Molecular, Cellular, and Tissue Engineering Commons^™

Bioengineered Cell Niche For Skeletal Muscle Regeneration, Nicole M. Whittern, Naagarajan Narayanan, Chunhui Jiang, Owen Jones, Jay Gilbert, Michael Whittern, Shihuan Kuang, Meng Deng

The Summer Undergraduate Research Fellowship (SURF) Symposium

Skeletal muscles can self-repair minor strains, lacerations, and contusions; however, in cases of volumetric muscle lossand muscle degenerative diseases, tissue fails to regenerate. Current cell-based therapies, such as myoblast transplantation, have significant drawbacks of low survival rates and engraftment efficacy, mainly due to the absence of supportive cell microenvironment. Scaffolds that mimic the natural cell microenvironment provide a robust platform to support cell adhesion, migration, proliferation, and differentiation. Electrospinning is a versatile technology platform used for fabricating the fiber scaffold that mimics the extracellular matrix. Thus, we aim to reconstitute the cell microenvironment through development of aligned fiber scaffolds by …

Viewing The Extracellular Matrix: An Imaging Method For Tissue Engineering, Michael Drakopoulos, Sarah Calve

The Summer Undergraduate Research Fellowship (SURF) Symposium

The field of regenerative medicine seeks to create replacement tissues and organs, both to repair deficiencies in biological function and to treat structural damage caused by injury. Scaffoldings mimicking extracellular matrix (ECM), the structure to which cells attach to form tissues, have been developed from synthetic polymers and also been prepared by decellularizing adult tissue. However, the structure of ECM undergoes significant remodeling during natural tissue repair, suggesting that ECM-replacement constructs that mirror developing tissues may promote better regeneration than those modeled on adult tissues. This work investigated the effectiveness of a method of viewing the extracellular matrix of developing …

Selective Protein Labelling To Visualize Cellular Differentiation, Andrew J. Witten, Tamara L. Kinzer-Ursem

The Summer Undergraduate Research Fellowship (SURF) Symposium

Protein post-translational modifications serve to give proteins new cellular function, spatial localization, or enzymatic activity. Myristoylation is a common post-translational modification where the enzyme N-myristoyltransferase adds myristic acid onto the N-terminus of a variety of proteins. In this work we use a myristic acid analog, 12-azidododecanoic acid (12ADA) to facilitate the implementation of azide-alkyne cycloaddition reactions on myristoylated proteins. Selective protein labeling methods such as these are useful in research because they can be used to help determine the biological function of this protein lipid modification and can be extended to study disregulated protein myristoylation in disease states. To validate …

Proteoglycan Mimic Of The Glycocalyx To Treat Endothelial Dysfunction, Victoria Messerschmidt, Alyssa Panitch, James R. Wodicka

The Summer Undergraduate Research Fellowship (SURF) Symposium

Patients with kidney failure usually undergo hemodialysis, a process by which toxins produced by the body are filtered from the blood, in order to survive. The preferred form for vascular access is called an arteriovenousfistula (AVF), a surgically created connection between an artery and vein that is utilized to undergo dialysis. However, AVFs have a failure rate of 50-60%. One of the contributions to AVF failure is endothelial cell dysfunction and loss of glycocalyx, which allows neutrophils and other native cells into the media of the vessel, which causes an inflammatory response. Our lab addresses endothelial dysfunction by mimicking the …

Buckling-Driven Force Generation Of Cell Cortex, Pranith Lomada, Wonyeong Jung, Taeyoon Kim

The Summer Undergraduate Research Fellowship (SURF) Symposium

Actomyosin cortex, a thin network underlying cell membrane, is known to generate a large portion of tensile forces required for various cellular processes. Recently, theoretical studies predicted that buckling of actin filaments breaks symmetry between tensile and compressive forces developed by myosin motors, resulting in tensile stress at a network level. However, the significance of the filament buckling of the cortex has yet to be demonstrated either computationally or experimentally. Here, buckling-dependent stress generation of the cortex-like actomyosin network was investigated using an agent-based computational model consisting of actin filaments, actin cross-linking proteins (ACPs), and molecular motors. First, a wide …

Harnessing Notch Signaling For Biomaterial Scaffold-Based Bone Regeneration, Helena P. Lysandrou, Chunhui Jiang, Naagarajan Narayanan, Shihuan Kuang, Meng Deng

The Summer Undergraduate Research Fellowship (SURF) Symposium

Bone fracture has recently become prevalent, especially with an increasingly aging population. Current bone grafts procedures, including autografts and allografts, are hindered by multiple factors, such as limited supplies and inconsistent bone healing. Scaffold-based bone tissue engineering emerges as a prospective strategy to aid in bone regeneration through delivery of growth factors such as bone morphogenic proteins (BMPs). However, the use of BMPs suffers from several drawbacks such as protein instability and immunogenicity. Therefore, there exists a great need for the development of novel therapies to promote bone healing. Notch signaling, a pathway critical for cell-fate determination has been shown …

Breast Cancer/Stromal Cells Coculture On Polyelectrolyte Films Emulates Tumor Stages And Mirna Profiles Of Clinical Samples, Amita Daverey, Karleen M. Brown, Srivatsan Kidambi

Department of Chemical and Biomolecular Engineering: Faculty Publications

In this study, we demonstrate a method for controlling breast cancer cells adhesion on polyelectrolyte multilayer (PEM) films without the aid of adhesive proteins/ ligands to study the role of tumor and stromal cell interaction on cancer biology. Numerous studies have explored engineering coculture of tumor and stromal cells predominantly using transwell coculture of stromal cells cultured onto coverslips that were subsequently added to tumor cell cultures. However, these systems imposed an artificial boundary that precluded cell−cell interactions. To our knowledge, this is the first demonstration of patterned coculture of tumor cells and stromal cells that captures the temporal changes …

Monitoring The Progression Of Spontaneous Articular Cartilage Healing With Infrared Spectroscopy, Megan P. O'Brien, Madhuri Penmatsa, Uday Palukuru, Paul West, Xu Yang, Mathias P. G. Bostrom, Theresa Freeman, Nancy Pleshko

Publications and Research

Objective. Evaluation of early compositional changes in healing articular cartilage is critical for understanding tissue repair and for therapeutic decision-making. Fourier transform infrared imaging spectroscopy (FT-IRIS) can be used to assess the molecular composition of harvested repair tissue. Furthermore, use of an infrared fiber-optic probe (IFOP) has the potential for translation to a clinical setting to provide molecular information in situ. In the current study, we determined the feasibility of IFOP assessment of cartilage repair tissue in a rabbit model, and assessed correlations with gold-standard histology.

Design. Bilateral osteochondral defects were generated in mature white New Zealand rabbits, and …

Characterization Of Human Muscle Extracellular Matrix, Lauren Klaire Wilson

Graduate Theses and Dissertations

The performance of extracellular matrix (ECM) biological scaffolds for the treatment of volumetric muscle loss (VML) has shown promising results with regenerating native muscle in animal models and human patients. However, the limitations of these scaffolds include non-specific characteristics based on the original parameters of the muscle that is lost and this is why our group chose to characterize human skeletal ECM. By understanding the characteristics of the native human skeletal ECM, more desired queues for the biological scaffolds being used to treat VML can be provided. Upper limb and lower limb skeletal muscles were commercially obtained, decellularized using common …

Capillary Electrophoresis For Analysis Of Aβ1-42 Adsorption And Photo-Crosslinking, Jennifer Lynn Kurtz

Graduate Theses and Dissertations

The accumulation of amyloid-beta (Aβ) oligomers is believed to be the driving force behind Alzheimer’s disease (AD) pathogenesis. Due to the metastable nature of Aβ oligomers, the knowledge of Aβ aggregation and accumulation is not well understood. Here, we use capillary electrophoresis (CE) and photo-induced crosslinking of unmodified proteins (PICUP) to learn about the aggregation of Aβ.

First, we explore the effect of capillary coating on Aβ1-42 protein loss using CE. The dynamic coatings tested were PHEA and 50 kDa, 2000 kDa, 5000 kDa, and 8000 kDa PEO. The covalent coating tested was PVA. The results indicated that 2000 kDa …

Design And Development Of Two Component Hydrogel Ejector For Three-Dimensional Cell Growth, Thomas Dunkle, Jessica Deschamps, Connie Dam

Honors Scholar Theses

Hydrogels are useful in wound healing, drug delivery, and tissue engineering applications, but the available methods of injecting them quickly and noninvasively are limited. The medical industry does not yet have access to an all-purpose device that can quickly synthesize hydrogels of different shapes and sizes. Many synthesis procedures that have been developed result in the formation of amorphous hydrogels. While generally useful, amorphous hydrogels exhibit limited capability in tissue engineering applications, especially due to their viscous properties. This endeavor aims to modulate the appropriate gelation parameters, optimize the injection process, and create a prototype that allows for the extrusion …

Finite Element Analysis Of The Application Of Ultrasound-Generated Acoustic Radiation Force To Biomaterials, Nicole J. Piscopo

Honors Scholar Theses

While most bone fractures can heal simply by being stabilized, others can take a longer time to rejoin or they could fail to merge back together completely. Numerous studies have shown the positive effects that ultrasonic therapy have had on delayed-union and non-union bone fracture repair but little is known as to what specific biological mechanisms are at play. Ultrasound may be a valuable tool for bone tissue regeneration at these fracture sites using a tissue engineering approach, however, more must be understood about its impact on stimulating tissues to heal before this can be a reality. For that reason, …

Selection Methods For Genetically-Modified T Cells: In Support Of Translational Therapy, David Rushworth

Dissertations and Theses (Open Access)

T cells are blood cells which organize the immune system of the host. These cells are necessary for the host to respond appropriately to threats from foreign organisms and cancerous growth. However, in the case of certain infections and cancer, T cells are unable to respond appropriately to a threat and establish immunity. This leads to disease when the infection or cancer is not sufficiently eliminated. On the other hand, T cells can lack tolerance for healthy tissue and perceive healthy tissue as infected. The ensuing over-reactive immune response also leads to disease. A delicate balance must exist between immunity …

Mitochondrial Dysfunction And Loss Of Glutamate Uptake In Primary Astrocytes Exposed To Titanium Dioxide Nanoparticles, Christina L. Wilson

Department of Chemical and Biomolecular Engineering: Dissertations, Theses, and Student Research

The proper function of astrocytes is critical for brain health as they are the most abundant cells in the brain which monitor ion homeostasis, recycle neurotransmitters and respond to tissue damage therefore disruption in astrocyte function can result in overall detrimental effects and has been linked with neurodegenerative diseases. Titanium Dioxide (TiO₂) nanoparticles are currently the second most produced engineered nanomaterial in the world with vast usage in consumer products leading to recurrent human exposure. Animal studies indicate significant nanoparticle accumulation in the brain while cellular toxicity studies demonstrate negative effects on neuronal cell viability and function. However, …

Man Made Organs: Technology Made It Possible But Can The Fda Keep Up?, Amanda Chatman

North Carolina Central University Science & Intellectual Property Law Review

No abstract provided.

Altered Cd161bright Cd8+ Mucosal Associated Invariant T (Mait)-Like Cell Dynamics And Increased Differentiation States Among Juvenile Type 1 Diabetics., Robert Z. Harms, Kristina M. Lorenzo, Kevin P. Corley, Monina S. Cabrera, Nora Sarvetnick

Journal Articles: Regenerative Medicine

Type 1A diabetes (T1D) is believed to be caused by immune-mediated destruction of β-cells, but the immunological basis for T1D remains controversial. Microbial diversity promotes the maturation and activation of certain immune subsets, including CD161bright CD8+ mucosal associated invariant T (MAIT) cells, and alterations in gut mucosal responses have been reported in type 1 diabetics (T1Ds). We analyzed T cell populations in peripheral blood leukocytes from juvenile T1Ds and healthy controls. We found that proportion and absolute number of MAIT cells were similar between T1Ds and controls. Furthermore, while MAIT cell proportions increased with age among healthy controls, this trend …

Multiplexed Loop-Mediated Isothermal Amplification Of The 16s Rrna Gene For The Diagnosis Of Neonatal Sepsis In Resource-Limited Environments, Griffin T. Sonaty

Inquiry: The University of Arkansas Undergraduate Research Journal

Sepsis, or dysregulated inflammation caused by bacterial infection, places a disproportionately high burden on newborns in developing countries. This is due in part to a lack of diagnostic tools suitable for sustainable use in resource-limited nurseries. One potential vehicle for a new diagnostic assay is loop-mediated isothermal amplification (LAMP), a high-yield DNA amplification method. LAMP has previously been used to detect genes from single species of bacteria in blood serum samples to aid in sepsis diagnosis. LAMP could be adapted to detect a broad set of bacteria, while retaining a degree of specificity that allows clinicians to begin directed antimicrobial …

Fluorescent Biosensors To Measure Endothelial Cell Responses To Fluid Shear Stress, Natalie Noll

UROP Posters

The response of endothelial cells, innermost layer of blood vessels, to blood flow is thought to be critical in the initiation and progression of atherosclerosis. Atherosclerosis in the human body is non-random and is highly correlated to vessel sites which experience oscillatory and reversing blood flow. Endothelial cells (ECs), the inner most cell layer of blood vessels are highly responsive to the drag force from blood flow, known as shear stress. To study endothelial cell responses to shear stress we used a parallel plate flow chamber in which we exposed endothelial cells to defined fluid shear stress. Using fluorescence resonance …

The Effect Of Cholesterol On The Osteoblast Responsiveness To Hydrodynamic Pressure Stimulation, Kristen Lough

Theses and Dissertations--Biomedical Engineering

Hypercholesterolemia is a risk factor for osteoporosis but the underlying mechanism is unknown. Previous evidence suggests that osteoporosis results from an impaired regulation of osteoblasts by fluid pressure fluctuations in the bone matrix. Recently, our laboratory showed that enhanced cholesterol in the cell membrane, due to hypercholesterolemia, alters leukocyte mechanosensitivity. We predict a similar link between osteoblasts and hypercholesterolemia leading to osteoporosis. Specifically, we hypothesize that extracellular cholesterol modifies the osteoblast sensitivity to pressure. MC3T3-E1 cells were exposed to hydrodynamic pressures regimes (mean=40mmHg, amplitude=0-20mmHg, frequency=1Hz) for 1-12 hours. To assess the impact of membrane cholesterol enrichment, cells were pre-treated with …

Up Regulation Of Heat Shock Protein 70b (Hsp70b) And Ssa1 In Chlamydomonas Reinhardtii Via Hsp70a-Rbcs2 And Psad Promoter, B. Kirtley Amos

Theses and Dissertations--Biosystems and Agricultural Engineering

Fabrication of effective algae cultivation systems adjacent to coal-fired power plants to fixate waste CO₂ would represent a sizable step towards achieving a carbon neutral energy cycle. However, emission gas would elevate the algal cultivation system temperature and decreases its pH without expensive preprocessing. Increased temperature and acidity constitutes a profound stress on the algae. Although stressed algae produce heat shock proteins (HSPs) that promote protein folding and protect against stress, the ordinary biological response is insufficient to protect against coal flue gas. Experimental upregulation of HSPs could make algae respond to the stress caused by high temperatures and …