<p> The tumor microenvironment has been demonstrated to be a key determinant in the progression of cancer. Unfortunately, the mechanisms behind the different microenvironments (cytokine gradients, hypoxia, hypoglycemia, etc) have not been fully elucidated. Identifying these mechanisms can lead to targeted, individualized therapy to prevent metastasis. The Nano Intravital Device (NANIVID) is a microfabricated, implantable device designed to initiate specific microenvironments <i> in vivo</i> so that the time course of the effects can be observed. With both spatial and temporal control over the induced environments, the affected regions of the tumor can be compared to the rest of the tumor. The NANIVID was first used to establish cytokine gradients to monitor the migration of invasive cancer cells. The three projects that comprise this work expand the applications of the NANIVID to establish the device as a robust platform for investigating tumor microenvironment interactions. The first project released chemical mimics from the device to induce the cellular hypoxic response in tumors to determine how hypoxia affects the fate of disseminated tumor cells. The second project used the NANIVID in combination with an atomic force microscope to investigate the altered mechanics of migrating invasive cancer cells. The final project was to develop a cell counter to monitor the isolation of the invasive subpopulation of cells that were drawn into the device using a chemoattractant. These three projects demonstrate the potential of the NANIVID as a platform for investigating the tumor microenvironment.</p>

Thesis (Ph.D.)--University of California, San Francisco with the University of California, Berkeley, 2010. / Source: Dissertation Abstracts International, Volume: 71-02, Section: B, page: . Adviser: Francis C. Szoka.

<p> Magnetic nanoparticle hyperthermia (mNPH) is an emerging cancer therapy which has shown to be most effective when applied in the adjuvant setting with chemotherapy, radiation or surgery. Although mNPH employs heat as a primary therapeutic modality, conventional heat may not be the only cytotoxic effect. As such, my studies have focused on the mechanism and use of mNPH alone and in conjunction with cisplatinum chemotherapy in murine breast cancer cells and a related in vivo model. MNPH was compared to conventional microwave tumor heating, with results suggesting that mNPH (mNP directly injected into the tumor and immediately activated) and 915 MHz microwave hyperthermia, at the same thermal dose, result in similar tumor regrowth delay kinetics. However, mNPH shows significantly less peri-tumor normal tissue damage. MNPH combined with cisplatinum also demonstrated significant improvements in regrowth delay over either modality applied as a monotherapy. Additional studies demonstrated that a relatively short tumor incubation time prior to AMF exposure (less than 10 minutes) as compared to a 4-hour incubation time, resulted in faster heating rates, but similar regrowth delays when treated to the same thermal dose. The reduction of heating rate correlated well with the observed reduction in mNP concentration in the tumor observed with 4 hour incubation. The ability to effectively deliver cytotoxic mNPs to metastatic tumors is the hope and goal of systemic mNP therapy. However, delivering relevant levels of mNP is proving to be a formidable challenge. To address this issue, I assessed the ability of cisplatinum to simultaneously treat a tumor and improve the uptake of systemically delivered mNPs. Following a cisplatinum pretreatment, systemic mNPs uptake was increased by 3.1 X, in implanted murine breast tumors. Additional in vitro studies showed the necessity of a specific mNP/ Fe architecture and spatial relation for heat-based cytotoxicity in cultured cells.</p>

<p> PAMAM dendrimers are a class of well-defined, hyperbranched polymeric nanocarriers that are being investigated for ocular drug and gene delivery. Their favorable properties such as small size, multivalency and water solubility can provide significant opportunities for many biologically unstable drugs and allows potentially favorable ocular biodistribution. This work exploits hydroxyl terminated dendrimers (G4-OH) as drug/gene delivery vehicles that can target retinal microglia and pigment epithelium via systemic delivery with improved efficacy at much lower concentrations without any side effects. </p><p> Two different drugs Triamcinolone acetonide (TA) and N-Acetyl Cysteine (NAC) conjugated to G4-OH dendrimers showed tailorable sustained release in physiological relevant solutions and were evaluated in-vitro and in-vivo. Dendrimer-TA conjugates enhanced the solubility of TA and were 100 fold more effective at lower concentrations than free TA in its anti-inflammatory activity in activated microglia and in suppressing VEGF production in hypoxic RPE cells. Dendrimers targeted activated microglia/macrophages and RPE and retained for a period of 21 days in I/R mice model. The relative retention of intravitreal and intravenous dendrimers was comparable, if a 30-fold intravenous dose is used; suggesting intravenous route targeting retinal diseases are possible with dendrimers. D-NAC when injected intravenously attenuated retinal and choroidal inflammation, significantly reduced (&sim;73%) CNV growth at early stage of AMD in rat model of CNV. A combination therapy of D-NAC + D-TA significantly suppressed microglial activation and promoted CNV regression in late stages of AMD without causing side-effects. </p><p> G4-OH was modified with linker having minimal amine groups and incorporation of TA as a nuclear localization enhancer resulted in compact gene vectors with favorable safety profile and achieved high levels of transgene expression in hard to transfect human retinal pigment epithelial cells (hRPE). Prepared dendrimer-gene complexes were non-toxic and achieved significant cell uptake and safe delivery of gene in to the nucleus. Further, polyethylene glycol (PEG) surface coating enhanced colloidal stability in physiological relevant solutions without affecting its transfection efficacy.</p>

<p> Chitosan is a naturally derived polymer, which represents one of the most technologically important classes of active materials with applications in a variety of industrial and biomedical fields. Polymeric materials can be regarded as promising candidates for next generation devices due to their low energy payback time. These devices can be fabricated by high-throughput processing methodologies, such as spin coating, inkjet printing, gravure and flexographic printing onto flexible substrates. However, the extensive applications of polymeric films are still limited because of disadvantages such as poor electromechanical properties, high brittleness with a low strain at break, and sensitivity to water. For certain critical applications the need for modification of physical, mechanical and electrical properties of the polymer is essential. When blends of polymer films with other materials are used, as is commonly the case, device performance directly depends on the nanoscale morphology and phase separation of the blend components. To prepare nanocomposite thin films with the desired functional properties, both the film composition and microstructure have to be thoroughly characterized and controlled.</p><p> Chitosan reinforced bio-nanocomposite films with varying concentrations of gold nanoparticles were prepared through a solution casting method. Gold nanoparticles (&sim; 32 nm diameter) were synthesized <i> via</i> a citrate reduction method from chloroauric acid and incorporated in the prepared Chitosan solution. Uniform distribution of gold nanoparticles was achieved throughout the chitosan matrix and was confirmed by SEM images. Synthesis outcomes and prepared nanocomposites were characterized using TEM, SAED, SEM, EDX, XRD, UV-Vis, particle size analysis, zeta potential and FT-IR for their physical, morphological and structural properties. Nanoscale mechanical properties of the nanocomposite films were characterized at room temperature, human body temperatures and higher temperatures using instrumented indentation techniques. The obtained films were confirmed to be biocompatible by their ability to support the growth and proliferation of human tissue cells <i> in vitro.</i> Statistical analysis on mechanical properties and biocompatibility results, were conducted. Results revealed significant enhancement on both the mechanical properties and cell adherence and proliferation. The results will enhance our understanding of the effect of nanostructures reinforcement on these important functional polymeric thin films for potential biomedical applications.</p>