Molecular, Cellular, and Tissue Engineering Commons^™

Cartilage Engineering: Optimization Of Media For Chondrogenic Differentiation In Vitro, Evan Surma, Sherry L. Harbin, Hongji Zhang, Stacy Halum

The Summer Undergraduate Research Fellowship (SURF) Symposium

Lower back pain from intervertebral disc injury affects around 84% of the population at some point in their life, which at its worst may cause total immobilization. This pain can only be temporarily relieved by spinal fusion or intervertebral disc replacement; however, both of these cause loss of natural motion in patients by removing damaged fibrocartilage discs. While these techniques help mitigate pain briefly, no permanent solution exists currently to both relieve pain and preserve natural motion. My work may be a solution by eventually providing patient-specific implants that resemble native tissue in the regeneration process that could be absorbed …

Lysis And Amplifciation Of Neonatal Sepsis Causing Pathogens, Gregory Berglund, Elizabeth A. Phillips, Jacqueline C. Linnes

The Summer Undergraduate Research Fellowship (SURF) Symposium

Neonatal sepsis, resulting from a bloodstream infection within the first few weeks of life, is the leading cause of newborn deaths worldwide. The gold standard of neonatal sepsis diagnosis requires a blood culture to identify the infecting bacteria, however require days of incubation, expensive equipment, and expertise. Any delay in diagnosis is critical, as the condition can be treated easily if appropriate antibiotics are administered promptly. A low-cost, rapid, and sensitive diagnostic test would enable more timely treatment and lead to better patient outcomes with fewer required resources. Point-of-care, nucleic acid amplification assays are a promising alternative to blood culture …

Alginate Hydrogels As Three-Dimensional Scaffolds For In Vitro Culture Models Of Growth Plate Cartilage Development And Porcine Embryo Elongation, Taylor D. Laughlin

Department of Agricultural and Biological Systems Engineering: Dissertations, Theses, and Student Research

The establishment of in vitro culture models utilizes tissue engineering principles to design functional mimics of in vivo environments in vitro. Advantages for the use of in vitro culture models include ethical alleviation of animal models for therapeutic testing, cost efficiency, and a greater ability to study specific mechanisms via a systematic, ground-up approach to development. In this thesis, alginate hydrogels are utilized in the development of in vitro culture models of porcine embryo elongation and growth plate cartilage development. First, the effect of scaffold and modifications to the scaffold were explored in both projects. In order to modulate …

Automated Microscopy Platform For High-Throughput Analysis Of Cellular Characteristics, Hussam Ibrahim

Physics: Student Scholarship & Creative Works

Existing microscopy platforms allow analysis post-hoc, but not in real time. This is an issue in the world of Bioengineering because you are limited to performing further analysis on specimen. The aim of my research was to design a sophisticated system whereby information can be exchanged between the software which acquires images and software that analyzes the images immediately after acquisition. In this system, images would be acquired by the microscope and analyzed by customized scripts (MATLAB, Mathworks) in real time. Specifically, MATLAB would wait for new images to be saved on the hard drive, import these images, and perform …

Hydrothermally Processed 1d Hydroxyapatite: Mechanism Of Formation And Biocompatibility Studies, Zoran Stojanović, Nenad Ignjatović, Victoria M. Wu, Vojca Žunič, Ljiljana Veselinović, Srečo D. Škapin, Miroslav Miljković, Vuk Uskoković, Dragab Uskoković

Pharmacy Faculty Articles and Research

Recent developments in bone tissue engineering have led to an increased interest in one-dimensional (1D) hydroxyapatite (HA) nano- and micro-structures such as wires, ribbons and tubes. They have been proposed for use as cell substrates, reinforcing phases in composites and carriers for biologically active substances. Here we demonstrate the synthesis of 1D HA structures using an optimized, urea-assisted, high-yield hydrothermal batch process. The one-pot process, yielding HA structures composed of bundles of ribbons and wires, was typified by the simultaneous occurrence of a multitude of intermediate reactions, failing to meet the uniformity criteria over particle morphology and size. To overcome …

Calcium Phosphate As A Key Material For Socially Responsible Tissue Engineering, Vuk Uskoković, Victoria M. Wu

Pharmacy Faculty Articles and Research

Socially responsible technologies are designed while taking into consideration the socioeconomic, geopolitical and environmental limitations of regions in which they will be implemented. In the medical context, this involves making therapeutic platforms more accessible and affordable to patients in poor regions of the world wherein a given disease is endemic. This often necessitates going against the reigning trend of making therapeutic nanoparticles ever more structurally complex and expensive. However, studies aimed at simplifying materials and formulations while maintaining the functionality and therapeutic response of their more complex counterparts seldom provoke a significant interest in the scientific community. In this review …

Hiv Vaccines: Progress, Limitations And A Crispr/Cas9 Vaccine, Omar A. Garcia Martinez

Biology: Student Scholarship & Creative Works

ABSTRACT: The HIV-1 pandemic continues to thrive due to ineffective HIV-1 vaccines. Historically, the world’s most infectious diseases, such as polio and smallpox, have been eradicated or have come close to eradication due to the advent of effective vaccines. Highly active antiretroviral therapy is able to delay the onset of AIDS but can neither rid the body of HIV-1 proviral DNA nor prevent further transmission. A prophylactic vaccine that prevents the various mechanisms HIV-1 has to evade and attack our immune system is needed to end the HIV-1 pandemic. Recent advances in engineered nuclease systems, like the CRISPR/Cas9 system, have …

Interaction Of Fibrinogen With Fibronectin: Purification And Characterization Of A Room Temperature-Stable Fibrinogen-Fibronectin Complex From Normal Human Plasma, Ayman E. Ismail

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

A fibrinogen-fibronectin complex (γγ’pdFI-pdFN) was purified from normal human plasma using a sequence of cryoprecipitation, ammonium sulfate fractionation, and DEAE Sepharose chromatography. Sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) under reducing condition showed both a 1:1 stoichiometric ratio of fibrinogen (FI) to fibronectin (FN) as well as a stoichiometric ratio of 1:1 of γg to gγ’. The γγ’pdFI-pdFN complex was non-covalent in nature as it was disrupted by affinity adsorption to Gelatin Sepharose where pdFN bound strongly and the disrupted γγ’pdFI fell through the chromatographic column. Surprisingly, the purified γγ’pdFI-pdFN complex was more broadly thermally stable than plasma FI (pdFI) …

Isolation Of Metallic Single-Walled Carbon Nanotubes For Electrically Conductive Tissue Engineering Scaffolds, Jakob Hockman

Biomedical Engineering Undergraduate Honors Theses

Metallic single-walled carbon nanotubes (m-SWNTs) were separated from pristine SWNTs using affinity chromatography for use in electrically conductive tissue engineering scaffolds. Approximately one third of SWNTs have metallic properties. Separations were achieved using a protocol modified from Liu & coworkers (2011) in order to improve the method for cell culture environments. Samples enriched in m-SWNTs were isolated and characterized. However, challenges still remain for the complete separation of m-SWNTs from their semiconducting counterpart (s-SWNTs) using this protocol. Approaches to improve separation and reduce the difficulties associated with processing the nanotubes were suggested. One of the ultimate destinations of these nanotubes …

Exploring Biomarkers For Point Of Care Bladder Cancer Detection, Cassandra M. Walker

Biomedical Engineering Undergraduate Honors Theses

Bladder cancer is the 5th most common non-cutaneous human cancer in the United States. While effective methods of detecting bladder cancer are currently practiced, they are often expensive and invasive. There is a need for a noninvasive detection method that can be used in areas with few medical resources. Cell free DNA in urine is normally present only in very low concentrations. Abnormally high levels of cell free DNA in urine could be indicative of disease. This study tests the hypothesis that DNA present in urine can be used as a biomarker for bladder cancer before hematuria is seen in …

A 'Tissue Model' To Study The Barrier Effects Of Living Tissues On The Reactive Species Generated By Surface Air Discharge, Tongtong He, Dingxin Liu, Han Xu, Zhichao Liu, Dehui Xu, Dong Li, Qiosong Li, Mingzhe Rong, Michael G. Kong

Bioelectrics Publications

Gelatin gels are used as surrogates of human tissues to study their barrier effects on incoming reactive oxygen and nitrogen species (RONS) generated by surface air discharge. The penetration depth of nitrite into gelatin gel is measured in real time during plasma treatment, and the permeabilities of nitrite, nitrate, O₃ and H₂O₂ through gelatin gel films are quantified by measuring their concentrations in the water underneath such films after plasma treatment. It is found that the penetration speed of nitrite increases linearly with the mass fraction of water in the gelatin gels, and the permeabilities of …

Sustained Cell Differentiation Of 2d H9 Human Embryonic Stem Cells Into Mesenchymal Stem Cells, Hannah M. Christian

UCARE: Research Products

This experiment consisted of the controlled differentiation of H9 embryonic stem cells to mesenchymal stem cells. Though this experiment was repeated twice and improvement was seen in these repetitions, the cells were only able to be partially differentiated. However, the morphology of the differentiated cells is similar to those of healthy adult mesenchymal stem cells.

The progression of the differentiation can be seen in the microscope slide photos below. Throughout the differentiation, there occurred a decrease in cell survival and reduction of cell growth, but an increase in mesenchymal stem cell morphology. Throughout the last week of the differentiation, very …

Hyaluronic Acid-Conjugated Liposome Nanoparticles For Targeted Delivery To Cd44 Overexpressing Glioblastoma Cells, Stephen L. Hayward, Christina L. Wilson, Srivatsan Kidambi

Department of Chemical and Biomolecular Engineering: Faculty Publications

Glioblastoma Multiforme (GBM) is a highly prevalent and deadly brain malignancy characterized by poor prognosis and restricted disease management potential. Despite the success of nanocarrier systems to improve drug/gene therapy for cancer, active targeting specificity remains a major hurdle for GBM. Additionally, since the brain is a multi-cell type organ, there is a critical need to develop an approach to distinguish between GBM cells and healthy brain cells for safe and successful treatment. In this report, we have incorporated hyaluronic acid (HA) as an active targeting ligand for GBM. To do so, we employed HA conjugated liposomes (HALNPs) to study …

Patterned Alginate Hydrogels To Induce Chondrocyte Alignment, Jordan Catherine Verplank, Taylor D. Laughlin, Angela K. Pannier

UCARE: Research Products

The growth plate has an intricate architecture, and this architecture is necessary for directional growth of bones. Specifically, the cells align in longitudinal columns. As the growth plate expands with this pattern, the bone elongates with the same alignment pattern. The purpose of this research is to mimic this single celled, columnar alignment in vitro. In developing this alignment in vitro, this research will contribute to the overall study of growing growth for the development of improved therapeutic treatments and engineered tissues for transplants.

Membrane Channel Gene Expression In Human Costal And Articular Chondrocytes, A. Asmar, R. Barrett-Jolley, A. Werner, R. Kelly Jr., M. Stacey

Bioelectrics Publications

Chondrocytes are the uniquely resident cells found in all types of cartilage and key to their function is the ability to respond to mechanical loads with changes of metabolic activity. This mechanotransduction property is, in part, mediated through the activity of a range of expressed transmembrane channels; ion channels, gap junction proteins, and porins. Appropriate expression of ion channels has been shown essential for production of extracellular matrix and differential expression of transmembrane channels is correlated to musculoskeletal diseases such as osteoarthritis and Albers-Sch€onberg. In this study we analyzed the consistency of gene expression between channelomes of chondrocytes from human …

Equilibrium Testing Of Rat Tail Tendon: An Analysis Of The Viscoelastic Properties Of Collagen Under Different Strain Points, Joshua C. Witt

Graduate Theses - Biology & Biomedical Engineering

Instantaneous tensile testing and stress-relaxation testing are forms of mechanical testing used to determine the elastic and viscoelastic properties of biological tissue. Equilibrium testing is a form of testing that combines both of these testing approaches at different strain points to determine the elastic properties of a material and also assess their viscoelastic properties in the same test. This testing method is commonly used on highly viscoelastic materials such as cartilage but has never been fully described in dense collagenous materials such as tendon or ligament. This analysis utilizes different strain points selected to capture the classic non-linear behavior of …

Pch1 Integrates Circadian And Light- Signaling Pathways To Control Photoperiod-Responsive Growth In Arabidopsis, He Huang, Chan Yul Yoo, Rebecca Bindbeutel, Jessica Goldsworthy, Allison Tielking, Sophie Alvarez, Michael J. Naldrett, Bradley S. Evans, Meng Chen, Dmitri A. Nusinow

Nebraska Center for Biotechnology: Faculty and Staff Publications

Plants react to seasonal change in day length through altering physiology and development. Factors that function to harmonize growth with photoperiod are poorly understood. Here we characterize a new protein that associates with both circadian clock and photoreceptor components, named PHOTOPERIODIC CONTROL OF HYPOCOTYL1 (PCH1). pch1 seedlings have overly elongated hypocotyls specifically under short days while constitutive expression of PCH1 shortens hypocotyls independent of day length. PCH1 peaks at dusk, binds phytochrome B (phyB) in a red light-dependent manner, and co-localizes with phyB into photobodies. PCH1 is necessary and sufficient to promote the biogenesis of large photobodies to maintain an …

Quantitative Comparison Of A Nanoengineered Alumina Coated Cnt Arrays To Sio2 Coated Cnts And Solution Based Delivery System, Shree Aier

Undergraduate Research & Mentoring Program

To meet the growing need for nanoengineered biocompatible materials to serve as drug delivery platforms, in this research, carbon nanotube arrays were fabricated by chemical vapor deposition, followed by an alumina coating by the high yielding, tightly controlled atomic layer deposition. This nanoengineered vertically aligned alumina nanowire array serves as a platform for delivering antigens, which act as cancer adjuvants. The physicochemical characteristics of the nanowires (NWs) can significantly influence the delivery of a biomolecule to immune cells. To investigate the material characteristics, the delivery efficiency of the antigen using NWs was quantitatively assessed by flow cytometry. Further, the mechanism …

Design And Production Of A Hydrogel Forming Polypeptide: Engaging High School Students In Protein Design, James K. Deyling

ETD Archive

Bioinks are a class of hydrogel that have the potential to be the ink used in the creation of printed organs, connective tissue, and other important structures within the body. One class of material that may be a suitable bioink hydrogel is elastin-like polypeptides (ELPs), which are synthetic biopolymers inspired by the naturally existing connective tissue elastin. ELPs consist of a repeat pentapeptide sequence (GXGVP)n, where X is any of the 20 naturally existing amino acids other than proline. These biomolecules are capable of exhibiting environmental responsiveness when exposed to certain stimulus such as salt concentration, temperature, and pH, depending …

Role Of E-Cadherin Force In The Spatial Regulation Of Cell Proliferation, Abhinav Mohan

Theses and Dissertations

Cell proliferation and contact inhibition play a major role in maintaining epithelial cell homeostasis. A hallmark of epithelial cells is strong cell-cell junctions. These junctions include E-Cadherin, a type of adherens junction that is critical for both barrier function and contact inhibition. Prior experiments by other groups have shown that adherens junctions are subject to mechanical tension. Externally applied forces (e.g. stretch) results in changes in E-Cadherin forces that coordinate proliferation. My current work tests the hypothesis that E-Cadherin forces mediate the spatial regulation of cell proliferation even in the absence of externally applied forces.