Shell effects on acoustic performance of a drug-delivery system activated by ultrasound.

The composition of microcapsules designed for drug delivery significantly impacts their properties. Ultrasound contrast agents, consisting of stabilized microbubbles (MBs), have emerged as versatile potential drug delivery vehicles to both image and overcome challenges associated with systemic chemotherapy. In our development of polylactic acid MBs decorated with immune-shielding polyethylene glycol chains, we have shown that the balance between acoustic behavior and immune avoidance was scalable and amenable to two distinct PEGylation methods, either incorporation of 5 wt% PEGylated PLA or insertion of 1 wt% PEGylated lipid (LipidPEG) in the polymeric shell. Here we describe the effects of shell compositions on MB functionalization for use in targeted cancer therapy. We chose tumor necrosis factor-related apoptosis inducing ligand (TRAIL) as the targeting ligand, motivated by the ability to both target cells and selectively induce tumor cell death upon binding. Additionally, the MBs were designed to co-encapsulate the chemotherapeutic doxorubicin (Dox) within the shell that works with TRAIL to sensitize resistant cells. We have previously shown that the MBs shatter in response to ultrasound focused at the tumor site, delivering drug-eluting fragments. This study demonstrates the effect of shell characteristics and MB functionalization (TRAIL-ligated and Dox-loaded MBs) on the acoustic response of MBs, and the cumulative effect of shell type. © 2017 Wiley Periodicals, Inc. J Biomed Mater Res Part A: 105A: 3189-3196, 2017.

Cellular signal transduction can be induced by TRAIL conjugated to microcapsules.

The extracellular agent tumor necrosis factor-related apoptosis-inducing ligand (TRAIL) can induce apoptosis in tumor cells but spare normal cells. Ligation of TRAIL to a nanoparticle would serve to facilitate targeting to an extravascular site. Polymeric ultrasound contrast agents (UCA) (microencapsulated gas bubbles) can be tracked by ultrasound imaging, and fragmented into nanoparticles by focused ultrasound. This tumor-targeted delivery system has been shown to deliver more efficiently than solid nanoparticles. Additionally, small molecule inhibitors such as bortezomib, shown to sensitize TRAIL-resistant cells, could be co-administered within these UCA. In this pilot study, TRAIL was conjugated to UCA while preserving the agent's sensitivity to ultrasound. Human cancer cell lines, OVCAR-3 and A2058, were bathed with the TRAIL-UCA with and without the addition of bortezomib. Apoptosis was quantified using flow cytometry. OVCAR-3 treated with TRAIL-UCA exhibit significant (p < 0.05) apoptotosis compared to unmodified UCA, equal to positive controls, but no synergistic effect when combined with bortezomib. A2058 cells treated with TRAIL-UCA also exhibited significant apoptosis (p < 0.01) compared to unmodified UCA, similar to positive controls and bortezomib significantly increased apoptosis in combination with TRAIL-UCA. We conclude that TRAIL-ligated UCA show exciting potential as a new therapy.

Doxorubicin and paclitaxel loaded microbubbles for ultrasound triggered drug delivery.

A polymer ultrasound contrast agent (UCA) developed in our lab has been shown to greatly reduce in size when exposed to ultrasound, resulting in nanoparticles less than 400 nm in diameter capable of escaping the leaky vasculature of a tumor to provide a sustained release of drug. Previous studies with the hydrophilic drug doxorubicin (DOX) demonstrated enhanced drug delivery to tumors when triggered with ultrasound. However the therapeutic potential has been limited due to the relatively low payload of DOX. This study compares the effects of loading the hydrophobic drug paclitaxel (PTX) on the agent's acoustic properties, drug payload, tumoricidal activity, and the ability to deliver drugs through 400 nm pores. A maximum payload of 129.46 ± 1.80 µg PTX/mg UCA (encapsulation efficiency 71.92 ± 0.99%) was achieved, 20 times greater than the maximum payload of DOX (6.2 µg/mg), while maintaining the acoustic properties. In vitro, the tumoricidal activity of paclitaxel loaded UCA exposed to ultrasound was significantly greater than controls not exposed to ultrasound (p<0.0016). This study has shown that PTX loaded UCA triggered with focused ultrasound have the potential to provide a targeted and sustained delivery of drug to tumors.

Disposition of ultrasound sensitive polymeric drug carrier in a rat hepatocellular carcinoma model.

RATIONALE AND OBJECTIVES: A doxorubicin-loaded microbubble has been developed that can be destroyed with focused ultrasound resulting in fragments, or "nanoshards" capable of escaping through the leaky tumor vasculature, promoting accumulation within the interstitium. This study uses a rat liver cancer model to examine the biodistribution and tumoral delivery of this microbubble platform compared with de novo drug-loaded polymer nanoparticles and free doxorubicin. MATERIALS AND METHODS: Microbubbles (1.8 µm) and 217-nm nanoparticles were prepared containing 14-C labeled doxorubicin. Microbubbles, nanoparticles, a combination of the two, or free doxorubicin were administered intravenously in rats bearing hepatomas, concomitant with tumor insonation. Doxorubicin levels in plasma, organs, and tumors were quantified after 4 hours and 7 and 14 days. Tumors were measured on sacrifice and evaluated with autoradiography and histology. RESULTS: Animals treated with microbubbles had significantly lower plasma doxorubicin concentrations (0.466 ± 0.068%/mL) compared with free doxorubicin (3.033 ± 0.612%/mL, P = .0019). Drug levels in the myocardium were significantly lower in animals treated with microbubbles compared to free doxorubicin (0.168%/g tissue vs. 0.320%/g, P = .0088). Tumors treated with microbubbles showed significantly higher drug levels than tumors treated with free doxorubicin (2.491 ± 0.501 %/g vs. 0.373 ± 0.087 %/g, P = .0472). These tumors showed significantly less growth than tumors treated with free doxorubicin (P = .0390). CONCLUSIONS: Doxorubicin loaded microbubbles triggered with ultrasound provided enhanced, sustained drug delivery to tumors, reduced plasma and myocardium doxorubicin levels, and arresting tumor growth. The results suggest that in situ generation of nano particles provides a superior treatment over injection of free drug and also de novo synthesized nanoparticles.