Molecular, Cellular, and Tissue Engineering Commons^™

Engineering Next-Generation Cardiovascular Therapies, Ryan Walker Barrs

All Dissertations

Cardiovascular diseases are the leading cause of death worldwide. Heart disease, in particular, accounted for 1 in every 5 deaths in the United States in 2021, representing a substantial public health burden that will require more effective treatment options to alleviate. Heart transplant surgery is currently the only option to completely restore cardiac function, but many patients die while waiting for a transplant due to a longstanding organ donor shortage. Thus, there is a dire need for accessible treatment options that can repair damaged hearts. Recent advances in tissue engineering and regenerative medicine have accelerated the development of therapies for …

Experimental And Computational Platforms For Studying Systems Mechanobiology, Brendyn Miller

All Dissertations

Mechanical stimulation through physical activity has been shown to play an important role in treating and preventing several non-communicable diseases such as hypertension, lower back pain (LBP), type-2 diabetes mellitus, and several cancers. This is accomplished through the regulation of cellular behavior and tissue remodeling within the body at both the micro- and macro-scale levels. The goal of mechanobiology research is to gain in-depth knowledge and understanding of how cells sense physical forces in conjunction with other biochemical cues and translate those factors into important biological functions that either maintain tissue homeostasis or lead to pathological states. Understanding these processes …

Integrative Transcriptomics And Proteomics Profiling Of Arabidopsis Thaliana Elucidates Novel Mechanisms Underlying Spaceflight Adaptation, Gbolaga O. Olanrewaju, Natasha J. Haveman, Michael J. Naldrett, Anna-Lisa Paul, Robert J. Ferl, Sarah E. Wyatt

Nebraska Center for Biotechnology: Faculty and Staff Publications

Spaceflight presents a unique environment with complex stressors, including microgravity and radiation, that can influence plant physiology at molecular levels. Combining transcriptomics and proteomics approaches, this research gives insights into the coordination of transcriptome and proteome in Arabidopsis’ molecular and physiological responses to Spaceflight environmental stress. Arabidopsis seedlings were germinated and grown in microgravity (μg) aboard the International Space Station (ISS) in NASA Biological Research in Canisters – Light Emitting Diode (BRIC LED) hardware, with the ground control established on Earth. At 10 days old, seedlings were frozen in RNA-later and returned to Earth. RNA-seq transcriptomics and TMTlabeled LC-MS/MS proteomic …

Computational Modeling Using A Novel Continuum Approach Coupled With Pathway-Informed Neural Networks To Optimize Dynein-Mediated Centrosome Positioning In Polarized Cells, Arkaprovo Ghosal, Padmanabhan Seshaiyar Dr., Adriana Dawes Dr., General Genomics Inc.

Annual Symposium on Biomathematics and Ecology Education and Research

No abstract provided.

Development Of An Angiogenic Tissue-On-A-Chip Microenvironment, Eric Stuehr

Master's Theses

Preclinical testing is necessary to investigate the safety and efficacy of novel therapeutics before moving to clinical trials, yet approximately 90% of these therapies fail once tested in humans. This has led to increased interest in developing robust preclinical models that accurately mimic the complex human in vivo physiology. Microfluidic devices that can introduce dynamic conditions to 3D cell/organoid cultures, also known as tissue-on-a-chip, have emerged as physiologically relevant in vitro preclinical models that can achieve high throughput screening of therapeutics. The research presented here aimed to develop an angiogenic environment within a novel microfluidic device to stimulate formation of …

Characterization Of Influenza-Specific Antibodies, Xinping Ran

ENGS 86 Independent Projects (AB Students)

Influenza viruses cause respiratory illness that affects 8% of the U.S. population annually. However, current influenza vaccines have a limited efficacy of only 40-60%. This highlights the need for a universal vaccine design that can generate broadly neutralizing antibodies against influenza surface protein hemagglutinin (HA). Previous work by our group identified HA-specific monoclonal antibodies (mAbs) from serum samples through high-throughput antibody profiling. In this study, three of the identified mAbs were generated using mammalian cell expression and their binding affinity against three HA antigens was characterized via Enzyme-Linked Immuno-Sorbent Assay (ELISA). The ELISA results demonstrated specific binding affinity towards B/Yamagata/18/1998 …

Guar-Based Injectable Hydrogel For Drug Delivery And In Vitro Bone Cell Growth, Humandra Poudel, Ambar R. Rangumagar, Pooja Singh, Adeolu Oluremi, Nawab Ali, Fumiya Watanabe, Joseph Batta-Mpouma, Jin-Woo Kim, Ahona Ghosh, Anindya Ghosh

Chemistry & Biochemistry Faculty Publications and Presentations

Injectable hydrogels offer numerous advantages in various areas, which include tissue engineering and drug delivery because of their unique properties such as tunability, excellent carrier properties, and biocompatibility. These hydrogels can be administered with minimal invasiveness. In this study, we synthesized an injectable hydrogel by rehydrating lyophilized mixtures of guar adamantane (Guar-ADI) and poly-β-cyclodextrin (p-βCD) in a solution of phosphate-buffered saline (PBS) maintained at pH 7.4. The hydrogel was formed via host-guest interaction between modified guar (Guar-ADI), obtained by reacting guar gum with 1-adamantyl isocyanate (ADI) and p-βCD. Comprehensive characterization of all synthesized materials, including the hydrogel, was performed using …

The Effects Of Mechanical Strain On Vascular Calcification And The Canonical Wnt Pathway, Hannah E. Douglas

Theses and Dissertations

Cardiovascular disease is a significant health crisis, representing 32% of deaths worldwide in 2019. Vascular calcification (VC), a major contributor to cardiovascular disease, is a regulated biomineralization process whose exact mechanisms are unknown. Additionally, vascular smooth muscle cells (VSMCs) significantly contribute to VC by undergoing a phenotypic switch and differentiating into osteoblast-like cells. When factors like hypertension cause disturbed laminar flow in the body’s vasculature, the mechanical stress promotes the phenotypic switch and calcification of VSMCs via mechanotransduction. VC is also induced by the Wnt pathway, which is activated via mineral imbalance and mechanical stimulation. However, the exact mechanisms behind …

Development Of Air-Jetting System For Bioprinting Of Alginate Micro-Droplets, Nathan Lingenfelter

Electronic Theses and Dissertations

Transplantation of organoids has been demonstrated to be a promising cure to many diseases including Type-1 diabetes. However, achieving consistent and robust derivations of mature organoids from human pluripotent stem cells (hPSCs) is challenging. Cellular mechano-transduction, where mechanical stimuli is sensed and converted to biochemical signaling, has been proposed to overcome this challenge. This pathway relies on encapsulating hPSCs in a small volume of a biomaterial such as alginate, which has been widely used as cell encapsulation materials due to its biocompatibility and selective permeability. In this project, we focused on developing an air-jetting based technique to generate small and …

Tandem Peptides Targeting Her2 For Delivery Of Cd44 Sirna Into Her2+ Breast Cancer Cells In Vitro, James Kalogeros

All Theses

More than 300,000 women will be diagnosed with breast cancer, and approximately 48,000 women will die this year due to the disease in the United States. HER2+ breast cancer accounts for 20-25% of invasive breast cancer, and it has a worse prognosis and higher early-stage mortality rate than other breast cancer subtypes. Common treatments for HER2+ breast cancer involve surgical removal of the breast or systemic therapy with chemotherapeutic drugs, both of which cause damage to healthy tissue. Due to the lack of a natural ligand for the HER2 receptor, there is a need for improved targeting strategies for the …

Prevention And Reversal Of Elastin-Associated And Phosphate-Dependent Arterial Calcification Using Targeted Nanotherapeutics In Pre-Clinical Models, Fatema Tuj Zohora

All Dissertations

Vascular calcification can be life-threatening, involving calcium phosphate crystal deposits in the blood vessels. The calcification in the intimal layer of the arterial wall is usually associated with atherosclerotic plaque build-up that, in the advanced stage, harbors ‘dispersed’ and ‘spotty’ deposits of calcium phosphate crystals in the necrotic core of the plaque. In contrast, medial arterial calcification (MAC) can form independently of atherosclerosis and appears as ‘Railroad-like’ calcific deposits along degraded elastin lamellae in the medial layer of arteries. The mechanisms of such elastin-associated medial arterial calcification and potential therapies are the key concerns of this dissertation. MAC is common …

Investigating The Potential Of A Cell-Based Gene Editing Therapy For Inherited Metabolic Liver Disease, Ilayda Ates

All Dissertations

Inherited metabolic diseases (IMDs) affecting the liver are relatively rare but collectively have a prevalence of 1 in 800 live births. These diseases result from autosomal recessive single-gene mutations, leading to organ dysfunction and potentially fatal consequences if left untreated. One potential therapeutic strategy for IMDs of the liver involves using CRISPR-Cas9-induced loss of function mutations. However, translating this approach into the clinic is limited by the need for safe and effective CRISPR delivery methods. Adeno-associated viral vectors (AAVs), commonly used for CRISPR delivery, are associated with significant safety and efficacy concerns, including risks for immunogenicity, off-target mutagenesis, and genotoxicity …

Protein L Functionalized Microparticles For Early Detection Of Surface Markers In Heterogeneous Colorectal Lesions, Saleh Ramezani

Dissertations and Theses (Open Access)

Colorectal cancer (CRC) is the third leading cause of cancer-related deaths among both men and women worldwide, a statistic that can be improved with early detection. Increased understanding of the tumor landscape has identified surface antigens that can be utilized to distinguish normal and cancerous tissue lining the colonic lumen. Use of antibodies conjugated to nano/micromaterials offers a means to detect these antigens, however, the heterogeneous nature of tumors makes it unlikely that antibodies against a single antigen will have sufficient sensitivity and specificity to detect all tumors. A versatile micromaterial able to be functionalized with more than one targeting …

In Vitro And In Vivo Diabetic Models For Assessment Of Tissue Engineered Vascular Grafts, Juan Carlos Carrillo Garcia

All Dissertations

Diabetes has become one of the leading causes of lower-limb loss worldwide. Every 30 seconds, a person loses a limb due to diabetic-related vascular complications. About one-third of patients needing lower-limb bypass surgery have debilitated autologous vessels unsuitable for use, and no other good long-term options are available. These detrimental effects on the vasculature are caused mainly by the hyperglycemic and hyperlipidemic conditions derived from diabetes. Under these conditions, an increase in advanced glycation end products (AGEs) and reactive oxygen species leads to irreversible crosslinks of extracellular matrix proteins, accelerating vascular pathology through vascular stiffening, endothelial dysfunction, inflammation, atherosclerosis, fibrosis, …

Editorial: Intervertebral Disc Degeneration And Osteoarthritis: Mechanisms Of Disease And Functional Repair., Graciosa Q Teixeira, Jana Riegger, Raquel M Gonçalves, Makarand V. Risbud

Department of Orthopaedic Surgery Faculty Papers

No abstract provided.

Physiologically Relevant Fluid Shear Stress Induced Receptor Conversion In Hormone Receptor-Positive (Hr+) Breast Cancer, Jonathan James Cuccia

LSU Doctoral Dissertations

Breast cancer is the most common type of cancer among all people in the United States as of 2023 and often has devastating impacts on afflicted patients. Although some treatments show great efficacy against primary breast tumors, none have been able to significantly reduce the mortality rate of breast cancer once it has metastasized to distant sites. In primary, hormone receptor-positive (HR+) breast cancers, endocrine therapies are often used as first-line treatments and are generally well-tolerated. However, as the disease progresses, these previously sensitive cancers can become resistant to endocrine therapies and continue proliferating despite them. Therefore, there is a …

Mass Spectrometry Of The White Adipose Metabolome In A Hibernating Mammal Reveals Seasonal Changes In Alternate Fuels And Carnitine Derivatives, Frazer I. Heinis, Sophie Alvarez, Matthew T. Andrews

Nebraska Center for Biotechnology: Faculty and Staff Publications

Mammalian hibernators undergo substantial changes in metabolic function throughout the seasonal hibernation cycle. We report here the polar metabolomic profile of white adipose tissue isolated from active and hibernating thirteen-lined ground squirrels (Ictidomys tridecemlineatus). Polar compounds in white adipose tissue were extracted from five groups representing different timepoints throughout the seasonal activity-torpor cycle and analyzed using hydrophilic interaction liquid chromatography-mass spectrometry in both the positive and negative ion modes. A total of 224 compounds out of 660 features detected after curation were annotated. Unsupervised clustering using principal component analysis revealed discrete clusters representing the different seasonal timepoints throughout hibernation. One-way …

Control Of The Electroporation Efficiency Of Nanosecond Pulses By Swinging The Electric Field Vector Direction, Vitalii Kim, Iurii Semenov, Allen S. Kiester, Mark A. Keppler, Bennett L. Ibey, Joel N. Bixler, Ruben M. L. Colunga Biancatelli, Andrei G. Pakhomov

Bioelectrics Publications

Reversing the pulse polarity, i.e., changing the electric field direction by 180°, inhibits electroporation and electrostimulation by nanosecond electric pulses (nsEPs). This feature, known as “bipolar cancellation,” enables selective remote targeting with nsEPs and reduces the neuromuscular side effects of ablation therapies. We analyzed the biophysical mechanisms and measured how cancellation weakens and is replaced by facilitation when nsEPs are applied from different directions at angles from 0 to 180°. Monolayers of endothelial cells were electroporated by a train of five pulses (600 ns) or five paired pulses (600 + 600 ns) applied at 1 Hz or 833 kHz. Reversing …

Clinical Utility Of Mesenchymal Stem/Stromal Cells In Regenerative Medicine And Cellular Therapy, Vitali V. Maldonado, Neel H. Patel, Emma E. Smith, C. Lowry Barnes, Michael P. Gustafson, Raj R. Rao, Rebekah M. Samsonraj

Biomedical Engineering Faculty Publications and Presentations

Mesenchymal stem/stromal cells (MSCs) have been carefully examined to have tremendous potential in regenerative medicine. With their immunomodulatory and regenerative properties, MSCs have numerous applications within the clinical sector. MSCs have the properties of multilineage differentiation, paracrine signaling, and can be isolated from various tissues, which makes them a key candidate for applications in numerous organ systems. To accentuate the importance of MSC therapy for a range of clinical indications, this review highlights MSC-specific studies on the musculoskeletal, nervous, cardiovascular, and immune systems where most trials are reported. Furthermore, an updated list of the different types of MSCs used in …

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 …

Modeling Epithelial-Mesenchymal Transition In A 3d Multicellular Model Of Tgf-Β1 Signaling, Kristin Kim, Chris Lemmon

Biology and Medicine Through Mathematics Conference

No abstract provided.

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 …

The Development Of A Primer Payload With Microparticles For Uti Pathogen Identification Using Polythymidine- Modified Lamp Primers In Droplet Lamp, Jonas Otoo

KGI Theses and Dissertations

Nucleic acid amplification tests (NAATs) are among the diagnostic tests with the highest sensitivity and specificity. However, they are more complex to develop than other diagnostic tests such as biochemical tests and lateral flow immunoassay tests. Polymerase chain reaction (PCR) is the gold standard for NAATs. PCR requires thermal cycling to achieve clonal amplification of the target pathogen DNA for diagnosis. Thermal cycling poses a challenge in the development of PCR diagnostics for point-of-care (POC) settings. Loop-mediated isothermal amplification (LAMP) offers an isothermal method for NAATs diagnostics. The advancement of the microfluidics field significantly enhances the development of LAMP diagnostics …

Understanding Vascular Calcification Through The Lens Of Canonical Wnt Signaling, Karlee Mcneel

Theses and Dissertations

Every 37 seconds, someone in the United States dies from cardiovascular disease. Vascular calcification is one of the underlying causes of these fatal events. Medial calcification develops following arteriosclerosis, or hardening of the arteries. Medial calcification is characterized by the deposition of hydroxyapatite in the medial layer of the arteries after normal vascular smooth muscle cells undergo a phenotypic switch to resemble osteoblast-like cells. It is hypothesized that this switch is caused by the wingless related (WNT)-Signaling pathway. The WNT-Signaling pathway, upon activation, causes the upregulation of osteogenic markers for the development of osteoblast-like cells. Current treatments alleviate consequences of …

Growth Plate Cartilage: Understanding The Contribution Of Adhesion To Column Formation And Matrix Structure, Sydney E. Greer

Theses & Dissertations

Throughout fetal and adolescent development, bone growth is regulated by fine-tuned and controlled maturation of chondrocytes through a cartilaginous template called the growth plate. Bone growth rate is controlled through cell enlargement and extracellular matrix deposition, while the polarized arrangement of proliferative chondrocytes into columns aligned with the long axis of the bone potentiate growth. Chondrocytes are surrounded by a complex three-dimensional arrangement of matrix molecules, all of which are secreted by chondrocytes and assembled/remodeled to support the biological functions of the cell. Adhesion receptors found on the cell membrane of chondrocytes are crucial to the organization of matrix proteins …

Implementation Of Methods For Cell Therapy Research And Assessment Of The Impact Of Cryopreservation, Samantha Lynes

Master's Theses

Cell therapy is the administration of living cells into a patient to prevent or treat a variety of diseases and illnesses. The cell therapy industry is rapidly expanding, and continued research is necessary for manufacturing safe and effective therapies. Although cell therapy manufacturing generally involves cryopreservation processes for storage, there are limited standards for cryopreservation processes and assays required to evaluate cell therapies post-thaw, and limited understanding exists about how a recovery period post-thaw could impact cell health. The overall goal of this thesis was to evaluate the effect of cryopreservation and potential subsequent recovery time on cell viability.

Because …

Assesment Of Structure, Function, And Microevolutionary Dynamics Of Extrachromosomal Circular Dna In Chinese Hamster Ovary Cells, Dylan Chitwood

All Dissertations

Chinese hamster ovary (CHO) cell lines are among the most popular expression hosts used in biopharmaceutical manufacturing due to relative ease of culture, capacity to perform human-like post-translational modifications, and non-susceptibility to viruses. However, the intrinsic plasticity of the CHO genome can lead to undesired genetic rearrangements, phenotypic shifts, reduced product quality, and early culture termination that prevents continuous biomanufacturing. A characteristic of plastic and unstable genomes that is poorly understood in CHO cells is extrachromosomal circular DNA (eccDNA). EccDNAs are focal amplifications of the genome that reside in the extranuclear space. These plasmid-like entities are structurally complex and are …

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 …