allie Burgess

Biomedical Engineering Undergraduate Honors Theses

Deep vein thrombosis (DVT) is a condition where blood clots form in one or more of the deep veins of the body, causing leg pain and/or swelling, potentially leading to pulmonary embolism and/or stroke. The incidence of DVT among astronauts serving on the International Space Station has been shown to be greater than that of the general population, calling into question the effects of microgravity on human thrombolytic processes. Additionally, microgravity conditions have been shown to cause muscular atrophy and a decrease in muscle strength and mobility. Because collagen composes a large portion of muscle tissue, this study also assessed …

Hydrogel Drug Delivery Platform Integrity And Utilization Analysis, Joshua Carson Adams

Biomedical Engineering Undergraduate Honors Theses

Traumatic Spinal Cord Injury (SCI) generally results in permanent sensorimotor dysfunction. This is due to failed axonal regeneration at the site of injury. Recent studies have highlighted the use of stem-cell based therapies embedded in hydrogel to treat SCI. Embedding cell therapies in hydrogel provides a platform that increases the survival of both integrated grafted cells and stem cells/stem cell progeny. There are a variety of drug delivery agents for treating SCI such as neuro-regenerative cells and Schwann cells, however, single agents are not promising for functional regeneration. We propose a combinatorial approach using sciatic nerve (SN) extracellular matrix (ECM), …

Characterizing The Expression Pattern And Function Of Tartan During Drosophila Development, Alaina Baggett

Biomedical Engineering Undergraduate Honors Theses

The development of complex structures and organs by multicellular organisms relies on the ability of epithelial cells to self-organize. Epithelia are sheets of connected cells, and compartment boundaries are formed between certain epithelial cells to create distinct tissue compartments. Compartment boundaries are specialized cell-cell interfaces that are enriched for the cytoskeletal proteins actin and myosin, leading to straight cell edges under relatively high tension that act as fences keep cells from moving between compartments. In the model organism Drosophila melanogaster (fruit fly), compartment boundaries in the early embryo are established in response to the non-uniform striped expression of the cell-surface …

Modifications Of A Machine To Produce Clinically-Sized Angioplasty Balloons For Bifurcations, Natalie Smith

Biomedical Engineering Undergraduate Honors Theses

Coronary artery disease is the most common form of heart disease and the leading cause of death in the U.S. ("Heart Disease Facts | cdc.gov", 2021). Typically, coronary artery disease is a result of coronary stenosis, constriction of the coronary artery, which can cause heart failure, arrhythmia, or heart attack ("Coronary Artery Disease: Prevention, Treatment and Research", 2021). Angioplasty balloons are a common treatment method for coronary stenosis and work by placing a stent at the lesion site to widen the vessel and promote blood flow. A type of lesion which remains particularly difficult and relatively common to treat is …

Investigating The Impact Of Hypoxia On Reactive Oxygen Species Generation Within Murine Breast Cancer Cells, Jared Mcpeake

Biomedical Engineering Undergraduate Honors Theses

When cancer metastasizes from a primary tumor site to secondary site through the bloodstream or lymph, the cancer becomes more difficult to treat. For this reason, it is vital to study what indicates the metastatic potential of a tumor. Current research has shown that cell lines with high metastatic potential display increased levels of metabolic adaptability over their nonmetastatic counterparts after undergoing hypoxic conditions. One method of assessing this adaptability is to measure the concentration of reactive oxygen species (ROS) produced by the cells while undergoing oxidative stress. In highly adaptable metastatic cells, an increase of ROS buildup within the …

Erythrocyte Deformability In Response To Glucose Using Liquid Crystals, Jayden Goff

Biomedical Engineering Undergraduate Honors Theses

The worldwide prevalence of diabetes mellitus is rapidly increasing with about 9.3% of the adult population living with the disease. People with diabetes have trouble regulating their blood glucose levels which typically leads to hyperglycemia. Under normal physiological conditions, erythrocytes can undergo deformations in response to shear stress when passing through capillaries with a smaller diameter. Poorly managed hyperglycemia can lead to the glycosylation of erythrocyte membrane proteins and hemoglobin. This glycosylation leads to increased rigidity of the cells along with decreased deformability in response to mechanical stress; therefore, these cells have a higher susceptibility of getting stuck in the …

Clot Analogs For The Development Of Improved Treatment Methods In Ischemic Stroke, Charles A. Rieth

Biomedical Engineering Undergraduate Honors Theses

According to the CDC, nearly 800,000 individuals experience stroke each year in the United States [1]. Greater than 70% of strokes are of ischemic etiology and involve the occlusion of key arteries in the cardiovascular system [2]. Tissue plasminogen activator (IV-tPA) is the current gold standard for thrombolytic approaches [3]; however, this therapeutic is only effective between 3 and 4.5 hours from the patient’s last know well [4]. As a result, less than 5% of acute ischemic stroke patients receive IV-tPA. In order to increase the viability of AIS treatment as a whole, it is essential that thrombolytic techniques are …

The Influence Of Mmp14 On Angiogenesis In Chemotherapy-Treated Tumors, Abdussaboor Muhammad

Biomedical Engineering Undergraduate Honors Theses

Colorectal cancer is the third most common cancer in the world, and it is the fourth most common cause of cancer related death (1). There have been many significant advancements regarding the treatment of cancer which aim to shrink the size of tumors in patients. However, there is still more that needs to be understood about the many different factors that play a role in colorectal cancer development.

Angiogenesis is the process of forming new blood vessels from existing ones and it requires breaking down and remodeling of the extracellular matrix (ECM) in order to allow endothelial cells to migrate …

Tissue Engineering And The Effect Of Volumetric Muscle Loss On Muscle Performance, Alex Pemberton

Biomedical Engineering Undergraduate Honors Theses

Trauma, for so many people, creates life altering injuries. One of the major injuries that results from trauma is volumetric muscle loss (VML), which is the loss of muscle tissue that cannot be naturally regenerated and results in chronic loss of function. When a small amount of damage is done to a muscle, but the underlying extracellular matrix of the cell is still intact, the muscle can regenerate much of its volume and function. If the injury damages the extracellular matrix, the injury site is instead filled with scar tissue, which cannot regenerate muscle mass or function. Seen widely in …

Guide Rnas Preparation For In-Vitro Crispr-Cas9 Complex Delivery Targeting Genes That Affect Wound Healing., Prashant Khatiwada

Biomedical Engineering Undergraduate Honors Theses

CRISPR-Cas9 technology has widely been used as a viable genome engineering platform to make site-specific insertion, deletion, and breaks. The nuclease dead version of Cas9 or dCas9 can be used for the activation and repression of target gene sites using specific activation or repression domains. In this study, CRISPR guide RNAs were designed for a CRISPR inhibition approach to repress the transcriptional activity of the target genes. An expression plasmid vector composed of a U6 promoter sequence, BbsI restriction sites, and a chimeric gRNA sequence was digested, and the phosphorylated forward and reverse gRNAs were ligated with the plasmid vector. …

Evaluating Cellular Response Of Different Valve Interstitial Cell (Vic) Phenotypes To Angiotensin I, Smit Patel

Biomedical Engineering Undergraduate Honors Theses

Calcific Aortic Valve Disease (CAVD), one of the leading causes of death in the United States, is characterized by diminished functioning and limited movement of the aortic valve (AV) due to increased thickening, disorganized extracellular matrix elastin fibers, increased collagen content, and abnormal deposition and formation of calcium nodules on the AV [1, 2]. There are two types of CAVD: i) Aortic valve sclerosis (AVS), in which patients suffer from tissue hardening, fibrosis, and early calcification, and ii) calcific aortic stenosis (CAS), representative by excessive calcification on the AV and reduced AV opening, are the two main forms of CAVD …

Involvement Of The Ino80 Chromatin Remodeling Complex In Cell Division And Genomic Stability, Ethan Chen

Biomedical Engineering Undergraduate Honors Theses

Cell division is a vital biological process for growth and development in both single and multi-cellular organisms—whereby the cell must duplicate its organelles and genome in entirety and appropriately distribute the copied contents to the daughter cells. Cells undergo a cycle of two distinct phases: interphase and mitosis. During interphase, the cell replicates its genomic DNA (in the form of chromosomes) located within the nucleus. DNA replication is carried out in a euchromatin state, where the chromosome structure is loose and easily accessible by DNA polymerase and other replication enzymes. Upon the completion of replication, chromatin is condensed into highly …

Mapping The Domain Of Rna Helicase Ddx1 That Interacts With The Ns1 Of Influenza A Viruses, Elise Wyatt

Biomedical Engineering Undergraduate Honors Theses

Of the known influenza virus strains, influenza A viruses (IAVs) are the most catastrophic due to their highly contagious nature and possibly life-threatening symptoms. IAVs invade a host cell and enter the nucleus where they use the cell’s nuclear machinery for replication of their viral RNA genome which is then exported to neighboring cells to establish a productive infection. Host pathogen recognition receptors (PRRs) respond to viral infection by signaling the cell to produce type I interferons (IFNs) to fight the foreign invader. Non-structural protein 1 (NS1) of IAVs, a major component in the regulation of viral replication, opposes host …

Developing Aligned Nerve Scaffolds In A 3d Type-I Collagen Gel, Gabriel David

Biomedical Engineering Undergraduate Honors Theses

Despite significant progress in the field of peripheral nerve repair, clinical success is still limited, leaving millions to suffer from peripheral neuropathy with billions spent every year for treatment. Nerve repair methods that are capable of maximizing the regenerative properties of peripheral nerves are greatly desired in the field of medical science. This research aims to fill the gap between modern methods and the future of nerve repair by creating type-I collagen scaffolds with aligned degradation pores that will assist and nurture nerves growing through them. This is achieved by incorporating adipose stem cells into type-I collagen hydrogels and aligning …