Epithelial-mesenchymal transition (EMT) regulates the cellular processes of migration, growth, and proliferation - as well as the collective cellular process of tissue remodeling - in response to mechanical and chemical stimuli in the cellular microenvironment. Cells of the epithelium form cell-cell junctions with adjacent cells to function as a barrier between the body and its environment. By distributing localized stress throughout the tissue, this mechanical coupling between cells maintains tensional homeostasis in epithelial tissue structures and provides positional information for regulating cellular processes. Whereas in vitro and in vivo models fail to capture the complex interconnectedness of EMT-associated signaling networks, previous computational models have succinctly reproduced components of the EMT program. In this work, we have developed a computational framework to evaluate the mechanochemical signaling dynamics of EMT at the molecular, cellular, and tissue scale. First, we established a model of cell-matrix and cell-cell feedback for predicting mechanical force distributions within an epithelial monolayer. These findings suggest that tensional homeostasis is the result of cytoskeletal stress distribution across cell-cell junctions, which organizes otherwise migratory cells into a stable epithelial monolayer. However, differences in phenotype-specific cell characteristics led to discrepancies in the experimental and computational observations. To better understand the role of mechanical cell-cell feedback in regulating EMT-dependent cellular processes, we introduce an EMT gene regulatory network of key epithelial and mesenchymal markers, E-cadherin and N-cadherin, coupled to a mechanically-sensitive intracellular signaling cascade. Together these signaling networks integrate mechanical cell-cell feedback with EMT-associated gene regulation. Using this approach, we demonstrate that the phenotype-specific properties collectively account for discrepancies in the computational and experimental observations. Additionally, mechanical cell-cell feedback suppresses the EMT program, which is reflected in the gene expression of the heterogeneous cell population. Together, these findings advance our understanding of the complex interplay in cell-cell and cell-matrix feedback during EMT of both normal physiological processes as well as disease progression.

Optimal management of fractures, post-traumatic arthritis and instability of the distal radioulnar joint (DRUJ) requires an understanding of the forces existing across this joint as a function of the activities of daily living. However, such knowledge is currently incomplete. The goal of this research was to quantify the loads that occur at the DRUJ during forearm rotation and to determine the effect that individual muscles have on those loads. Human and cadaver studies were used to analyze the shear (A-P), transverse (M-L) and resultant forces at the DRUJ and to determine the role that 15 individual muscles had on those forces. Data for scaling the muscles forces came from EMG analysis measuring muscle activity at nine positions of forearm rotation in volunteers during isometric pronation and supination. Muscle orientations were determined from the marked muscle origin and insertion locations of nine cadaveric arms at various stages of forearm rotation. The roles that individual muscles played in DRUJ loading were analyzed by removing the muscle of interest from the analysis and comparing the results. The EMG portion of this study found that the pronator quadratus, pronator teres, brachioradialis, flexor carpi radialis and palmaris longus contribute significantly to forearm pronation. The supinator, biceps brachii, and abductor pollicis longus were found to contribute significantly to supination. The results of the DRUJ analysis affirm that large transverse forces pass from the radius to the ulnar head at all positions of forearm rotation during pronation and supination (57.5N-181.4N). Shear forces exist at the DRUJ that act to pull the radius away from the ulna in the AP direction and are large enough to merit consideration when examining potential treatment options (7.9N-99.5N). Individual muscle analysis found that the extensor carpi radialis brevis, extensor pollicis longus, extensor carpi ulnaris, extensor indicis and palmaris longus had minimal effect on DRUJ loading. Other than the primary forearm rotators (pronator quadratus, pronator teres, supinator, biceps brachii), the muscles that exhibited the largest influence on DRUJ loading were the abductor pollicis longus, brachialis, brachioradialis, extensor carpi ulnaris, flexor carpi radialis, and flexor carpi ulnaris.

Valve replacement for critical heart valve diseases is in many cases not an option. Our clinical experience in pediatric compassionate care has shown robust function of porcine small intestinal submucosa (PSIS) valves. We assessed functional effectiveness of 4ply (~320µm) and 2ply (~166µm) PSIS mitral valves under pediatric-relevant hemodynamic pulsatile conditions. Key conclusions: (i)PSIS valves demonstrated statistically similar acute functionality in comparison to a commercially available valve. (ii)Energy losses were similar (p>0.05) under pediatric conditions which was not the case under adult aortic conditions. (iii)2ply valves were observed to be superior to 4ply, based on the robust hydrodynamic data, the mechanical properties suitable for pediatric applications and de-novo tissue replacement potential with less demand on the body. Demonstrating somatic growth, valve tissue filling matching PSIS degradation and PSIS-valve fatigue assessment are critical endeavors that need to be carried out to ensure mid to long term function of these bioscaffold mitral valves.

Low back pain in collegiate athletes has been reported at a rate of 37% from a wide array of sports including soccer, volleyball, football, swimming, and baseball. Whereas, in a military population the prevalence of low back pain is 70% higher than the general population. Compensatory movement strategies are often used as an attempt to reduce pain. Though compensatory movement strategies may effectively reduce pain, they are often associated with altered lower extremity loading patterns. Those who suffer from chronic low back pain tend to walk and run slower and with less trunk and pelvis coordination and variability. Individuals with low back pain also tend to run with more stiffness in their knees. Moving with less joint coordination and more stiffness are potential compensatory movement patterns acting as a guarding mechanism for pain. Overall the purpose of this project was to determine how chronic low back pain influences lower extremity biomechanics and shock attenuation in active individuals compared to healthy individuals and examine how the altered lower extremity biomechanics are related to clinical outcome measures. We hypothesized that individuals who present with chronic low back pain are more likely to exhibit higher vertical ground reaction forces and less knee flexion excursion during landing, compared to healthy individuals. We also hypothesized that individuals with chronic low back pain will have a reduced ability to attenuate shock during landing compared to the healthy individuals. This study was a case control design in which physically active individuals suffering from chronic low back pain were matched to healthy controls. All participants reported for one testing session to assess self-perceived knee function in the form of the Knee Osteoarthritis Outcomes Score (KOOS), lower extremity strength and mechanics during three landing tasks. Isometric strength was assessed using an isokinetic dynamometer during hip abduction, hip extension, and knee extension. The landing tasks included a drop vertical jump, a single leg hop, and a crossover hop. A three-dimensional motion analysis system with two in-ground force plates and four inertial measurement units were used to assess lower extremity mechanics during the landing tasks. Individuals with low back pain presented with reduced KOOS scores compared to healthy individuals in four of the five subscales, including Symptoms (p=0.007), Pain (p=0.002), Activities of Daily Living (p=0.021), and Quality of Life (p=0.003). Alternatively, while there were some strength, kinematic, and kinetic between limb asymmetries noted in the low back pain group, there were not between group differences with the healthy individuals. In the low back pain group, individuals presented with greater dominant limb knee extension strength (p=0.039) and greater dominant limb ankle plantarflexion at initial contact during the drop vertical jump, compared to the non-dominant limb (p=0.022). Individuals with low back pain also presented with greater non-dominant limb tibia impact during the single limb hop (p=0.008). While we did not identify any mechanical differences between individuals suffering from chronic low back pain and those who do not, we did identify that an active population suffering from low back pain does present with decreased self-perceived knee function compared to active individuals without low back pain. As these groups biomechanically perform similarly, they do not clinically perform the same, specifically, in terms of the KOOS. Such differences should not be overlooked when treating active populations with low back pain. If this population is presenting with altered self-perceived knee function at a young age, it is likely that it will continue to decline and negatively affect their function.

A new detection reagent that could possibly augment or replace antibodies research and diagnosis methods are aptamers. Aptamers are ssDNA, RNA or polypeptide constructs that function like active antibodies. Antibodies and aptamers both specifically bind to selected target molecules, and as such they enable the detection or targeting of the presence or absence of a specific antigen. In order to ensure that ssDNA aptamers perform similarly to antibodies, anti-VEGF-A polyclonal antibody and anti-VEGF-A ssDNA aptamer were evaluated against vascular endothelial growth factor A (VEGF-A) using Surface Plasmon Resonance (SPR). It was hypothesized that the anti-VEGF-A aptamer had the same, if not better, binding kinetics than the anti-VEGF-A polyclonal antibody, and as such offers an ideal replacement for use in in field, real-time testing assays. SPR revealed that both the polyclonal antibody and ssDNA aptamer bound the target antigen, VEGF-A. Additionally, from the SPR kinetic analysis, the anti-VEGF-A aptamer had KD values of 20-28 nM and the anti-VEGF-A antibody had KD values of 16-127 uM. The binding efficacy of the aptamer was several orders of magnitude better than that of the antibody. The aptamer was also stable in solution for a longer amount of time than the antibody, which denatured in solution after two weeks.

Transfemoral amputees around the world experience increased difficulty in climbing stairs due to lack of muscle, balance, and other factors. The loss of a lower limb greatly diminishes the amount of natural force generation provided that is necessary to propel oneself up stairs. This study investigated possible solutions to the problem of stair ascension for transfemoral amputees by the means of designing and developing an externally attachable device to a prosthesis. The number of amputations from military service has greatly increased since 2008, which shows there is a clear need for assistive devices (Wenke, Krueger, & Ficke, 2012). With the number of amputations rising and no current externally attachable products on the market to aid in stair ascension for transfemoral amputees, the need for this specific device has become more prominent. Research, previous work, and preliminary testing provided a basis for design and development of a new prototype. Bench top testing was conducted to review concepts in the prototype and provide data for further modifications. Results from testing of previous work, as well as testing of new concepts and modifications, provided a framework for designing a new externally attachable device for assistance in stair ascension. A new prototype was then designed, manufactured, and tested with bench models as well as real-time testing with amputees. Success of the device’s performance was based on bench top results and feedback from amputees, noting both the advantages and shortcomings of the new prototype. Testing provided results and feedback that the device was well built and functioned properly, but did not perform satisfactorily, particularly in the categories of force generation and balance.

Cerebral ischemia leading to an ischemic stroke is a possible complication of a transcatheter aortic valve replacement (TAVR) procedure. This is because embolic debris can become dislodged and travel through the aortic arch, where they either continue to the descending aorta and join the systemic circulation or travel into the cerebral vasculature through the three daughter vessels that branch off the top of the aortic arch. These three vessels are the brachiocephalic artery, the left subclavian artery, and the left common carotid artery. These three vessels lead either directly or indirectly to the cerebral vasculature, where the diameter of vessels become very small. If a large enough embolus travels into the cerebral vasculature, it can become stuck in the small cerebral vessels, blocking blood flow and cutting off the supply of oxygen to brain cells. The purpose of this study is to expand upon previous work in order to 1) create a more accurate physics simulation of blood and debris flow through the aortic arch 2) report on embolic debris distribution through the aortic arch and 3) analysis on which physical parameters affect embolic debris distribution. The physical parameters analyzed were particle diameter and particle density. This study was performed by creating a finite element model in COMSOL Multiphysics™ using a SolidWorks model of an aortic arch, with dimensions taken from a patient’s CT scan. Computational fluid dynamics was performed using a pulsatile pressure waveform throughout the aortic arch with a non-constant viscosity model. Once the velocity profile through the aortic arch matched with value ranges from literature, the particle tracing study was implemented. Both a pulsatile pressure waveform and a constant pressure model were analyzed, as well as a constant viscosity model and a non-constant viscosity model. The pulsatile pressure waveform influenced particle distribution and is recommended for future studies since this model leads to pulsatile flow, which is representative of flow through the aorta. It was seen that the non-constant viscosity model did not have a large effect on the velocity profile, but more than doubled the surface average value of viscosity. It also had an effect on the particle distribution through the aortic arch. Small diameter emboli were more likely to flow into the descending aorta, the brachiocephalic artery, and the left subclavian artery; larger emboli were more likely to flow into the left common carotid. Lower density emboli were more likely to flow into the descending aorta and the brachiocephalic artery. Averaging all densities and sizes, it was determined 44.8% of emboli flow into the three daughter vessels, but ultimately only 30.61% of emboli flow into the cerebral vasculature and have the potential to cause an ischemic stroke.

Tooth cusp radius of curvature (RoC) has been hypothesized to play an important role in food item breakdown, but has remained largely unstudied due to difficulties in measuring and modeling RoC in multicusped teeth. We tested these hypotheses using a parametric model of a four cusped, maxillary, bunodont molar in conjunction with finite element analysis. When our data failed to support existing hypotheses, we put forth and tested the Complex Cusp Hypothesis which states that, during brittle food items breakdown, an optimally shaped molar would be maximizing stresses in the food item while minimizing stresses in the enamel. After gaining support for this hypothesis, we tested the effects of relative food item size on optimal molar morphology and found that the optimal set of RoCs changed as relative food item size changed. However, all optimal morphologies were similar, having one dull cusp that produced high stresses in the food item and three cusps that acted to stabilize the food item. We then set out to measure tooth cusp RoC in several species of extant apes to determine if any of the predicted optimal morphologies existed in nature and whether tooth cusp RoC was correlated with diet. While the optimal morphologies were not found in apes, we did find that tooth cusp RoC was correlated with diet and folivores had duller cusps while frugivores had sharper cusps. We hypothesize that, because of wear patterns, tooth cusp RoC is not providing a mechanical advantage during food item breakdown but is instead causing the tooth to wear in a beneficial fashion. Next, we investigate two possible relationships between tooth cusp RoC and enamel thickness, as enamel thickness plays a significant role in the way a tooth wears, using CT scans from hundreds of unworn cusps. There was no relationship between the two variables, indicating that selection may be acting on both variables independently to create an optimally shaped tooth. Finally, we put forth a framework for testing the functional optimality in teeth that takes into account tooth strength, food item breakdown efficiency, and trapability (the ability to trap and stabilize a food item).

This research developed methods for culturing self-assembling capillaries in an injectable gel as a potential method for vascularizing tissue-on-a-chip models to mimic physiological drug delivery. Additionally, a mathematical model was developed as a tool for understanding nutrient delivery and comparison of potential delivery systems. Organs-on-a-chip provide novel platforms for studying biology and physiology in 3D, allow exploration of tissue engineering on a manageable scale, and serve as models for drug screening and drug-delivery testing. Methods were first developed for co-culture of endothelial cells and fibroblasts (3T3s or HDFs) in 2D, evaluating culture time, seeding density and ratio of HUVECs and fibroblasts, and immunostaining with a HUVEC-specific marker. Cells formed large sheets with no signs of vessel formation in 2D; therefore, the setup was translated to 3D culture to further induce stress and release of angiogenetic factors, using fibrin gel to suspend cells in 3D. After 9 days of culture, HUVECs had extensive network formation with a high degree of complexity in the experimental cell ratios (especially with 5:1 HUVECs:HDFs). Therefore, these parameters can be used as a starting point for further development of vascularized tissue constructs. A mathematical model was also successfully developed to assess the impact of cell concentration, consumption, and mode of nutrient delivery on 3D cellular constructs which can be used to predict the spatial distribution of glucose over time. Although the model shows flow introduced through a device is sufficient to maintain nutrient levels for cell growth, developing perfusable capillaries is still a critical part of creating physiologically representative tissues.

Ion sensitive field effect transistors (ISFETs) are semiconductor sensors that have the capability to determine the selected concentration of a specific ion in a solution. Most modern ISFETs utilize their ion selective properties for glucose monitors for diabetics. However, in this thesis, the ISFET fabricated is for the selective detection of K+. The goals of this thesis are to develop a functioning ion-selective polymer membrane, manufacture a working FET device, and implement the two aspects together into a working bench-top K+ selective ISFET device. Properties of a polymer composed of 33 wt.% polyvinyl chloride (PVC) 66 wt.% dioctyl sebacate (DOS) and 1 wt.% valinomycin applied to an ion-sensitive electrode (ISE) were investigated. The membrane generated a sensitivity value of -9.864E-08 Ω/log10(CK). Though this data set was affected by both the maximum resolution of the I-V curve tracing device and the thin-membrane effect. Selectivity tests following the IUPAC two-solution method in the presence of Na+ as the interfering ion, provided selectivity values of 0.228 and 0.443 with higher ratios of primary ion to interfering ion resulting in higher selectivity coefficients. Additionally, utilizing an illumination test, dielectric constants of 17.71 and 10.88 were calculated dependent on the amount of solvent used during formulation. Fabrication of the FET device also resulted in developments in metal contact materials, nitride film processing, and physical vapor deposition (PVD) processes. With further improvements, it is possible to fabricate a biocompatible, wearable K+-selective monitor for continuously testing dialysis patients.