Molecular, Cellular, and Tissue Engineering Commons^™

Functional Characterization Of Petiolule-Like Pulvinus (Plp) Gene In Abscission Zone Development In Medicago Truncatula And Its Application To Genetic Improvement Of Alfalfa, Juan Du, Shaoyun Lu, Maofeng Chai, Chuanen Zhou, Liang Sun, Yuhong Tang, Jin Nakashima, Jaydeep Kolape, Zhaozhu Wen, Marjan Behzadirad, Tianxiu Zhong, Juan Sun, Yunwei Zhang, Zeng-Yu Wang

Nebraska Center for Biotechnology: Faculty and Staff Publications

Alfalfa (Medicago sativa L.) is one of the most important forage crops throughout the world. Maximizing leaf retention during the haymaking process is critical for achieving superior hay quality and maintaining biomass yield. Leaf abscission process affects leaf retention. Previous studies have largely focused on the molecular mechanisms of floral organ, pedicel and seed abscission but scarcely touched on leaf and petiole abscission. This study focuses on leaf and petiole abscission in the model legume Medicago truncatula and its closely related commercial species alfalfa. By analysing the petiolule-like pulvinus (plp) mutant in M. truncatula at phenotypic level (breakstrength and …

The Impact Of Aging And Mechanical Injury On Alveolar Epithelial And Macrophage Responses In Acute Lung Injury And Inflammation, Michael S. Valentine

Theses and Dissertations

Patients with severe lung pathologies, such as Acute Respiratory Distress Syndrome (ARDS), often require mechanical ventilation as a clinical intervention; however, this procedure frequently exacerbates the original pulmonary issue and produces an exaggerated inflammatory response that potentially leads to sepsis, multisystem organ failure, and mortality. This acute lung injury (ALI) condition has been termed Ventilator-Induced Lung Injury (VILI). Alveolar overdistension, cyclic atelectasis, and biotrauma are the primary injury mechanisms in VILI that lead to the loss of alveolar barrier integrity and pulmonary inflammation. Stress and strains during mechanical ventilation are believed to initiate alveolar epithelial mechanotransduction signaling mechanisms that contribute …

A Framework For Heterologous Biosynthesis Of Natural Products In Mammalian Cells Via Polymer-Mediated Transfections, Logan Warriner

Theses and Dissertations--Chemical and Materials Engineering

With the promise to treat a multi-faceted list of serious inherited and acquired diseases, such as cancer, neurodegenerative and infectious diseases, and inherited genetic indications, gene therapy has continued to push the boundaries of traditional medicine since its earliest implementation. While much progress has been made, clinical success has largely remained elusive. Immunogenicity, difficulty producing commercially relevant quantities, and having a limited genetic payload still limits the ability of viruses to act as directed delivery agents for genetic material. As such, researchers have turned to cationic synthetic materials as a means of delivering nucleic acids, which can circumvent the immune …

All-Wheel-Ukraine, Sofiya Rakovska, Hannah Mcculloch, Andrey Garasimchuk, Ahmad Alsaihati

Williams Honors College, Honors Research Projects

We are working with a non-profit organization in Ukraine that provides wheelchairs/strollers to families of disables children who cannot afford them. Their current preferred solution is a baby stroller. The organization asked us to create a wheelchair that is cost efficient, light and collapsible to be used for children ages up to thirteen years old. We will be creating an alpha prototype wheelchair by modifying a basic wheelchair that we purchased. This modifications include adding a headrest that is adjustable and provides a sufficient support for children within the age group given to us and diverse levels of disabilities. We …

Higher Tensile Forces Across Cellular Junctions And An Intact Nuclear Linc Complex Is Required For Epithelial Function And Stability, Fnu Vani Narayanan

Theses and Dissertations

Recent advances in three-dimensional (3D) cell culture systems have provided key insights into the understanding of biochemical and physiological states of native tissue. A significant progress in the field of mechanobiology involves measuring cellular traction forces in a more native 3D environment. However, the effects of mechanical forces exerted across cellular junctions and the nuclear LINC complex, in an organized 3D system has not been investigated thus far. Epithelial cells spontaneously form acini (also known as cysts or spheroids) with a single, fluid-filled central lumen, when grown in 3D matrices. The size of the lumen is dependent on apical secretion …

Artificial Synthetic Scaffolds For Tissue Engineering Application Emphasizing The Role Of Biophysical Cues, Samerender Nagam Hanumantharao

Dissertations, Master's Theses and Master's Reports

The mechanotransduction of cells is the intrinsic ability of cells to convert the mechanical signals provided by the surrounding matrix and other cells into biochemical signals that affect several distinct processes such as tumorigenesis, wound healing, and organ formation. The use of biomaterials as an artificial scaffold for cell attachment, differentiation and proliferation provides a tool to modulate and understand the mechanotransduction pathways, develop better in vitro models and clinical remedies. The effect of topographical cues and stiffness was investigated in fibroblasts using polycaprolactone (PCL)- Polyaniline (PANI) based scaffolds that were fabricated using a self-assembly method and electrospinning. Through this …

Electrospun Nanofiber Scaffolds As A Platform For Breast Cancer Research, Carolynn Que

Dissertations, Master's Theses and Master's Reports

Tumorigenesis is a complex process involving numerous cellular signaling cascades and environmental factors. Here, we report the fabrication of 3D scaffolds with different morphologies obtained by to study cancer cell proliferation and migration. Using an FDA approved, biocompatible and biodegradable polymer Polycaprolactone (PCL), we electrospun nanofiber scaffolds having mesh, aligned, and honeycomb morphologies. The role of the morphology and cellular preferences to nutrition in cell adhesion and proliferation was assessed using scaffolds obtained by electrospinning PCL with fluorescent fructose-like molecular probes. Cell viability, cell morphology, localized cellular growth as related to scaffold morphology and availability of the fructose-like molecular probes …

Nanofiber Scaffolds As 3d Culture Platforms, Stephanie Bule

Dissertations, Master's Theses and Master's Reports

Cell culture has become the basis for understanding the fundamental mechanisms of cell, tissue and organ function. Although major advancements in uncovering the underlying processes and mechanisms of normal and diseased cell biology have been made by using two-dimensional (2D) cell culture, there is a recent shift in moving towards three-dimensional (3D) culture platforms. The motivation is to better recapitulate the microenvironment of cells in vivo to obtain results that are more indicative to actual cellular processes. In this study, electrospun nano-fibrous scaffolds made of polycaprolactone were used as a 3D culture tool to investigate difference in cell behavior and …

Cerebro-Vascular Disruption Mediated Initiation And Propagation Of Traumatic Brain Injury In A Fluid Percussion Injury Model, Xiaotang Ma

Dissertations

Traumatic brain injury (TBI) is a major health problem for over 3.17 million people in the US. There is no FDA-approved drug for the treatment because the injury mechanisms have not been clearly identified. The knowledge gap is addressed here by the lateral fluid percussion injury (FPI) rat model, through the understanding of layer-structured mechanisms from physical vascular rupture to acute necrosis, as well as biochemical changes in perivascular space as secondary events.

Firstly, the cerebrovascular hemorrhage and related infarct volume are investigated as the primary events in moderate FPI, which is found to be increased with injury severity in …

Controlling Strain Energy Density In 3d Cellular Collagen Constructs During Complex Loads, Katherine Hollar

Boise State University Theses and Dissertations

Mechanical stimulation applied to damaged soft tissues, such as ligament, can promote tissue remodeling to accelerate healing. To help identify treatments that encourage ligament healing, bioreactors have been designed to subject 3D cellularized constructs to various loading conditions in order to determine the mechanical mechanisms that trigger cell-mediated repair. An innovative approach is to use a bioreactor to apply controlled states of biaxial stress to study the effects of strain energy density and distortion energy on cell activity. Tissue distortion has been linked to changes in the structure and function of ligament, yet the specific impact of distortion energy on …

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 …

Effects Of Cryopreservation And Next Generation Sequencing Of Adipose Derived Stem Cells, Mulla Shahensha Shaik

LSU Doctoral Dissertations

The research in this thesis is divided into two areas. 1) Studying the effects of cryopreservation and developing freezing protocols suitable for adipose derived stem cells and 2) Identifying the underlying mechanisms involved in the differentiation of ASCs especially into osteogenesis and endothelial-genesis using next generation sequencing.

In the part one (Chapter 2 and 3), a novel protocol to freeze ASCs and the effects of long-term cryopreservation on ASCs were studied. Heat shocking ASCs at 43°C for 1 hour resulted in the temporal expression of HSPs particularly HSP70 and HSP32 to reach maximum expression at 3 hours post heat shock …

Plant Cell-Derived Growth Factors For Ex Vivo Mass Production Of Red Blood Cells, Xiaoting Wang

Student Theses and Dissertations

Generation of red blood cells from hematopoietic stem cells (HSCs) used for blood transfusion represents one of the focus in regenerative medicine. Erythropoietic growth factors (eGFs), including stem cell factor (SCF), erythropoietin (EPO) and interleukin 3 (IL-3) are vital for proliferation and differentiation of HSCs into mature red blood cells (RBCs) ex vivo. However, massive production of RBCs requires significant quantity and high quality of eGFs, making manufacturing at large scale cost prohibitive. Plant cell culture is proposed to be a promising bioproduction platform for growth factors based on the advantages of, fast growing, no risk of human pathogen, and …

Synthesis And Secretome Release By Human Bone Marrow Mesenchymal Stem Cell Spheroids Within Three-Dimensional Collagen Hydrogels: Integrating Experiments And Modelling, Jyotsna Joshi, Mohammadreza Dehghan Abnavi, Chandrasekhar R. Kothapalli

Chemical & Biomedical Engineering Faculty Publications

Myocardial infarction results in loss of cardiac cell types, inflammation, extracellular matrix (ECM) degradation, and fibrotic scar. Transplantation of bone marrow-derived mesenchymal stem cells (BM-MSCs) is being explored as they could differentiate into cardiomyocyte-like cells, integrate into host tissue, and enhance resident cell activity. The ability of these cells to restore lost ECM, remodel the inflammatory scar tissue, and repair the injured myocardium remains unexplored. We here elucidated the synthesis and deposition of ECM (e.g., elastin, sulfated glycosaminoglycans, hyaluronan, collagen type III, laminin, fibrillin, lysyl oxidase, and nitric oxide synthases), matrix metalloproteinases (MMPs) and their inhibitors (TIMPs), and other secretome …

Identifying Extracellular Matrix Protein Turnover Rates For Tissue Engineers, Alita F. Miller

The Journal of Purdue Undergraduate Research

No abstract provided.

Genome-Edited Animals Are Not Transgenic Animals: Moving Toward Responsible Research And Innovation With New Biotechnologies, Yvie Yao

Minnesota Journal of Law, Science & Technology

No abstract provided.

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 …

Mechanisms Of Reduced Vascular Tone Following Arteriogenesis Induced By Femoral Artery Ligation, Christopher Hatch

Biomedical Engineering

The presence of a developed, native collateral network can decrease the severity of ischemic injury proceeding arterial occlusion. The collateral network must under arteriogenesis to enlarge and increase blood flow to the ischemic region. Although there has been tremendous effort attempting to understand the mechanisms of arteriogenesis, no therapies have been successful in improving patient outcome. To better understand the mechanisms involved in arteriogenesis, the effect of nitric oxide production, myogenic tone, and a-adrenergic receptors were evaluated as these have been identified as playing an important role in vascular injury. Arteriogenesis was induced by ligating the femoral artery between the …

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 …