Molecular, Cellular, and Tissue Engineering Commons^™

Development Of A Long-Read Sequencing Protocol To Assess The Precision And Efficacy Of Gene Editing For Duchenne Muscular Dystrophy, Landon Andrew Burcham

Graduate Theses and Dissertations

This work establishes a method for assessing on-target precision due to CRISPR-Cas9 gene editing, especially within the context of exon skipping therapy for Duchenne Muscular Dystrophy. The proposed method utilizes an Oxford nanopore long-read sequencing approach to sequence amplified regions of DNA that have been edited using CRISPR-Cas9. NIH3T3 and C2C12 cell lines were treated with a dual-guide CRISPR-Cas9 system, that targets and deletes exon 23 from the DMD gene in mouse samples. Deletion PCR revealed deletion of exon 23 in both DNA and cDNA samples. Additionally, sequencing using Oxford Nanopore revealed targeted exon 23 deletion as the most prevalent …

Design Of A Four Channel Pulsatile Perfusion Bioreactor For Ex-Vivo Study Of Vascular Grafts, Thomas M. Fair

All Theses

Cardiovascular diseases are the leading cause of death in the United States. Atherosclerosis in peripheral arteries is a major contributing factor. Autologous saphenous vein grafts are the most common bypass grafts for treating peripheral artery diseases. A quarter of these grafts fail within a year, and around half have failed within 10 years of the initial surgery. Graft failure is attributed to the development of intimal hyperplasia indicated by the migration and proliferation of vascular smooth muscle cells, fibroblasts, and the deposition of extracellular proteins.

Increases in flow, pressure, the pulse frequency, and the differential of pressure that model an …

In Vitro Mitral Valve Model With Unrestricted Ventricular Access: Using Vacuum To Close The Valve And Enable Static Trans-Mitral Pressure, Sam E. Stephens, Alexander J. Kammien, Jacob C. Paris, Alexis P. Applequist, Neil B. Ingels, Hanna K. Jensen, Drew E. Rodgers, Charles R. Cole, Jonathan F. Wenk, Morten O. Jensen

Biomedical Engineering Faculty Publications and Presentations

Current in vitro models of the left heart establish the pressure difference required to close the mitral valve by sealing and pressurizing the ventricular side of the valve, limiting important access to the subvalvular apparatus. This paper describes and evaluates a system that establishes physiological pressure differences across the valve using vacuum on the atrial side. The subvalvular apparatus is open to atmospheric pressure and accessible by tools and sensors, establishing a novel technique for experimentation on atrioventricular valves. Porcine mitral valves were excised and closed by vacuum within the atrial chamber. Images were used to document and analyze closure …

Diagnosis Of Melanoma Disease State From Patient Blood Samples Using Photoacoustic Flow Cytometry, Jacob Salvatore

Electronic Theses and Dissertations

Approximately 1 in 50 Americans develop melanoma in their lifetime. Early detection of melanoma is pivotal to the survival of the patient, with a 99% survival rate for 5 or more years after an early diagnosis. Metastasis, or the spread of cancer, increases the chances of cancerous growth in other parts of the body. In this study, we propose the use of photoacoustic flow cytometry as a diagnostic of patient disease state, by closely monitoring the amount of circulating melanoma cells (CMCs) in a patient’s blood before and after chemotherapy treatments. Using this patient data, we were able to make …

Development Of An Integrated Salt Cartridge-Reverse Electrodialysis (Red) Device To Increase Electrolyte Concentrations Of Human Blood Flow To Power Biomedical Devices, Caroline Campbell

Chemical Engineering Undergraduate Honors Theses

Emerging technologies in nanotechnology and biomedical sciences have led to an increase in biomedical implantable devices including cardiac pacemakers, artificial organs, drug pumps, and sensors. These devices require continuous stable and reliable power to operate, which creates the demand for the need to find a safe, reliable, and stable power source. A promising avenue for a power source for these devices is a miniaturized reverse electrodialysis (RED) biopower cell design that utilizes the salinity differences between bloodstreams that flow inside the human body. Initial results of the RED system demonstrate that higher gradient salinity differences between streams lead to a …

Development And Applications Of In-Vitro And In-Silico Models Of The Cardiovascular System To Study The Effects Of Mechanical Circulatory Support., Moustafa Meki

Electronic Theses and Dissertations

Cardiovascular diseases (CVDs) are the leading cause of mortality globally. With ongoing interest in CVDs treatment, preclinical models for drug/therapeutic development that allow for fast iterative research are needed. Owing to the inherent complexity of the cardiovascular system, current in-vitro models of the cardiovascular system fail to replicate many of the physiological aspects of the cardiovascular system. In this dissertation, the main concern is with heart failure (HF). In advanced HF, patients may receive Left Ventricular Assist Devices (LVADs) as a bridge to transplant or destination therapy. However, LVADs have many limitations, including inability to adapt to varying tissue demand …

Development Of Light Actuated Chemical Delivery Platform On A 2-D Array Of Micropore Structure, Hojjat Rostami Azmand, Hojjat Rostami Azmand

Dissertations and Theses

Localized chemical delivery plays an essential role in the fundamental information transfers within biological systems. Thus, the ability to mimic the natural chemical signal modulation would provide significant contributions to understand the functional signaling pathway of biological cells and develop new prosthetic devices for neurological disorders. In this paper, we demonstrate a light-controlled hydrogel platform that can be used for localized chemical delivery in a high spatial resolution. By utilizing the photothermal behavior of graphene-hydrogel composites confined within micron-sized fluidic channels, patterned light illumination creates the parallel and independent actuation of chemical release in a group of fluidic ports. The …

Improve The Prototype Of Low-Cost Near-Infrared Diffuse Optical Imaging System, Chen Xu, Mohammed Z. Shakil

Publications and Research

Diffuse Optical Tomography (DOT) and Optical Spectroscopy using near-infrared (NIR) diffused light has demonstrated great potential for the initial diagnosis of tumors and in the assessment of tumor vasculature response to neoadjuvant chemotherapy. The aims of this project are 1) to test the different types of LEDs in the near-infrared range, and design the driving circuit, and test the modulation of LEDs at different frequencies; 2) to test the APDs as a detector, and build the receiver system and compare efficiency with pre-built systems. In this project, we are focusing on creating a low-cost infrared transmission system for tumor and …

Bacteria Analysis By Using A Supervised Machine Learning Algorithm Based On Droplet Microfluidics, Yulder Daniel Angarita

Electronic Theses and Dissertations

Sepsis is a major medical problem and massive resources have been invested in developing and evaluating alternative treatments. Statistics indicate that sepsis causes between one third and one half of all hospital deaths in the United States. Sepsis has a high impact on health care in the US, with direct sepsis costs in 2009 exceeding $15.4 billion. A research study found that a 1-hour delay in appropriate antimicrobial care resulted in a 7% - 10% rise in mortality. Several professional societies seek to reduce sepsis mortality by targeting the timely use of diagnostic tests and antimicrobial therapy. The diagnostic instruments …

3-Dimensional Carbon Micro-Electro Mechanical System (Mems) Electrode For Potential Dielectrophoretic Hepatic Cell Patterning Application, Wan Yahya Wan Nurlina

Student Works (2020-2029)

The maintenance of liver tissue unique pattern remains a great challenge in the generation of functional engineered liver tissue in vitro. This thesis presents a carbon dielectrophoresis (DEP) based cell patterning for generating hepatic cell constructs, mimicking biological hepatic lobule. A new design of electrode named interdigitated radiating-strips electrode (IRSE) was simulated and optimized using COMSOL Multiphysics 4.2a. The simulation result proved that IRSE is capable to provide uniform distribution of electric field for generating three-dimensional (3D) DEP force. The use of carbon electrodes has been an alternative to traditional metal electrodes and insulator structures for DEP applications since more …

Hydrolytic Degradation Study Of Polyphosphazene-Plga Blends, Riley Blumenfield

Honors Scholar Theses

The synthesis and in vitro degradation analysis of thin films of poly[(glycineethylglycinato)₇₅(phenylphenoxy)₂₅phosphazene]
(PNGEG₇₅PhPh₂₅) and poly[(ethylphenylalanato)₂₅(glycine-
ethylglycinato)₇₅phosphazene] (PNEPA₂₅GEG₇₅) blended with poly(lactic-co-glycolic acid) (PLGA) was conducted to determine the blends’ potential for use as scaffolding materials for tissue regeneration applications. The samples were synthesized with glycylglycine ethyl ester (GEG) acting as the primary substituent side group, with cosubstitution by phenylphenol (PhPh) and phenylalanine ethyl ester (EPA) to make the final product [1]. Blends of 25% polyphosphazene, 75% PLGA and 50% polyphosphazene, 50% PLGA were …

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 …

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 …

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 …

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 …

An Engineer’S Take On The Bone-Ligament Interface: Utilizing Novel Technology To Improve Clinical Outcomes, Emma Garcia

Shared Knowledge Conference

Ligament repair is a common surgical practice with a significant lack of viable replacements. The current gold standard for repair is the use of tendon grafts from cadavers or from another place in the patient’s body; however, these often cause more problems than they solve including immune responses or a lack of mobility in another place in the body. Synthetic replacements are of growing interest, though the ability to mimic the complex structure of the ligament and how it connects to the bone remains an obstacle. Our lab built a 3D bioprinter combined with an electrospinner to address this complicated …

Hyperspectral Imaging For Characterizing Single Plasmonic Nanostructure And Single-Cell Analysis, Nishir Sanatkumar Mehta

LSU Master's Theses

Orientation of plasmonic nanostructures is an important feature in many nanoscale applications such as photovoltaics, catalyst, biosensors DNA interactions, protein detections, hotspot of surface-enhanced Raman spectroscopy (SERS), and fluorescence resonant energy transfer (FRET) experiments. Silver nanocubes with significant spectral signatures between 400-700 nm are observed in this experimental research. Whereas study of single cells will enable the analysis of cell-to-cell variations within a heterogeneous population. These variations are important for further analysis and understanding of disease propagation, drug development, stem cell differentiation, embryos development, and how cells respond to each other and their environment. Adipose-derived mesenchymal stem cells possess the …

Developing Droplet Based 3d Cell Culture Methods To Enable Investigations Of The Chemical Tumor Microenvironment, Jacqueline Alexandria De Lora

Biomedical Sciences ETDs

Adaptation of cancer cells to changes in the biochemical microenvironment in an expanding tumor mass is a crucial aspect of malignant progression, tumor metabolism, and drug efficacy. In vitro, it is challenging to mimic the evolution of biochemical gradients and the cellular heterogeneity that characterizes cancer tissues found in vivo. It is well accepted that more realistic and controllable in vitro 3D model systems are required to improve the overall cancer research paradigm and thus improve on the translation of results, but multidisciplinary approaches are needed for these advances. This work develops such approaches and demonstrates that new droplet-based cell-encapsulation …

Preparation For The Study Of Gene Expression Aboard The International Space Station, Amber Paturzo

STAR Program Research Presentations

WetLab-2 performs quantitative polymerase chain reaction (qPCR) aboard the micro-gravity environment of the International Space Station in order to research how the stress of micro-gravity affects an organism’s RNA production and gene expression. This on-station qPCR allows researchers to gather information earlier and thereby allow them to modify experiments in a matter of hours rather than wait until the next flight opportunity. After a successful validation flight on SpaceX-8 and follow-up investigation on SpaceX-9, system components must now be adjusted for a new type of biological sample. Specifically, the WetLab-2 facility includes a commercial qPCR instrument (RNA SmartCycler), a sample …

Detection Of Leukocytes Stained With Acridine Orange Using Unique Spectral Features Acquired From An Image-Based Spectrometer, Courtney J. Hunter

Biomedical Engineering Undergraduate Honors Theses

A leukocyte differential count can be used to diagnosis a myriad blood disorders, such as infections, allergies, and efficacy of disease treatments. In recent years, attention has been focused on developing point-of-care (POC) systems to provide this test in global health settings. Acridine orange (AO) is an amphipathic, vital dye that intercalates leukocyte nucleic acids and acidic vesicles. It has been utilized by POC systems to identify the three main leukocyte subtypes: granulocytes, monocytes, and lymphocytes. Subtypes of leukocytes can be characterized using a fluorescence microscope, where the AO has a 450 nm excitation wavelength and has two peak emission …

Development And Evaluation Of A Biocompatible Electroactive Sensor For Continuous Blood Pressure Measurement., Scott D. Cambron

Electronic Theses and Dissertations

Piezo-active composites have been implemented for sensing and transduction for decades. The 0-3 ceramic/polymer composite is one of the most common composite types used for sensing applications, owing to their tailorable properties of the two-phase composition, consisting of a three-dimensionally connected polymer/rubber matrix (inactive phase) with a dispersion of isolated piezo-ceramic particles (active phase). This thesis describes a method to develop novel biocompatible perivascular band comprised of a two-phase piezo-active composite to be fabricated using simple manufacturing processes. Biomaterials such as tissue scaffolds comprised of silk fibroin (SF) and chitosan (CS), and biocompatible soft rubbers will be implemented as the …

Development Of An Electrospun And 3d Printed Cellular Delivery Device For Dermal Wound Healing, Ryan M. Clohessy

Theses and Dissertations

The goal of this research was to develop a system of individualized medicine that could be applied to dermal wounds serving as a wound dressing and synthetic extracellular matrix while delivering stem cells to the wound bed. First, fabrication parameters for electrospinning polymer fibers were determined. This involved evaluating fiber morphology with respect to polymer selection and solution concentration. Next, construct fabrication was examined to produce an integrated void space, or cargo area, suitable to maintain stem cells. In vitro studies to ensure stem cell viability and phenotype were conducted, and results supported the notion that cells could be administered …

Surface Engineering Of Titanium Alloy To Stimulate The Bone Regeneration, Yubin Meng, Zhenduo Cui, Zhaoyang Li, Xue Li, Jin Zhao, Xubo Yuan, Xianjin Yang

The 8th International Conference on Physical and Numerical Simulation of Materials Processing

No abstract provided.

Breast Cancer/Stromal Cells Coculture On Polyelectrolyte Films Emulates Tumor Stages And Mirna Profiles Of Clinical Samples, Amita Daverey, Karleen M. Brown, Srivatsan Kidambi

Department of Chemical and Biomolecular Engineering: Faculty Publications

In this study, we demonstrate a method for controlling breast cancer cells adhesion on polyelectrolyte multilayer (PEM) films without the aid of adhesive proteins/ ligands to study the role of tumor and stromal cell interaction on cancer biology. Numerous studies have explored engineering coculture of tumor and stromal cells predominantly using transwell coculture of stromal cells cultured onto coverslips that were subsequently added to tumor cell cultures. However, these systems imposed an artificial boundary that precluded cell−cell interactions. To our knowledge, this is the first demonstration of patterned coculture of tumor cells and stromal cells that captures the temporal changes …

Design And Development Of Two Component Hydrogel Ejector For Three-Dimensional Cell Growth, Thomas Dunkle, Jessica Deschamps, Connie Dam

Honors Scholar Theses

Hydrogels are useful in wound healing, drug delivery, and tissue engineering applications, but the available methods of injecting them quickly and noninvasively are limited. The medical industry does not yet have access to an all-purpose device that can quickly synthesize hydrogels of different shapes and sizes. Many synthesis procedures that have been developed result in the formation of amorphous hydrogels. While generally useful, amorphous hydrogels exhibit limited capability in tissue engineering applications, especially due to their viscous properties. This endeavor aims to modulate the appropriate gelation parameters, optimize the injection process, and create a prototype that allows for the extrusion …

Characterization Of Bone Material Properties And Microstructure In Osteogenesis Imperfecta/Brittle Bone Disease, John Robert Jameson

Dissertations (1934 -)

Osteogenesis imperfecta (OI) is a genetic disorder primarily associated with mutations to type I collagen and resulting in mild to severe bone fragility. To date, there is very little data quantifying OI cortical bone mechanics. The purpose of this dissertation was to investigate bone microstructure, mineralization, and mechanical properties in adolescents with OI. Characterization studies were performed on small osteotomy specimens obtained from the extremities during routine corrective surgeries. Nanoindentation was used to examine the longitudinal elastic modulus and hardness at the material level for mild OI type I vs. severe OI type III. Both modulus and hardness were significantly …

Utilizing Fast Spin Echo Mri To Reduce Image Artifacts And Improve Implant/Tissue Interface Detection In Refractory Parkinson’S Patients With Deep Brain Stimulators, Subhendra N. Sarkar, Pooja R. Sarkar, Efstathios Papavassiliou, Rafael Rojas

Publications and Research

Introduction. In medically refractory Parkinson’s disease (PD) deep-brain stimulation (DBS) is an effective therapeutic tool. Postimplantation MRI is important in assessing tissue damage and DBS lead placement accuracy. We wanted to identify which MRI sequence can detectDBS leads with smallest artifactual signal void, allowing better tissue/electrode edge conspicuity.

Methods. Using an IRB approved protocol 8 advanced PDpatientswere imagedwithinMRconditional safety guidelines at lowRF power (SAR ≤ 0.1 W/kg) in coronal plane at 1.5T by various sequences.The image slices were subjectively evaluated for diagnostic quality and the lead contact diameters were compared to identify a sequence least affected by metallic leads.

Results …

Method And Apparatus For Electrical Stimulation Of Tissues Using Signals That Minimize The Effects Of Tissue Impedance, Jeffrey Hargrove

Mechanical Engineering Patents

A tissue stimulation system that generates an electrical tissue stimulation signal configured to reduce tissue impedance and increase depth of signal penetration. The use of leads is dynamically controlled and altered between conducting biopotential voltages, conducting electrical tissue stimulation signals, and grounding, in response to a computational analysis of biopotential data acquired from a region of tissue to be stimulated.

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, …