Ph-Sensitive Oxygen Release Microspheres To Enhance Cell Survival In Ischemic Condition, Zhongting Liu

McKelvey School of Engineering Theses & Dissertations

Ischemic diseases such as myocardial infarction, stroke and limb ischemia are severe cardiovascular diseases with high rate of death and millions of people suffered from these diseases. Under ischemic environment, cells die due to deficient supply of nutrient and oxygen. To regenerate ischemic tissues, stem cell therapy is a promising approach because stem cells can differentiate into cells necessary for the regeneration. However, stem cell therapy has limitations. For example, few cells can survive under harsh ischemic environment. To enhance stem cells survival, implantation of oxygen release microspheres to sustained supply cells with oxygen represents an effective strategy. Previously, our …

The Co-Delivery Of Syngeneic Adipose-Derived Stromal Cells And Macrophages On Decellularized Adipose Tissue Bioscaffolds For In Vivo Soft Tissue Regeneration, Hisham A. Kamoun

Electronic Thesis and Dissertation Repository

Decellularized adipose tissue (DAT) bioscaffolds are a promising platform for the delivery of pro-regenerative cell populations with the goal of promoting adipose tissue regeneration. The current study investigated the effects of seeding DAT bioscaffolds with syngeneic bone marrow-derived macrophages and/or adipose-derived stromal cells (ASCs) on in vivo soft tissue regeneration. Methods were established to derive the macrophages from MacGreen mice, which were dynamically seeded onto the DAT scaffolds alone or in combination with ASCs. Seeded and unseeded scaffolds were implanted subcutaneously into C57Bl/6 mice. At 2 and 4 weeks, cell infiltration, angiogenesis, and adipogenesis were analyzed through histology and immunohistochemistry. …

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 …

Optimized Production And Evaluation Of Cellulose Nanocrystals Derived From Pre-Extracted Kraft Pulp Of Different Wood Species, Gurshagan Kandhola

Graduate Theses and Dissertations

Production of nanocellulose from a variety of naturally abundant, locally available and industrially significant wood species provides an opportunity for diversifying the portfolio of traditional pulp and paper industries. The U.S. has a prolific forest products industry with a well-established infrastructure that could be utilized for optimized and customized production of cellulose nanomaterials. However, to achieve that, it is important to a) understand how biorefining strategies for complete fractionation of biomass affect the downstream processing of pulp into nanocellulose, b) maximize the yields of cellulose nanocrystals and nanofibers (CNCs and CNFs) from pretreated raw materials, and c) evaluate if the …

Bioluminescence Resonance Energy Transfer (Bret) - Based Nanostructured Biosensor For Detection Of Glucose, Eugene Hwang

Electronic Thesis and Dissertation Repository

Bioluminescence resonance energy transfer (BRET) is a distance dependent, non-radiative energy transfer, which uses a bioluminescent protein to excite an acceptor through resonance energy transfer. In this thesis, BRET technology is incorporated into a sensor comprised of a recombinant protein and quantum dots. The recombinant protein, which includes the bioluminescent protein, Renilla luciferase (Rluc), is used as the donor molecule and cadmium tellurium quantum dots as the acceptor molecules. Separating the donor-acceptor pair is a recombinant protein, glucose binding protein, which changes conformation upon binding glucose and brings the pair closer together, thus allowing BRET to occur. Optimization of the …

High-Throughput Fabrication Of Drug-Loaded Core-Shell Tablets With Adjustable Release Profiles From Surface-Erodible And Photocrosslinkable Polyanhydrides, Armin Geraili Nejadfomeshi

Electronic Thesis and Dissertation Repository

Controlled-release tablets enhance the effectiveness of therapies for various clinical conditions. Photocrosslinkable polyanhydrides that undergo surface erosion were recently introduced as suitable materials for manufacturing tablets with tunable release profiles. However, their erosion behavior has not been comprehensively studied. In this thesis, the erosion kinetics of photocrosslinkable polyanhydrides was studied by exploring the impact of different parameters (the polymer composition and geometry, as well as the temperature, pH, and shaking rate of the solution during the in vitro experiments) on their mass loss profiles, followed by a release kinetic model fitting. The results indicate that the temperature was the only …

Bioinspired Complex Nanoarchitectures By Dna Supramolecular Polymerization, Laura A. Lanier

Doctoral Dissertations

Bioinspired nanoarchitectures are of great interest for applications in fields such as nanomedicine, tissue engineering, and biosensing. With this interest, understanding how the physical properties of these complex nanostructures relate to their function is increasingly important. This dissertation describes the creation of complex nanoarchitectures with controlled structure and the investigation of the effect of nanocarrier physical properties on cell uptake for applications in nanomedicine. DNA self-assembly by supramolecular polymerization was chosen to create complex nanostructures of controlled architectures. We demonstrated that the supramolecular polymerization of DNA known as hybridization chain reaction (HCR) is in fact a living polymerization. The living …

Engineering Nanomaterials For Imaging And Therapy Of Bacteria And Biofilm-Associated Infections, Akash Gupta

Doctoral Dissertations

Infections caused by multidrug-resistant (MDR) bacteria pose a serious global burden of mortality, causing thousands of deaths each year. The “superbug” risk is further exacerbated by chronic infections generated from antibiotic-resistant biofilms that are highly resistant to available treatments. Synthetic macromolecules such as polymers and nanoparticles have emerged as promising antimicrobials. Moreover, ability to modulate nanomaterial interaction with bacterial cellular systems plays a pivotal role in improving the efficacy of the strategy. In the initial studies on engineering nanoparticle surface chemistry, I investigated the role played by surface ligands in determining the antimicrobial activity of the nanoparticles. In further study, …

Design Of Cell-Instructive Biomaterial Scaffolds For Intervertebral Disc Regeneration, Nadia Sharma

Electronic Thesis and Dissertation Repository

Biomaterials-based therapies targeting the nucleus pulposus (NP) have the potential to promote regeneration and restore mechanical function to the intervertebral disc. This study developed composite hydrogels incorporating decellularized NP (DNP) and assessed its effects on viability, retention and differentiation of U-CH1 cells, an NP progenitor-like cell line. A minimal protocol was developed to decellularize bovine NP that reduced nuclear content while preserving key extracellular matrix components predicted to be favourable for bioactivity. The resulting DNP demonstrated cell-instructive effects, supporting U-CH1 viability and retention within the hydrogels, and promoted the differentiation of the progenitor-like cells towards an NP-like phenotype. These studies …

Formation Of A Vascular Regenerative Microenvironment Within Implantable Human Decellularized Adipose Tissue Bioscaffolds, Christopher Leclerc

Electronic Thesis and Dissertation Repository

Cellular therapies targeted at stimulating therapeutic angiogenesis in individuals with critical limb ischemia (CLI) have been under intense investigation. Hematopoietic progenitor cells (HPC) derived from umbilical cord blood have been previously shown to support limb revascularization in animal models of CLI, despite limited cell survival at the site of ischemia. This study attempted to improve HPC survival after transplantation and prolong pro-angiogenic function using human decellularized adipose tissue (hDAT) as a novel cell delivery platform. Compared to HPC conventionally grown on tissue-cultured plastic, hDAT scaffolds were shown to promote viability and proliferation of seeded HPC, and had cell- instructive effects …

Characterization Of Nano-Cellulose Based Composites For Biomedical Applications, Mitchell P. Chesley

Electronic Theses and Dissertations

The number of orthopedic surgeries performed globally has steadily increased over the past decade due to the standardization of procedures as well as technological advancements. During this time orthopedic devices have been composed predominantly of metals, such as Titanium, Vanadium, Molybdenum, and Stainless steel, as well as their alloys, due to the high strength and durability of these materials. However, metals may, in fact, be suboptimal for orthopedic devices. For example, metals exhibit Young’s modulus much greater than the surrounding bone, inducing localized stress-shielding promoting cortical atrophy, which can lead to osteoporosis. In recent years polymers have been successfully explored …

Scaffold Design Considerations For Soft Tissue Regeneration, Madeleine M. Di Gregorio

Electronic Thesis and Dissertation Repository

Tissue engineering has emerged as a promising strategy for the replacement of degenerating or damaged tissues in vivo. Also known as regenerative medicine, integral to this therapeutic strategy is biomimetic scaffolds and the biomaterial structural components used to form them. In this study, three different biomaterial scaffolds for tissue engineering applications were fabricated: three-dimensional reverse embedded collagen scaffolds, polymer fusion printed polycaprolactone (PCL) scaffolds, and electrospun gelatin scaffolds. Three-dimensional collagen and PCL scaffolds promoted human adipose-derived stem/stromal cell (ASC) spreading, proliferation, and fibronectin deposition in vitro. Secondly, this study investigated the efficacy of exogenous galectin-3 delivery as a …

Assessment Of The Biocompatibilty, Stability, And Suitability Of Natural And Synthetic Polymers And Drugs In The Fda Approval Process, Phuong Anh Hoang Nguyen

Biomedical Engineering ETDs

Regulation of the development, production, marketing, and sales of medical pharmaceuticals and devices in the United States fall under the regulatory functions of the Food and Drug Administration (FDA). The current FDA approval process takes an average of 10 years from start to completion, and costs over $100 million. As a result, companies use many different methods to find additional use of their drugs through marketing directly to the physician, or recycling of previously approved drug moieties. In this work, an evaluation of the in vitro and ex vivo biocompatibility of polymers and drugs in different phases of FDA approval …

Design, Construction And Application Of A Home-Built, Two-Photon Microscope, William P. Breeding

Electronic Theses and Dissertations

Two-photon microscopy (TPM) is a powerful, versatile imaging modality for the study of biological systems. This thesis overviews the relevant physics involved in TPM, design considerations and process of constructing a home-built, two-photon microscope, and provides a set of procedures to operate the system. Furthermore, this work explores several applications of TPM through the study of single-cell metabolism and imaging the cellular-material interface. Explored in particular depth was the imaging of cellulose nanofiber (CNF) materials, with the goal of understanding the three-dimensional nature of fibroblast cell growth when embedded within the materials. This work uncovered several optical properties of CNF, …

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 …

Compositional Optimization Of Amyloid-Graphene Oxide Nanohybrids For Biomaterials, Claire L. Drewery

Materials Engineering

Amyloid nanofibrils are natural materials capable of self-assembling into precise structures with tunable functionalities, while exhibiting excellent mechanical properties. In combination with highly conductive graphene oxide (GO), the 1-D amyloid nanofibrils and 2-D nanosheets of GO can produce a robust and bio-functional nanohybrid, hypothesized to exhibit multi-domain functional properties useful for enzyme sensing, water purification, drug delivery, and tissue scaffolding applications. Here, we examine the properties of an amyloid-graphene oxide nanohybrid film made with amyloids derived from hen egg white lysozymes in an attempt to explore the diverse toolbox of amyloid derivatives and establish ideal fabrication methods and formulations of …

Applying An Innovative Idea To Kidney Stone Removal, Jack Beaty

Honors Theses

Renal lithiasis, commonly referred to as “kidney stones” is a condition in which the urine composition crystallizes and cannot pass through the ureter, causing pain and discomfort. Renal lithiasis can be caused by diet, infection, reduced water retention, or hereditary disorders; each of which results in a distinct stone composition [1]. Stone compositions can vary between calcium oxalate, struvite, uric acid, and cystine[2]. Each stone type, however, provides the same pain to the patient and the same potential risk of blocking the urinary system.

Kidney stone symptoms may include but are not limited to: pain in the lower back, the …

The Theia Soteria: Alternative Design For Safer Initial Entry During Laparoscopic Procedures, Kayla Dubois, Patrick Ryan, Madelyn Joanis

Honors Theses

Laparoscopic procedures account for 15 million surgeries worldwide [1], with the initial entry into the peritoneal cavity accounting for 33-50% of all major laparoscopic complications [7]. This initial entry is the most dangerous as surgeons must enter the cavity using a sharp object with no visibility and space between the outer surface of the cavity and internal tissues. During the initial entry into the peritoneal cavity, the patients undergoing laparoscopic procedures are at a high risk for damage to internal organs and vasculature, necessitating the development of a device to protect these internal tissues and increase patient safety.

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 …

Two And Three-Dimensional Models For Material And Cells Interaction, Nam H. Nguyen

Doctoral Dissertations

Three-dimensional (3D) cell spheroid model has been long considered a better model to mimic in vivo physiology compared to two-dimensional (2D) cell culture model. Traditional 2D cell models provide a simple, convenient and quick technique for drug screening but fail to simulate the complexity and heterogeneity of cells in the in vivo environment. The last few decades have remarked substantial progress toward the advancement of three-dimensional (3D) cell cultures as systems which better mimic cellcell and cell-matrix interaction in the in vivo physiology. Nowadays, 3D cell models have been emerging, not only as an important approach in drug discovery and …

A Ph-Sensitive Delivery System For The Prevention Of Dental Caries Using Salivary Proteins, Yi Zhu

Electronic Thesis and Dissertation Repository

Dental caries remains one of the most common chronic diseases worldwide. Salivary proteins such as histatins have demonstrated biological functions directly related to tooth homeostasis and prevention of dental caries. However, histatins are susceptible to the high proteolytic activities in the oral environment. Therefore, pH-sensitive chitosan nanoparticles (CNs) have been proposed as potential carriers to target major oral diseases that occur under acidic conditions (e.g. dental caries and dental erosion). Four different types of chitosan polymers were investigated and the optimized CNs successfully loaded histatin 3 and released it selectively under acidic conditions. Through loading the survival time of histatin …

Cellular Modification And Artificial Cell Construction, Jimin Guo

Chemical and Biological Engineering ETDs

With all its complexities and different biofunctionalities, a cell is the basic structural and functional unit of all living organisms with the exception of viruses. In this dissertation, I demonstrated gain of function strategies of modifying mammalian cells using silicification and modular nanoparticle (NP) exoskeletons. In addition, I used a modular design concept to build a multifunctional artificial RBC system which can mimic the unique characteristics (e.g. shape, flexibility, the ability to carry oxygen, and long circulation times) of the native RBCs. Together, these cell modification or artificial cell construction strategies are expected to generate new mammalian cell-like structures with …

Fabrication And Characterization Of Collagen-Polypyrrole Constructs Using Direct-Ink Write Additive Manufacturing, Rooshan Arshad

Electronic Thesis and Dissertation Repository

Current efforts in the tissue engineering field are being directed towards the creation of platforms which will facilitate in instructing cells towards biologically relevant outcomes such as stem cell differentiation and disease pathophysiology. Traditional fabrication methods serve as a limiting factor for the production of such platforms as they lack feature and geometric complexity. Additive Manufacturing (AM) offers advantage over said methods by affording designers creative freedom and great control over printed constructs. Such constructs can then be used to create appropriate models for study- ing a plethora of tissues and structures. An AM methodology for Direct-Ink Write (DIW) printing …

Design Of Microporous Membranes For The Development Of Brain-On-Chip Devices, Andre Figueroa Milla

Biomedical Engineering Undergraduate Honors Theses

Traumatic brain injuries (TBIs) are a major global health concern that have an economic impact of $60 billion in the United States in related costs annually. Developing drugs for TBI treatment is an approach that currently faces limitations involving the permeability of the blood-brain barrier (BBB). The BBB naturally limits molecules from reaching the brain as a protective mechanism against disease, acting as a barrier during drug delivery. Understanding the BBB mechanically and chemically following a TBI could potentially assist future studies to alleviate the symptoms and long-term effects of TBI by pharmaceuticals. The Mechanobiology and Soft Materials Laboratory (MSML) …

Effect Of Polymer Composition Of Injectable Hydrogels On Programmable Release Of Methylene Blue, Mary Brandecker

Biomedical Engineering Undergraduate Honors Theses

Temperature sensitive hydrogels have been used as injectable hydrogels because the polymer solutions can be liquid at room temperature and gel at body temperature. Injectable hydrogels have been used in biomedical applications to deliver drugs and other small molecules throughout the body. Methylene Blue was used as the drug in this study for its antioxidant and neuroprotective properties. PluronicⓇ F127 (PF127) is a copolymer consisting of repeating units of polyethylene oxide and polypropylene oxide in the form PEO-PPO-PEO. PF127 in solution is a temperature sensitive hydrogel that transitions to a gel at body temperature at specific polymer compositions. A standard …

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 …

Tissue Equivalent Gellan Gum Gel Materials For Clinical Mri And Radiation Dosimetry, Pawel Brzozowski

Electronic Thesis and Dissertation Repository

Hydrogels contain high amount of water allowing their use as surrogates to human tissues with specific properties that can be tuned by additives. Gellan gum is a gel-forming material of interest and is a replacement for other common gelling agent with limited use as a tissue phantom. Therefore, this thesis examines the application of gellan gum gels as a novel magnetic resonance imaging (MRI) phantom with a design of experiments model to obtain tunable properties. The analysis was extended to include mechanical and electrical properties with a separate design of experiment. Gels doped with synthesized superparamagnetic iron oxide nanoparticles (SPIONs) …

Design Of Tissue-Specific Cellular Microenvironments For Adipose-Derived Stromal Cell Culture And Delivery, Arthi Shridhar

Electronic Thesis and Dissertation Repository

The development of in vitro cell culture models that investigate tissue-specific effects of the extracellular matrix (ECM) on stem/progenitor cell lineage-commitment can contribute towards the design of improved cell delivery strategies. This thesis developed processing methods that conserved ECM bioactivity to generate well-characterized 2- and 3-D culture platforms that facilitated the evaluation of ECM composition on the adipogenic and osteogenic differentiation of human adipose-derived stromal cells (ASCs). Initial work compared α-amylase and pepsin digestion as methods to fabricate ECM coatings. The effects of enzyme processing and ECM composition were explored using human decellularized adipose tissue (DAT) and bovine tendon collagen …

Suture-Less Trocar Site Closure Clip, Hares A. Patel

Electronic Theses and Dissertations

Introduction: Following laparoscopic surgery, there is a need, in many cases, to close trocar sites to prevent hernias. Currently, devices that exist on the market are suture based, but the lack of standardization in the suturing techniques together with the time-consuming nature of the procedure leads to the need for improvement in trocar site closure products. Trocar closure sites do not need to be fully closed on the fascial layer; rather, sufficiently blocking the hole at the abdominal wall can significantly reduce post-operative herniation. A retrospective study on trocar site herniation (TSH) after laparoscopic surgery indicates a TSH incidence …

The Effect Of Defects And Surface Modification On Biomolecular Assembly And Transport, Haneen Martinez

Nanoscience and Microsystems ETDs

Nanoscale transport using the kinesin-microtubule (MT) biomolecular system has been successfully used in a wide range of nanotechnological applications including self-assembly, nanofluidic transport, and biosensing. Most of these applications use the ‘gliding motility geometry’, in which surface-adhered kinesin motors attach and propel MT filaments across the surface, a process driven by ATP hydrolysis. It has been demonstrated that active assembly facilitated by these biomolecular motors results in complex, non-equilibrium nanostructures currently unattainable through conventional self-assembly methods. In particular, MTs functionalized with biotin assemble into rings and spools upon introduction of streptavidin and/or streptavidin-coated nanoparticles. Upon closer examination of these structures …