Stem Cell Regulation Using Nanofibrous Membranes with Defined Structure and Pore Size

Electrospun nanofibers have been researched extensively in the culturing of stem cells to understand their behavior since electrospun fibers mimic the native extracellular matrix (ECM) in many types of mammalian tissues. Here, electrospun nanofibers with defined structure (orientation/alignment) and pore size could significantly modulate human mesenchymal stem cell (hMSC) behavior. Controlling the fiber membrane pore size was predominantly influenced by the duration of electrospinning, while the alignment of the fiber membrane was determined by parallel electrode collector design. Electric field simulation data provided information on the electrostatic interactions in this electrospinning apparatus.hMSCs on small-sized pores (~3-10 µm²) tended to promote the cytoplasmic retention of Yes-associated protein (YAP), while larger pores (~30-45 µm²) promoted the nuclear activation of YAP. hMSCs also displayed architecture-mediated behavior, as the cells aligned along with the fiber membranes orientation. Additionally, fiber membranes affected nuclear size and shape, indicating changes in cytoskeletal tension, which coincided with YAP activity. The mechanistic understanding of hMSC behavior on defined nanofiber structures seeks to advance their translation into more clinical settings and increase biomanufacturing efficiencies.
Date: August 2022
Creator: Blake, Laurence A
System: The UNT Digital Library

Development and Characterization of Compliant Bioelectronic Devices for Gastrointestinal Stimulation

In this research, we aimed to develop thin-film devices on a polymer substrate and an alternative 3D-printed device with macroelectrodes for treating gastrointestinal (GI) conditions. First, the fabrication of thin-film devices was demonstrated on a softening thiol-ene/acrylate polymer utilizing titanium nitride (TiN) as electrode material. This was achieved by utilizing cleanroom fabrication processes such as photolithography, wet and dry etching. The functionality of the device was shown by performing electrochemical characterization tests, mainly cyclic voltammetry, electrochemical impedance spectroscopy, and voltage transient. We synthesized a novel thiol-ene/acrylate polymer based on 1,3,5-triallyl-1,3,5-triazine-2,4,6(1H,3H,5H)-trione (TATATO), trimethylolpropanetris (3-mercaptopropionate) (TMTMP), and polyethylene glycol diacrylate (PEGDA). We show that this stretchable shape memory polymer substrate is well suited for cleanroom processes. Finally, for the high throughput of the wearable devices with electrodes size 10 mm in diameter, we implemented single electrode fabrication using printed circuit boards (PCBs) and depositing gold (Au) and TiN on the plated side of PCBs utilizing the sputtering tool. This step was followed by the assembly of those single electrodes on the flexible 3D printed device. We showed that the TiN electrode material performed better in terms of charge storage capacity and charge injection capacity than the widely used stainless steel electrode material …
Date: December 2023
Creator: Chitrakar, Chandani
System: The UNT Digital Library
Engineering Whole Cell-Based Biosensors for Heavy Metal Detection Using Metalloregulatory Transcriptional Repressors of the SmtB/ArsR Family (open access)

Engineering Whole Cell-Based Biosensors for Heavy Metal Detection Using Metalloregulatory Transcriptional Repressors of the SmtB/ArsR Family

This study focuses on engineering whole cell-based biosensors for heavy metal detection. Through the exploitation of metalloregulatory proteins, fabrication of metal ion-responsive biosensors is achieved. Metalloregulatory proteins of the SmtB/ArsR family including arsenite-responsive ArsR, cadmium-responsive CadC, zinc-responsive CzrA, and nickel-responsive NmtR were evaluated as biosensor sensing modules. Characterization of these four metal sensing modules was accomplished through quantification of a reporter green fluorescence protein (gfp) gene. As such, biosensors pCTYC-r34ArsR-pL(ArsOvN)GFP and pCTYC-r34CadC-pL(CadOv1)GFP displayed excellent gfp expression and sensitivity to As(III) and Cd (II), respectively. These two biosensors were consequently selected and successfully implemented in soil bacterium Pseudomonas putida. Lastly, a proof of concept arsenite-responsive genetic toggle switch is proposed utilizing PurRcelR467 (PC47), a cellobiose-responsive gene, and an LAA degradation tag. Overall, this study expands the bank of metalloregulatory bioparts for heavy metal sensing in the aim of constructing an optimized water monitoring system.
Date: May 2022
Creator: Draeger, Alison
System: The UNT Digital Library
Parkinson's Disease and UPDRS-III Prediction Using Quiet Standing Data and Applied Machine Learning (open access)

Parkinson's Disease and UPDRS-III Prediction Using Quiet Standing Data and Applied Machine Learning

Parkinson's disease (PD) is a neurodegenerative disease that affects motor abilities with increasing severity as the disease progresses. Traditional methods for diagnosing PD require specialists scoring qualitative symptoms using the motor subscale of the Unified Parkinson's Disease Rating Scale (UPDRS-III). Using force-plate data during quiet standing (QS), this study uses machine learning to target the characterization and prediction of PD and UPDRS-III. The purpose of predicting different subscores of the UPDRS-III is to give specialists more tools to help make an informed diagnosis and prognosis. The classification models employed classified PD with a sensitivity of 87.5% and specificity of 83.1%. Stepwise forward regression indicated that features correlated with base of support were most useful in the prediction of head rigidity (r-square = .753). Although there is limited data, this thesis can be used as an exploratory study that evaluates the predictability of UPDRS-III subscores using QS data. Similar prediction models can be implemented to a home setting using low-cost force plates as a novel telemedicine technique to track disease progression.
Date: May 2021
Creator: Exley, Trevor Wayne
System: The UNT Digital Library
Effects of Attachment Height and Rail Material of Resistance Training Sled on Trunk Lean and Jerk During Linear Acceleration Training (open access)

Effects of Attachment Height and Rail Material of Resistance Training Sled on Trunk Lean and Jerk During Linear Acceleration Training

Sprint acceleration training has been highly researched and found that resistance sleds are one of the most effective tools for maximizing training adaptations. The resistance sled is being used by many of the world leaders in athletic training but has yet to be researched for the kinetic and kinematic effects some of its key components cause. The aim of this study was to better understand the effects of the attachment height on the sled and sled rail material on the user's trunk lean and jerking effect caused by the sled. This was done because it was hypothesized that the attachment height has a direct impact on trunk lean and sled rail material has a direct impact on jerk caused by the sled. To test these assumptions, experimental and theoretical data was collected using a single subject study analyzing trunk lean and acceleration values of the sled. The results presented a significant decrease in trunk lean (more horizontal line of action) when the attachment height was raised. Additionally, no significant values were attained to support the assumption that by modifying the sled rail material, jerking effects will decrease. The results indicate that there is a direct correlation between attachment height and …
Date: May 2021
Creator: Fitzgerald, Sean
System: The UNT Digital Library
Bio-Inspired Designs to Reduce Human-Exoskeleton Interaction to Prevent Falls in an Aging Population (open access)

Bio-Inspired Designs to Reduce Human-Exoskeleton Interaction to Prevent Falls in an Aging Population

As a large generation ages, the collective financial and ethical responsibility to prevent egregious bodily harm through fall prevention and gait assistant exoskeleton devices increases. Risk for falls increases with age and the severity of the fall does as well. To support this elderly population, motorized exoskeletons can both increase stability as well as respond faster to fall scenarios, but current models do not more around the existing biological framework. Giving participants a range of motion in key pelvic areas can closely approximate synchronous rotation around the femoral head, while limiting an increase in their sagittal profile. Utilizing 3D printed components while incorporating existing orthic methods provide short production times on modular designs. Although primarily mechanically based, these designs consider electronic requirements and are capable for supporting movement for a 200 lbs. user at a brisk walking pace for 1 hour.
Date: August 2021
Creator: Gates, Edward Sean
System: The UNT Digital Library

Engineering Cardiac Organoid Vascularization via Fluid Shear Stress and Vascular-Promoting Growth Factors

Cardiovascular disease (CVD) is the leading cause of death internationally. Efforts to decrease CVD death has been explored through stem cell technology, specifically organoid formation. Current cardiac organoid models lack the vascular networks for nutrient supply and maturation. In this study, pillar perfusion technology is used to fabricate cardiac organoids and induce vascularization via dynamic culturing and the addition of vascular promoting growth factors (GFs). In addition to this study, a millifluidic chip is engineered for shear stress application via flow simulations and experimental flow analysis. We successfully optimized the millifluidic chip to achieve fluid shear stress of 20mPa and validated through particle tracking velocimetry using 0.1um diameter beads under flow. The results of cardiac organoids displayed contraction and growth of endothelial cells (ECs) under dynamic flow with GFs. In addition, smooth muscle cells (SMCs) displayed growth via GFs in both dynamic and static culturing. Although vascular networks were not present in all conditions of this experiment, this thesis can serve a basis for searching other methods of inducing vascularization.
Date: August 2022
Creator: Huerta Gomez, Angello
System: The UNT Digital Library

Novel Phase-Chance Soft Actuators Controlled via Peltier

Soft actuation methods are a developing field of robotics deemed suitable for physical human-robot interactions due to the adaptability of materials and compliant structures. Thermo-active soft actuators are a subset of these which convert thermal energy to mechanical work in the form of elongation, bending, or twisting to conform to the environment. This study is divided into three major studies that all use actuators with a working principle of phase-change fluid vaporizing for expansion with applied heat from a Peltier. The first study evaluates the bandwidth and efficiency between (i) traditional Joule heating, and (ii) Peltier heating, finding that Peltier heating can considerably improve the operational bandwidth of the actuator. The second study uses a thin membrane actuator placed in a braided mesh to form a McKibben muscle capable of lifting 5N, and formed into a gripper capable of manipulating objects within the environment. The third study uses actuators of a solid, hollow and flexible Peltier embedded silicone structure and are evaluated and optimized in order to increase actuation speed, finding that the embedded flexible Peltier design was able to elongate over 50% of its original height in 20 seconds. The overall aim of all of these studies was to …
Date: July 2023
Creator: Johnson, Daniel Cody
System: The UNT Digital Library
Viability Study of Nylon-12 Carbon Fiber Filaments for Use in the Construction of a Powered Lower Body Exoskeleton via Fused Deposition Modeling by Means of Computer Simulation (open access)

Viability Study of Nylon-12 Carbon Fiber Filaments for Use in the Construction of a Powered Lower Body Exoskeleton via Fused Deposition Modeling by Means of Computer Simulation

Members of the elderly population is disproportionately prone to experiencing mobility impairment due to their aging bodies and as a result have frail bodies that are at a higher risk of grave injury due to falling. In order to combat this assistive mobility devices such as exoskeletons have been developed to help patients enhance their range of motion. With additive manufacturing techniques, such as fused deposition modeling (FDM), becoming a more mainstream form of design, the inclusion of lightweight polymers such as nylon 12 as primary construction materials for these devices has increased. In this thesis computer aided design (CAD) software was used to design a prototype lower body exoskeleton and simulation software was used to give the device the characteristics of Stratasys' nylon 12 carbon fiber FDM material to verify it if could be used as the primary construction material for this device when extruded from a FDM printer on either the XZ or ZX printing plane. From the simulations it was found that the material printed along the XZ plane could create a device that could withstand the weight of an average elderly male patient (200 lbs.) as well as the 35 lbs. of force applied to the …
Date: May 2021
Creator: Joiner, Michael Andrew Lown
System: The UNT Digital Library

Mesenchymal Stem Cells Encapsulated and Aligned in Self-Assembling Peptide Hydrogels

This study presents a viable strategy using fmoc-protected peptides hydrogels, to encapsulate and stretch mesenchymal stem cells (MSC). To explore the peptide hydrogel potential, a custom mechanical stretching device with polydimethylsiloxane (PDMS) chambers were used to stretch MSCs encapsulated in Fmoc hydrogels. We investigated the impact of fmoc- FF prepared in dimethyl sulfoxide (DMSO), 1,1,1,3,3,3-hexafluoro-2-propanol (HFP) and deionizied water in the self-assembly, and mechanical properties of the gels. The peptide hydrogel is formed through molecular self-assembly of peptide sequence into β-sheets that are connected with the π-π aromatic stacking of F-F groups. The hydrogels provided a stiff, hydrated gel with round nanofiber morphology representing an elastic modulus of 174-266 KPa. MSCs cultured on peptide hydrogels undergo viability, morphology, and alignment evaluations using MTT, live/dead, and phalloidin (F-actin) staining. The F-actins of 3D- cultured MSCs in Fmoc-FF/HFP, and Fmoc-FF/DMSO followed by mechanical stretching showed elongated morphology with defined microfilament fibers compared to the round and spherical F-actin shape of the control cells. Peptide gels with 5mM concentration preserved 100% viability of MSC. Results reveals the feasibility and conditions for successful cell encapsulation and alignment within peptide hydrogels. Encapsulation of MSC in peptide nanofiber followed by a stretching process present a promising …
Date: December 2023
Creator: Kasani, Yashesh Varun
System: The UNT Digital Library

Engineering a Microfluidic Blood-Brain Barrier on a Silicon Chip

The blood-brain barrier (BBB) is composed of brain microvascular endothelial cells (BMECs), pericytes, and astrocytic endfeet, which regulate the transport of molecules into and out of the brain. BMECs possess intrinsic barrier properties that limit the passage of approximately 98% of small molecules into the brain in healthy individuals. However, in some brain diseases, the BBB undergoes structural and functional alterations, which can contribute to disease progression. In this study, we aimed to investigate the BBB by exploring the effects of endothelial cell stretching and the optimal dimensionality of stretching to enhance endothelium barrier tightness in Chapter 2. Subsequently, we developed an endothelium gradient stretching device to further examine the stretching effect in Chapter 3. Additionally, we investigated the promotion of endothelium tightness through the use of electrospun fibers, wherein we controlled the pore size. Based on these findings, we designed and fabricated an organ chip model that incorporates mechanical stretching, microfluidic techniques, electrospun fibers, and hydrogel extracellular matrix (ECM). The results of permeability testing demonstrated that this chip significantly improved the tightness of microvascular selective transport ability and has the potential to be used in drug sorting for central nervous system (CNS) diseases.
Date: July 2023
Creator: Liu, Jiafeng
System: The UNT Digital Library

Development of Biomimetic Human Lung Alveolus Chip

The potential of physiologically relevant in vitro cell culture models for studying physiological and pathophysiological phenomena has been widely recognized as replacements for animal and conventional in vitro models. To create models that accurately replicate the structure and function of tissues and organs, it is essential to comprehend the biophysical and mechanical features of the extracellular matrix (ECM) and incorporate them into the in vitro cell culture models. Therefore, we first aimed to investigate how nanotopography can modulate cell behaviors by studying cell behaviors on nanostructures of various aspect ratios on a cobalt-chromium-molybdenum (CoCrMo) alloy surface. We also explored the impact of nanofibrous membranes on the formation of alveolar epithelium, which is critical for lung alveolar interstitium chips. In addition, we investigated the effect of mechanical stretch on cell behaviors and focused on how the dimensionality of the stretch affects cell behaviors. To create physiologically relevant in vitro models based on our findings, we engineered a stem cell niche using a combination of nanofibrous membranes, mechanical stretch, and a soft substrate, and evaluated its impact on stem cell behaviors. Finally, we created a biomimetic human lung interstitium chip for application in physiological and pathophysiological in vitro studies.
Date: May 2023
Creator: Man, Kun
System: The UNT Digital Library

Nanolithographic Approaches to Probing Cell Membrane Modulation

Metastatic cancer is more dangerous and difficult to treat than pre-metastatic cancer. Ninety percent of cancer-related deaths are caused by metastatic cancer. When cells go through metastases, they go through changes that allow them to break away from the primary tumor and invade secondary tissues. These changes, in lipid membrane composition and cellular glycocalyx, make the cell more resistant to therapeutics. Actin cytoskeleton contractility plays a major role in these changes, as increased contractility has been linked to upregulation of phosphoinositides and production of glycoproteins. Light induced molecular adsorption of proteins (LIMAP) was used to control the actin arrangement and cell shape in order to mimic and study metastatic cells. Negatively charged proteins electrostatically adhere to the surface in order to create patterns for the cells to stick. Neutravidin was conjugated to poly(glutamic acid) to improve attachment to the surface. We observed differences in cell shape and phosphoinositide behavior based on LIMAP patterning. Additionally, expression of key glycoproteins related to cancer metastasis increased with increased actin contractility. The actin cytoskeleton was the main driver of changes to the cell membrane and glycocalyx.
Date: May 2022
Creator: Mathis, Katelyn
System: The UNT Digital Library

Magnetoresponsive Layer-by-Layer (LBL) Polyelectrolyte Microcapsules Exposed to Low Frequency Alternating Magnetic Field for Drug Delivery to Breast Cancer Cells

Layer-by-layer (LBL) polyelectrolyte capsules can be modified to incorporate stimuli such as superparamagnetic nanoparticles which respond to a magnetic field only when it is turned on. Thus, they can act as a switch to load or unload their drug cargo on demand. Specifically, magnetite is incorporated into bilayer capsules made of alternating poly(allylamine hydrochloride) (PAH) and poly(sodium-p-styrenesulfonate) (PSS) which surrounds calcium carbonate core. The core is then dissolved using ethylenediaminetetraacetic acid (EDTA). These capsules are loaded with at FITC-BSA conjugate and examined with fluorescence to show the unloading of the FITC-BSA from capsules as it brightens the entire field of view of the microscope. The results suggest that we can next load and unload an anticancer drug such as doxorubicin using the combination of microcapsule and alternating magnetic field (AMF) to treat the cancer cells. Preliminary data interprets that the low frequency AMF we use has little to no adverse effect cells viability. This coincides with the general thought that low frequency AMF signals are not harmful to humans. Therefore, as an alternative to hyperthermia methods which use heat, it may be possible to deliver the anticancer drugs specifically to the cells when and where it is needed.
Date: December 2023
Creator: Powell, Robert Darrel
System: The UNT Digital Library
Modeling of Hypertrophic Cardiomyopathy Using hiPSC-Derived Cardiomyocytes with Static Mechanical Stretching (open access)

Modeling of Hypertrophic Cardiomyopathy Using hiPSC-Derived Cardiomyocytes with Static Mechanical Stretching

The heart is a dynamic environment that is constantly experiencing some degree of remodeling from the point of development, all the way through adulthood. While many genetic components may contribute to the overall presentation of hypertrophic cardiomyopathy (HCM), mutations occurring in sarcomere components such as myosin binding protein C3 (MYBPC3) are of the greatest popularity for study. Aiming to understand the mechanisms underlying heart diseases and to develop effective treatments that circumvent the need for direct patient study, we investigated the use of a platform to mimic the unique physiological conditions of HCM within an in-vitro setting. Following the induction of mechanical stretch on three human induced pluripotent stem cell derived cardiomyocyte (hiPSC-CM) cell lines containing mutations for MYBPC3 (WT, HET, HOM), all displayed HCM like reactions in calcium waveform. In conclusion, this system demonstrated the potential to apply a constant, static strain to healthy and mutated hiPSC-CMs for the MYBPC3 protein to model HCM in-vitro.
Date: May 2023
Creator: Rogozinski, Nicholas
System: The UNT Digital Library

Differentiating Cardiac Organoids with Chamber Formations

Considering that both cardiovascular disease (CVD) and congenital heart diseases (CHD) are still the leading cause of morbidity and mortality worldwide, there is a need for a robust and reliable cardiac model. Cardiac organoids are complex, three-dimensional cellular constructs that recapitulate the processes of the human embryonic heart. However, certain vital morphological features within the fetus are not yet replicable with cardiac organoids. Here we report our investigation to generate cardiac organoids with chamber formations. Our method involves modulating the Wnt pathway at two different instances while also implementing two cell seeding densities, all to determine the most optimized that to produce chamber formations within cardiac organoids.
Date: July 2023
Creator: Seddoh, Percyval Prince-Danny
System: The UNT Digital Library
In Vitro Electrochemical Evaluation of Bioelectronic Arrays (open access)

In Vitro Electrochemical Evaluation of Bioelectronic Arrays

In this paper, I sought to identify and develop a protocol on electrode arrays as a result of rapid aging by applying rapid current over time. We, however, apply a different approach by using phosphate buffer solution (PBS) to mimic the conditions of the body. Here we have established an in vitro protocol for accelerated aging, a process that involves testing in extreme conditions such as oxygen, heat, sunlight, humidity, and vibration aimed at speeding the normal aging process of items; on commercially available shape memory polymer electrode arrays from Qualia over a period of 30 days in PBS. Two electrode arrays were placed in 37°C and 2 were placed in 57°C. Open lead electrochemical impedance spectroscopy (EIS) was conducted on the electrode arrays. Overall, the results showed there were differences in average impedance during this accelerated aging protocol. At 37°C we see that the average impedance values increased as the electrodes were aged at 1kHz from an average of 4.15E6 to 9.14E6 Ohms. At 57°C electrode arrays 4 and 5 showed strong P values well above 0.05, but average impedance increased drastically from 3.27E6 to 9.97E6 and P value of 0.04 from measurement day 24 to day 30. This …
Date: December 2021
Creator: Singh, Sukhpreet
System: The UNT Digital Library
Modeling Hypertrophic Cardiomyopathy Using Genome-Edited Human Induced Pluripotent Stem Cell-Derived Cardiomyocytes in Response to Dynamic Mechanotransduction (open access)

Modeling Hypertrophic Cardiomyopathy Using Genome-Edited Human Induced Pluripotent Stem Cell-Derived Cardiomyocytes in Response to Dynamic Mechanotransduction

Familial hypertrophic cardiomyopathy (HCM) is a genetic disease largely caused by a mutation in myosin binding protein C (MYBPC3) and it affects about 1:500 population leading to arrhythmic sudden death, heart failure, and atrial fibrillation. MYBPC3 activates calcium-induced actin-myosin filament sliding within the cardiac sarcomere, creating the force necessary for heart contraction. The underlying molecular mechanisms causing HCM phenotype remain elusive, therefore, there is an urgent need for a reliable in vitro human HCM model to investigate the pathogenesis of HCM. This study utilized isogenic human induced pluripotent stem cell-derived cardiomyocytes (hiPSC-CMs) with MYBPC3 gene mutation (wildtype, heterozygous, homozygous) and further micropatterned them into fiber-like structures on polyacrylamide hydrogels of physiological and fibrotic-like stiffnesses. Cells were cultured for an extended culture time up to 60 days and their morphology/attachment, contractility, and calcium transient were extensively and carefully evaluated. It was found that MYBPC3 knockout cells maintained the highest contraction amplitude, but had increased contraction, and relaxation durations, decreased calcium transient amplitude, as well as time to peak and decay times over the culture period in comparison to the isogenic wildtype. Overall, this study demonstrates that hiPSC-CMs can be successfully patterned and cultured for an extended time on hydrogels forming end-to-end …
Date: May 2022
Creator: Strimaityte, Dovile
System: The UNT Digital Library
Integrating a Brain Control Interface towards the Development of a Retrofitted, Low-Cost, Open Sourced, Electric Wheelchair (open access)

Integrating a Brain Control Interface towards the Development of a Retrofitted, Low-Cost, Open Sourced, Electric Wheelchair

The Emotiv Insight is a commercially available, low-cost, mobile EEG device that is commonly overshadowed by its costlier counterpart, the Emotiv EPOC. The purpose of this report is to investigate if the Emotiv Insight is a suitable headset to be used as a controlling factor in conjunction with an Arduino microcontroller and various electrical components that are used towards the development of an open-sourced, affordable electric wheelchair with the primary goal of providing those who either do not have the financial resources or the physical capability to operate a traditional wheelchair due to their disability a viable option to improve their quality of life. All of the C++ code, STL files used to fabricate the 3d-printed components are uploaded to a GitHub repository as open sourced files to allow individuals with access to a 3d-printer to either build the open sourced wheelchair for their personal use, or refine the design to suit their needs.
Date: August 2020
Creator: Ta, Christopher Ian
System: The UNT Digital Library