Chitosan is widely investigated for biomedical applications due to its excellent properties, such as biocompatibility, biodegradability, bioadhesivity, antibacterial, etc. In the field of neural engineering, it has been extensively studied in forms of film and hydrogel, and has been used as scaffolds for nerve regeneration in the peripheral nervous system and spinal cord. One of the main issues in neural engineering is the incapability of neuron to attach on biomaterials. The present study, from a new aspect, aims to take advantage of the bio-adhesive property of chitosan to develop chitosan-based materials for neural engineering, specifically in the fields of brain repair and neuroprosthetics. Neuronal responses to the developed biomaterials will also be investigated and discussed.In the first part of this study (Chapter II), chitosan was blended with a well-studied hydrogel material (agarose) to form a simply prepared hydrogel system. The stiffness of the agarose gel was maintained despite the inclusion of chitosan. The structure of the blended hydrogels was characterized by light microscopy and scanning electron microscopy. In vitro cell studies revealed the capability of chitosan to promote neuron adhesion. The concentration of chitosan in the hydrogel had great influence on neurite extension. An optimum range of chitosan concentration in agarose hydrogel, to enhance neuron attachment and neurite extension, was identified based on the results. A “steric hindrance” effect of chitosan was proposed, which explains the origin of the morphological differences of neurons in the blended gels as well as the influence of the physical environment on neuron adhesion and neurite outgrowth. This chitosan-agarose (C-A) hydrogel system and its multi-functionality allow for applications of simply prepared agarose-based hydrogels for brain tissue repair.In the second part of this study (Chapter III), chitosan was blended with graphene to form a series of graphene-chitosan (G-C) nanocomposites for potential neural interface applications. Both substrate-supported coatings and free standing films could be prepared by air evaporation of precursor solutions. The electrical conductivity of graphene was maintained after the addition of chitosan, which is non-conductive. The surface characteristic of the films was sensitively dependent on film composition, and in turn, influenced neuron adhesion and neurite extension. Biological studies showed good cytocompatibility of graphene for both fibroblast and neuron. Good cell-substrate interactions between neurons and G-C nanocomposites were found on samples with appropriate compositions. The results suggest this unique nanocomposite system may be a promising substrate material used for the fabrication of implantable neural electrodes. Overall, these studies confirmed the bio-adhesive property of chitosan. More importantly, the developed chitosan-based materials also have great potential in the fields of neural tissue engineering and neuroprosthetics.

Planar supported, polymerized phospholipid bilayers (PPBs) composed of 1,2-bis[10-(2’,4’-hexadienoyloxy)decanoyl]-sn-glycero-3-phosphocholine (bis-SorbPC or BSPC) were generated by a redox polymerization method. The PPBs were supported by a silicon substrate. The PPBs were characterized and tested for uniformity and stability under physiological conditions. The PPBs were analyzed in vitro with murine derived cells that are pertinent to the host response. Cellular attachment and phenotypic changes in RAW 264.7 macrophages and NIH 3T3 fibroblasts were investigated on PPBs and compared to bare silicon controls. Fluorescent and SEM images were used to observe cellular attachment and changes in cellular behavior. The PPBs showed much lower cellular adhesion for both cell lines than bare silicon controls. Of the cells that attached to the PPBs, a very low percentage showed the same morphological expressions as seen on the controls. The hypothesis generated from this work is that defects in the PPBs mediated the cellular attachment and morphological changes that were observed. Finally, a layer-by-layer (LbL) deposition of a poly(acrylic acid) (PAA) and poly(N-vinylpyrrolidone) (PNVP) alternating bilayer was attempted as a proof of concept for future modification of this system.

Polymers have deep roots as drug delivery tools, and are widely used in clinical to private settings. Currently, however, numerous traditional therapies exist which may be improved through use of polymeric biomaterials. Through our work with infectious and oxidative stress disease prevention and treatment, we aimed to develop application driven, enhanced therapies utilizing new classes of polymers synthesized in-house. Applying biodegradable poly(β-amino ester) (PBAE) polymers, covalent-addition of bioactive substrates to these PBAEs avoided certain pitfalls of free-loaded and non-degradable drug delivery systems. Further, through variation of polymer ingredients and conditions, we were able to tune degradation rates, release profiles, cellular toxicity, and material morphology. Using these fundamentals of covalent drug-addition into biodegradable polymers, we addressed two problems that exist with the treatment of patients with high-risk wound-sites, namely non-biodegradability that require second-surgeries, and free-loaded antibiotic systems where partially degraded materials fall below the minimum inhibitory concentration, allowing biofilm proliferation. Our in situ polymerizable, covalently-bound vancomycin hydrogel provided active antibiotic degradation products and drug release which closely followed the degradation rate over tunable periods. With applications of antioxidant delivery, we continued with this concept of covalent drug addition and modified a PBAE, utilizing a disulfide moiety to mimic redox processes which glutathione/glutathione disulfide performs. This material was found to not only be hydrolytically biodegradable, but tunable in reducibility through cleavage of the disulfide crosslinker, forming antioxidant groups of bound-thiols, similar to drugs currently used in radioprotective therapies. The differential cellular viability of degradation products containing disulfide or antioxidant thiol forms was profound, and the antioxidant form significantly aided cellular resistance to a superoxide attack, similar to that of a radiation injury. Pathophysiological oxidation in the form of radiation injury or oxidative stress based diseases are often region specific to the body and thus require specific targeting, and nanomaterials are widely researched to perform this. Utilizing a tertiary-amine base-catalyst, we were able to synthesize a high drug content (20-26 wt%) version of the disulfide PBAE previously unattainable. The reduced version of this material created a linear-chain polymer capable of single-emulsion nanoparticle formulation for use with intravenous antioxidant delivery applications instead of local.

Common biodegradable polyesters such as poly(lactic acid) (PLA), poly(lactic-co-glycolic acid) (PLGA) and poly(ε-caprolactone) (PCL) are used as drug delivery vehicles for tissue regenerative applications. However, they are typically bioinert, with drug loading limitations. Polymerizing the active agent or precursor into its respective biodegradable polymer would control drug loading via molar ratios of drug to initiator used for synthesis. Simvastatin was chosen due to its favorable anti-inflammatory, angiogenic, and osteogenic properties. In addition, its lactone ring lends itself to ring-opening polymerization and, consequently, the synthesis of poly(simvastatin) with controlled simvastatin release. Simvastatin was first polymerized with a 5kDa methyl-terminated poly(ethylene glycol) (mPEG) initiator and catalyzed via stannous octoate to form poly(simvastatin)-block-poly(ethylene glycol). Molecular weights ranged from 9.5kDa, with a polydispersity index (PDI) of 1.1 at 150 °C, to 75kDa with a PDI of 6.9 at 250 °C. First-order propagation rates were seen. Infrared spectroscopy showed carboxylic and methyl ether stretches unique to simvastatin and mPEG in the copolymer, respectively. Slow degradation was seen in neutral and alkaline conditions, with simvastatin, simvastatin-incorporated macromolecules, and mPEG identified as degradation products. Alternatively, triazabicyclodecene (TBD) was used to mediate simvastatin polymerization. A lower temperature of 150°C led to successful polymerization using 5kDa mPEG, compared to at least 200 °C via stannous octoate. TBD was also successful for reactions using 2 or 0.55kDa mPEG. The biodegradability of poly(simvastatin)-block-poly(ethylene glycol) via TBD improved, losing twice more mass in phosphate-buffered saline, pH 7.4, than the copolymer synthesized via stannous octoate. Release rates of three different copolymers synthesized demonstrated tunable simvastatin release. To further modulate degradation, poly(simvastatin)-block-poly(ethylene glycol) was blended with 5, 2, or 0.55kDa mPEG-initiated PLA copolymers. The blends showed a compressive elastic modulus ranging from 26 to 44MPa, within the magnitude of trabecular bone (approximately 50MPa). Tunability in mass loss and release was also seen due to varied ratios of incorporated PLA copolymers. Lastly, copolymer degradation byproducts inhibited HMG-CoA reductase and showed possible enhancement of osteoblastic activity in vitro. A pilot study using a rodent calvarial onlay model showed tolerability of the polymers and potential for long-term evaluations of bioactivity. Poly(simvastatin) may be useful in regenerative applications.

Gene therapy involves the use of nucleic acids, either DNA or RNA for the treatment, cure, or prevention of human diseases. Synthetic cationic polymers are promising as a tool for gene delivery because of their high level of design flexibility for biomaterial construction and are capable of binding and condensing DNA through electrostatic interactions. Our lab has developed a novel polymer (poly (polyethylene glycol-dodecanoate) (PEGD), a polyester of polyethylene glycol (PEG) and dodecanedioic acid (DDA). PEGD is a linear viscous polymer that self-assembles into a vesicle upon immersion in an aqueous solution. A copolymer of dodecanedioc acid and polyethylene glycol (PEG) was synthesized at a 1:1 ratio. Furmaric (FA) or itaconic acid (IA) was used to suppress DDA in the PEGD copolymer at an 80:20 ratio (DDA: furmaric/itaconic acid) to form the PEGDF/I variant. PEGDF/I are then modified through the Michael addition of Protamine Sulfate (PEGDF/I-PS) and Cys-Arg8 (PEGDF/I-CA) peptide to the carbon-carbon double bond on the polymer backbone to introduce a positive charge. The modified PEGDF/I polymers were capable of binding and condensing DNA. Transfection of HEK 293 cells with pTurboGFP plasmid using modified PEGDF/I polymers was successful but showed varied efficiency. The PEGDF/I-CA polymer had around 30% transfection efficiency and was shown to be non-cytotoxic.

This thesis focuses on the electrical output of polyaniline films subjected to uniaxial strain in hydrochloric acid solutions. Polyaniline belongs to novel class of materials known as conducting polymers. Alternating single and double bonds in the backbone of conducting polymers allow them to transmit electric charge when they are doped with negatively charged ions. Modifying the degree of doping and other electrical/chemical treatments allow conducting polymers to exhibit conducting, semi-conducting, or insulating electrical properties. Resilient mechanical properties, good processability, and low cost make conducting polymers good candidates for applications traditionally held by metals and semi-conductors. When tensile strain is applied to polyaniline in an electrolyte solution, the material selectively absorbs negatively charged ions. This charge imbalance produces a measurable electrical output. Theoretical models based on Fick’s second law of diffusion were compared against experimental results to determine fundamental material properties such as diffusivity and ion solubility in polyaniline. These properties were used to quantify polyaniline as a sensor material based on characteristics including sensitivity, accuracy, precision, range, linearity, and error. Films were cast from solutions of polyaniline powder (Mn = 65,000) in N-methyl-2-pyrrolidinone solvent, with thicknesses ranging from 2.72 to 158 µm.

Conjugated polymers are semiconducting materials that are currently being researched for numerous applications from chemical and biological sensors to electronic devices, including photovoltaics and transistors. Much of the novel research on conjugated polymers is performed in academic settings, where scientists are working to prepare conjugated polymers for commercially viable applications. By offering numerous advantages, inherent in macromolecular materials, conjugated polymers may hold the key to cheap and environmentally friendly manufacturing of future electronic devices. Mechanical flexibility, and solvent-based coating processes are two commonly cited advantages. Transitions in the backbone conformation of polythiophenes (PT) in organic solvents have been widely observed to influence thin-film morphology. However, conformational transitions of water-soluble PT derivatives, with respect to their intramolecular versus intermolecular origin, remain largely obscure. Here, conformational transitions of a water- soluble polythiophene in aqueous ionic surfactants are investigated by means of Fourier transform infrared (FTIR) spectroscopy, differential scanning calorimetry (DSC), polarizing optical microscopy (POM), ultraviolet-visible (UV-Vis) absorption and fluorescence spectroscopy, and various X-ray scattering techniques. As-prepared complexes exist as stable hydrogels. Upon dilution, a significant time-dependent chromism occurs spontaneously. A coil-to-rod conformational transition is identified in this mechanism and verified using small-angle x-ray scattering (SAXS). Study into the corresponding kinetics demonstrates an inverse first-order rate law. It is found that the conformational transition is thermally reversible and concentration-independent. The critical transition temperature is largely dependent on the surfactant formulation. A theoretical model is presented to explain this new phenomenon and the mechanisms behind its influence on the optoelectronic and solid-state morphological properties. A relationship between the dilute-solution processing with surfactants and the final properties of the system is substantiated.

There is a need for a minimally invasive delivery method to enable cell therapies to combat peripheral artery occlusive disease (PAOD) in end stage patients. Myoblasts show promise as a cell mediated therapy but warrant an improved delivery method to increase cell retention in the region of interest because of their adherent nature, relative to previously used BM-MNC’s that are non-adherent. Contemporary issues with achieving successful cell therapies of vasculature can be mainly characterized by the lack of clinical translation from promising animal studies and absence of cell delivery scaffolding. Naturally, polymers have been widely experimented with as grafts to both culture and implant cells into tissue with recognizable success due to their analogous physical properties, such as stiffness, hydrophilicity, & surface energy, that mimic tissue conditions. Polymers having similar mechanical properties to anatomical structures are conducive to cell integration & retention, making polymers an effective biomaterial choice as a cell delivery vehicle. This thesis will evaluate the application of N-isopropylacrylamide (NIPAM) based copolymers as a biomaterial scaffold for myoblast delivery, as it is one of the most widely used biocompatible polymers with thermoreversible properties that is non-toxic and has manipulatable mechanical properties. We hypothesized that fluctuations in polymer construct stiffness, surface energy, and water retention affect myoblast proliferation & viability within the cell delivery vehicle. After measuring the physical properties and cellular proliferation in for each polymer composition, the goal of this thesis was to establish a statistical model to characterize the effect of polymer material properties on myoblast behavior and create a predictive model to optimize further iterations of NIPAM-based copolymers for cell delivery.

Intravascular devices, such as stents, must be rigorously tested before they can be approved by the FDA. This includes bench top in vitro testing to determine biocompatibility, and animal model testing to ensure safety and efficacy. As an intermediate step, a blood vessel mimic (BVM) testing method has been developed that mimics the three dimensional structure of blood vessels using a perfusion bioreactor system, human derived endothelial cells, and a biocompatible polymer scaffold used to support growth of the blood vessel cells. The focus of this thesis was to find an in-house fabrication method capable of making cellular scaffolding for use in the BVM. Research was conducted based on three aims. The first aim was to survey possible fabrication methods to choose a technique most appropriate for producing BVM scaffolding. The second aim was to set up the selected fabrication method (electrospinning) in-house at Cal Poly and gain understanding of the process. The third aim was to evaluate consistency of the technique. The work described in this thesis determined that electrospinning is a viable fabrication technique for producing scaffolding for BVM use. Electrospun scaffolding is highly tailorable, and a structure that mimics the natural organization of nano sized collagen fibers is especially desirable when culturing endothelial cells. An electrospinning apparatus was constructed in house and a series of trial experiments was conducted to better understand the electrospinning process. A consistency study evaluated scaffold reproducibility between different spins and within individual spins while setting a baseline that can be used for comparison in future work aimed at electrospinning.

Abstract: Under normal sealing and storage conditions, Nylon-6, poly-caprolactam-based plastic laminates may release impurities to packaged foods and liquids and the application of heat for cooking often increases the rate of migration. Epsilon-caprolactam is one of the main contaminates found to migrate from a Nylon-6 poly-caprolactam plastic film. The objectives of this study were to determine the effects of solvent, transportation and storage time on the migration of ε-caprolactam from a Nylon-6-based lidding material into water or a white wine substitute (12% ethanol). Polypropylene plastic cups were filled, sealed, packaged, stacked and exposed to a simulated 3-day cross-country shipment. Cups were sampled with or without simulated shipment after 0, 7, 14 and 28 days at 20.6°C. Epsilon-caprolactam was determined using a GC equipped with FID and a Restek Rtx 1301 megabore column. Results of the study indicated migration of ε-caprolactam into containers at the time of sealing with significantly higher levels (4.42 ppm average) occurring in cups containing 12% ethanol vs. water (0.01 ppm average). After the cups were sealed, neither simulated cross-country shipment nor storage increased levels of ε-caprolactam in either solvent. The results indicate that wine sealed in packages lined with Nylon-6-based plastic could contain significant amounts of ε-caprolactam. However, it is not understood how the alcohol, whether as liquid or vapor, interacted with the lidding material to increase migration at the time of sealing. No delamination of the polypropylene layer in the lidding material was observed after sealing. Future research needs to be conducted to study the effects of alcohol, alcohol vapor, sealing time and temperature on ε-caprolactam migration.