Molecular, Cellular, and Tissue Engineering Commons^™

Development Of An Integrated Salt Cartridge-Reverse Electrodialysis (Red) Device To Increase Electrolyte Concentrations Of Human Blood Flow To Power Biomedical Devices, Caroline Campbell

Chemical Engineering Undergraduate Honors Theses

Emerging technologies in nanotechnology and biomedical sciences have led to an increase in biomedical implantable devices including cardiac pacemakers, artificial organs, drug pumps, and sensors. These devices require continuous stable and reliable power to operate, which creates the demand for the need to find a safe, reliable, and stable power source. A promising avenue for a power source for these devices is a miniaturized reverse electrodialysis (RED) biopower cell design that utilizes the salinity differences between bloodstreams that flow inside the human body. Initial results of the RED system demonstrate that higher gradient salinity differences between streams lead to a …

Evaluating The Effects Of Wood Source On The Physicochemical Properties Of Crosslinked Cellulose Nanocrystals, Helena Tchoungang Nkeumen

Graduate Theses and Dissertations

Cellulose is an abundant and naturally occurring biopolymer that has been used by humans for food, shelter, and clothing for about two centuries now. Highly crystalline nanoparticles derived from cellulose, called cellulose nanocrystals (CNCs), show great potential to meet the rising need for sustainable and nontoxic materials for biomedical applications. However, multiple biomedical applications of CNCs, such as those involving their use in tissue engineering scaffolds, require CNC-based structures to be stable in aqueous environments, a property that native CNCs do not possess due to their inherent hydrophilicity. Chemical crosslinking of CNCs addresses this issue by providing aqueous stability to …

Quantifying The Effect Of Intrinsic Aging On Skin Microstructure And Mechanical Function Using Multiphoton Microscopy, Alan Edward Woessner

Graduate Theses and Dissertations

Over the next couple of decades, the number of elderly individuals is expected to double, bringing increased healthcare spending due to seemingly minor injuries to skin. In skin, the 3D dermal collagen fiber network, which is the primary load-bearing structure, undergoes changes in organization and composition due to intrinsic aging. However, the relationships between altered microstructure and mechanical function is not well understood. Quantitative imaging techniques have been used in the past to link skin structure to mechanical function, but previous analysis has been limited to 2D assessments. Multiphoton microscopy (MPM) is a non-destructive imaging technique with intrinsic depth-sectioning capabilities, …

Oxone® Mediated Tempo-Oxidized Cellulose Nanomaterials: Material Characterization, Ultrafiltration Membrane Separations, And Thin Film Composite Gas Transport Analysis, John Phillips Moore

Graduate Theses and Dissertations

Cellulose nanomaterials (CNMs) are derived from plant matter and are comprised of nanoscopic cellulose crystals and fibers. They have a diverse set of applications, from cosmetics to oil recovery. This study focuses on the properties of Oxone® mediated TEMPO-oxidized cellulose nanomaterials (OTO-CNMs) and their use in controlling the transport properties of polymeric substrates. Synthesis and characterization of cellulosic nanoparticles have resulted in the creation of OTO-CNMs with properties that increase hydrophilicity. With added hydrophilicity, OTO-CNMs possess lower fouling propensity, making them ideal membrane additive for transport limited separations such as hemodialysis.

To utilize the material and unique properties thereof, this …

Peptoid-Based Microsphere Coatings For Biomaterial Applications, Jesse Leland Roberts

Graduate Theses and Dissertations

Peptoids are peptidomimetic oligomers that predominantly harness similarities to peptides for biomimetic functionality. The incorporation of chiral, aromatic side chains in the peptoid sequence allows for the formation of distinct secondary structures and self-assembly into supramolecular assemblies, including microspheres. Peptoid microspheres can be coated onto substrates for potential use in biosensor technologies, tissue engineering platforms, and drug-delivery systems. They have the potential for use in biomedical applications due to their resistance to proteolytic degradation and low immunogenicity. This dissertation focuses on the physical characteristics and robustness of the peptoid microsphere coatings in various physiological conditions, along with their ability to …

Autofluorescence Spectral Unmixing For Quantitative Metabolic Imaging Within Tissues, Lucy Woodbury

Biomedical Engineering Undergraduate Honors Theses

Label-free metabolic imaging through quantification of NADH and FAD autofluorescence has become a powerful and efficient tool for non-invasive measurements of cell metabolism. It has applications in a variety of fields including diagnostic and therapeutic monitoring. However, NADH and FAD imaging typically selects specific single excitation and emission bands where it is assumed that these fluorophores are individually isolated. In some cases of isolated cell imaging, this assumption holds true, but for samples with other intrinsic fluorophores present (i.e. collagen and elastin), it can interfere with quantitative results. Elastin and collagen autofluorescence prevent the broader application of these optical metabolic …

Stirred Suspension Bioreactor Differentiation Of Human Mesenchymal Stem Cells Into Smooth Muscle Cells, Chris Slavin

Biomedical Engineering Undergraduate Honors Theses

Human mesenchymal stem cells (hMSCs) are a promising candidate for cellular therapies due to their multipotency, self-renewal capacity, and immunomodulatory properties. However, their isolation is a difficult and potentially painful process with very low yield, and traditional static mammalian cell culture techniques are too slow and expensive for large scale growth and differentiation of stem cells to be practical. Current research is focused on improving methods for cultivating hMSCs in stirred suspension bioreactors (SSBs), but little work has been done with regard to their differentiation in dynamic conditions such as those in SSBs. Differentiation at a large scale would increase …

The Utilization Of Autofluorescence To Study The Effects Of L-Buthionine-Sulfoximine On Cellular Metabolism In Vitro, Madison Belew

Biomedical Engineering Undergraduate Honors Theses

Oxidative damage and stress impacts metabolic activity and has been implicated in a number of diseases . These medical conditions have been found to be associated with a lack of glutathione (GSH). There is a need to study the effects that the absence of glutathione has in order to expand knowledge of indicators that cause conditions like cataracts, cancer, or impaired wound healing. L-buthionine-sulfoximine (BSO) has been used in past studies to hinder GSH production and induce oxidative stress within a cell. This study explores the effects of BSO and induced oxidative stress on the metabolic activity and processes within …

Cell Line-Specific Differences In Guided Differentiation Of Adipose-Derived Mesenchymal Stem Cells Towards Smooth Muscle Cells, Sherly Makar

Biomedical Engineering Undergraduate Honors Theses

Mesenchymal stem cells (MSCs) are a population of stromal cells found traditionally in the bone marrow and adipose tissues. They can also be found in other tissues including fallopian tube, core blood, peripheral blood, fetal liver, and lungs. Mesenchymal stem cells have profound effects in regenerative engineering, tissue repair and drug discovery owing to the excellent properties such as proliferation, self-renewal, and multipotency generating multiple cell types including adipocytes, osteocytes, cardiomyocytes (CMs), pericytes (PCs), and chondrocytes. MSCs are used as immunomodulators in generating progenitor cells to be used for transplantation, creating engineered organs, and preventing graft vs. host disease (GVHD). …

Comparative Genomics Of Human Mesenchymal Stem Cells And Human Mesenchymal Stem Cells Derived Vascular Smooth Muscle Cells, Samia Ismail

Biomedical Engineering Undergraduate Honors Theses

As of 2017, vascular diseases contributed to 23.1% of all deaths in America. To address the need for more effective and sustainable treatment options for these ailments, stem cell differentiation and implantation has emerged as a viable alternative to standard bypass and graft insertions. A completely autologous treatment can be achieved by extracting adult stem cells, differentiating them into vascular smooth muscle cells (VSMCs), and then reimplanting these cells at the affected tissue site. This study aims to investigate the efficiency of the VSMC differentiation from human mesenchymal stem cells (hMSCs) by comparing 4 cell lines of untreated hMSCs with …

Production And Purification Of Basic Fibroblast Growth Factor Fused To Two Collagen Binding Domains Expressed In E. Coli Bl21 Using Flask And Fed-Batch, Hazim Aljewari

Graduate Theses and Dissertations

Delivering effective and non-toxic doses of bioactive materials that can aid in activating tissue regeneration to wounded tissue has proven to be an enormous challenge. This study was designed to produce a potential therapeutic recombinant protein by fusing two collagen binding domains to basic fibroblast growth factors (bFGF) through a collagenase cleavage site linker, so it can release the bFGF in a wound site by the action of this enzyme. The novel fusion protein was expressed in Escherichia coli BL-21 (E. coli) using traditional flask shaker and fed-batch cultivation. Cell lysate was purified by FPLC using Immobilized metal affinity chromatography …

In Vivo Metabolic And Vascular Response To Hypoxia In Twist Knockdown Murine Breast Cancer, Brandon Sturgill

Graduate Theses and Dissertations

Twist transcription factor is often overexpressed in aggressive tumors. Although needed in early embryonic development for organogenesis, Twist is known to induce an epithelial to mesenchymal transition in cells. In cancer, epithelial to mesenchymal transitions can lead to increased motility and invasiveness. It has also been linked to metabolic reprogramming and increased metastatic risk. Furthermore, metabolic preferences can increase proliferation, enhance metastatic potential, and influence the site of metastasis. We hypothesize that Twist directly affects the metabolism of cancer cells. We expect to see in vivo what we have seen in vitro; Twist overexpression should promote a shift away from …

Generation Of A Ccl2 Knockout Using Crispr/Cas9 And Lipid Mediated Transfection In Ct-26 Murine Colon Carcinoma Cells, Emma Sullivan

Biomedical Engineering Undergraduate Honors Theses

CCL2 is an inflammatory mediator that is released by tumor cells to activate and direct immune cell species, especially macrophages, to inflammatory sites within the body. The goal of this project was to successfully generate knockout the CCL2 ligand gene using a CRISPR/Cas9 complex delivered via lipid mediated transfection. The sgRNA and Cas9 mRNA were introduced into the cells via lipid-mediated transfection. The cells were incubated for 4 days, before being analyzed using PCR and gel electrophoresis. We expected to see one band on the first gel and two bands on the second gel. Two bands appeared on the first …

A Study Of Protein And Peptide-Directed Nanoparticle Synthesis For Catalytic Materials, Abdollah Mosleh

Graduate Theses and Dissertations

Nanoparticles have received much attentions due to their unique properties that makes them suitable candidates for a broad range of applications. As the size of particles decreases, their surface area-to-volume ratio would increase which is the main cause of much attention. In addition to the size, their morphologies and compositions may also play important roles for defining unique properties. Nanoparticle synthesis include both bottom-up and top-down strategies. To control the process of inorganic nanoparticles synthesis one could follow the bottom-up approach to have atom-level control over their compositions, morphologies, phases, and sizes which is the subject of this work. Due …

Bioengineering Extracellular Matrix Scaffolds For Volumetric Muscle Loss, Kevin Roberts

Graduate Theses and Dissertations

Volumetric muscle loss overwhelms skeletal muscle’s ordinarily capable regenerative machinery, resulting in fibrosis and severe functional deficits which have defied clinical repair strategies. My work spans the design and preclinical evaluation of implants intended to drive the cell community of injured muscle toward a regenerative state, as well as the development of an understanding of the molecular responses of this cell community to biomaterial interventions. I demonstrate a new class of biomaterial by leveraging the productive capacity of sacrificial hollow fiber membrane cell culture; I show specifically that unique threads of whole extracellular matrix can be isolated by solvent degradation …

Investigation Of The Effect Of Age On Regenerative Outcomes Following Treatment Of Volumetric Muscle Loss Injuries, John Taehwan Kim

Graduate Theses and Dissertations

Volumetric muscle loss (VML) is a traumatic injury in skeletal muscle resulting in the bulk loss of more than 20% of the muscle’s volume. Included in the bulk loss of muscle is the skeletal muscle niche comprised of nerve bundles, vasculature, local progenitor cells, basal lamina, and muscle fibers, overwhelming innate repair mechanisms. The hallmark of VML injury is the excessive accumulation of non-contractile, fibrotic tissue and permanent functional deficits. Though predominant in the younger demographic, the elderly population is also captured within VML injuries. There are many factors that change with aging in skeletal muscle that may further hinder …

Visualizing Ischemic Skin Flap Necrosis Through Phasor Analysis Of Autofluorescence Lifetime Images, Hallie Ramser Toomer

Graduate Theses and Dissertations

Necrotic skin flaps are difficult to predict and treat due to the lack of quantitative biomarkers. Label-free multiphoton microscopy is well suited for non-invasively monitoring skin metabolism through NAD(P)H and other intrinsic fluorophores, and offers immediate future directions for assessing necrosis in the clinic. The objective of this study was to assess whether phasor FLIM could be used to evaluate skin flap status and treatment efficacy in ex vivo skin sections. Phasor maps revealed differences in growth factor treatment and region, but changes in skin flap autofluorescence at 755nm excitation and 460nm emission were not just related to NAD(P)H. A …

Investigating Virus Clearance Via Ph Inactivation During Biomanufacturing, Wenbo Xu

Biomedical Engineering Undergraduate Honors Theses

In the processing of biopharmaceuticals, viral clearance and viral safety are important for the development of monoclonal antibodies. Murine xenotropic leukemia virus (XMuLV) is one of the retroviruses, recommended by Food and Drug Administration (FDA) as a model virus for viral clearance via inactivation from therapeutics derived from Chinese hamster ovary cells (CHO). A robust and effective method was investigated to clear or inactivate endogenous viruses by low pH inactivation. The effects of different conductivity and inactivated time on XMuLV clearance was determined. Acetate buffer was prepared with different conductivity, and 2% XMuLV was spiked into acetate buffer. XMuLV virus …

Phenotypic Characterization Of Differentiation Of Human Mesenchymal Stem Cells To Smooth Muscle Cells, Jackson Hedrick

Biomedical Engineering Undergraduate Honors Theses

Abdominal aortic aneurysms (AAAs) are expansions in the aortic wall with an increased tendency to rupture. These expansions are triggered by the breakdown of key aortic extracellular matrix (ECM) proteins—elastin and collagen. The degradation of aortic ECM proteins is the consequence of overexpressed matrix metalloproteases (MMPs)–MMP-2 and MMP-9. Adult vascular smooth muscle cells (SMCs) lack the ability to repair the elastic matrix assembly; thus, it is imperative to find a solution to both restore the imbalance of essential ECM proteins in the aorta and mitigate the effects of MMPs. Previous studies have revealed that bone marrow derived mesenchymal stem cells …

Comparing The Effects Of Fibroblast Growth Factors On Growth Rate Of Human Fibroblast Cell Lines, Tasha Repella

Biomedical Engineering Undergraduate Honors Theses

Numerous studies have demonstrated the usefulness of growth factors in medium formulations to promote proliferation and viability of human fibroblasts in laboratory settings. However, due to the differences in the source and age of fibroblasts, the ability of growth factors like hFGF1 to promote proliferation varies. Equally difficult is the ability of hFGF1 to promote proliferation of diseased fibroblasts due to complexities associated with specific mitochondrial diseases. In this context, we decided to evaluate the potential for novel hyper stable growth factors super hFGF and hFGF1 (shFGF and hFGF1 respectively), to contribute to proliferation of different normal and diseased fibroblasts. …

Role Of Local Renin-Angiotensin System In Potentiating Early Calcific Aortic Valve Disease, Jessica Perez

Graduate Theses and Dissertations

Angiotensin-II (Ang-II), a peptide hormone, is a potent vasoconstrictor and cell mitogen. It has been implicated in the development of hypertension as well as atherosclerosis. Recent work has shown that sclerotic aortic valves possess expression of angiotensin type I receptor (AT-1R) and angiotensin converting enzyme (ACE), suggesting altered angiotensin signaling during disease. The role of altered angiotensin signaling on aortic valve mechanics, however, is not clearly understood. We seek to understand the direct effects of the renin angiotensin signaling (RAS) system on the biological and biomechanical properties of aortic valve tissue and develop a finite constitutive model that mimics the …

Diffuse Reflectance Spectroscopy To Quantify In Vivo Tissue Optical Properties: Applications In Human Epithelium And Subcutaneous Murine Colon Cancer, Gage Joseph Greening

Graduate Theses and Dissertations

Colorectal cancer is the 4th most common and 2nd deadliest cancer. Problems exist with predicting which patients will respond best to certain therapy regimens. Diffuse reflectance spectroscopy has been suggested as a candidate to optically monitor a patient’s early response to therapy and has been received favorably in experimentally managing other cancers such as breast and skin. In this dissertation, two diffuse reflectance spectroscopy probes were designed: one with a combined high-resolution microendoscopy modality, and one that was optimized for acquiring data from subcutaneous murine tumors. For both probes, percent errors for estimating tissue optical properties (reduced scattering coefficient and …

Impedance Sensing Of Cancer Cells Directly On Sensory Bioscaffolds Of Bioceramics Nanofibers, Hanan Alismail

Graduate Theses and Dissertations

Cancer cell research has been growing for decades. In the field of cancer pathology, there is an increasing and long-unmet need to develop a new technology for low-cost, rapid, sensitive, selective, label-free (i.e. direct), simple and reliable screening, diagnosis, and monitoring of live cancer and normal cells in same shape and size from the same anatomic region. For the first time on using an impedance signal, the breast cancer and normal cells have been thus screened, diagnosed and monitored on a smart bioscaffold of entangled nanowires of bioceramics titanate grown directly on the surface of implantable Ti-metal and characterized by …

Gold Nanoparticles For Stem Cell Delivery In Myocardial Infarction: Analysis Of Toxicity And Development Of A Cell Culture Platform For Use In An In-Vitro Model Of Mi-Associated Ischemia/Reperfusion, Jeffrey Curran Henson

Graduate Theses and Dissertations

In this work, the potential for PEGylated gold nanoparticles for use as a platform for stem cell delivery in treatment of myocardial infarction (MI) is preliminarily investigated. Cardiovascular disease is currently the leading cause of death worldwide, with majority of mortality resulting from coronary artery disease and associated MI. The ensuing ischemic conditions resulting from MI cause substantial heart muscle tissue loss and scarring in the heart. Adverse tissue remodeling creates a significant loss in heart function that can result in the formation of cardiac hypertrophy, ventricular dilation and arrythmias. The long-term prognosis of patients who have suffered MI is …

Effects Of Gsk101 Exposure On Astrocyte Mechanical Properties, Collagen Production, And Viability, Ethan Echols

Biomedical Engineering Undergraduate Honors Theses

Transient receptor potential vanilloid 4 (TRPV4) is a cation channel protein which is thought to facilitate extracellular matrix (ECM) modification and glial scar formation following traumatic brain injury (TBI). Activation of TRPV4 has been shown to correlate with an increase in intracellular Ca²⁺ concentration. TRPV4 is subject to both chemical and mechanical activation. This study involved chemical activation of TRPV4 in cultured mouse astrocytes using GSK101, the primary agonist of TRPV4, and subsequent evaluation of mechanical and chemical changes to the ECM. Prior to GSK101 exposure, the cells were seeded into Matrigel Matrix, a gelatinous protein mixture intended to …

Quantification Of Tumor-Associated Macrophages Following Immunomodulation Therapy In A Murine Allograft Model Of Colorectal Carcinoma, Caroline Spainhour

Biomedical Engineering Undergraduate Honors Theses

Colorectal Carcinoma (CRC) is one of the deadliest cancers in the world, with 150,000 new cases annually in the United States. Traditional treatments include chemotherapy and invasive surgery; however, research has shown that only 25% of patients that undergo traditional treatment have a positive result. Immunotherapy is an emerging form of cancer treatment that utilizes the patients’immune system to fight cancer cells by targeting inflammation, which plays a large role in the proliferation and metastasis of cancer cells.

Tumor-associated macrophages (TAMs) are immune cells that affect the inflammatory microenvironment of tumors. TAMs are M1 in the early stages of tumors, …

Structure And Property Of Polymers And Biopolymers From Molecular Dynamic Simulations, Xiaoquan Sun

Graduate Theses and Dissertations

Natural and synthetic polymers and biopolymers have been studied for a variety of applications in food emulsion, biopharmaceutical purification, tissue engineering, and biosensor. The structure and property of polymers and biopolymers are critically important to determine their functions. Molecular dynamics (MD) simulations have a unique advantage to explore the structure and property of polymers and biopolymers from the molecular level. In the dissertation, MD simulations were conducted to study the mechanisms of various biological and chemical processes controlled by polymers and biopolymers based on real-world experimental results.

Seven heptapeptides have been screened from a peptide library in our earlier study …

Novel Methodology For Emboli Analog Production And In-Vitro Simulation Of Acute Ischemic Stroke, Anne Preut

Graduate Theses and Dissertations

Stroke is the leading cause of disability and a primary cause of mortality, resulting in an estimated $33 billion dollars spent on healthcare, rehabilitation and lost productivity in the United States each year. The most prevalent cause of stroke incidents are acute ischemic events, manifested as blood clots in the vasculature supplying the brain. Current gold standard treatments have improved since 2006 with the introduction of mechanical stent retrievers; however, several issues with the current treatments to acute ischemic stroke remain. Thrombolysis of the clot with a tissue plasminogen activator may lead to weakening the vessel wall and consequently, hemorrhaging. …

Analysis Of Biological Response To Ecm Hydrogel Injection, Grady Dunlap

Biomedical Engineering Undergraduate Honors Theses

Abstract

Under normal circumstances, skeletal muscle possesses the capacity to regenerate and heal via inflammatory and myogenic pathways. In cases of severe tissue loss or certain diseases, this capacity is lost, often resulting in loss of tissue function. Extracellular matrix (ECM), the protein scaffold which houses cells in physiological tissue, has been shown to have structural and chemical properties which influence cell migration and phenotype. This results in ECM’s capacity to encourage a regenerative response when implanted into severely damaged skeletal muscle. Additional advantages are apparent when an ECM scaffold is digested into a hydrogel, namely less invasive implantation via …

Mitochondrial Mrna Translation Is Required For Maintenance Of Oxidative Capacity, David Lee

Graduate Theses and Dissertations

Oxidative metabolism is required to produce adequate energy to sustain human life. A primary example of deteriorating oxidative capacity is seen in the cardiac musculature during chronic heart failure. This suggests that by improving oxidative potential, chronic heart disease could be mitigated and one approach to accomplish this may be through targeting the mt-mRNA translation system. Purpose: This investigation’s purpose was to characterize disruptions in mt-mRNA translation machinery in multiple forms of cardiomyopathy and to determine if mitochondrial mRNA translation initiation factor (mtIF2) is necessary to maintain oxidative capacity in cardiomyocytes. Methods Using a combination of animal and cell culture …