Contrast Ultrasound Targeted Treatment of Gliomas in Mice via Drug-Bearing Nanoparticle Delivery and Microvascular Ablation
Summary
Insonation of microbubbles is a promising strategy for tumor ablation at reduced time-averaged acoustic powers, as well as for the targeted delivery of therapeutics. The purpose of the present study is to develop low duty cycle ultrasound pulsing strategies and nanocarriers to maximize non-thermal microvascular ablation and payload delivery to subcutaneous C6 gliomas.
Abstract
We are developing minimally-invasive contrast agent microbubble based therapeutic approaches in which the permeabilization and/or ablation of the microvasculature are controlled by varying ultrasound pulsing parameters. Specifically, we are testing whether such approaches may be used to treat malignant brain tumors through drug delivery and microvascular ablation. Preliminary studies have been performed to determine whether targeted drug-bearing nanoparticle delivery can be facilitated by the ultrasound mediated destruction of “composite” delivery agents comprised of 100nm poly(lactide-co-glycolide) (PLAGA) nanoparticles that are adhered to albumin shelled microbubbles. We denote these agents as microbubble-nanoparticle composite agents (MNCAs). When targeted to subcutaneous C6 gliomas with ultrasound, we observed an immediate 4.6-fold increase in nanoparticle delivery in MNCA treated tumors over tumors treated with microbubbles co-administered with nanoparticles and a 8.5 fold increase over non-treated tumors. Furthermore, in many cancer applications, we believe it may be desirable to perform targeted drug delivery in conjunction with ablation of the tumor microcirculation, which will lead to tumor hypoxia and apoptosis. To this end, we have tested the efficacy of non-theramal cavitation-induced microvascular ablation, showing that this approach elicits tumor perfusion reduction, apoptosis, significant growth inhibition, and necrosis. Taken together, these results indicate that our ultrasound-targeted approach has the potential to increase therapeutic efficiency by creating tumor necrosis through microvascular ablation and/or simultaneously enhancing the drug payload in gliomas.
Protocol
1. Microbubble Production To prepare albumin microbubbles ( MBs), place a 1% solution of serum albumin in normal saline in a flask with a blanket of gas (octafluoropropane) above the aqueous phase. Briefly sonicate the solution (30 sec) with an ultrasound disintegrator equipped with an extended ½” titanium probe. This formulation is similar to Optison (GE Heathcare), which is provided in a concentration range of 0.5- 1.2 x 109 MBs/ml. Determine mean MB diameter with a Multisizer Coulter …
Discussion
Critical Steps
Cannulation of mouse tail vein:
Intravenous injection into the mouse tail vein can be a challenging procedure. However, a tail vein catheter can greatly improve the likelihood of a successful injection. To make the catheter, repeatedly bend back and forth a 25 gauge needle until it breaks from the hub. Insert the blunt end into the end of PE 20 tubing and seal the connection with silicon glue. To prepare the catheter for cannulation, attach a syringe …
Declarações
The authors have nothing to disclose.
Acknowledgements
Supported by NIH R01 HL74082, the Hartwell Foundation, and the Focused Ultrasound Surgery Foundation.
Materials
| Material Name | Tipo | Company | Catalogue Number | Comment |
|---|---|---|---|---|
| ApoptTag kit | Intergen Co. | S7110 | ||
| un-capped 85:15 poly(lactic-co-glycolic acid) (PLAGA) | Lakeshore Biomaterials | Custom | ||
| Vivo Tag 680 | VisEn Medical | 10120 | Used to Tag BSA | |
| Poly(vinyl alcohol) | Sigma-Aldrich | 363136 | ||
| MicroTip Sonicator | Misonix | S-4000 | ||
| Sequoia | Simons Medical | P.O.A | Equipped with CPS | |
| FreeZone 2.5 | Labconco | 7670020 | Equipped with Nitrogen Trap | |
| Methylene chloride (CH2Cl2) | Fisher Scientific | D37-500 | ||
| FMT 250 | VisEn Medical | P.O.A | ||
| F-12K Nutrient Mixture | Gibco | 21127-022 | ||
| polyethyleneglycol-40 stearate | Sigma Chemical | 9004-99-3 | ||
| distearoyl phosphatidylcholine | Avanti Polar Lipids | 770365 | ||
| Multisizer Coulter Counter | Beckman Coulter | P.O.A | ||
| Waveform Generator | Tektronix, Inc. | AFG-310 | ||
| water-based ultrasound gel | Parker Laboratories | Aquasonic 100 | ||
| Infusion pump | Harvard Apparatus | Harvard Apparatus PHD 2000 | ||
| 1-Ethyl-3-(3-dimethylaminopropyl)-carbodiimide (EDC) | Pierce Biotechnology | 25952-53-8 | ||
| N-hydroxysulfosuccinimide (Sulfo-NHS) | Pierce Biotechnology | 106627-54-7 | ||
| Succinic anhydride | Sigma Aldrich | 603902 | ||
| Power Amplifier | Electronic Navigation Industries | ENI 3100LA | ||
| Needle Thermocouple Probe | Omega | HYP1-30-1/2-T-G-60-SMPW-M | ||
| BioGel (P100, medium) | Bio-Rad | 150-4170 | ||
| .75’’ diameter 1 MHz unfocused transducer | Panametrics | A314S |
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Tags
Contrast UltrasoundTargeted TreatmentGliomasMiceDrug-bearing NanoparticlesMicrovascular AblationMinimally-invasiveContrast Agent MicrobubbleUltrasound Pulsing ParametersMalignant Brain TumorsDrug DeliveryComposite Delivery AgentsPoly(lactide-co-glycolide) NanoparticlesAlbumin Shelled MicrobubblesSubcutaneous C6 GliomasNanoparticle DeliveryMicrobubble-nanoparticle Composite Agents (MNCAs)Cancer ApplicationsTumor MicrocirculationTumor HypoxiaApoptosisNon-thermal Cavitation-induced Microvascular Ablation
Citar este artigo
Burke, C. W., Price, R. J. Contrast Ultrasound Targeted Treatment of Gliomas in Mice via Drug-Bearing Nanoparticle Delivery and Microvascular Ablation. J. Vis. Exp. (46), e2145, doi:10.3791/2145 (2010).