Molecular, Cellular, and Tissue Engineering Commons^™

A Morphological Analysis Of A Hybrid Swarm Of Native Ulmus Rubra And Introduced U. Pumila (Ulmaceae) In Southeastern Nebraska, Christian Elowsky, Ingrid E. Jordon-Thaden, Robert B. Kaul

Nebraska Center for Biotechnology: Faculty and Staff Publications

The parental species and hybrid swarm of native Ulmus rubra Muhl. and the introduced, naturalized and weedy U. pumila L. were investigated in a 65-km transect in Lancaster, Saunders, and Butler counties in Nebraska. Thirty-two trees of U. rubra, 32 of U. pumila, and 50 of the hybrid swarm were sampled for leaves and buds and subsampled for flowers and fruits. Leaves were measured for petiole length, blade length, width, primary and secondary teeth per cm, number of secondary teeth per primary tooth, and texture. Buds were scored for color and distribution of trichomes. Flowers were sampled for stamen counts …

Shelf Life Study Of Electrospun Plga Copolymers, Sean Youra, Nick Hudson

Biomedical Engineering

Poly(lactic-co-glycolic acid) (PLGA) is one of the most commonly used copolymers for electrospinning in tissue engineering applications. However, most research has not focused on the copolymer itself in regards to how long it can be used effectively and if varying the concentrations of polylactic acid (PLA) and polyglycolic acid (PGA) affect the resulting properties. Electrospinning is the method we use to create the three-dimensional constructs, or “scaffolds”, for the blood vessel mimic (BVM) in the tissue engineering lab. The aim of our project was to investigate if the morphology and mechanical properties of the scaffolds changed over time when they …

Matrix Remodeling Accompanies In Vitro Articular Cartilage Spherical Shaping, Nathan Thomas Balcom

Master's Theses

Introduction: Articular cartilage (AC) is a low friction load bearing material found in synovial joints. The natural repair of damaged tissue is difficult and often requires surgical intervention. With large defects it becomes necessary to match the original tissue geometry. We hypothesized that localized collagen (COL) and/or proteoglycan (PG) remodeling occurs during AC spherical reshaping. The objective of this study was to determine the presence, magnitude and depth dependence of COL and PG remodeling that accompanies AC reshaping. Methods: Full thickness AC blocks (7x7 mm² surface area) were harvested from the ridges of the patellofemoral groove of immature (1-3 …

Delayed Thrombus Resolution And Fibroproliferative Vascular Wound Healing From Deficiency Of Type Iii Collagen: A Paradoxical Mechanism For Tissue Fragility, Amy J. Reid

Dissertations and Theses (Open Access)

Vascular Ehlers-Danlos syndrome is a heritable disease of connective tissue caused by mutations in COL3A1, conferring a tissue deficiency of type III collagen. Cutaneous wounds heal poorly in these patients, and they are susceptible to spontaneous and catastrophic rupture of expansible hollow organs like the gut, uterus, and medium-sized to large arteries, which leads to premature death. Although the predisposition for organ rupture is often attributed to inherent tissue fragility, investigation of arteries from a haploinsufficient Col3a1 mouse model (Col3a1^+/-) demonstrates that mutant arteries withstand even supraphysiologic pressures comparably to wild-type vessels. We hypothesize that injury …

Improvements In Microdialysis Sampling Extraction Efficiency Using Recycled Flow, Justin Cole Deaton

Graduate Theses and Dissertations

Incomplete recovery during microdialysis sampling is hindering important research in neurology, proteomics, and immunology. Although the current generalized solution, decreasing volumetric flow rates (Q), has been and will remain to be a useful strategy it has reached it's a physical limitation due to evaporation at the collection site. Consequently, many important signaling molecules, such as signaling proteins, remain difficult to study. It is more fundamental to consider relative recovery as a function of the interaction time between the perfusate and the environment surrounding the probe.

In this work an increase in relative recovery was predicted by a mathematical model. Using …

Investigating The Reproducibility Of The Current Bvm Protocol, Corey Gross

Biomedical Engineering

Coronary Artery Disease (CAD) is responsible for 1 death every minute in the US. Angioplasty with the implantation of stents is a common treatment method for CAD. Although there is a variety of stents currently on the market, there is still a need to develop new types for different pathologic conditions. Preliminary assessment of the physiological response to new stents is needed as they are being developed. The FDA approval process implemented today is a long, tedious path with a range of testing methods that include static in vitro testing and high-cost animal testing. Tissue engineered blood vessels have been …

The Effects Of Hydrostatic Pressure On Early Endothelial Tubulogenic Processes, Ryan M. Underwood

Theses and Dissertations--Biomedical Engineering

The effects of mechanical forces on endothelial cell function and behavior are well documented, but have not been fully characterized. Specifically, fluid pressure has been shown to elicit physical and chemical responses known to be involved in the initiation and progression of endothelial cell-mediated vascularization. Central to the process of vascularization is the formation of tube-like structures. This process—tubulogenesis—is essential to both the physiological and pathological growth of tissues. Given the known effects of pressure on endothelial cells and its ubiquitous presence in the vasculature, we investigated pressure as a magnitude-dependent parameter for the regulation of endothelial tubulogenic activity. To …

Growth Plate Regeneration Using Polymer-Based Scaffolds Releasing Growth Factor, Amanda Clark

Theses and Dissertations--Biomedical Engineering

Currently growth plate fractures account for nearly 18.5% of fractures in children and can lead to stunted bone growth or angular deformation. If the body is unable to heal itself a bony bar forms, preventing normal bone growth. Clinical treatment involves removing the bony bar and replacing it with a filler substance, which causes poor results 60% of the time.

Using primarily poly(lactic-co-glycolic acid) (PLGA) as the scaffold material, the goal was to develop an implant that would support to the implant site, allow for cell ingrowth, and degrade away over time. Porous scaffolds were fabricated from PLGA microspheres using …

Surface- And Hydrogel-Mediated Delivery Of Nucleic Acid Nanoparticles, Angela K. Pannier, Tatiana Segura

Department of Agricultural and Biological Systems Engineering: Faculty Publications

Gene expression within a cell population can be directly altered through gene delivery approaches. Traditionally for nonviral delivery, plasmids or siRNA molecules, encoding or targeting the gene of interest, are packaged within nanoparticles. These nanoparticles are then delivered to the media surrounding cells seeded onto tissue culture plastic; this technique is termed bolus delivery. Although bolus delivery is widely utilized to screen for efficient delivery vehicles and to study gene function in vitro, this delivery strategy may not result in efficient gene transfer for all cell types or may not identify those delivery vehicles that will be efficient in vivo. …

Bile-Acid-Mediated Decrease In Endoplasmic Reticulum Stress: A Potential Contributor To The Metabolic Benefits Of Ileal Interposition Surgery In Ucd-T2dm Rats, Bethany P. Cummings, Ahmed Bettaieb, James L. Graham, Jaehyoung Kim, Fangrui Ma, Noreene Shibata, Kimber L. Stanhope, Cecilia Giulivi, Frederik Hansen, Jacob Jelsing, Niels Vrang, Mark Kowala, Michael L. Chouinard, Fawaz G. Haj, Peter J. Havel

Nebraska Center for Biotechnology: Faculty and Staff Publications

Post-operative increases in circulating bile acids have been suggested to contribute to the metabolic benefits of bariatric surgery; however, their mechanistic contributions remain undefined. We have previously reported that ileal interposition (IT) surgery delays the onset of type 2 diabetes in UCD-T2DM rats and increases circulating bile acids, independently of effects on energy intake or body weight. Therefore, we investigated potential mechanisms by which post-operative increases in circulating bile acids improve glucose homeostasis after IT surgery. IT, sham or no surgery was performed on 2-month-old weight-matched male UCD-T2DM rats. Animals underwent an oral fat tolerance test (OFTT) and serial oral …

A Dna Computer For Glioblastoma Multiforme Diagnosis And Drug Delivery, Sumaiya F. Hashmi

CMC Senior Theses

Glioblastoma multiforme (GBM) is a debilitating malignant brain tumor with expected patient survival of less than a year and limited responsiveness to most treatments, often requiring biopsy for diagnosis and invasive surgery for treatment. We propose a DNA computer system, consisting of input, computation, and output components, for diagnosis and treatment. The input component will detect the presence of three GBM biomarkers: vascular endothelial growth factor (VEGF), caveolin-1α (CAV), and B2 receptors. The computation component will include indicator segments for each of these genes, and ensure that output is only released if all the biomarkers are present. The output component …

A Depolymerization Based Model Of Neuron Growth Cone Shape And Motility, Neerajha Nagarajan

Master's Theses

Cell migration is a ubiquitous process underlying critical biological mechanisms like wound healing, cancer metastasis and even neuron growth cone motility. It is a critical process for the living organisms as it ensures proper functioning of the system, for example, crawling fibroblasts endure closure of wounds during wound healing. Here we focus our interest on neuron growth cones, a sensory and motile organelle present at the tip of extending neurites, like the axons, in neurons. These are responsible for neuron pathfinding onto specific targets and synapses, in responses to various guidance cues. Interestingly, the motility of the growth cones is …

Stem Cell Based Tissue Engineering And Regenerative Medicine: A Review Focusing On Adult Stem Cells, Jordan Greenberg, Veronica R. Fortino, Daniel Pelaez, Herman S. Cheung

Biology Faculty Articles

Tissue engineering has emerged as a field that attempts to harness the bodies' own developmental and repair features to treat diseases and illnesses. Many of these illnesses are caused by necrosis or loss of functionality of complete organs or specific cell types. Early discoveries in embryonic stem cells fueled a wave of research that led to claims about possibly regenerating nonfunctioning organs. Although we are still far away from being able to grow functional organs in a Petri dish, the field continues to progress forward, and new clinical trials have been approved for using both embryonic and adult stem cell …

Investigation Of A New Material For Heart Valve Tissue Engineering, Claire Brougham, Gerard Cooney, Thomas Flanagan, Stefan Jockenhoevel, Fergal O'Brien

Conference Papers

No abstract provided.

Characterization Of Printed Skin Equivalents, Mayela R, Aldaz Cervantes, Maria Yanez, Thomas Boland

COURI Symposium Abstracts, Summer 2012

Severe burns, ulcerations, and cuts are some of the reasons people need bioengineered skin as a treatment for full-thickness wounds. When patients run out of donor sites in their own body, using the patient’s skin as a graft is no longer an option, and tissue-engineered constructs are the only possible replacement for damaged skin. The goal of this research is to develop a hydrogel-based skin graft that will be able to function as a skin replacement in full-thickness wounds while minimizing the possibility of rejection, being cost-effective, and possessing a longer shelf-life than the grafts currently in the market. Endothelial …

Tissue Engineering A Blood Vessel Mimic While Monitoring Contamination Through Sterility Assurance Testing, Navid Djassemi

Master's Theses

Tissue Engineering A Blood Vessel Mimic While Monitoring Contamination Through Sterility Assurance Testing

Navid Djassemi

Tissue engineering blood vessel mimics has been proposed as a method to analyze the endothelial cell response to intravascular devices that are used in today’s clinical settings for the treatment of cardiovascular disease. Thus, the development of in vitro blood vessel mimics (BVMs) in Cal Poly’s Tissue Engineering Lab has introduced the possibility of assessing the characteristics of cellular response to past, present, and future intravascular devices that aim at treating coronary artery disease.

This thesis aimed at improving the methods and procedures utilized in …

Development Of In Vitro Tissue Engineered Blood Vessel Mimics In Complex Geometries For Coronary Stent Testing, Robert Dalton Chavez

Master's Theses

Coronary heart disease is the leading cause of death in the United States and occurs when plaque occludes coronary arteries. Coronary stents, which may be used to treat coronary occlusions, are small metal tubes that are implanted in coronary arteries to restore blood flow. After stent implantation, endothelial cells grow over the stent so that blood contacts the endothelial cells instead of the stent surface; this event is known as re-endothelialization. Re-endothelialization prevents blood from clotting on the stent surface and is a good predictor of stent success. Blood vessel mimics (BVMs) are in vitro tissue engineered models of human …

Characterization Of Tight Junction Formation In An In-Vitro Model Of The Blood-Brain Barrier, Michael Robert Machado

Master's Theses

Active and passive transport of substances between the microcirculation in the brain and the central nervous system is regulated by the Blood-Brain Barrier (BBB). This barrier allows for chronic and acute modulation of the CNS microenvironment, and protects the brain from potentially noxious compounds carried in the circulatory system. In-vitro modeling of the BBB has become the target of much research over the past decade, as there are many unanswered questions regarding modulations in the permeability of this barrier. Additionally, the development of a practical and inexpensive model of the BBB would facilitate a much more efficient drug development process. …

Development And Characterization Of An In-House Custom Bioreactor For The Cultivation Of A Tissue Engineered Blood-Brain Barrier, Amin Hadi Mirzaaghaeian

Master's Theses

The development of treatments for neurological disorders such as Alzheimer’s and Parkinson’s disease begins by understanding what these diseases affect and the consequences of further manifestation. One particular region where these diseases can produce substantial problems is the blood-brain barrier (BBB). The BBB is the selective diffusion barrier between the circulating blood and the brain. The barrier’s main function is to maintain CNS homeostasis and protect the brain from the extracellular environment. The progression of BBB research has advanced to the point where many have modeled the BBB in vitro with aims of further characterizing and testing the barrier. Particularly, …

Viscoelastic Anisotropic Finite Element Mixture Model Of Articular Cartilage Using Viscoelastic Collagen Fibers And Validation With Stress Relaxation Data, Matthew Alexander Griebel

Master's Theses

Experimental results show that collagen fibers exhibit stress relaxation under tension and a highly anisotropic distribution. To further develop the earlier model of Stender [1], the collagen constituent was updated to reflect its intrinsic viscoelasticity and anisotropic distribution, and integrated with an existing mixture model with glycosaminoglycans and ground substance matrix. A two-term Prony series expansion of the quasi-linear viscoelastic model was chosen to model the viscoelastic properties of the collagen fibers. Material parameters were determined by using the simplex method to minimize the sum of squared errors between model results and experimental stress relaxation data of tissue in tension. …

Design Of Experimentation To Systematically Determine The Interaction Between Electrospinning Variables And To Optimize The Fiber Diameter Of Electrospun Poly (D, L-Lactide-Co-Glycolide) Scaffolds For Tissue Engineered Constructs, Yvette S. Castillo

Master's Theses

Cardiac disease causes approximately a third of the deaths in the United States. Furthermore, most of these deaths are due to a condition termed atherosclerosis, which is a buildup of plaque in the coronary arteries, leading to occlusion of normal blood flow to the cardiac muscle. Among the methods to treat the condition, stents are devices that are used to restore normal blood flow in the atherosclerotic arteries. Before advancement can be made to these devices and changes can be tested in live models, a reliable testing method that mimics the environment of the native blood vessel is needed. Dr. …

Development Of A Protocol To Measure Gene Expression In The Mouse Tibia, Daniel Hoover

Biomedical Engineering

Numerous molecular factors active in bone tissue direct fracture repair and remodeling which can be altered by disease conditions such as Peripheral Arterial Disease (PAD) and Osteoporosis. Methods of molecular biology are commonly applied to investigate the expression and role of these molecular factors. This project presents a robust three-step protocol for examining gene expression in the mouse tibia. The protocol begins with isolating RNA from a flash frozen tibia sample. The isolated RNA is reverse transcribed into cDNA. Finally, PCR is performed to indentify expressed genes. Establishing this protocol will allow further research into the mechanisms of bone remodeling …

Development And Characterization Of Plga And Eptfe Blood Vessel Mimics Using Gene Expression Analysis, Michael Gibbons, Sarah Ur

Biomedical Engineering

Tissue engineered blood vessels (TEBV’s) have the potential to act not only as a replacement for diseased vessels, but also as a testing platform for intravascular devices such as stents. To this end, the goal of this study was to develop protocols for the construction of TEBV’s composed of human vascular cells and either expanded polytetrafluoroethylene (ePTFE) or poly-lactic-co-glycolic acid (PLGA), as well as a protocol for gene expression in those TEBV’s. Initial experiments involved only human umbilical vein endothelial cells (HUVEC’s), but after low cell confluency and spreading in single-sodded vessels a second cell type, human umbilical vein smooth …

Growth And Expression Of Halorhodopsin For Application In A Protein-Based Artificial Retina, Megan Ryan Gillespie

Honors Scholar Theses

Halorhodopsin (HR), a light-activated chloride ion pump, demonstrates potential for use as the scaffolding in an artificial retina. Retinal implants are needed to restore vision to people afflicted with ophthalmic diseases, such as age-related macular degeneration (AMD) and retinitis pigmentosa (RP). A protein-based chloride ion-patch would be utilized by the eye to create an influx of chloride ions, similar to ion concentrations in healthy retinas during the conversion of light stimuli to electrochemical signals. This protein-based retinal prosthesis will directly stimulate the bipolar cells of the retina, replacing the function of damaged photoreceptor cells. Other alternative treatments for AMD and …

Electrospun Polycaprolactone Nanofiber Scaffolds For Tissue Engineering, Andreas Haukas

Graduate Theses and Dissertations

Stem cell and tissue engineering offer us with a unique opportunity to research and develop new therapies for treating various diseases that are otherwise incurable using traditional medicines. However, development of these new therapies replies upon the establishment of in vitro cell culture and differentiation systems that mimic in vivo microenvironments required for cell-cell and cell-ECM interaction. The development of these cell culture systems depends upon the identification of appropriate biomaterials and cell sources. Biomaterials should be carefully selected and fabricated into scaffolds for supporting cell growth and differentiation. In this study, we explored the fabrication of 3D electrospun nanofiber …

Development Of A Microfluidic Device Coupled To Microdialysis Sampling For The Pre-Concentration Of Cytokines, Randy Francisco Espinal Cabrera

Graduate Theses and Dissertations

A proof-of-concept microfluidic device combined with heparin-immobilized magnetic beads was created to concentrate cytokine proteins collected from microdialysis samples. Cytokines are known to be related to several diseases such as cancer, and Parkinson's diseases, so to be able to develop more effective diseases treatments their interactions have to be well understood. Amine-functionalized polystyrene and carboxyl-functionalized magnetic microspheres of ~6.0 ìm in diameter were used to immobilize heparin. The amount of heparin immobilized on polystyrene beads was 5.82 x 10-8 ± 0.36 x 10-8 M per 1.0 x 106 beads and for magnetic beads was 0.64 x 10-8 ± 0.01 x …

Evaluation Of Decellularization Procedures For Porcine Arteries, Charles Clapp

Biomedical Engineering

Coronary artery disease has become the leading cause of death in the United States, with over 425,000 deaths in 2006. Stenting has evolved into the preferred preventative technique for myocardial infarction by opening up an occluded artery, due to its low invasiveness compared to the alternative of coronary artery bypass grafting. Bare metal stents have been improved by coating with anti-proliferative drugs to advance their effects, but even drug eluting stents still have a risk of restenosis, thrombus formation, and necessary revascularization. Continual advancement in stent design necessitates faster, effective pre-clinical evaluation techniques. Kristen Cardinal, Ph.D., developed the blood vessel …

Biomolecular Mechanisms And Functional Characterization Of Nanomaterials In Programmed Cancer Cell Death For Potential Applications In Medicine, Biomedical Sciences, And Bioengineering, Md. Zakir Hossain

Theses and Dissertations

Cancer bionanotechnology is an interdisciplinary area of research in applied biosciences, biomedical engineering, and nano-medicine. Currently, apoptotic cell death inducers and regulators are considered to have significant potential for cancer therapy. The ability to escape apoptosis is one of the key properties of human cancer. Pancreatic cancer that has the highest fatality rates of all cancers is prompt to evade apoptosis. Nanomaterials like Ni NWs have various novel properties that could be very useful in bio-medical applications at the molecular and cellular levels. The overall objective of this research was to investigate the antiproliferative and apoptogenic properties of Ni NWs …

Methamphetamine Administration Targets Multiple Immune Subsets And Induces Phenotypic Alterations Suggestive Of Immunosuppression., Robert Z. Harms, Brenda M. Morsey, Craig W. Boyer, Howard S. Fox, Nora E. Sarvetnick

Journal Articles: Regenerative Medicine

Methamphetamine (Meth) is a widely abused stimulant and its users are at increased risk for multiple infectious diseases. To determine the impact of meth on the immune system, we utilized a murine model that simulates the process of meth consumption in a typical addict. Our phenotypic analysis of leukocytes from this dose escalation model revealed that meth affected key immune subsets. Meth administration led to a decrease in abundance of natural killer (NK) cells and the remaining NK cells possessed a phenotype suggesting reduced responsiveness. Dendritic cells (DCs) and Gr-1(high) monocytes/macrophages were also decreased in abundance while Gr-1(low) monocytes/macrophages appear …

The Use Of An In Vitro 3d Melanoma Model To Predict In Vivo Plasmid Transfection Using Electroporation, Benadette Marrero, Richard Heller

Bioelectrics Publications

A large-scale in vitro 3D tumor model was generated to evaluate gene delivery procedures in vivo. This 3D tumor model consists of a "tissue-like" spheroid that provides a micro-environment supportive of melanoma proliferation, allowing cells to behave similarly to cells in vivo. This functional spheroid measures approximately 1 cm in diameter and can be used to effectively evaluate plasmid transfection when testing various electroporation (EP) electrode applicators. In this study, we identified EP conditions that efficiently transfect green fluorescent protein (GFP) and interleukin 15 (IL-15) plasmids into tumor cells residing in the 3D construct. We found that plasmids …