Molecular, Cellular, and Tissue Engineering Commons^™

Multipolymer Microsphere Delivery Of Sars-Cov-2 Antigens, Farah Shahjin, Milankumar Patel, Jatin Machhi, Jacob D. Cohen, Mohammad Ullah Nayan, Pravin Yeapuri, Chen Zhang, Emiko Waight, Mahmudul Hasan, Mai Mohamed Abdelmoaty, Prasanta K. Dash, You Zhou, Irene Andreu, Howard E. Gendelman, Bhavesh D. Kevadiya

Nebraska Center for Biotechnology: Faculty and Staff Publications

Effective antigen delivery facilitates antiviral vaccine success defined by effective immune protective re- sponses against viral exposures. To improve severe acute respiratory syndrome coronavirus-2 (SARS-CoV- 2) antigen delivery, a controlled biodegradable, stable, biocompatible, and nontoxic polymeric micro- sphere system was developed for chemically inactivated viral proteins. SARS-CoV-2 proteins encapsulated in polymeric microspheres induced robust antiviral immunity. The viral antigen-loaded microsphere sys- tem can preclude the need for repeat administrations, highlighting its potential as an effective vaccine.

Development Of A Crosslinked Osteochondral Xenograft And A Collagen Stabilizing Intra-Articular Injection To Remediate Cartilage Focal Lesions To Prevent Osteoarthritis, Mark Lewis Mosher

Theses and Dissertations

Osteoarthritis is one of the most common causes of disability in adults in America. It is a progressive and degenerative disease where the articular cartilage is broken down and lost from the surfaces of bones causing chronic pain and swelling in the joints, and currently has no cure. The most commonly osteoarthritis starts from a focal lesion on the cartilage surface, which will expand on the surface and downwards through the thickness of the tissue. The current gold standard for correcting cartilage focal lesions is the osteochondral autograft/allograft transplantation (OAT), which replaces the defect with a fresh osteochondral graft. The …

In Vitro Bioreactor For Mechanical Control And Characterization Of Tissue Constructs, Samuel Coeyman

All Dissertations

Heart failure (HF) currently affects over 6 million Americans, 50% of whom die within 5 years of their initial diagnosis. A major contributor to the onset of HF is cardiac fibrosis in the myocardium, which arises when fibroblasts (FBs) are activated in response to heightened mechanical stress from overload conditions like hypertension. Activated FBs remodel the extracellular matrix (ECM) and secrete ECM proteins including collagen. FB remodeling has been studied in the past by applying forces and/or deformations to three-dimensional, cell-seeded gels and tissue constructs in vitro. Unfortunately, previous stretching platforms have traditionally not enabled mechanical property assessment to be …

Ultrasound Mediated Tissue Growth In Bioinspired Scaffolds, Marco Fielder

Graduate Theses and Dissertations

Porous scaffolds mechanically stimulated with ultrasound are used for the treatment of bone conditions such as osteoporosis or traumatic bone injury. However, one current issue in these treatment techniques is designing a scaffold structure with a high structural integrity that also does not impede tissue growth. A larger ultrasound wave transmission would also allow more of a scaffold to receive mechanical stimulation from ultrasound. Controlling mesenchymal stem cell seeding of tissue scaffolds also has potential for developing optimal seeding patterns to maximize or control tissue growth in the scaffold based on an individual’s physiology. There is also a need for …

Engineer And Test A Biodegradable Microneedle Patch To Deliver Meloxicam For Managing Pain In Cattle, Katherine Alejandra Miranda Munoz

Graduate Theses and Dissertations

Microneedle patches are a promising source for transdermal diffusion of macromolecules and are designed to painlessly penetrate the skin. In this study, a biodegradable chitosan microneedle patch to deliver meloxicam for managing pain in cattle was tested. The potential of reuse of the polymeric solution to fabricate the patches, optimization of fabrication, morphological analysis of the microneedle patch and analysis of preservation of the chemical composition after sterilization were evaluated. In-vitro analysis consisted of studying in-vitro penetration mechanical properties, compression testing analysis of microneedle patch, and in-vitro drug release analysis. In-vivo studies were performed to analyze the dissolution capability of …

Evaluation Of Cellulose Nanocrystal Inks And Their Structural Characteristics For 3d Bioprinting Of Customized Scaffolds, Patrick William Kuczwara

Graduate Theses and Dissertations

3D bioprinting of biological scaffolds requires control of the physicochemical properties of each unique structures. A promising material for control of properties is hydrogels, which can help create biomimetic scaffolds with controlled spatial arrangement of materials by integrating biological materials directly into layers during the bioprinting process. Nanocellulose offers a unique combination of properties including mechanical, biomimetic, and biocompatibility. These properties offer flexibility over the types, shapes, and applications of their printed hydrogel scaffolds, (i.e., tissue, drug, encapsulation). However, 3D bioprinting of nanocellulose-based hydrogels requires high loading percentages (i.e., >10 wt%) or chemical crosslinkers (i.e., bis(acyl)phosphane oxides (BAPO)). High solid …

Development Of A Tissue Engineered Cardiac Patch, Howard Herbert

All Dissertations

Cardiovascular Disease(CVD) is the leading cause of mortality in the developed world. CVD is most commonly manifested as atherosclerosis of the coronary arteries leading to Myocardial Infarction(MI). After MI, fibrosis of the ventricular wall leads to heart failure(HF), a pandemic affecting 26 million people globally. While therapies are continuously developed to combat HF, the treatment of choice, whole heart transplant, is limited by the availability of donor hearts. It is clear that there is a need to develop a long-term solution to combat HF and its enormous economic burden. Tissue Engineering and Regenerative Medicine holds promise as a possible solution …

Establishing The Efficacy Of Non-Cellular Components Of Adipose-Derived Stromal Vascular Fraction In Promoting Angiogenesis., Daniel Benson

Electronic Theses and Dissertations

Microvascular disease is hallmarked by pathophysiological conditions such as endothelial senescence, intimal thickening which impairs vasodilation, and regression of the capillary beds causing tissue ischemia in the myocardium or in peripheral vascular networks. Adipose-derived stromal vascular fraction (SVF) has previously demonstrated the ability to revascularize tissue. Increasing evidence shows that regenerative cells elicit their therapeutic benefit by paracrine mechanisms, leaving open extracellular vesicles (EVs) as a potential crux of the cell therapy paradigm. To test this idea, three types of gelatin methacrylate hydrogels were employed: SVF gels, EV gels derived from SVF, and blank control gels, which were used in-vitro …

Enhancing Human Schwann Cells Reparative Behavior Using Heparin/Collagen Layer-By-Layer Coatings, Luis Carlos Pinzon-Herrera

Graduate Theses and Dissertations

When a peripheral nerve injury (PNI) occurs, the gold standard for tissue regeneration is the use of autografts. However, due to the secondary effects produced by multiple surgeries involved in the removal and implantation of autografts for very small lesions, it is possible to replace them with the use of Nerve Guide Conduits (NGCs). However, NGCs are limited to short lesions (less than 1 cm). This limitation is caused by the absence of compounds in the extracellular matrix (ECM) that autografts can provide. Since much of the regenerative process takes place on the NGC surface, our work aims to modify …

Investigating Spatial Heterogeneity In Myocardial Wound Environments To Improve Therapy, Michael Potter

All Dissertations

Heart failure is a broad pathology manifestation categorized by an inability of the heart to successfully pump blood throughout the vast vessel network of the body. Within the United States, heart failure is projected to increase by approximately 46% from 2012 to 2030. Modalities of heart failure are generally related to wall mechanics that are impacted following myocardial infarction events. Interplay exists between the wall mechanics, responding cell populations, and the spatial heterogeneities in the resultant scar. This interplay directs the myocardium towards heart failure modalities governed by overly stiff or compliant states. It is essential to elucidate details underlying …

Piglet Cardiopulmonary Bypass Induces Intestinal Dysbiosis And Barrier Dysfunction Associated With Systemic Inflammation, Jeffrey D. Salomon, Haowen Qiu, Dan Feng, Jacob Owens, Ludmila Khailova, Suzanne Osorio Lujan, John Iguidbashian, Yashpal S. Chhonker, Daryl J. Murry, Jean-Jack Riethoven, Merry L. Lindsey, Amar B. Singh, Jesse A. Davidson

Nebraska Center for Biotechnology: Faculty and Staff Publications

The intestinal microbiome is essential to human health and homeostasis, and is implicated in the pathophysiology of disease, including congenital heart disease and cardiac surgery. Improving the microbiome and reducing inflammatory metabolites may reduce systemic inflammation following cardiac surgery with cardiopulmonary bypass (CPB) to expedite recovery postoperatively. Limited research exists in this area and identifying animal models that can replicate changes in the human intestinal microbiome after CPB is necessary. We used a piglet model of CPB with two groups, CPB (n=5) and a control group with mechanical ventilation (n=7), to evaluate changes to the microbiome, …

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 …

Cellular Bioenergetics: Experimental Evidence For Alcohol-Induced Adaptations, Liz Simon, Patricia E. Molina

School of Medicine Faculty Publications

At-risk alcohol use is associated with multisystemic effects and end-organ injury, and significantly contributes to global health burden. Several alcohol-mediated mechanisms have been identified, with bioenergetic maladaptation gaining credence as an underlying pathophysiological mechanism contributing to cellular injury. This evidence-based review focuses on the current knowledge of alcohol-induced bioenergetic adaptations in metabolically active tissues: liver, cardiac and skeletal muscle, pancreas, and brain. Alcohol metabolism itself significantly interferes with bioenergetic pathways in tissues, particularly the liver. Alcohol decreases states of respiration in the electron transport chain, and activity and expression of respiratory complexes, with a net effect to decrease ATP content. …

Labeling Melanoma Cells With Black Microspheres For Improved Sensitivity In Detection Via Photoacoustic Flow Cytometry, Tori Kocsis

Electronic Theses and Dissertations

Melanoma is an aggressive form of skin cancer known for developing into metastatic disease. Current clinical diagnostics, including medical imaging and tissue biopsy, provide a poor prognosis since the cancer is in the late stages of disease progression. In recent years, photoacoustic flow cytometry has allowed for the detection of circulating melanoma cells within patient blood samples in vitro. Although this method exploits the naturally-produced melanin within the cells, it has only successfully detected highly-pigmented melanoma cell lines. Since various forms of melanoma exist, each with varying melanin concentrations, this research aims to provide a novel method for detecting lightly-pigmented …

Optimizing Crispr/Cas9-Mediated Knockdown Of Angptl3 In Liver Cell Lines And Mouse Hepatocytes, Meredith Reeves

All Theses

Familial hypercholesterolemia (FH) is a genetic condition characterized by elevated levels of low-density lipoprotein cholesterol (LDL-C) that leads to an increased risk of developing cardiac disease early in life (Shah et al., 2020). Current treatments such as statins and PCSK9 inhibitors have helped lower LDL-C levels, however they require repeated administration every 4-6 weeks to remain effective (Raal et al., 2018). Angiopoietin-like 3 (ANGPTL3) is an inhibitor of plasma lipid metabolism that has become a promising molecular target for the treatment of FH. Individuals with non-functional copies of ANGPTL3 demonstrate low levels of plasma LDL-C and triglycerides, indicating a protective …

A 3d Tissue Engineering Model To Study Mitral Valve Annulus Calcification Under Diabetic Conditions, Erin James

All Theses

The most complex heart valve is the mitral valve (MV). Many pathologies can affect the MV, including stenosis, regurgitation, prolapse, and mitral annulus calcification (MAC). MAC is chronic degeneration of the annulus, which is the fibrous, saddle-shaped “ring” that can contract and relax with the myocardium. The prevalence of MAC is around 15% but increases in patients with other cardiovascular diseases and risk factors. It is also thought to increase in patients with type 2 diabetes, but MAC has not been properly characterized within this population because of confounding factors such as cardiac disease and kidney disease. The goal of …

Mitral Valve Tissue Engineering - A Dynamic Model For Investigating The Mechanism Of Valvular Pathology, Collin Owens

All Dissertations

Heart valve disease affects an average of 2.5% of the population in the United States. The mitral valve (MV) is the most complex of the heart’s four valves and is most associated with the disease by exhibiting altered extracellular matrix (ECM) which translates into stenosis or regurgitation. These diseases are typically degenerative in nature and can be accelerated by risk factors such as diabetes and hypertension. With diabetes and hypertension affecting 425 million and 1.39 billion people worldwide, further investigation into these risk factors is warranted. This study aims to develop and test an in vitro model of MV disease. …

Engineering Of Ideal Systems For The Study And Direction Of Stem Cell Asymmetrical Division And Fate Determination, Martina Zamponi

Biomedical Engineering Theses & Dissertations

The cellular microenvironment varies significantly across tissues, and it is constituted by both resident cells and the macromolecules they are exposed to. Cues that the cells receive from the microenvironment, as well as the signaling transmitted to it, affect their physiology and behavior. This notion is valid in the context of stem cells, which are susceptible to biochemical and biomechanical signaling exchanged with the microenvironment, and which plays a fundamental role in establishing fate determination and cell differentiation events. The definition of the molecular mechanisms that drive stem cell asymmetrical division, and how these are modulated by microenvironmental signaling, is …

Functional Requirements For A Samd14-Capping Protein Complex In Stress Erythropoiesis, Suhita Ray, Linda Chee, Yichao Zhou, Meg A. Schaefer, Michael J. Naldrett, Sophie Alvarez, Nicholas T. Woods, Kyle J. Hewitt

Nebraska Center for Biotechnology: Faculty and Staff Publications

Acute anemia induces rapid expansion of erythroid precursors and accelerated differentiation to replenish erythrocytes. Paracrine signals—involving cooperation between stem cell factor (SCF)/Kit signaling and other signaling inputs—are required for the increased erythroid precursor activity in anemia. Our prior work revealed that the sterile alpha motif (SAM) domain 14 (Samd14) gene increases the regenerative capacity of the erythroid system in a mouse genetic model and promotes stress-dependent Kit signaling. However, the mechanism underlying Samd14’s role in stress erythropoiesis is unknown. We identified a protein-protein interaction between Samd14 and the α- and β-heterodimers of the F-actin capping protein (CP) complex. Knockdown of …

A Novel Electroconductive Nanofibrous Scaffold For Bone Regeneration, Mitchell Kenter

Medical Engineering Theses

The goal of this study was to develop a biodegradable and conductive scaffold to mimic the piezoelectric properties of bone and the architecture of the extracellular matrix. Poly(3,4- ethylenedioxythiophene) (PEDOT) is a conductive polymer of great interest in tissue engineering due to excellent electrical stability and biocompatibility. To enhance its conductivity, dopants such as dimethyl sulfoxide (DMSO) can be added. Engineered graphene oxide (GO) can also be introduced as oxidant to enhance conductivity and mechanical properties. PEDOT nanocomposites were synthesized by oxidative polymerization of 3, 4-Etylenedioxythiophene monomer (EDOT) in the presence of GO, DMSO, ferric chloride and various solvents. The …

A Microfluidic Assay For Single Cell Bacterial Adhesion Studies Under Shear Stress, Amanda Trusiak

Electronic Theses and Dissertations

The study of bacterial adhesion to host cells is important in understanding bacterial pathogenesis and developing new therapeutic approaches. Here, we studied bacterial adhesion under shear stress using a novel microfluidic method. Specifically, the adhesion of a uropathogenic E. coli strain (FimHOn, ATCC 700928/CFT073) to mannose-modified substrates was studied under flow conditions. The FimHOn E. coli strain expresses FimH which is a mannose-specific adhesin found on the fimbriae that binds to glycoproteins on the epithelium. We developed a microfluidic method that mimics bacterial adhesion to urothelial cells. First, the microfluidic channels were modified by sequentially adsorbing BSA-mannose and BSA onto …

Investigating The Effect Of Dissolved Oxygen-Assisted Corneal Cross-Linking (Cxl) On Porcine Corneas, Julianni Dar

Electronic Theses and Dissertations

Corneal cross-linking is a clinical procedure that is known to stop the progression of keratoconus, an eye disease that affects the cornea’s structure, ultimately leading to vision loss in its advanced stages. The typical treatment plan includes riboflavin and UV-A exposure in the hope to increase the mechanical properties of the cornea. There are two types of CXL pathways, with Type-II CXL requiring oxygen. Naturally, the dissolved oxygen is limited in the cornea; therefore, limiting the effect of Type-II CXL. This study proposes to improve the Type-II CXL contribution by integrating dissolved oxygen during the standard CXL treatment used in …

Designing A Self-Regulating And Portable Heating Device For A Microfluidic Based Biosensor, Riya Mahajan

Discovery Undergraduate Interdisciplinary Research Internship

Paper-based biosensors are powerful microfluidic analytical devices that are potentially useful for a wide range of applications, ranging from medical diagnostics to agricultural and environmental monitoring. Molecular diagnostics have limitations because they need to send samples back to a centralized laboratory, which increases the cost and turnaround time of the test. This project aims to create a simple-to-use, low-cost, and portable heating system that would facilitate the creation of a field-deployable paper-based analytical device that can incubate the sample at elevated temperatures for conducting isothermal molecular assays. Our design aims to miniaturize a commercial water bath and will be fabricated …

A Versatile Python Package For Simulating Dna Nanostructures With Oxdna, Kira Threlfall

Computer Science and Computer Engineering Undergraduate Honors Theses

The ability to synthesize custom DNA molecules has led to the feasibility of DNA nanotechnology. Synthesis is time-consuming and expensive, so simulations of proposed DNA designs are necessary. Open-source simulators, such as oxDNA, are available but often difficult to configure and interface with. Packages such as oxdna-tile-binding pro- vide an interface for oxDNA which allows for the ability to create scripts that automate the configuration process. This project works to improve the scripts in oxdna-tile-binding to improve integration with job scheduling systems commonly used in high-performance computing environments, improve ease-of-use and consistency within the scripts compos- ing oxdna-tile-binding, and move …

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), …