Gold Nanorod Mediated Mild Hyperthermia and PEGylated Human Serum Albumin Drug Delivery for Cancer Therapies
Abstract
Photothermally active gold nanorods were used to sensitize cells to chemotherapeutic agents by producing mildly hyperthermic effects (42-43 °C). We examined the synergistic effects of GNR-mediated mild hyperthermia (MHT) on cisplatin (CP) activity against SKOV3 ovarian cancer cells. In vitro studies were performed using CP at cytostatic concentrations (5 &mgr;M) and mPEG-stabilized GNRs (λmax 815 nm) with near-infrared laser excitation for MHT (or external heating as a positive control), followed by 72 hours incubation at 37 °C. The amount of PEG-GNRs needed for GNR-mediated MHT was determined to be 1 &mgr;g/mL, several times lower than the loadings used in tumor tissue ablation. A cell viability assay indicated 80% enhancement in CP-mediated cytotoxicity 3 days after GNR-mediated MHT relative to the projected additive effect. A pilot in vivo study showed preliminary results that cisplatin chemotherapy can be developed in combination with low loadings of GNR-mediated MHT for localized MHT to treat tumors. Stable aqueous dispersions of citrate-stabilized gold nanorods were prepared in scalable fashion by surfactant exchange from cetyltrimethylammonium bromide (CTAB)-stabilized GNRs, using sodium polystyrenesulfonate (PSS) as a detergent. Nanoparticle tracking analysis (NTA) was used to measure the size of the Cit-GNR dispersions, which provides particle sizing resolution several times better than that of dynamic light scattering (DLS). Cit-GNRs were further functionalized with human serum albumin (HSA) and thiols and dithiocarbamates (DTCs) of varying molecular weights. The quality of the Cit-GNR dispersions allows us to address fundamental questions relating GNR stabilization to surface adsorption, including insights into the formation of the protein corona in serum-containing media. Mono-PEGylated human serum albumin was synthesized to investigate its ability to improve the bioavailability of the ability of paclitaxel (PTX), a poorly soluble drug. Matrix assisted laser desportion/ionization mass spectrometry was used to confirm the formation of the mono-PEGylated adduct, and HPLC analysis revealed that 70% of native HSA was converted to HSA-mPEG. A cell viability assay with MCF-7 breast cancer cells was used to measure the enhancement of the therapeutic efficacy of a PTX formulation with HSA, HSA-mPEG 5 kDa, or HSA-mPEG 20 kDa compared to PTX in PBS in the absence of HSA. We observed that the therapeutic efficacy of PTX was maximized when it was formulated in a 10:1 molar ratio with HSA, and for intermediate PTX doses (3.3 and 6.6 nM) the therapeutic efficacy of PTX with HSA-mPEG 20 kDa was greater than that for HSA-mPEG 5 kDa. These results suggest that PTX can bind to PEGylated HSA, and that most of the drug can be released.
Degree
Ph.D.
Advisors
Wei, Purdue University.
Subject Area
Nanotechnology
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