Local Intratracheal Delivery of Perfluorocarbon Nanoparticles to Lung Cancer Demonstrated with Magnetic Resonance Multimodal Imaging.

Eighty percent of lung cancers originate as subtle premalignant changes in the airway mucosal epithelial layer of bronchi and alveoli, which evolve and penetrate deeper into the parenchyma. Liquid-ventilation, with perfluorocarbons (PFC) was first demonstrated in rodents in 1966 then subsequently applied as lipid-encapsulated PFC emulsions to improve pulmonary function in neonatal infants suffering with respiratory distress syndrome in 1996. Subsequently, PFC nanoparticles (NP) were extensively studied as intravenous (IV) vascular-constrained nanotechnologies for diagnostic imaging and targeted drug delivery applications. Methods: This proof-of-concept study compared intratumoral localization of fluorescent paramagnetic (M) PFC NP in the Vx2 rabbit model using proton (1H) and fluorine (19F) magnetic resonance (MR) imaging (3T) following intratracheal (IT) or IV administration. MRI results were corroborated by fluorescence microscopy. Results: Dynamic 1H-MR and 19F-MR images (3T) obtained over 72 h demonstrated marked and progressive accumulation of M-PFC NP within primary lung Vx2 tumors during the first 12 h post IT administration. Marked 1H and 19F MR signal persisted for over 72 h. In contradistinction, IV M-PFC NP produced a modest transient signal during the initial 2 h post-injection that was consistent circumferential blood pool tumor enhancement. Fluorescence microscopy of excised tumors corroborated the MR results and revealed enormous intratumor NP deposition on day 3 after IT but not IV treatment. Rhodamine-phospholipid incorporated into the PFC nanoparticle surfactant was distributed widely within the tumor on day 3, which is consistent with a hemifusion-based contact drug delivery mechanism previously reported. Fluorescence microscopy also revealed similar high concentrations of M-PFC NP given IT for metastatic Vx2 lung tumors. Biodistribution studies in mice revealed that M-PFC NP given IV distributed into the reticuloendothelial organs, whereas, the same dosage given IT was basically not detected beyond the lung itself. PFC NP given IT did not impact rabbit behavior or impair respiratory function. PFC NP effects on cells in culture were negligible and when given IV or IT no changes in rabbit hematology nor serum clinical chemistry parameters were measured. Conclusion: IT delivery of PFC NP offered unique opportunity to locally deliver PFC NP in high concentrations into lung cancers with minimal extratumor systemic exposure.

Surface passivation of carbon nanoparticles with branched macromolecules influences near infrared bioimaging.

A superior and commercially exploitable 'green synthesis' of optically active carbon nanoparticle (OCN) is revealed in this work. The naked carbon particles (<20 nm) were derived from commercial food grade honey. The fluorescence properties of these particles were significantly enhanced by utilizing hyberbranched polymer for surface passivation. A dramatic increase in near infrared emission was achieved compared to a linear polymer (PEG) coated carbon nanoparticles. Interestingly, as passivating agent becomes more extensively branched (pseudo generation 2 to 4), the average radiant efficiency amplifies considerably as a direct result of the increasing surface area available for light passivation. The particles showed negligible loss of cell viability in presence of endothelial cells in vitro. Preliminary in vivo experiment showed high contrast enhancement in auxiliary lymphnode in a mouse model. The exceptionally rapid lymphatic transport of these particles suggests that such an approach may offer greater convenience and reduced procedural expense, as well as improved surgical advantage as the patient is positioned on the table for easier resection.

A Green Synthesis of Carbon Nanoparticle from Honey for Real-Time Photoacoustic Imaging.

Imaging sentinel lymph nodes (SLN) could provide us with critical information about the progression of a cancerous disease. Real-time high-resolution intraoperative photoacoustic imaging (PAI) in conjunction with a near infrared (NIR) probe may offer the opportunities for the immediate imaging for direct identification and resection of SLN or collecting tissue samples. In this work a commercially amenable synthetic methodology is revealed for developing luminescent carbon nanoparticles with rapid clearance properties. A one-pot "green" technique is pursued, which involved rapid surface passivation of carbon nanoparticles with organic macromolecules (e.g. polysorbate, polyethyleneglycol) in a solvent free condition. Interestingly, the naked carbon nanoparticles are derived for the first time, from commercial food grade honey. Surface coated particles are markedly smaller (~7 nm) than the previously explored particles (gold, SWNT, copper) for SLN imaging. Results indicate an exceptionally rapid signal enhancement (~2 min) of the SLN. Owing to their strong optical absorption in the near infrared region, tiny size and rapid lymphatic transport, this platform offers great potential for faster resection of SLN and may lower complications caused by axillary investigation for mismarking with dyes or low-resolution imaging techniques.

Rapid synthesis of near infrared polymeric micelles for real-time sentinel lymph node imaging.

In this manuscript a synthetic methodology for developing sub 20 nm sized polymeric micellar nanoparticles designed for extravascular imaging and therapy is revealed. A simple, one-pot method is followed, which involves a rapid co-self-assembly of an amphiphilic diblock copolymer (PS-b-PAA) and polyoxyethylene (80) sorbitan monooleate in water. Sorbitan monooleate imparts stability to the micelles and helps to drive down the particle size below 20 nm. The particles are incorporated with a water soluble dye ADS832WS, which absorbs in the near infrared range (λ(ex) = 832 nm) for sensitive detection with optical and photoacoustic imaging techniques. A candidate lipophilic anti-angiogenic therapeutic agent fumagillin was also incorporated with high entrapment (>95%) efficiency. The effectiveness of this theranostic platform for real-time, high-resolution intraoperative photoacoustic imaging for facilitating direct assessment of the sentinel lymph nodes (SLN) in breast cancer staging is demonstrated. The technique offers huge potential providing faster resection of SLN and may minimize complications caused by axillary exploration due to mismarking with dyes or low-resolution imaging techniques. Finally, the biodistribution and organ accumulation of the intravenously and intradermally injected particles are studied in a rodent model by optical imaging. Data suggest that intraveneously injected NIR-polymeric nanoparticles follow a typical bio-distribution clearance path through the reticuloendothelial (RES) system. For the intradermally injected particles, a slower mechanism of clearance is noticed.

Second Generation Gold Nanobeacons for Robust K-Edge Imaging with Multi-Energy CT.

Spectral CT is the newest advancement in CT imaging technology, which enhances traditional CT images with the capability to image and quantify certain elements based on their distinctive K-edge energies. K-edge imaging feature recognizes high accumulations of targeted elements and presents them as colorized voxels against the normal grayscale X-ray background offering promise to overcome the relatively low inherent contrast within soft tissue and distinguish the high attenuation of calcium from contrast enhanced targets. Towards this aim, second generation gold nanobeacons (GNB(2)), which incorporate at least five times more metal than the previous generation was developed. The particles were synthesized as lipid-encapsulated, vascularly constrained (>120 nm) nanoparticle incorporating tiny gold nanoparticles (2-4 nm) within a polysorbate core. The choice of core material dictated to achieve a higher metal loading. The particles were thoroughly characterized by physicochemical techniques. This study reports one of the earlier examples of spectral CT imaging with gold nanoparticles demonstrating the potential for targeted in vitro and in vivo imaging and eliminates calcium interference with CT. The use of statistical image reconstruction shows high SNR may allow dose reduction and/or faster scan times.

Sensitive biological detection with a soluble and stable polymeric paramagnetic nanocluster.

We describe the design, synthesis, and biological characterization of manganese oxocluster-based "single molecule magnets (SMMs)". We demonstrate that polymeric micellar nanoparticles can serve as a carrier and help to stabilize delicate SMM molecules from breaking down easily and thus prevent their property loss. Concentrating thousands of Mn-clusters per micelle provided a high ionic and per-particle relaxivity allowing sensitive MR imaging in vivo. This reports one of the earliest examples of in vivo imaging of a rationally designed polymeric micelle that features SMM.

An early investigation of ytterbium nanocolloids for selective and quantitative "multicolor" spectral CT imaging.

We report a novel molecular imaging agent based on ytterbium designed for use with spectral "multicolor" computed tomography (CT). Spectral CT or multicolored CT provides all of the benefits of traditional CT, such as rapid tomographic X-ray imaging, but in addition, it simultaneously discriminates metal-rich contrast agents based on the element's unique X-ray K-edge energy signature. Our synthetic approach involved the use of organically soluble Yb(III) complex to produce nanocolloids of Yb of noncrystalline nature incorporating a high density of Yb (>500K/nanoparticle) into a stable metal particle. The resultant particles are constrained to vasculature (∼200 nm) and are highly selective for binding fibrin in the ruptured atherosclerotic plaque. Nanoparticles exhibited excellent signal sensitivity, and the spectral CT technique uniquely discriminates the K-edge signal (60 keV) of Yb from calcium (bones). Bioelimination and preliminary biodistribution reflected the overall safety and defined clearance of these particles in a rodent model.

Synthesis of NanoQ, a copper-based contrast agent for high-resolution magnetic resonance imaging characterization of human thrombus.

A new site-targeted molecular imaging contrast agent based on a nanocolloidal suspension of lipid-encapsulated, organically soluble divalent copper has been developed. Concentrating a high payload of divalent copper ions per nanoparticle, this agent provides a high per-particle r1 relaxivity, allowing sensitive detection in T1-weighted magnetic resonance imaging when targeted to fibrin clots in vitro. The particle also exhibits a defined clearance and safety profile in vivo.