Avinash Murthy

Controlled Assembly of Biodegradable Plasmonic Nanoclusters for Near-Infrared Imaging and Therapeutic Applications

Metal nanoparticles with surface plasmon resonance (SPR) in the near-infrared region (NIR) are of great interest for imaging and therapy. Presently, gold nanoparticles with NIR absorbance are typically larger than 50 nm, above the threshold size of approximately 5 nm required for efficient renal clearance. As these nanoparticles are not biodegradable, concerns about long-term toxicity have restricted their translation into the clinic. Here, we address this problem by developing a flexible platform for the kinetically controlled assembly of sub-5 nm ligand-coated gold particles to produce metal/polymer biodegradable nanoclusters smaller than 100 nm with strong NIR absorbance for multimodal application. A key novel feature of the proposed synthesis is the use of weakly adsorbing biodegradable polymers that allows tight control of nanocluster size and, in addition, results in nanoclusters with unprecedented metal loadings and thus optical functionality. Over time, the biodegradable polymer stabilizer degrades under physiological conditions that leads to disassembly of the nanoclusters into sub-5 nm primary gold particles which are favorable for efficient body clearance. This synthesis of polymer/inorganic nanoclusters combines the imaging contrast and therapeutic capabilities afforded by the NIR-active nanoparticle assembly with the biodegradability of a polymer stabilizer.

Kinetic assembly of near-IR-active gold nanoclusters using weakly adsorbing polymers to control the size.

Clusters of metal nanoparticles with an overall size of less than 100 nm and high metal loadings for strong optical functionality are of interest in various fields including microelectronics, sensors, optoelectronics, and biomedical imaging and therapeutics. Herein we assemble approximately 5 nm gold particles into clusters with controlled size, as small as 30 nm and up to 100 nm, that contain only small amounts of polymeric stabilizers. The assembly is kinetically controlled with weakly adsorbing polymers, PLA(2K)-b-PEG(10K)-b-PLA(2K) or PEG (MW = 3350), by manipulating electrostatic, van der Waals (VDW), steric, and depletion forces. The cluster size and optical properties are tuned as a function of particle volume fractions and polymer/gold ratios to modulate the interparticle interactions. The close spacing between the constituent gold nanoparticles and high gold loadings (80-85 w/w gold) produce a strong absorbance cross section of approximately 9 x 10(-15) m(2) in the NIR at 700 nm. This morphology results from VDW and depletion attractive interactions that exclude the weakly adsorbed polymeric stabilizer from the cluster interior. The generality of this kinetic assembly platform is demonstrated for gold nanoparticles with a range of surface charges from highly negative to neutral with the two different polymers.

Equilibrium gold nanoclusters quenched with biodegradable polymers

Although sub-100 nm nanoclusters of metal nanoparticles are of interest in many fields including biomedical imaging, sensors, and catalysis, it has been challenging to control their morphologies and chemical properties. Herein, a new concept is presented to assemble equilibrium Au nanoclusters of controlled size by tuning the colloidal interactions with a polymeric stabilizer, PLA(1k)-b-PEG(10k)-b-PLA(1k). The nanoclusters form upon mixing a dispersion of ~5 nm Au nanospheres with a polymer solution followed by partial solvent evaporation. A weakly adsorbed polymer quenches the equilibrium nanocluster size and provides steric stabilization. Nanocluster size is tuned from ~20 to ~40 nm by experimentally varying the final Au nanoparticle concentration and the polymer/Au ratio, along with the charge on the initial Au nanoparticle surface. Upon biodegradation of the quencher, the nanoclusters reversibly and fully dissociate to individual ~5 nm primary particles. Equilibrium cluster size is predicted semiquantitatively with a free energy model that balances short-ranged depletion and van der Waals attractions with longer-ranged electrostatic repulsion, as a function of the Au and polymer concentrations. The close spacings of the Au nanoparticles in the clusters produce strong NIR extinction over a broad range of wavelengths from 650 to 900 nm, which is of practical interest in biomedical imaging.

Utility of biodegradable plasmonic nanoclusters in photoacoustic imaging

Plasmonic metal nanoparticles are used in photoacoustic imaging as contrast agents because of their resonant optical absorption properties in the visible and near-IR regions. However, the nanoparticles could accumulate and result in long-term toxicity in vivo, because they are generally not biodegradable. Recently, biodegradable plasmonic gold nanoclusters, consisting of sub-5 nm primary gold nanoparticles and biodegradable polymer stabilizer, were introduced. In this Letter, we demonstrate the feasibility of biodegradable nanoclusters as a photoacoustic contrast agent. We performed photoacoustic and ultrasound imaging of a tissue-mimicking phantom with inclusions containing nanoclusters at various concentrations. The results indicate that the biodegradable gold nanoclusters can be used as effective contrast agents in photoacoustic imaging.

Quenched Assembly of NIR-Active Gold Nanoclusters Capped with Strongly Bound Ligands by Tuning Particle Charge via pH and Salinity

Gold nanospheres coated with a binary monolayer of bound citrate and cysteine ligands were assembled into nanoclusters, in which the size and near-infrared (NIR) extinction were tuned by varying the pH and concentration of added NaCl. During full evaporation of an aqueous dispersion of 4.5 ± 1.8 nm Au primary particles, the nanoclusters were formed and quenched by the triblock copolymer polylactic acid (PLA)(1K)-b-poly(ethylene glycol) (PEG)(10K)-b-PLA(1K), which also provided steric stabilization. The short-ranged depletion and van der Waals attractive forces were balanced against longer ranged electrostatic repulsion to tune the nanocluster diameter and NIR extinction. Upon lowering the pH from 7 to 5 at a given salinity, the magnitude of the charge on the primary particles decreased, such that the weaker electrostatic repulsion increased the hydrodynamic diameter and, consequently, NIR extinction of the clusters. At a given pH, as the concentration of NaCl was increased, the NIR extinction decreased monotonically. Furthermore, the greater screening of the charges on the nanoclusters weakened the interactions with PLA(1K)-b-PEG(10K)-b-PLA(1K) and thus lowered the amount of adsorbed polymer on the nanocluster surface. The generalization of the concept of self-assembly of small NIR-active nanoclusters to include a strongly bound thiol and the manipulation of the morphologies and NIR extinction by variation of pH and salinity not only is of fundamental interest but also is important for optical biomedical imaging and therapy.

Thermal stability of biodegradable plasmonic nanoclusters in photoacoustic imaging.

The photothermal stability of plasmonic nanoparticles is critically important to perform reliable photoacoustic imaging and photothermal therapy. Recently, biodegradable nanoclusters composed of sub-5 nm primary gold particles and a biodegradable polymer have been reported as clinically-translatable contrast agents for photoacoustic imaging. After cellular internalization, the nanoclusters degrade into 5 nm primary particles for efficient excretion from the body. In this paper, three different sizes of biodegradable nanoclusters were synthesized and the optical properties and photothermal stability of the nanoclusters were investigated and compared to that of gold nanorods. The results of our study indicate that 40 nm and 80 nm biodegradable nanoclusters demonstrate higher photothermal stability compared to gold nanorods. Furthermore, 40 nm nanoclusters produce higher photoacoustic signal than gold nanorods at a given concentration of gold. Therefore, the biodegradable plasmonic nanoclusters can be effectively used for photoacoustic imaging and photothermal therapy.

Biodegradable plasmonic nanoclusters as contrast agent for photoacoustic imaging

Metallic nanoparticles have been widely used in a variety of imaging and therapeutic applications due to their unique optical properties in the visible and near-infrared (NIR) regions - for example, various plasmonic nanoparticles are used for molecular photoacoustic imaging and photothermal therapy. However, there are concerns that these agents may not be safe under physiological conditions, because these nanoparticles are not biodegradable, could accumulate and, therefore, could be toxic long-term. We investigate the feasibility of using biodegradable gold nanoclusters as a contrast agent for highly sensitive photoacoustic imaging. The size of these biodegradable nanoclusters, consisting of sub-5 nm primary gold particles and a biodegradable polymer binder, is less than 100 nm. Due to plasmon coupling, these nanoclusters are characterized by a broad extinction spectrum that extends to the near infrared (NIR) spectral range. Photoacoustic imaging of tissue models containing inclusions with different concentrations of nanoparticles was performed using a tunable pulsed laser system. The results indicate that the biodegradable nanoclusters, comprised of small gold nanoparticles, can be used as contrast agents in photoacoustic imaging.

Plasmonic biodegradable gold nanoclusters with high NIR-absorbance for biomedical imaging

Gold plasmonic nanoparticles are receiving attention for a variety of types of NIR optical biomedical imaging including photoacoustic imaging. Herein we present a novel method to assemble equilibrium gold nanoclusters from 5 nm primary gold nanospheres, which exhibit high near-infrared (NIR) absorbance and subsequently fully dissociate back to primary particles, which has the potential to enable renal clearance. The nanoparticle assembly is manipulated via controlling colloidal interactions, specifically electrostatic repulsion and depletion attraction. The charge on the primary ~5 nm gold nanospheres is tailored via place exchange reactions with a variety of biocompatible ligands such as citrate, lysine and cysteine. The primary particles form clusters upon addition of a biodegradable polymer, PLA(1k)-b- PEG(10k)-b-PLA(1k), followed by controlled solvent evaporation. The cluster size may be tuned from 20-40 nm in diameter by manipulating the gold and polymer concentrations along with the solvent evaporation extent. Salt is also added to increase the NIR absorbance and reduce the nanocluster size by reducing polymer adsorption. The adsorption of the polymer onto the Au surfaces effectively quenches the nanoclusters. High NIR absorption facilitates photoacoustic imaging, even for the small cluster sizes. In response to acidic cellular pH environments, the polymer degrades and the clusters dissociate back to primary particle on the order of 5 nm, which are small enough for renal clearance.

Biodegradable Plasmonic Nanoparticles: Overcoming Clinical Translation Barriers

We present biodegradable gold nanoparticles with plasmon resonances in the NIR region that can provide a crucial link between the enormous potential of metal nanoparticles for cancer imaging and therapy and translation into clinical practice.

Photoacoustic imaging with biodegradable plasmonic nanoclusters

Plasmonic nanoparticles have been widely used for various biomedical applications such as biological imaging, sensing, and cancer therapy. Specifically, gold nanospheres, nanorods, and nanoshells are used as contrast agents for photoacoustic imaging due to their strong absorptive property. However, there are concerns about using these nanoparticles in-vivo because they are not biodegradable and cannot be cleared from the body. Recently, biodegradable nanoclusters have been reported. The biodegradable nanoparticles are composed of primary 4-nm gold nanoparticles and stabilized by a biodegradable polymer binder. In this study, we demonstrated the utility of biodegradable nanoclusters as a contrast agent in photoacoustic imaging. The tissue mimicking phantoms were used for ultrasound and photoacoustic imaging. The results show that the biodegradable plasmonic nanoclusters can be used as photoacoustic contrast agent.