Research Article
Pharmaceutical Nanotechnology
Combined Treatment of Tyrosine Kinase Inhibitor–Labeled Gold Nanorod Encapsulated Albumin With Laser Thermal Ablation in a Renal Cell Carcinoma Model

https://doi.org/10.1016/j.xphs.2015.11.017Get rights and content

Abstract

To characterize and evaluate human serum albumin–encapsulated nanoparticles for drug delivery of a tyrosine kinase inhibitor combined with induction of photothermal ablation combination therapy of renal cell carcinoma (RCC), nanoparticles of varying preparations and concentrations were characterized via zeta potential, drug loading, and release profile. Cytotoxicity and uptake trials were also studied using clear cell RCC cell line RCC 786-0, a human metastatic carcinoma. Target temperatures of >50°C were consistently attained by 0.1 and 0.05 μM concentrations of irradiated human serum albumin nanoparticle-gold nanorods (HSAP-AuNRs). Irradiated trials of HSAP-AuNRs demonstrated significantly decreased cell viabilities compared with nonirradiated “dark” controls (p < 0.01). Increasing loaded masses of sorafenib (SRF) also significantly decreased relative cell viability of RCC (p < 0.05). Photothermal ablation using HSAP-AuNRs is capable of inducing significant hyperthermia while the loading of SRF further enhances cytotoxicity relative to treatment with HSAP-AuNRs alone. HSAP-AuNR-SRFs have the potential to be an effective, novel combination treatment for advanced RCC.

Introduction

Nanoparticles (NPs) have a diversity of clinical applications including diagnostic imaging, chemotherapeutic drug delivery, and photothermal ablation (PTA). Gold nanorods (AuNRs) are effective in all 3 of these applications.1 In clinical use, AuNRs have been conjugated to biomarkers and chemotherapeutic drugs, coated to reduce toxicity, and irradiated to produce hyperthermia.2 Previous studies show human serum albumin proteins (HSAPs) can encapsulate iron oxide particles or act as a vector to improve delivery of chemotherapeutic drugs such as sorafenib (SRF) and paclitaxel.3, 4

Renal cell carcinoma (RCC) is among the most common adult cancers, representing approximately 3% of all malignant tumors in the United States and 85%-90% of all renal malignancies. An estimated 15%-20% of patients with RCC present with metastatic or locally advanced disease.5, 6 The metastatic clear-cell subtype of RCC has an especially poor prognosis, with approximate 5-year survival rates of less than 10%.7 However, in the past 15 years, agents that inhibit vascular endothelial growth factor and mammalian target of rifampin pathways have enhanced treatment options for metastatic and advanced RCC.5 Among them is SRF, a Food and Drug Administration–approved tyrosine kinase inhibitor. Because of the hydrophobic nature and resultant low bioavailability of SRF, high doses may be necessary to reach therapeutic levels. This can lead to systemic complications including hypertension and gastrointestinal hemorrhage.8 NPs offer a potential solution as a drug delivery platform, which could isolate chemotherapeutic payloads from the systemic environment until arrival at target tissues. In addition, because nascent tumor vasculature typically exhibits abnormal fenestrations of 100-200 nm, particles on the nanoscale are capable of preferentially accumulating in neoplastic solid tumors–a phenomenon that has been dubbed the “enhanced permeability and retention” (EPR) effect.9

AuNRs are also capable of PTA, in which laser radiation delivered to excitable particles is converted into thermal energy sufficient to cause hyperthermia, protein denaturation, and cell lysis.10 Generally, target temperatures >50°C are required to induce coagulative necrosis,11 critical in an organ as vascular as the kidney. Studies using AuNRs to induce PTA have achieved these target temperatures both in vitro and in vivo.12, 13

This study used NPs of HSAPs loaded with AuNRs (HSAP-AuNRs) as a vehicle for inducing a synergistic response with photothermic ablation. A variety of mass–ratio preparations of HSA:AuNRs and concentrations of the completed HSAP-AuNR NPs in media were tested for their ability to achieve target temperatures >50°C and to induce irradiation-triggered reduction in cell viabilities. These studies also loaded HSAP-AuNRs with SRF (HSAP-AuNR-SRF). Our hypothesis was that concentrated release of the tyrosine kinase inhibitor from the gold rod would affect cellular viability, when combined with photothermic ablation. Other models have used albumin-loaded AuNRs for delivery such as paclitaxel.14 In this study, our goal was to characterize the loading and irradiation-triggered release of SRF and the effects of combination therapy with PTA on a human metastatic RCC cell line.

Section snippets

Materials

SRF (Nexovar) was purchased from Cayman Chemical (Ann Arbor, MI). Bare AuNRs with aspect ratio of 4.3, diameters of 10 nm, lengths of 43 nm (surface plasmon resonance ∼808-829 nm,) were purchased from Nanopartz Inc. (Loveland, CO). HSA, lyophilized powder ≥97%, coumarin-6, Acrodisc syringe filters with nylon membrane (13 mm diameter, pore size 0.2 μm), 8% aqueous glutaraldehyde, and potassium bromide were purchased from Sigma-Aldrich (St. Louis, MO). Ethanol (200-proof, ACS/USP grade) was

Particle Characteristics

Spherical HSAP-AuNRs achieved varying diameters as measured by field emission-SEM, LEO 1530VP (LEO Elektronenmikroskopie GmbH, Oberkochen, Germany) depending on the mass of HSA used in synthesis (Fig. 5). HSA:AuNR mass ratios of 1.61 × 1013, 8.06 × 1013, and 3.22 × 1014 resulted in HSAP-AuNR diameters of 409 ± 224, 294 ± 83, and 167 ± 4 nm and zeta potentials of −39 ± 3, −35 ± 4, and −33 ± 3 mV, respectively (Table 1). Percent encapsulation of SRF by HSAP-AuNR-SRFs was determined using HPLC

Discussion

In this study, we characterized several NP platforms and evaluated their ability to induce PTA, to load and release SRF, and to reduce relative cell viabilities of metastatic, advanced RCC. The goal was to develop an irradiation-triggered combination therapy of hyperthermia and chemotherapy. We specifically targeted preparations and concentrations of NPs capable of producing peak temperatures in excess of 50° Celsius, which would be capable of causing coagulative necrosis in vivo.11 In

Conclusion

In conclusion, we have demonstrated that these NPs are capable of irradiation-triggered release of SRF and PTA combination therapy to reduce viability of RCC 768-0 at certain preparations, concentrations, and loadings of SRF. Specifically, preparations of HSAP-AuNRs (0.1/0.05 μM) and HSAP-AuNR-SRFs (0.1 μM) reliably and consistently achieved target temperatures >50°C and resulted in significant cell death of RCC 786-O when irradiated. These NPs show promising in vitro results and in the future

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