Myocardial infarction (MI) remains the leading cause of mortality and morbidity in the U.S., accounting for nearly 140,000 deaths per year. Heart transplantation and implantation of mechanical assist devices are the options of last resort for intractable heart failure, but these are limited by lack of organ donors and potential surgical complications. In this regard, there is an urgent need for developing new effective therapeutic strategies to induce regeneration and restore the loss contractility of infarcted myocardium. Over the past decades, regenerative medicine has emerged as a promising strategy to develop scaffold-free cell therapies and scaffold-based cardiac patches as potential approaches for MI treatment. Despite the progress, there are still critical shortcomings associated with these approaches regarding low cell retention, lack of global cardiomyocytes (CMs) synchronicity, as well as poor maturation and engraftment of the transplanted cells within the native myocardium. The overarching objective of this dissertation was to develop two classes of nanoengineered cardiac patches and scaffold-free microtissues with superior electrical, structural, and biological characteristics to address the limitations of previously developed tissue models. An integrated strategy, based on micro- and nanoscale technologies, was utilized to fabricate the proposed tissue models using functionalized gold nanomaterials (GNMs). Furthermore, comprehensive mechanistic studies were carried out to assess the influence of conductive GNMs on the electrophysiology and maturity of the engineered cardiac tissues. Specifically, the role of mechanical stiffness and nano-scale topographies of the scaffold, due to the incorporation of GNMs, on cardiac cells phenotype, contractility, and excitability were dissected from the scaffold’s electrical conductivity. In addition, the influence of GNMs on conduction velocity of CMs was investigated in both coupled and uncoupled gap junctions using microelectrode array technology. Overall, the key contributions of this work were to generate new classes of electrically conductive cardiac patches and scaffold-free microtissues and to mechanistically investigate the influence of conductive GNMs on maturation and electrophysiology of the engineered tissues.

Cooperativity can be used to manipulate binding affinities of DNA biosensors – improving specificity without sacrificing sensitivity; examples include tentacle probes (TPs) and cooperative primers (CPs). This thesis body of work: (1) used TPs to develop a rapid, low-cost diagnostic for detecting the point mutation leading to Navajo Neurohepatopathy (NNH) and (2) used CPs to amplify a symmetric bowtie-barcoded origami with captured t-cell receptor (TCR) α and β mRNA of a single cell.

NNH (affecting 1-in-1600 Navajo babies) is a fatal genetic disorder often caused by 149G>A mutation and is characterized by brain damage and liver disease/failure. Phoenix Children’s Hospital currently uses gene sequencing to identify the 149G>A mutation. While this process is conclusive, there are limitations, as it requires both time (3-4 weeks) and money (>$700). Ultimately, these factors create barriers that can directly impact a patient’s quality of life. Assessment of the developed TP diagnostic, using genomic DNA derived from FFPE patient liver samples, suggests nearly 100% specificity and sensitivity while reducing cost to ~$250 (including cost of labor) and providing a diagnosis within 48 hours.

TCR specificity is dependent on V(D)J recombination as well as pairing of the αβ chains. Drs. Schoettle and Blattman have developed a solution in which a bowtie-barcoded origami strand nanostructure is transfected into individual cells of a heterogeneous cell population to capture and protect αβ mRNA. When PCR of the origami template is performed with Vα, X, Vβ, and Y primers, the α and β gene segments cannot be tied back to a barcode – and paired. Assessment of the developed CPs for PCR suggests correct individual amplification using (1) Va + Xcp and (2) Vβ + Ycp primers, whereas combination of all the primers (Va, Xcp, Vb, and Ycp) suggests hybridization of the Vα + Xcp and Vβ + Ycp products due to the origami target symmetry.

With an increased demand for more enzyme-sensitive, bioresorbable and more biodegradable polymers, various studies of copolymers have been developed. Polymers are widely used in various applications of biomedical engineering such as in tissue engineering, drug delivery and wound healing. Depending on the conditions in which polymers are used, they are modified to accommodate a specific need. For instance, polymers used in drug delivery are more efficient if they are biodegradable. This ensures that the delivery system does not remain in the body after releasing the drug. It is therefore crucial that the polymer used in the drug system possess biodegradable properties. Such modification can be done in different ways including the use of peptides to make copolymers that will degrade in the presence of enzymes. In this work, we studied the effect of a polypeptide GAPGLL on the polymer NIPAAm and compare with the previously studied Poly(NIPAAm-co-GAPGLF). Both copolymers Poly(NIPAAm-co-GAPGLL) were first synthesized from Poly(NIPAAm-co-NASI) through nucleophilic substitution by the two peptides. The synthesis of these copolymers was confirmed by 1H NMR spectra and through cloud point measurement, the corresponding LCST was determined. Both copolymers were degraded by collagenase enzyme at 25 ° C and their 1H NMR spectra confirmed this process. Both copolymers were cleaved by collagenase, leading to an increase in solubility which yielded a higher LCST compared to before enzyme degradation. Future studies will focus on evaluating other peptides and also using other techniques such as Differential Scanning Microcalorimetry (DSC) to better observe the LCST behavior. Moreover, enzyme kinetics studies is also crucial to evaluate how fast the enzyme degrades each of the copolymers.

This study demonstrates that a polymer and drug conjugate can be tattooed onto tissue and deliver drug in a sustained manner. A number of polymers and drugs were investigated in this study in the aims of developing a formulation that could achieve sustained drug delivery for 1-2 weeks. The polymers selected for testing were PDLG 5004, PDLLA-Glycerol, and PEG-PLA, and the drugs used in conjunction with these polymers were rifampicin, moxifloxacin, and dexamethasone. Varying formulas containing these polymer and drug combinations were tattooed onto three different tissue types: bovine pericardial tissue, porcine corneal tissue, and porcine sclera tissue. The drug release rates from these tattoos were determined and characterized after studying the release for up to 20 days. The release rate of dexamethasone from both PDLG 5004 and PDLLA-Glycerol when tattooed onto bovine pericardial tissue demonstrated the best release rate of the formulations tested, with up to 14 days of sustained release. This preliminary research into tattoo-based, polymeric drug delivery is promising, and has the possibility to be developed into a beneficial form of ophthalmic drug delivery that could be expanded to other areas of treatment as well.

Biofilm derived orthopedic infections are increasingly common after contamination of an open bone fracture or the surgical site pre- and post-orthopedic prosthetic insertion or removal. These infections are usually difficult to eradicate due to the resistant nature of biofilms to antimicrobial therapy. Difficulty of treatment of biofilm derived infections is also partly due to the presence of persister cells in the biofilm matrix. Persister cells are tolerant to antimicrobial therapy delivered via the systemic route. It is thus possible for these cells to repopulate their environment once systemic antimicrobial delivery is discontinued. The antimicrobial concentration required to eradicate bacterial biofilms, minimum biofilm eradication concentration (MBEC), can be determined in vitro by exposing biofilms to different regimens of antimicrobial solutions. Previous studies have demonstrated that values of the MBEC vary depending on the material and surface the biofilm grows on. This study investigated the relationship between antimicrobial susceptibility and antimicrobial exposure time, and the effects of surface material type on the antimicrobial susceptibility of staphylococcal biofilms. It was concluded that antimicrobial susceptibility increases with increased antimicrobial exposure time, and that the investigated surface and material properties did not have an effect on the susceptibility of staphylococcal biofilms to antimicrobial therapy. Further investigation is however necessary to confirm these results due to some inconsistent data obtained over the course of the trials.

A self-stirring syringe pump was created in order to fill a void in the market for a medical device that could perform a lengthy drug infusion. This was accomplished by using a magnetic field mechanism that enclosed the body of a syringe. A stator was created in order to facilitate the induction of magnetic fields around the syringe body. A flexible magnetic stir bar was created to rotate within the syringe body while also being able to conform to the syringe plunder as an infusion occurred. In order for the stator with the syringe to fit onto a conventional syringe pump, a mount had to be made. This mount was removable to ensure easy access to the syringe once an infusion had occurred. A study was performed to determine whether or not the self-stirring syringe pump could keep a suspension homogenous over a lengthy infusion. It was found that the self-stirring syringe pump was able to accomplish this task.

Rupture of intracranial aneurysms causes a subarachnoid hemorrhage, which is often lethal health event. A minimally invasive method of solving this problem may involve a material, which can be administered as a liquid and then becomes a strong solid within minutes preventing flow of blood in the aneurysm. Here we report on the development of temperature responsive copolymers, which are deliverable through a microcatheter at body temperature and then rapidly cure to form a highly elastic hydrogel. To our knowledge, this is the first physical-and chemical-crosslinked hydrogel capable of rapid crosslinking at temperatures above the gel transition temperature. The polymer system, poly(N-isopropylacrylamide-co-cysteamine-co-Jeffamine® M-1000 acrylamide) and poly(ethylene glycol) diacrylate, was evaluated in wide-neck aneurysm flow models to evaluate the stability of the hydrogels. Investigation of this polymer system indicates that the Jeffamine® M-1000 causes the gels to retain water, resulting in gels that are initially weak and viscous, but become stronger and more elastic after chemical crosslinking.

Revision hip procedures represent a large financial burden on hospitals and the problem will continue to worsen as the baby boomer generation ages and life expectancy goes up. The future problem is a complex issue that bridges scientific and anecdotal evidence and must be solved. A review of the current total hip arthroplasty procedure in regards to the physical properties of the materials used for hip prostheses is given. Revision procedures can be caused by infection or basic wear and tear from the stress that that implant is subjected to daily. Infections on these implants often present themselves as medical biofilms. The mechanisms of biofilm formation include a complex system of enzymes that work to initiate a phenotypic response based on an established quorum sensing within the colony of bacteria. Surgical methods to treat infection include irrigation and debridement as well as loading drug cement spacers with antimicrobial in hopes of delivering the antibiotic locally. Research is being done to better model the transport of drug through the tissue surrounding the implant, and will hopefully one day be available for use in individual patients.

This report provides information concerning qualities of methylcellulose and how those properties affect further experimentation within the biomedical world. Utilizing the compound’s biocompatibility many issues, ranging from surgical to cosmetic, can be solved. As of recent, studies indicate, methylcellulose has been used as a physically cross-linked gel, which cannot sustain a solid form within the body. Therefore, this report will ultimately explore the means of creating a non-degradable, injectable, chemically cross-linking methylcellulose- based hydrogel. Methylcellulose will be evaluated and altered in experiments conducted within this report and a chemical cross-linker, developed from Jeffamine ED 2003 (O,O′-Bis(2-aminopropyl) polypropylene glycol-block-polyethylene glycol-block-polypropylene glycol), will be created. Experimentation with these elements is outlined here, and will ultimately prompt future revisions and analysis.