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Abstract
The aim of the research performed was to increase research potential in the field of cell stimulation by developing a method to adhere human neural progenitor cells (hNPC’s) to a sterilized stretchable microelectrode array (SMEA). The two primary objectives of our research were to develop methods of sterilizing the polydimethylsiloxane

Abstract
The aim of the research performed was to increase research potential in the field of cell stimulation by developing a method to adhere human neural progenitor cells (hNPC’s) to a sterilized stretchable microelectrode array (SMEA). The two primary objectives of our research were to develop methods of sterilizing the polydimethylsiloxane (PDMS) substrate being used for the SMEA, and to derive a functional procedure for adhering hNPC’s to the PDMS. The proven method of sterilization was to plasma treat the sample and then soak it in 70% ethanol for one hour. The most successful method for cell adhesion was plasma treating the PDMS, followed by treating the surface of the PDMS with 0.01 mg/mL poly-l-lysine (PLL) and 3 µg/cm2 laminin. The development of these methods was an iterative process; as the methods were tested, any problems found with the method were corrected for the next round of testing until a final method was confirmed. Moving forward, the findings will allow for cell behavior to be researched in a unique fashion to better understand the response of adherent cells to physical stimulation by measuring changes in their electrical activity.
ContributorsBridgers, Carson (Co-author) / Peterson, Mara (Co-author) / Stabenfeldt, Sarah (Thesis director) / Graudejus, Oliver (Committee member) / Harrington Bioengineering Program (Contributor) / School of Human Evolution and Social Change (Contributor) / Barrett, The Honors College (Contributor)
Created2016-05
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Description
Oxygen delivery is crucial for the development of healthy, functional tissue. Low tissue oxygenation, or hypoxia, is a characteristic that is common in many tumors. Hypoxia contributes to tumor malignancy and can reduce the success of chemotherapy and radiation treatment. There is a current need to noninvasively measure tumor oxygenation

Oxygen delivery is crucial for the development of healthy, functional tissue. Low tissue oxygenation, or hypoxia, is a characteristic that is common in many tumors. Hypoxia contributes to tumor malignancy and can reduce the success of chemotherapy and radiation treatment. There is a current need to noninvasively measure tumor oxygenation or pO2 in patients to determine a personalized treatment method. This project focuses on creating and characterizing nanoemulsions using a pO2 reporter molecule hexamethyldisiloxane (HMDSO) and its longer chain variants as well as assessing their cytotoxicity. We also explored creating multi-modal (MRI/Fluorescence) nanoemulsions.
ContributorsGrucky, Marian Louise (Author) / Kodibagkar, Vikram (Thesis director) / Rege, Kaushal (Committee member) / Stabenfeldt, Sarah (Committee member) / Barrett, The Honors College (Contributor) / Harrington Bioengineering Program (Contributor)
Created2013-05
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Description
For my honors thesis, I developed a proof of concept alpha prototype of a biomedical device for detection of cerebrospinal fluid leaks during spinal surgery. Cerebrospinal fluid leaks are a consequence of tears in the dura mater of the spinal cord and can result in potentially life-threatening conditions and are

For my honors thesis, I developed a proof of concept alpha prototype of a biomedical device for detection of cerebrospinal fluid leaks during spinal surgery. Cerebrospinal fluid leaks are a consequence of tears in the dura mater of the spinal cord and can result in potentially life-threatening conditions and are overall a large burden not only on the patient but upon the clinical teams managing the patient postoperatively. What I created was an optical sensor that I programmed to be sensitive to detecting green wavelength light. The device would ideally be attached to surgical drain tubing and used in conjunction with fluorescein (a green fluorescent dye) infused lumbar punctures into the spinal canal of patients. As the dye circulates through the spinal cord, any tears in the dura mater would cause the fluorescein to leak out with cerebrospinal fluid into the incision site. This fluid may then be collected by the surgical drain where the sensor may detect the fluorescein, triggering a buzzer response that would notify the patient or the surgeons of an ongoing leak that requires repair. The time I spent on my thesis involved sensor validation to ensure it could differentiate between colors, testing the sensor's color sensitivity by performing a fluorescein aliquot, and running proof of concept testing that could show the sensor can detect fluorescein drain tubing and provide an adequate response. The sensor was able to differentiate between varying concentrations of fluorescein in solution and provided exceptional results in its proof-of-concept testing. Next steps will be to re-run the sensor validation study with different dyes as well as consolidating the device's electrical hardware onto a single circuit board as development of beta and gamma prototypes move forward.
ContributorsAlam, Framarz (Author) / Muthuswamy, Jitendran (Thesis director) / Bohl, Michael (Committee member) / Harrington Bioengineering Program (Contributor) / Barrett, The Honors College (Contributor)
Created2018-05
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Description
Abstract Modern imaging techniques for sciatic nerves often use imaging techniques that can clearly find myelinated axons (Group A and Group B and analyze their properties, but have trouble with the more numerous Remak Fibers (Group C). In this paper, Group A and B fibers are analyzed while also analyzing

Abstract Modern imaging techniques for sciatic nerves often use imaging techniques that can clearly find myelinated axons (Group A and Group B and analyze their properties, but have trouble with the more numerous Remak Fibers (Group C). In this paper, Group A and B fibers are analyzed while also analyzing Remak fibers using osmium tetroxide staining and imaging with the help of transmission electron microscopy. Using this method, nerves had various electrical stimuli attached to them and were analyzed as such. They were analyzed with a cuff electrode attached, a stimulator attached, and both, with images taken at the center of the nerve and the ends of them. The number and area taken by the Remak fibers were analyzed, along with the g-ratios of the Group A and B fibers. These were analyzed to help deduce the overall health of the fibers along with vacuolization, and mitochondria available. While some important information was gained from this evaluation, further testing has to be done to improve the myelin detection system, along with analyzing the proper and necessary Remak fibers and the role they play. The research tries to thoroughly look at the necessary material and find a way to use it as a guide to further experimentation with electrical stimuli, and notes the differences found within and without various groups, various points of observation, and various stimuli as a whole. Nevertheless, this research allows a strong look into the benefits of transmission electron microscopy and the ability to assess electrical stimulation from these points.
ContributorsNambiar, Karthik (Author) / Muthuswamy, Jitendran (Thesis director) / Towe, Bruce (Committee member) / Harrington Bioengineering Program (Contributor) / School of Mathematical and Statistical Sciences (Contributor) / Barrett, The Honors College (Contributor)
Created2018-05
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Description
Traumatic brain injury (TBI) is a major concern in public health due to its prevalence and effect. Every year, about 1.7 million TBIs are reported [7]. According to the According to the Centers for Disease Control and Prevention (CDC), 5.5% of all emergency department visits, hospitalizations, and deaths from 2002

Traumatic brain injury (TBI) is a major concern in public health due to its prevalence and effect. Every year, about 1.7 million TBIs are reported [7]. According to the According to the Centers for Disease Control and Prevention (CDC), 5.5% of all emergency department visits, hospitalizations, and deaths from 2002 to 2006 are due to TBI [8]. The brain's natural defense, the Blood Brain Barrier (BBB), prevents the entry of most substances into the brain through the blood stream, including medicines administered to treat TBI [11]. TBI may cause the breakdown of the BBB, and may result in increased permeability, providing an opportunity for NPs to enter the brain [3,4]. Dr. Stabenfeldt's lab has previously established that intravenously injected nanoparticles (NP) will accumulate near the injury site after focal brain injury [4]. The current project focuses on confirmation of the accumulation or extravasation of NPs after brain injury using 2-photon microscopy. Specifically, the project used controlled cortical impact injury induced mice models that were intravenously injected with 40nm NPs post-injury. The MATLAB code seeks to analyze the brain images through registration, segmentation, and intensity measurement and evaluate if fluorescent NPs will accumulate in the extravascular tissue of injured mice models. The code was developed with 2D bicubic interpolation, subpixel image registration, drawn dimension segmentation and fixed dimension segmentation, and dynamic image analysis. A statistical difference was found between the extravascular tissue of injured and uninjured mouse models. This statistical difference proves that the NPs do extravasate through the permeable cranial blood vessels in injured cranial tissue.
ContributorsIrwin, Jacob Aleksandr (Author) / Stabenfeldt, Sarah (Thesis director) / Bharadwaj, Vimala (Committee member) / Harrington Bioengineering Program (Contributor) / Barrett, The Honors College (Contributor)
Created2018-05
Description

Polymeric nanoparticles (NP) consisting of Poly Lactic-co-lactic acid - methyl polyethylene glycol (PLLA-mPEG) or Poly Lactic-co-Glycolic Acid (PLGA) are an emerging field of study for therapeutic and diagnostic applications. NPs have a variety of tunable physical characteristics like size, morphology, and surface topography. They can be loaded with therapeutic and/or

Polymeric nanoparticles (NP) consisting of Poly Lactic-co-lactic acid - methyl polyethylene glycol (PLLA-mPEG) or Poly Lactic-co-Glycolic Acid (PLGA) are an emerging field of study for therapeutic and diagnostic applications. NPs have a variety of tunable physical characteristics like size, morphology, and surface topography. They can be loaded with therapeutic and/or diagnostic agents, either on the surface or within the core. NP size is an important characteristic as it directly impacts clearance and where the particles can travel and bind in the body. To that end, the typical target size for NPs is 30-200 nm for the majority of applications. Fabricating NPs using the typical techniques such as drop emulsion, microfluidics, or traditional nanoprecipitation can be expensive and may not yield the appropriate particle size. Therefore, a need has emerged for low-cost fabrication methods that allow customization of NP physical characteristics with high reproducibility. In this study we manufactured a low-cost (<$210), open-source syringe pump that can be used in nanoprecipitation. A design of experiments was utilized to find the relationship between the independent variables: polymer concentration (mg/mL), agitation rate of aqueous solution (rpm), and injection rate of the polymer solution (mL/min) and the dependent variables: size (nm), zeta potential, and polydispersity index (PDI). The quarter factorial design consisted of 4 experiments, each of which was manufactured in batches of three. Each sample of each batch was measured three times via dynamic light scattering. The particles were made with PLLA-mPEG dissolved in a 50% dichloromethane and 50% acetone solution. The polymer solution was dispensed into the aqueous solution containing 0.3% polyvinyl alcohol (PVA). Data suggests that none of the factors had a statistically significant effect on NP size. However, all interactions and relationships showed that there was a negative correlation between the above defined input parameters and the NP size. The NP sizes ranged from 276.144 ± 14.710 nm at the largest to 185.611 ± 15.634 nm at the smallest. In conclusion, the low-cost syringe pump nanoprecipitation method can achieve small sizes like the ones reported with drop emulsion or microfluidics. While there are trends suggesting predictable tuning of physical characteristics, significant control over the customization has not yet been achieved.

ContributorsDalal, Dhrasti (Author) / Stabenfeldt, Sarah (Thesis director) / Wang, Kuei-Chun (Committee member) / Flores-Prieto, David (Committee member) / Barrett, The Honors College (Contributor) / Harrington Bioengineering Program (Contributor)
Created2023-05
Description
Recent studies in traumatic brain injury (TBI) have found a temporal window where therapeutics on the nanometer scale can cross the blood-brain barrier and enter the parenchyma. Developing protein-based therapeutics is attractive for a number of reasons, yet, the production pipeline for high yield and consistent bioactive recombinant proteins remains

Recent studies in traumatic brain injury (TBI) have found a temporal window where therapeutics on the nanometer scale can cross the blood-brain barrier and enter the parenchyma. Developing protein-based therapeutics is attractive for a number of reasons, yet, the production pipeline for high yield and consistent bioactive recombinant proteins remains a major obstacle. Previous studies for recombinant protein production has utilized gram-negative hosts such as Escherichia coli (E. coli) due to its well-established genetics and fast growth for recombinant protein production. However, using gram-negative hosts require lysis that calls for additional optimization and also introduces endotoxins and proteases that contribute to protein degradation. This project directly addressed this issue and evaluated the potential to use a gram-positive host such as Brevibacillus choshinensis (Brevi) which does not require lysis as the proteins are expressed directly into the supernatant. This host was utilized to produce variants of Stock 11 (S11) protein as a proof-of-concept towards this methodology. Variants of S11 were synthesized using different restriction enzymes which will alter the location of protein tags that may affect production or purification. Factors such as incubation time, incubation temperature, and media were optimized for each variant of S11 using a robust design of experiments. All variants of S11 were grown using optimized parameters prior to purification via affinity chromatography. Results showed the efficiency of using Brevi as a potential host for domain antibody production in the Stabenfeldt lab. Future aims will focus on troubleshooting the purification process to optimize the protein production pipeline.
ContributorsEmbrador, Glenna Bea Rebano (Author) / Stabenfeldt, Sarah (Thesis director) / Plaisier, Christopher (Committee member) / Harrington Bioengineering Program (Contributor, Contributor) / Barrett, The Honors College (Contributor)
Created2019-05
Description
Deep Brain Stimulation (DBS) is a stimulating therapy currently used to treat the motor disabilities that occur as a result of Parkinson’s disease (PD). Previous literature has proven the DBS to be an effective treatment in the effects of PD but the mechanism to validating this phenomenon is poorly understood.

Deep Brain Stimulation (DBS) is a stimulating therapy currently used to treat the motor disabilities that occur as a result of Parkinson’s disease (PD). Previous literature has proven the DBS to be an effective treatment in the effects of PD but the mechanism to validating this phenomenon is poorly understood. In this study, an evaluation of the DBS mechanism was analyzed in patients who received both contralateral and ipsilateral stimulation by the DBS electrode in relation to the recording microelectrode. I hypothesize that the data recorded from the neural tissue of the Parkinson’s patients will exhibit increased electromagnetic field (EMF) fall-off as spatial distance increases among the DBS lead and the microelectrode within the subthalamic nucleus (STN) as a result of the interaction between the EMF exuded by DBS and the neural tissue. Results depicted that EMF fall-off values increased with distance, observable upon comparing ipsilateral and contralateral patient data. The resulting analysis supported this phenomenon evidenced by the production of greater peak voltage amplitudes in ipsilateral patient stimulation with respect to time when compared to contralateral patient stimulation. The understanding of EMF strength and the associated trends among this data are vital to the progression and continued development of the DBS field relative to future research.
ContributorsKiraly, Alexis B (Author) / Greger, Bradley (Thesis director) / Muthuswamy, Jitendran (Committee member) / Harrington Bioengineering Program (Contributor) / Dean, W.P. Carey School of Business (Contributor) / Barrett, The Honors College (Contributor)
Created2020-12
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Description
Breast cancer can be imaged at greater depths using photoacoustic imaging to differentiate between cancerous and non-cancerous tissue. Current photoacoustic modalities struggle to display images in real-time because of the required image reconstruction. In this work, we aim to create a real-time photoacoustic imaging system where the photoacoustic effect is

Breast cancer can be imaged at greater depths using photoacoustic imaging to differentiate between cancerous and non-cancerous tissue. Current photoacoustic modalities struggle to display images in real-time because of the required image reconstruction. In this work, we aim to create a real-time photoacoustic imaging system where the photoacoustic effect is detected through changes in index of refraction. To reach this aim, two methods are applied to visualize the acoustic waves including Schlieren optics and differential interference contrast microscopy. This combined approach provides a new tool for the widespread application in clinical settings.
ContributorsSmetanick, Derek (Author) / Burgett, Joshua (Co-author) / Smith, Barbara (Thesis director) / Muthuswamy, Jitendran (Committee member) / Barrett, The Honors College (Contributor) / Harrington Bioengineering Program (Contributor) / School of Life Sciences (Contributor)
Created2022-05