Molecular, Cellular, and Tissue Engineering Commons^™

Plga-Modified Nanoparticles For The Treatment Of Hypo-Vascularized Hpv-Related Cervical Cancers., Lee B. Sims

Electronic Theses and Dissertations

A major challenge associated with delivery of active agents in the female reproductive tract (FRT) is the ability of agents to efficiently diffuse through the cervicovaginal mucosa (CVM) and reach the underlying sub-epithelial immune cell layer and vasculature. A variety of drug delivery vehicles have been employed to improve the delivery of agents across the CVM and offer the capability to increase the longevity and retention of active agents to treat infections of the female reproductive tract. Nanoparticles (NPs) have been shown to improve retention, diffusion, and cell-specific targeting via specific surface modifications, relative to other delivery platforms. In particular, …

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 …

Experimental And Model-Based Terahertz Imaging And Spectroscopy For Mice, Human, And Phantom Breast Cancer Tissues, Tyler Bowman

Graduate Theses and Dissertations

The goal of this work is to investigate terahertz technology for assessing the surgical margins of breast tumors through electromagnetic modeling and terahertz experiments. The measurements were conducted using a pulsed terahertz system that provides time and frequency domain signals. Three types of breast tissues were investigated in this work. The first was formalin-fixed, paraffin-embedded tissues from human infiltrating ductal and lobular carcinomas. The second was human tumors excised within 24-hours of lumpectomy or mastectomy surgeries. The third was xenograft and transgenic mice breast cancer tumors grown in a controlled laboratory environment to achieve more data for statistical analysis.

Experimental …

Eigenstrain As A Mechanical Set-Point Of Cells, Shengmao Lin, Marsha C. Lampi, Cynthia A. Reinhart-King, Gary C.P. Tsui, Jian Wang, Carl A. Nelson, Linxia Gu

Department of Mechanical and Materials Engineering: Faculty Publications

Cell contraction regulates how cells sense their mechanical environment. We sought to identify the set-point of cell contraction, also referred to as tensional homeostasis. In this work, bovine aortic endothelial cells (BAECs), cultured on substrates with different stiffness, were characterized using traction force microscopy (TFM). Numerical models were developed to provide insights into the mechanics of cell–substrate interactions. Cell contraction was modeled as eigenstrain which could induce isometric cell contraction without external forces. The predicted traction stresses matched well with TFM measurements. Furthermore, our numerical model provided cell stress and displacement maps for inspecting the fundamental regulating mechanism of cell …

Comparison Of Eosin And Fluorescein Conjugates For The Photoinitiation Of Cell-Compatible Polymer Coatings, Jacob L. Lilly, Anuhya Gottipati, Calvin F. Cahall, Mohamed Agoub, Brad J. Berron

Chemical and Materials Engineering Faculty Publications

Targeted photopolymerization is the basis for multiple diagnostic and cell encapsulation technologies. While eosin is used in conjunction with tertiary amines as a water-soluble photoinitiation system, eosin is not widely sold as a conjugate with antibodies and other targeting biomolecules. Here we evaluate the utility of fluorescein-labeled bioconjugates to photopolymerize targeted coatings on live cells. We show that although fluorescein conjugates absorb approximately 50% less light energy than eosin in matched photopolymerization experiments using a 530 nm LED lamp, appreciable polymer thicknesses can still be formed in cell compatible environments with fluorescein photosensitization. At low photoinitiator density, eosin allows more …

Preparation For The Study Of Gene Expression Aboard The International Space Station, Amber Paturzo

STAR Program Research Presentations

WetLab-2 performs quantitative polymerase chain reaction (qPCR) aboard the micro-gravity environment of the International Space Station in order to research how the stress of micro-gravity affects an organism’s RNA production and gene expression. This on-station qPCR allows researchers to gather information earlier and thereby allow them to modify experiments in a matter of hours rather than wait until the next flight opportunity. After a successful validation flight on SpaceX-8 and follow-up investigation on SpaceX-9, system components must now be adjusted for a new type of biological sample. Specifically, the WetLab-2 facility includes a commercial qPCR instrument (RNA SmartCycler), a sample …

Efficient One‑Step Fusion Pcr Based On Dual‑Asymmetric Primers And Two‑Step Annealing, Yilan Liu, Jinjin Chen, Anders Thygesen

Department of Chemical and Biomolecular Engineering: Faculty Publications

Gene splicing by fusion PCR is a versatile and widely used methodology, especially in synthetic biology. We here describe a rapid method for splicing two fragments by one-round fusion PCR with a dual-asymmetric primers and two-step annealing (ODT) method. During the process, the asymmetric intermediate fragments were generated in the early stage. Thereafter, they were hybridized in the subsequent cycles to serve as template for the target full-length product. The process parameters such as primer ratio, elongation temperature and cycle numbers were optimized. In addition, the fusion products produced with this method were successfully applied in seamless genome editing. The …

Wild-Type P53 Enhances Endothelial Barrier Function By Mediating Rac1 Signalling And Rhoa Inhibition, Nektarios Barabutis, Christiana Dimitropoulou, Betsy Gregory, John D. Catravas

Bioelectrics Publications

Inflammation is the major cause of endothelial barrier hyper-permeability, associated with acute lung injury and acute respiratory distress syndrome. This study reports that p53 "orchestrates" the defence of vascular endothelium against LPS, by mediating the opposing actions of Rac1 and RhoA in pulmonary tissues. Human lung microvascular endothelial cells treated with HSP90 inhibitors activated both Rac1- and P21-activated kinase, which is an essential element of vascular barrier function. 17AAG increased the phosphorylation of both LIMK and cofilin, in contrast to LPS which counteracted those effects. Mouse lung microvascular endothelial cells exposed to LPS exhibited decreased expression of phospho-cofilin. 17AAG treatment …

Alteration Of Metabolite Profiling By Cold Atmospheric Plasma Treatment In Human Myeloma Cells, Dehui Xu, Yujing Xu, Ning Ning, Qingjie Cui, Zhijie Liu, Xiaohua Wang, Dingxin Liu, Hailan Chen, Michael G. Kong

Bioelectrics Publications

BACKGROUND: Despite new progress of chemotherapy in multiple myeloma (MM) clinical treatment, MM is still a refractory disease and new technology is needed to improve the outcomes and prolong the survival. Cold atmospheric plasma is a rapidly developed technology in recent years, which has been widely applied in biomedicine. Although plasma could efficiently inactivate various tumor cells, the effects of plasma on tumor cell metabolism have not been studied yet.

METHODS: In this study, we investigated the metabolite profiling of He plasma treatment on myeloma tumor cells by gas-chromatography time-of-flight (GC-TOF) mass-spectrometry. Meanwhile, by bioinformatic analysis such as GO and …

Exogenous Fniii 12-14 Regulates Tgf-Β1-Induced Markers, Hilmi M. Humeid

Theses and Dissertations

The extracellular matrix protein Fibronectin (FN) plays an important role in cell contractility, differentiation, growth, adhesion, and migration. The 12th -14th Type III repeats of FN (FNIII 12-14), also referred to as the Heparin-II domain, comprise a highly promiscuous growth factor (GF) binding region. This binding domain aids in cellular signaling initiated from the ECM. Additionally, FN has the ability to assemble into fibrils under certain conditions, mostly observed during cell contractile processes such as those that initiate due to upregulation of Transforming Growth Factor Beta 1 (TGF-β1) [1], [2]. Previous work from our lab has shown that self-assembly of …

Decellularized Matrices Effect On The Adaptive Immune Response, Kegan Sowers

Theses and Dissertations

Decellularized extracellular matrices have been a growing area of interest in the biomedical engineering fields of tissue engineering and regenerative medicine.As these materials move toward clinical applications, the immune response to these materials will be a driving force toward their success in clinical approaches. Fully digested decellularized matrix constructs derived from porcine liver, muscle and lung were created to test the adaptive immune response. Hydrogel characterization ensured that the materials had relatively similar stiffness levels to reduce variability, and in vitro studies were conducted. Each individual construct as well as a gelatin control were plated with a co-culture of macrophages …

Characterization Of Electroactive Nanofibers For Nerve Regeneration, Jean-Yves Azar

Research Opportunities for Engineering Undergraduates (ROEU) Program 2017-18

In the event of peripheral nerve injury, loss of function may occur as the result of a severed nerve. Hollow nerve gap conduits are used to provide a protected environment for nerves to reconnect and regain function, although therapies for nerve gaps larger than 3 cm lack a stable environment for nerves to reconnect. Development of a nerve guide conduit filler has been proposed to mimic the cell’s microenvironment. This would lead to increased functional regeneration. We are aiming to create a biomaterial to act as a filler for the hollow nerve gap conduits to promote functional regeneration of the …

Developing Vascular Graft From Adipose-Derived Stem Cells, Ashley C. Apil

Research Opportunities for Engineering Undergraduates (ROEU) Program 2017-18

This project aims to differentiate adipose-derived stem cells into fibroblasts through the addition of platelet-derived growth factor (PDGF) into the culture media. Differentiation protocol will be optimized, then successful differentiation will be verified through PCR analysis. The ASC-fibroblasts will then be seeded into a ring construct as outlined by the protocol by Lam et al. The ring constructs will be tensile tested and analyzed through histology to characterize their strength and cellularity.

Developing A 3d In Vitro Model By Microfluidics, Hung-Ta Chien

Dissertations and Theses

In vitro tissue models play an important role in providing a platform that mimics the realistic tissue microenvironment for stimulating and characterizing the cellular behavior. In particular, the hydrogel-based 3D in vitro models allow the cells to grow and interact with their surroundings in all directions, thus better mimicking in vivo than their 2D counterparts. The objective of this thesis is to establish a 3D in vitro model that mimics the anatomical and functional complexity of the realistic cancer microenvironment for conveniently studying the transport coupling in porous tissue structures. We pack uniform-sized PEGDA-GelMA microgels in a microfluidic chip to …

Using Crosslinked Hyaluronic Acid (Ha) And Collagen Scaffolds With Sustained Brain-Derived Neurotrophic Factor (Bdnf) Release For Post-Sci Nerve Regeneration, Panth Doshi

Undergraduate Research Posters

Traumatic events resulting in spinal cord injuries (SCIs) often leave people paralyzed or with partial loss of motor function. The physical disabilities arising from traumatic events prevent people from functioning at the same level as pre-injury. My work aims to identify a plausible method to overcome the inhibitory post-SCI environment and to regenerate nervous tissue in order to restore neural function and, subsequently, motor function. I identified components of a new, hypothetical nerve scaffold based on the immune response after SCIs and the efficacy of currently used scaffolds for nerve regeneration. Hyaluronic acid (HA) polymer scaffolds and collagen-based scaffolds are …

Targeting Ovarian Cancer And Endothelium With An Allosteric Ptp4a3 Phosphatase Inhibitor, Kelley E. Mcqueeney, Joseph M. Salamoun, James C. Burnett, Nektarios Barabutis, Paula Pekic, Sophie L. Lewandowski, Danielle C. Llaneza, Robert Cornelison, Yunpeng Bai, Zhong-Yin Zhang, John D. Catravas

Bioelectrics Publications

Overexpression of protein tyrosine phosphatase PTP4A oncoproteins is common in many human cancers and is associated with poor patient prognosis and survival. We observed elevated levels of PTP4A3 phosphatase in 79% of human ovarian tumor samples, with significant overexpression in tumor endothelium and pericytes. Furthermore, PTP4A phosphatases appear to regulate several key malignant processes, such as invasion, migration, and angiogenesis, suggesting a pivotal regulatory role in cancer and endothelial signaling pathways. While phosphatases are attractive therapeutic targets, they have been poorly investigated because of a lack of potent and selective chemical probes. In this study, we disclose that a potent, …

A Purification And In Vitro Activity Assay For A (P)Ppgpp Synthetase From Clostridium Difficile, Astha Pokhrel, Asia Poudel, Erin B. Purcell

Chemistry & Biochemistry Faculty Publications

Kinase and pyrophosphokinase enzymes transfer the gamma phosphate or the beta-gamma pyrophosphate moiety from nucleotide triphosphate precursors to substrates to create phosphorylated products. The use of γ-³²-P labeled NTP precursors allows simultaneous monitoring of substrate utilization and product formation by radiography. Thin layer chromatography (TLC) on cellulose plates allows rapid separation and sensitive quantification of substrate and product. We present a method for utilizing the thin-layer chromatography to assay the pyrophosphokinase activity of a purified (p)ppGpp synthetase. This method has previously been used to characterize the activity of cyclic nucleotide and dinucleotide synthetases and is broadly suitable for …

Restoring Cholesterol Homeostasis In Familial Hypercholesterolemia Cell Line Model, Lubna Hindi

Electronic Theses and Dissertations

Familial hypercholesterolemia (FH) is an autosomal dominant disorder that results in elevated levels of low-density lipoprotein cholesterol (LDL-C). This increase in serum cholesterol level has been shown to result in premature coronary artery disease (CAD) with devastating symptoms from a young age. Today, the prevalence of heterozygous FH (HeFH) and homozygous FH (HoFH) is estimated to be 1 in 320 and 1 in 160,000 people, respectively. FH is referred to as an underdiagnosed disease due to the large number of mutations that continues to grow. These mutations often exist in one of or a combination of three genes: low-density lipoprotein …

Molded Features In Pdms For Fabricating Bacterial Cellulose For Various Geometries, Mitchell Habegger

Williams Honors College, Honors Research Projects

The purpose of producing features on bacterial cellulose (BC) is to facilitate the elongation and alignment for cells, in this case Normal Human Dermal Fibroblast (NHDF) cells. The elongated cells have applications in wound healing, tissue engineering, disease diagnostics, and many other fields. Experiments were run to test the effectiveness of transferring features to BC sheets from features induced by fracturing on polydimethylsiloxane (PDMS) and those duplicated from molds with existing features. The features were duplicated to BC sheets by either air drying or Guided Assembly-Based Biolithography (GAB). The research results showed that fracture inducing on PDMS produced very small …

Modification Of Bacterial Cellulose Using Organosilanes To Enhance Cell Adhesion And Growth, Kristi Ferrato

Williams Honors College, Honors Research Projects

In this project, drying bacterial cellulose (BC) to maintain its structural integrity, and treating BC with organosilanes 3-aminopropyltriethoxysilane (APTES) and (3-Mercaptopropyl)trimethoxysilane (MPTMS) as well as with various concentration of gelatin to increase the cells per area was studied. Drying BC with hexamethyldisilazane (HMDS) by soaking the BC in ascending concentrations of ethanol and HMDS did not protect the structural integrity of the BC grown in lab but did for the BC grown in Dr. Siriporn Taokaew’s lab. Freeze drying also did not fully protect the structural integrity as the BC was placed in a freezer for several hours at -20oC …