Biomedical Engineering and Bioengineering Commons^™

Design And Application Of Phase-Separating Peptides In Cytosolic Drug Delivery, Huihao Wang

McKelvey School of Engineering Graduate Student Theses & Dissertations

This research investigates the design and application of phase-separating peptides for direct cytosolic drug delivery. Novel peptides (GHGXY)4, with variable amino acids, were synthesized using microwave-assisted chemistry to enhance phase separation and membrane fusion. In vitro experiments utilizing advanced imaging techniques demonstrated the peptides' capability to form coacervates and facilitate direct cargo, DNA, mRNA, and protein, delivery into the cytosol, bypassing traditional cellular barriers. The study also explores the role of inhibitor Bafilomycin and NH4Cl in phase-separating peptides functionality, offering significant insights into their delivery mechanisms. This research underscores the potential of phase-separating peptides in improving therapeutic delivery strategies.

Environment And Response Of 3d-Encapsulated Mesenchymal Stem Cells To Mechanical Loading, Augustus Greenwood

McKelvey School of Engineering Graduate Student Theses & Dissertations

This thesis explores the micromechanical environment induced when cyclically compressing hydrogels via finite element modeling and experimentally on the impact of loading on mesenchymal stem cells (MSCs) when encapsulated withing 3D hydrogel matrices. Degenerative joint diseases, characterized by cartilage degradation, present significant challenges due to cartilage's limited self-repair capacity. Innovative approaches, including stem cell-based therapies and engineered biomaterials, have emerged as promising strategies for cartilage repair and regeneration. This work specifically investigates the calibration of a bioreactor, the uniformity of load response across the hydrogel constructs via finite element modeling (FEM), and the stress response of MSCs subjected to various …

Modeling Of Nk Cells In Pediatric Patients With Unusually Severe Or Recurrent Hsv Using High-Dimensional Flow Cytometry, Yunran Feng

McKelvey School of Engineering Graduate Student Theses & Dissertations

HSV infection is broadly spread all over the world with some patients having severe and/or recurrent HSV infections. Our lab studies human Natural Killer (NK) cells, which are important in innate immune responses to viral infections and tumors. A publication in 2013 by Ornstein et al from our lab studied HSV+ pediatric patients and found some associations between severe infection and defects in NK cytolytic function. PLCG2 haploinsufficient variants found in 2 HSV patients causing PLCγ2 hypophosphorylation, and loss of cytolytic function in NK cells is a novel finding recently published by Alinger et al from our lab in 2023. …

The Role Of Voluntary Descending Control In Enhancing Motor Function Via Transcutaneous Spinal Cord Stimulation, Yoon Kim

McKelvey School of Engineering Graduate Student Theses & Dissertations

Spinal cord injury (SCI) is a life-changing event that causes lasting motor impairments. Transcutaneous spinal cord stimulation (tSCS), a non-invasive form of neuromodulation in which electrodes are placed on the skin and used to stimulate the spinal circuits via an electrical current, has demonstrated positive effects on motor function recovery in individuals who have had SCIs. However, the precise mechanism of how tSCS interacts with voluntary descending drive remains poorly understood. This study aims to investigate the role of voluntary descending control in influencing reflex responses triggered by tSCS.

Electromyography (EMG) recordings were performed in ten unimpaired individuals while they …

Development Of A Wearable Short-Wave Infrared Photoplethysmography Device For Detection And Monitoring Of Hemodilution During Postpartum Hemorrhage, Hannah Gruensfelder

McKelvey School of Engineering Graduate Student Theses & Dissertations

ABSTRACT OF THE THESIS

A Thesis on the Development of a Wearable Short-Wave Infrared Photoplethysmography Device for Detection and Monitoring of Hemodilution During Postpartum Hemorrhage

by

Hannah Gruensfelder

Master of Science in Biomedical Engineering

Washington University in St. Louis, 2024

Professor Christine O’Brien, Chair

Postpartum hemorrhage (PPH), the leading cause of maternal death and morbidity, affects nearly 14 million people worldwide each year, disproportionally impacting racial minorities and people in low resource settings. A timely diagnosis of PPH is key in providing optimal patient care, as an estimated 90% of deaths due to PPH are preventable with early diagnosis and …

Low Impedance, Durable, Self-Adhesive Hydrogel Epidermal Electrodes For Electrophysiology Recording, Naiyan Wu

McKelvey School of Engineering Graduate Student Theses & Dissertations

Traditional electrodes used for electrophysiology recording, characterized by their hard, dry, and inanimate nature, are fundamentally mismatched with the soft, moist, and bioactive characteristics of biological tissues, leading to suboptimal skin-electrode interfaces. Hydrogel materials, mirroring the high water content and biocompatibility of biological tissues, emerge as promising candidates for epidermal electronic materials due to their adjustable physicochemical properties. However, challenges such as inadequate electrical conductivity, elevated skin impedance, unreliable adhesion in moist conditions, and performance decline from dehydration have significantly restricted the efficacy and applicability of hydrogel-based electrodes. In this thesis, we report a high-performance hydrogel epidermal electrode patch for …

Resolving The Nanoscale Structure Of Β-Sheet Peptide Self-Assemblies Using Single-Molecule Orientation–Localization Microscopy, Weiyan Zhou, Conor L. O'Neill, Tianben Ding, Oumeng Zhang, Jai S. Rudra, Matthew D. Lew

Electrical & Systems Engineering Publications and Presentations

Synthetic peptides that self-assemble into cross-β fibrils are versatile building blocks for engineered biomaterials due to their modularity and biocompatibility, but their structural and morphological similarities to amyloid species have been a long-standing concern for their translation. Further, their polymorphs are difficult to characterize using spectroscopic and imaging techniques that rely on ensemble averaging to achieve high resolution. Here, we utilize Nile red (NR), an amyloidophilic fluorogenic probe, and single-molecule orientation-localization microscopy (SMOLM) to characterize fibrils formed by the designed amphipathic enantiomers KFE8^L and KFE8^D and the pathological amyloid-beta peptide Aβ42. Importantly, NR SMOLM reveals the helical (bilayer) …

6d Single-Fluorogen Orientation-Localization Microscopy For Elucidating The Architecture Of Beta-Sheet Assemblies And Biomolecular Condensates, Tingting Wu, Weiyan Zhou, Jai S. Rudra, Rohit V. Pappu, Matthew D. Lew

Electrical & Systems Engineering Publications and Presentations

We develop six-dimensional single-molecule orientation-localization microscopy (SMOLM) to measure the 3D positions and 3D orientations simultaneously of single fluorophores. We show how careful optimization of phase and polarization modulation components can encode phase, polarization, and angular spectrum information from each fluorescence photon into a microscope’s dipole-spread function. We used the transient binding and blinking of Nile red (NR) to characterize the helical structure of fibrils formed by designed amphipathic peptides, KFE8^L and KFE8^D, and the pathological amyloid-beta peptide Aβ42. We also deployed merocyanine 540 to uncover the interfacial architectures of biomolecular condensates.

Engineering Mussel Foot Protein 5 (Mfp5) For Enhanced Adhesive Performance In Tendon And Bone Repair, Ziyue Zhu

McKelvey School of Engineering Graduate Student Theses & Dissertations

This study explores the development of Mussel Foot Protein 5 (MFP5) bioadhesives to improve tendon-to-tendon and tendon-to-bone adhesive outcomes. MFP5, known for its underwater adhesive properties, was synthesized using recombinant techniques and optimized through coacervation and cross-linking. Adhesion tests demonstrated the superior performance of the optimized MFP5 adhesive on the tendon and bone samples. Improved protein concentration testing and purification methods enhanced MFP5 yield and consistency. Future research will focus on further standardization, exploring alternative purification methods, and evaluating the adhesive's performance in animal models. This study lays a foundation for developing robust MFP5-based bioadhesives, potentially revolutionizing surgical repair techniques …

Design Of Human Serum Albumin And Adenovirus Conjugation Via Catcher/Tag Molecular Glue, Peijie Zhao

McKelvey School of Engineering Graduate Student Theses & Dissertations

Adenovirus (Ad) has been the ideal cargo delivery mechanism, and its moderate immunological response makes it ideal for in vivo gene therapies since its discovery in 1953. However, the robust immunogenicity of the Ad capsid and low vaccine absorption via mucous membranes and epithelium put a limit on the process of developing intranasal vaccines. Efforts are being made to enhance the effectiveness of Ad vectors and numerous studies have demonstrated the remarkable capacity of human serum albumin (HSA) to extend plasma half-life and facilitate targeted intranasal delivery. In this study, we devised an innovative method for employing the Catcher/Tag molecular …

Improving And Integrating Quantitative Single Cell Spatial Biology For Theranostic Medicine, Peng Lu

McKelvey School of Engineering Graduate Student Theses & Dissertations

Disease states are the result of a complex interplay of many different cell types interacting in close proximity in the context of often heterogeneous tissues. Alpha particles are drawing intense research and clinical interest because of their potent cytotoxic effects and their short path lengths. Analyzing the dose distribution and tissue micro-environment for alpha therapy plays a key role in predicting the efficacy of this targeted radiotherapy. However, to date there have been no direct on-tissue analytical methods for alpha dose distributions and the corresponding tissue microenvironments.

Therefore, we have developed a pipeline to overcome this limitation by utilizing quantitative …

Automatic Cardiac Mri Image Segmentation And Mesh Generation, Ziyuan Li

McKelvey School of Engineering Graduate Student Theses & Dissertations

Segmenting and reconstructing cardiac anatomical structures from magnetic resonance (MR) images is essential for the quantitative measurement and automatic diagnosis of cardiovascular diseases [1]. However, manual evaluation of the time-series cardiac MRI (CMRI) obtained during routine clinical care are laborious, inefficient, and tends to produce biased and non-reproducible results [2]. This thesis proposes an end-to-end pipeline for automatically segmenting short-axis (SAX) CMRI images and generating high-quality 2D and 3D meshes suitable for finite element analysis. The main advantage of our approach is that it can not only work as a stand-alone pipeline for the automatic CMR image segmentation and mesh …

Calculating The Difference In Stiffness Of Living T Cells Through Micropipette Aspiration, Minju Lee

McKelvey School of Engineering Graduate Student Theses & Dissertations

Cardiovascular disease (CVD) accounted for 17.9 million deaths in 2019, with fibrosis contributing to nearly a quarter of these fatalities [1,2]. Fibrosis, characterized by excessive connective tissue formation, has been strongly linked to T cells, essential components of the immune system. This study explores the mechanisms of T cell activation and the subsequent changes in biophysical properties like diameter, stiffness, and elasticity, aiming to develop therapeutic strategies for fibrosis-related diseases, including CVD. Utilizing the micropipette aspiration technique, we accurately assessed T cell stiffness and observed a change in bulk cell stiffness upon activation. The results demonstrated increased fluid-like behavior in …

A Novel Approach To Detect Modified Cytosines In A Nanopore Sequencer, Benjamin Davidorf

McKelvey School of Engineering Graduate Student Theses & Dissertations

Epigenetic changes, such as DNA methylation, have been seen in various types of cancers, diseases, and neurological disorders. A common form of methylation occurs when a methyl group is added to the C-5 position of a cytosine, which is called 5-methyl-cyosine (5- mC). Hydroxymethylated is a type of methylation where 5mC is oxidized to 5-hydroxy-methyl- cytosine (5hmC), which has been linked to normal brain development. Current methods to sequence DNA methylation patterns (5-mC and 5-hmC) at a genome-wide level have several limitations, so there remains a need for a method to efficiently map DNA methylation. The state- of-the-art method for …

The Effect Of Spinal Cord Stimulation And Video Games Training On Body-Machine Interface Control, Jie Fei

McKelvey School of Engineering Graduate Student Theses & Dissertations

Damage to the spinal cord causes long-lasting loss of motor and sensory function, and currently, there is no ‘cure’ for paralysis. However, even people with severe spinal cord injuries (SCI) have some residual mobility. Studies have shown that transcutaneous electrical spinal cord stimulation (tSCS) combined with functional training targeting residual mobility can further improve the motor function of individuals with SCI. In this study, we present a technical framework that aims to enhance rehabilitation outcomes by targeting residual mobility through a motor training-based approach. Our technical framework centers around a non-invasive body-machine interface (BoMI) that relies on the use of …

Novel Microfluidic Devices To Model The Interactions Between Lymphatics And Breast Cancer, Jade Weber

McKelvey School of Engineering Graduate Student Theses & Dissertations

The lymphatic system is responsible for immune circulation and fluid balance in the body. It accomplishes this by draining interstitial fluid from local tissue and transferring it to lymph nodes and back into blood circulation. However, this process is implicated in many pathologies, one of the most dangerous being breast cancer metastasis to the lymph nodes. The largest factor in breast cancer patient mortality is metastasis. Lymphangiogenesis, the growth of new lymphatic vessels, has been thought to play a dynamic role in aiding breast cancer metastasis. Breast cancer tumor cells have been shown to remodel the functionality of local lymph …

3-Dimensional Visualization Of Cardiac Plaque Mapping Data, Phan Ly Vy Nguyen

McKelvey School of Engineering Graduate Student Theses & Dissertations

Atrial Fibrillation (AF) of one of the most prevalent cardiac arrythmia in humans, and also the most studied arrythmias due to its high association with cardiovascular morbidity and mortality. Diagnosis of AF, which is highly dependent on the observation of the irregular signal in the atria, is often challenging since AF is often asymptomatic at the onset. There has been a lot of effort in exploring different cardiac mapping techniques to understand the dynamics of AF for better intervention. This study aims at developing a MATLAB interface that assists the development of a cardiac plaque mapping data acquisition system in …

Preparing Non-Human Primates To Study Hand-Eye Coordination In Frontal Eye Fields (Fef) During Delayed Movement Task, Juliusz Cydzik

McKelvey School of Engineering Graduate Student Theses & Dissertations

ABSTRACT OF THE THESIS Preparing Non-Human Primates to Study Hand-Eye Coordination in Frontal Eye Fields (FEF) During Delayed Movement Task by Juliusz Cydzik Master of Science in Biomedical Engineering Washington University in St. Louis, 2023 Professor Lawrence Snyder, Chair Hand-eye coordination enables humans and non-human primates to use their hands and eyes to perform various tasks. We are interested in coordination at the systems level, where saccades and reaches are encoded. The parietal reach region (PRR), situated at the posterior end of the intraparietal sulcus (IPS) and overlapping portions of the medial intraparietal area (MIP) and V6a, is commonly attributed …

A Novel Mrna Delivery Strategy Employing Adenovirus Piggyback Mrna Binders Via Catcher/Tag Molecular Glue, Kexin Geng

McKelvey School of Engineering Graduate Student Theses & Dissertations

mRNA-based therapeutics open a new era for the prevention and treatment of a wide range of diseases. However, existing mRNA delivery vehicles have limited repertoires for dendritic cell targeting and intranasal administration which are two paramount delivery pathways of significant advantages for treating cancer and infectious respiratory diseases. Adenovirus (Ad) with well-deciphered viral genomes and intensive-studied structure biology has been developed for DCs-targeted cancer vaccines and intranasal COVID vaccines. In light of these virtues, Ad presents as the “off-the-shelf” tool possessing well-defined manufacturable capabilities and translatable feasibility that perfectly complements the current limitations of mRNA delivery. In this study, we …

Reflectance Quantitative Polarized Light Imaging For Microstructural Evaluation Of Musculoskeletal Soft Tissues, Leanne Elizabeth Iannucci

McKelvey School of Engineering Graduate Student Theses & Dissertations

Musculoskeletal soft tissue injury is common and can lead to debilitating pain, loss of tissue function, and overall decrease in quality of life. Proper biomechanical function of musculoskeletal soft tissues is dictated by their hierarchically organized collagenous extracellular matrices, which are disrupted in periods of disease or injury. To best understand dynamic structure-function relationships of these tissues in normal and pathological conditions, it is imperative to be able to visualize collagen microstructure using a quantitative, high spatiotemporal resolution imaging modality. The innate birefringence associated with collagen fibers permits the use of biomedical polarimetry techniques to probe collagen in a specific …

Single-Molecule Super-Resolution Imaging Of Geobacter Sulfurreducens Under Anaerobic Conditions, Ziyi Hu

McKelvey School of Engineering Graduate Student Theses & Dissertations

Geobacter sulfurreducens are anaerobic bacteria capable of making electrical contacts with other organisms and extracellular electron acceptors. The challenge of imaging live Geobacter bacteria is maintaining anaerobic conditions during the imaging process. In this thesis, we augment a single-molecule localization microscope (SMLM) with a home-built anaerobic imaging chamber and use constant argon bubbling to maintain oxygen-free imaging conditions. To validate the imaging protocol, we use the transient binding of Nile red to resolve the spherical morphology of lipid-coated glass spheres with nanoscale resolution. However, when imaging Geobacter, the distribution of Nile red localizations is non-uniform, both between different cells …

A General-Purpose Software Platform For Closed-Loop Neuromodulation, William Engelhardt

McKelvey School of Engineering Graduate Student Theses & Dissertations

Implementing closed-loop neuromodulation therapies is a challenging and expensive endeavor. It requires developing software capable of acquiring signals from a bio-signal amplifier, analysis of these signals, and initiation of precisely timed stimulation, all of which need to be accomplished in real-time with very low latency. Developing this software is difficult, as it requires a wide range of expertise ranging from interfacing with hardware to real-time signal processing. Even when successfully implementing such a system for one set of hardware, it often then only works within the laboratory that conceived it. This is because of the inherent heterogeneity in the devices …

Dual Color Optogenetic Control For Analyzing Cardiac Function In Drosophila, Jiantao Zhu

McKelvey School of Engineering Graduate Student Theses & Dissertations

Prolonged consumption of carbohydrate-rich diets and immobile lifestyles frequently cause metabolic disorders and obesity and, as a result, may lead to progressive heart dysfunction among broad social groups of the population. Drosophila melanogaster serves as an essential model organism in cardiovascular disease research due to conserved physiological and genomic traits shared with humans, its genetic and molecular toolbox versatility, and cost-effective maintenance. Here, we combine optogenetics and optical coherence tomography to study cardiovascular function in D. melanogaster. A new optogenetic pacing system has been developed, employing a transgenic line carrying two opsins: ChR2 and NpHR2.0. A custom-built hardware setup …

Development Of A Multispectral Vis-Swir Imaging Modality For Cutaneous Water Assessment, Quinlan Mcgrath

McKelvey School of Engineering Graduate Student Theses & Dissertations

Inflammatory skin diseases are estimated to impact 20% of the global population and are the fourth leading cause of nonfatal disability worldwide. Diagnosis and management are predominantly based on clinician visual assessment of disease related changes in skin morphology. The qualitative nature of this method can result in misdiagnosis and underdiagnoses of treatable diseases. There persists systematic undertreatment of skin of color patients given the more subtle presentation of erythema against pigmented skin. There exists a clinical need for a quantitative and objective inflammation assessment tool that meets the needs of a diverse patient population. The accumulation of interstitial fluid …

Development Of Methods To Enhance Stem Cell Derived Islet Survival, Aining Fan

McKelvey School of Engineering Graduate Student Theses & Dissertations

No abstract provided.

Synthesis, Radiolabeling And Evaluation Of A Suite Of Tracers With 44Sc For Detecting Extracellular Dna, Zhiyao Li

McKelvey School of Engineering Graduate Student Theses & Dissertations

Neutrophil extracellular traps involve the rapid translocation of DNA to the outside of the cell under certain stimuli. This structure forms a fibrous network that is able to limit the spread of pathogens and to kill microorganisms. It has also been shown to be present in various pathological processes such as inflammation, autoimmune diseases, and cancer metastasis. Currently, the formation process of NETs in vivo is being extensively studied. However noninvasive detection and quantitation has yet to be achieved. A class of PET tracers are described here that consists of a DNA dye as the backbone that is labeled with …

A Magneto-Rheological Platform To Dynamically Control Afterload In Engineered Micro Heart Muscles, David Schuftan

McKelvey School of Engineering Graduate Student Theses & Dissertations

Cardiovascular disease is the leading cause of death is the United States, accounting for nearly 1 in 5 deaths. Many types of cardiovascular disease are linked to the mechanical forces placed on the heart. However, how these mechanical forces exactly affect the cellular biology of the heart is not well defined. In vitro models using cardiomyocytes derived from human induced pluripotent stem cells enable researchers to develop medium throughput systems to study cardiac mechanobiology at the cellular level. Previous models have been developed to enable the study of mechanical forces such as cardiac afterload. However, most of these models require …

Comparison Of In-Vitro 3d Human Embryoids With Current Models For Gastrulation, Jin Park

McKelvey School of Engineering Graduate Student Theses & Dissertations

Gastrulation is an early morphogenetic process that is conserved across most metazoans and lays out the future body plan through the formation and shaping of the three germ layers: endoderm, mesoderm, and ectoderm. Despite its importance, not much is known about the events surrounding human gastrulation that occurs in utero due to ethical and technical limitations on studying human embryos. Therefore, many researchers have devised protocols for creating in vitro models of gastrulation using embryonic stem cells. Initially starting with mouse embryonic stem cells, the field of in vitro embryo models has advanced rapidly, with protocols using human embryonic stem …

Six-Dimensional Single-Molecule Imaging With Isotropic Resolution Using A Multi-View Reflector Microscope, Oumeng Zhang, Zijian Guo, Yuanyuan He, Tingting Wu, Michael D. Vahey, Matthew D. Lew

Electrical & Systems Engineering Publications and Presentations

Imaging of both the positions and orientations of single fluorophores, termed single-molecule orientation-localization microscopy, is a powerful tool for the study of biochemical processes. However, the limited photon budget associated with single-molecule fluorescence makes high-dimensional imaging with isotropic, nanoscale spatial resolution a formidable challenge. Here we realize a radially and azimuthally polarized multi-view reflector (raMVR) microscope for the imaging of the three-dimensional (3D) positions and 3D orientations of single molecules, with precisions of 10.9 nm and 2.0° over a 1.5-μm depth range. The raMVR microscope achieves 6D super-resolution imaging of Nile red molecules transiently bound to lipid-coated spheres, accurately resolving …

The Influence Of Electrical Stimulation Pulse Frequency Of Macro-Sieve Electrode On Rat Sciatic Nerve Detection Threshold, Jingyuan Zhang

McKelvey School of Engineering Graduate Student Theses & Dissertations

No abstract provided.