Molecular, Cellular, and Tissue Engineering Commons^™

Bioproduction Of Adipic Acid Using Engineered Pseudomonas Putida Kt2440 From Lignin-Derived Aromatics, Howard Willett

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

Current industrial synthesis of adipic acid is nonrenewable and depends on a carcinogenic starting material, benzene. Biocatalysis with an engineered microorganism could turn a renewable feedstock into a value-added chemical such as adipic acid. Here we engineered P. putida KT2440 to transform lignin-derived aromatics, coumarate and ferulate, into adipic acid. Lignin is a recalcitrant plant biopolymer burned for thermal energy. Conversion of lignin into a value-added chemical will improve the efficiency of lignocellulose processing plants. The best performing engineered KT2440 strain produces 2.52 mM adipate at a 9.5% (mole/mole) yield. This was achieved by the genetic insertion of non-natural biosynthetic …

Developing A Control System To Better Understand The Effects Of Pyruvate Decarboxylase Activity On Clostridium Thermocellum Metabolism, Nicholas Cervenka

ENGS 88 Honors Thesis (AB Students)

In order for cellulosic biofuels from Clostridium thermocellum to be commercially viable, the ethanol yield and titer of the microbe must be increased. To accomplish this, it has been suggested to introduce the Pyruvate Decarboxylase (PDC) enzyme into C. thermocellum. In order to demonstrate effects on ethanol production by PDC prior to genetic modification, a cell free system (CFS) has been developed. A purified enzyme system was developed with the CFS to function as a control. Using the purified enzyme system, PDC from Saccharomyces cerevisiae was demonstrated to be a good candidate for further testing in the CFS.

Nano Fab Lab 63, Brian K. Deemer, Josh Clemons, Nick Brodine, Delaney Fitzsimmons

Mechanical Engineering

Lawrence Livermore National Laboratories (LLNL) has several steps in the production process for ceramic nanofiber tubes that they would like to improve - electrospinning, cutting, rolling, sealing and heat treating. We undertook the challenge to deliver LLNL with a semi-automated process that efficiently integrates the steps of cutting, rolling, and sealing to save time and improve control over end dimensions. In this document, we discuss the technical background of the manufacturing steps currently followed to create nanofiber tubes, identify which steps are incorporated in our prototype and explain how they will interface with one another, define the design requirements, present …

Evaluation Of Human Umbilical Vein Endothelial Cells In Blood Vessel Mimics Through Changes In Gene Expression And Caspase Activity, Conor Charles Hedigan

Master's Theses

Blood vessel mimics (BVMs) are simple tissue engineered blood vessel constructs intended for preclinical testing of vascular devices. This thesis developed and implemented methods to characterize two of these components. The first aim of this thesis investigated the effect of cell culture duration and flow conditions on endothelial cell gene expression, especially regarding endothelial-to-mesenchymal transition (EndMT). A trend of decreased endothelial marker gene expression and increased mesenchymal marker gene expression would indicate EndMT. qPCR analysis revealed that increased cell culture duration did not result in EndMT, and in fact increased endothelial marker expression as cell culture duration increased. Disturbed flow …

Predicting Tgf-Β-Induced Epithelial-Mesenchymal Transition Using Data Assimilation, Mario J. Mendez, Matthew J. Hoffman, Elizabeth M. Cherry, Dr. Christopher Lemmon, Seth Weinberg

Biology and Medicine Through Mathematics Conference

No abstract provided.

The Effect Of Heterobifunctional Crosslinkers On Hema Hydrogel Modulus And Toughness, Elizabeth M. Boazak, Vaughn K. Greene Jr., Debra T. Auguste

Publications and Research

The use of hydrogels in load bearing applications is often limited by insufficient toughness. 2-Hydroxyethyl methacrylate (HEMA) based hydrogels are appealing for translational work, as they are affordable and the use of HEMA is FDA approved. Furthermore, HEMA is photopolymerizable, providing spatiotemporal control over mechanical properties. We evaluated the ability of vinyl methacrylate (VM), allyl methacrylate (AM), and 3-(Acryloyloxy)-2-hydroxypropyl methacrylate (AHPM) to tune hydrogel toughness and Young’s modulus. The crosslinkers were selected due to their heterobifunctionality (vinyl and methacrylate) and similar size and structure to EGDMA, which was shown previously to increase toughness as compared to longer crosslinkers. Vinyl methacrylate …

Using The Xenopus Developmental Eye Regrowth Stystem To Distinguish The Role Of Developmental Versus Regenerative Mechanisms, Cindy X. Kha, Dylan J. Guerin, Kelly Ai-Sun Tseng

Life Sciences Faculty Research

A longstanding challenge in regeneration biology is to understand the role of developmental mechanisms in restoring lost or damaged tissues and organs. As these body structures were built during embryogenesis, it is not surprising that a number of developmental mechanisms are also active during regeneration. However, it remains unclear whether developmental mechanisms act similarly or differently during regeneration as compared to development. Since regeneration is studied in the context of mature, differentiated tissues, it is difficult to evaluate comparative studies with developmental processes due to the latter’s highly proliferative environment. We have taken a more direct approach to study regeneration …

Development Of A Sonically Powered Biodegradable Nanogenerator For Bone Regeneration, Avi S. Patel

University Scholar Projects

Background: Reconstruction of bone fractures and defects remains a big challenge in orthopedic surgery. While regenerative engineering has advanced the field greatly using a combination of biomaterial scaffolds and stem cells, one matter of difficulty is inducing osteogenesis in these cells. Recent works have shown electricity’s ability to promote osteogenesis in stem cell lines when seeded in bone scaffolds; however, typical electrical stimulators are either (a) externally housed and require overcomplex percutaneous wires be connected to the implanted scaffold or (b) implanted non-degradable devices which contain toxic batteries and require invasive removal surgeries.

Objective: Here, we establish a biodegradable, piezoelectric …

Development Of A Sonically Powered Biodegradable Nanogenerator For Bone Regeneration, Avi Patel

Honors Scholar Theses

Background: Reconstruction of bone fractures and defects remains a big challenge in orthopedic surgery. While regenerative engineering has advanced the field greatly using a combination of biomaterial scaffolds and stem cells, one matter of difficulty is inducing osteogenesis in these cells. Recent works have shown electricity’s ability to promote osteogenesis in stem cell lines when seeded in bone scaffolds; however, typical electrical stimulators are either (a) externally housed and require overcomplex percutaneous wires be connected to the implanted scaffold or (b) implanted non-degradable devices which contain toxic batteries and require invasive removal surgeries.

Objective: Here, we establish a biodegradable, piezoelectric …

Investigating Virus Clearance Via Ph Inactivation During Biomanufacturing, Wenbo Xu

Biomedical Engineering Undergraduate Honors Theses

In the processing of biopharmaceuticals, viral clearance and viral safety are important for the development of monoclonal antibodies. Murine xenotropic leukemia virus (XMuLV) is one of the retroviruses, recommended by Food and Drug Administration (FDA) as a model virus for viral clearance via inactivation from therapeutics derived from Chinese hamster ovary cells (CHO). A robust and effective method was investigated to clear or inactivate endogenous viruses by low pH inactivation. The effects of different conductivity and inactivated time on XMuLV clearance was determined. Acetate buffer was prepared with different conductivity, and 2% XMuLV was spiked into acetate buffer. XMuLV virus …

Phenotypic Characterization Of Differentiation Of Human Mesenchymal Stem Cells To Smooth Muscle Cells, Jackson Hedrick

Biomedical Engineering Undergraduate Honors Theses

Abdominal aortic aneurysms (AAAs) are expansions in the aortic wall with an increased tendency to rupture. These expansions are triggered by the breakdown of key aortic extracellular matrix (ECM) proteins—elastin and collagen. The degradation of aortic ECM proteins is the consequence of overexpressed matrix metalloproteases (MMPs)–MMP-2 and MMP-9. Adult vascular smooth muscle cells (SMCs) lack the ability to repair the elastic matrix assembly; thus, it is imperative to find a solution to both restore the imbalance of essential ECM proteins in the aorta and mitigate the effects of MMPs. Previous studies have revealed that bone marrow derived mesenchymal stem cells …

Diffuse Reflectance Spectroscopy To Quantify In Vivo Tissue Optical Properties: Applications In Human Epithelium And Subcutaneous Murine Colon Cancer, Gage Joseph Greening

Graduate Theses and Dissertations

Colorectal cancer is the 4th most common and 2nd deadliest cancer. Problems exist with predicting which patients will respond best to certain therapy regimens. Diffuse reflectance spectroscopy has been suggested as a candidate to optically monitor a patient’s early response to therapy and has been received favorably in experimentally managing other cancers such as breast and skin. In this dissertation, two diffuse reflectance spectroscopy probes were designed: one with a combined high-resolution microendoscopy modality, and one that was optimized for acquiring data from subcutaneous murine tumors. For both probes, percent errors for estimating tissue optical properties (reduced scattering coefficient and …

Comparing The Effects Of Fibroblast Growth Factors On Growth Rate Of Human Fibroblast Cell Lines, Tasha Repella

Biomedical Engineering Undergraduate Honors Theses

Numerous studies have demonstrated the usefulness of growth factors in medium formulations to promote proliferation and viability of human fibroblasts in laboratory settings. However, due to the differences in the source and age of fibroblasts, the ability of growth factors like hFGF1 to promote proliferation varies. Equally difficult is the ability of hFGF1 to promote proliferation of diseased fibroblasts due to complexities associated with specific mitochondrial diseases. In this context, we decided to evaluate the potential for novel hyper stable growth factors super hFGF and hFGF1 (shFGF and hFGF1 respectively), to contribute to proliferation of different normal and diseased fibroblasts. …

Role Of Local Renin-Angiotensin System In Potentiating Early Calcific Aortic Valve Disease, Jessica Perez

Graduate Theses and Dissertations

Angiotensin-II (Ang-II), a peptide hormone, is a potent vasoconstrictor and cell mitogen. It has been implicated in the development of hypertension as well as atherosclerosis. Recent work has shown that sclerotic aortic valves possess expression of angiotensin type I receptor (AT-1R) and angiotensin converting enzyme (ACE), suggesting altered angiotensin signaling during disease. The role of altered angiotensin signaling on aortic valve mechanics, however, is not clearly understood. We seek to understand the direct effects of the renin angiotensin signaling (RAS) system on the biological and biomechanical properties of aortic valve tissue and develop a finite constitutive model that mimics the …

Developing A Synthetic Model Of The Candida Albicans Cell Wall Using Self-Assembled Monolayers To Host Beta-Glucan As Ligands, Ushnik Ghosh

Biomedical Engineering ETDs

The goal of this investigation is to study mechanisms the immune cell receptor, Dectin-1, uses to identify the fungal cell species, Candida albicans. Dectin-1 identifies extracellular matrix polysaccharides that Candida albicans express known as β-glucan. To study the interaction of β-glucan – Dectin-1 at the nanoscopic scale, the investigators of this project have endeavored to model the Candida albicans cell wall with engineered Self-Assembled Monolayers presenting β-glucan as a ligand for immobilized Dectin-1 receptors. By engineering a simplified ex-vivo model of the Candida albicans cell wall, the investigators of this study aim to gain precise control of the composition …

Incorporation Of Egfr And Ron Receptors Into Nanodiscs, Cristina Flores-Cadengo

Biomedical Engineering ETDs

Understanding the structure-function relationship of membrane receptors is essential to comprehend the crosstalk between key signaling pathways. Aberrant trans-activation between receptors can lead to tumorigenesis. Two of these receptors known to be involved in cancer development are receptor tyrosine kinases (RTKs), RON (Recepteur d'Origine Nantais) and EGFR (Epidermal Growth Factor Receptor). There has been evidence of heterodimerization and crosstalk between these two receptors based on co-immunoprecipitation, however the structural requirements behind these interactions remain unknown. Structural studies could provide insights into these RTKs’ modes of dimerization and structure-function relationship. However, structural studies of full-length membrane proteins are often difficult due …

Plga Scaffold Device, Tyler D. Tackabery, Bryce Powada, Tate Morell

Biomedical Engineering

No abstract provided.

Fluid Delivery System For A Cell Culture On A Microfluidic Chip, Austin J. Roeder, Colleen A. Richards, Emily A. Matteson

Biomedical Engineering

This project report provides a description of the progress made in the development of a fluid delivery system for a microfluidic cell culture on a chip. The system is intended to be used in a humidified incubator in a university laboratory and the fluid delivery system is required to exist and operate within that incubator for extended periods of time. Therefore, the system will be gravity-driven and contain no electronic components. The key specification of the system is to provide fluid flow at a constant velocity.

After manufacturing and testing the device, all specifications were met except for the fluid …

Pressure Cylinder Controlled Release Valve, Alexa Dominique Balbuena

Biomedical Engineering

This project aims to integrate an automatic gas release system in a pre-existing scaffold fabrication process for tissue engineering applications.

To form the proper scaffold structure, the fabrication process is heavily influenced by the change in its surrounding pressure. The current production involves a pressure transducer and electric valve that is managed manually to create a suitable pressure environment for the scaffold. This method, although functional, proves to be ineffective when creating several batches; the user needs to constantly monitor the developing pressure profile and alter voltage parameters accordingly to create a linear gas release under a predetermined slope.

To …

Advanced Selection Methodologies For Dnazymes In Sensing And Healthcare Applications, Sandeep Kumar, Shikha Jain, Neeraj Dilbaghi, Amrik Singh Ahluwalia, Ashraf Aly Hassan, Ki-Hyun Kim

Department of Civil and Environmental Engineering: Faculty Publications

DNAzymes have been widely explored owing to their excellent catalytic activity in a broad range of applications, notably in sensing and biomedical devices. These newly discovered applications have built high hopes for designing novel catalytic DNAzymes. However, the selection of efficient DNAzymes is a challenging process but one that is of crucial importance. Initially, systemic evolution of ligands by exponential enrichment (SELEX) was a labor-intensive and time-consuming process, but recent advances have accelerated the automated generation of DNAzyme molecules. This review summarizes recent advances in SELEX that improve the affinity and specificity of DNAzymes. The thriving generation of new DNAzymes …