Biocompatible Gold Nanorod Conjugates for Preclinical Biomedical Research

Gold nanorods with a peak absorption wavelength of 760 nm were prepared using a seed-mediated method. A novel protocol has been developed to replace hexadecyltrimethylammonium bromide on the surface of the nanorods with 16-mercaptohexadecanoic acid and metoxy-poly(ethylene glycol)-thiol, and the monoclonal antibody HER2. The physical chemistry properties of the conjugates were monitored through optical and zeta-potential measurements to confirm surface chemistry changes. The efficiency of the modifications was quantified through measurement of the average number of antibodies per gold nanorod. The conjugates were investigated for different cells lines: BT-474, MCF7, MCF10, MDCK, and fibroblast. The results show successful cell accumulation of the gold nanorod HER2 conjugates in cells with HER2 overexpression. Incubation of the complexes in heparinized mouse blood demonstrated the low aggregation of the metallic particles through stability of the spectral properties, as verified by UV/VIS spectrometry. Cytotoxicity analysis with LDH release and MTT assay confirms strong targeting and retention of functional activity of the antibody after their conjugation with gold nanorods. Silver staining confirms efficient specific binding to BT-474 cells even in cases where the nanorod complexes were incubated in heparinized mouse blood. This is confirmed through in vivo studies where, following intravenous injection of gold nanorod complexes, silver staining reveals noticeably higher rates of specific binding in mouse tumors than in healthy liver. The conjugates are reproducible, have strong molecular targeting capabilities, have long term stability in vivo and can be used in pre-clinical applications. The conjugates can also be used for molecular and optoacoustic imaging, quantitative sensing of biological substrates, and photothermal therapy

Structure-Dependent Fluorescence Efficiencies of Individual Single-Walled Carbon Nanotubes

Single-nanotube photometry was used to measure the product of absorption cross-section and fluorescence quantum yield for 12 (n,m) structural species of semiconducting SWNTs in aqueous SDBS suspension. These products ranged from 1.7 to 4.5 x 10(-19) cm2/C atom, generally increasing with optical band gap as described by the energy gap law. The findings suggest fluorescent quantum yields of ~8% for the brightest, (10,2) species and introduce the empirical calibration factors needed to deduce quantitative (n,m) distributions from bulk fluorimetric intensities

Stepwise Quenching of Exciton Fluorescence in Carbon Nanotubes by Single Molecule Reactions

Single-molecule chemical reactions with individual single-walled carbon nanotubes were observed through near-infrared photoluminescence microscopy. The emission intensity within distinct submicrometer segments of single nanotubes changes in discrete steps after exposure to acid, base, or diazonium reactants. The steps are uncorrelated in space and time, and reflect the quenching of mobile excitons at localized sites of reversible or irreversible chemical attack. Analysis of step amplitudes reveals an exciton diffusional range of about 90 nanometers, independent of nanotube structure. Each exciton visits approximately 104 atomic sites during its lifetime, providing highly efficient sensing of local chemical and physical perturbations

Enabling in vivo measurements of nanoparticle concentrations with three-dimensional optoacoustic tomography

In this report, we demonstrate the feasibility of using optoacoustic tomography (OAT) to evaluate biodistributions of nanoparticles in animal models. The redistribution of single-walled carbon nanotubes (SWCNTs) was visualized in living mice. Nanoparticle concentrations in harvested organs were measured spectroscopically using the intrinsic optical absorption and fluorescence of SWCNTs. Observed increases in optoacoustic signal brightness in tissues were compared with increases in optical absorption coefficients caused by SWCNT accumulation. The methodology presented in this report can further be extended to calibrate the sensitivity of an optoacoustic imaging system for a range of changes in optical absorption coefficient values at specific locations or organs in a mouse body to enable noninvasive measurements of nanoparticle concentrations in vivo. Additionally, qualitative information provided by OAT and quantitative information obtained ex vivo may provide valuable feedback for advancing methods of quantitative analysis with OAT

Shear-strain step response in linear regime of dilute suspensions of naturally bent carbon nanotubes

Impressive enhancements of the storage modulus have been documented when low volume fractions of single wall carbon nanotubes (SWNTs) are added to a Newtonian solvent for obtaining dilute suspensions. The intrinsic bending dynamics of carbon nanotubes (CNTs) has been proposed to explain such elasticity. CNTs contain topological defects inducing naturally bent structures in absence of external forces and, hence, a semiflexible filament with a bent configuration at minimal internal-bending-energy is used for mimicking the structure of SWNTs in suspension. Previous continuous model is discretized as a non-freely jointed bead-rod chain with a naturally bent configuration for simulating the rheological behaviour after a shear-strain step in linear regime of SWNT dilute suspension by using a Brownian dynamics (BD) approach. In general, bead-rod chains exhibit an instantaneous relaxation after a high shear-strain step. Bending rigidity and number of constitutive rods are found to be determinant parameters in the internal-energy relaxation behaviour of non-freely jointed bead-rod chains in dilute solution. Proper comparisons between the BD simulation results and the experimental data for treated SWNT dilute suspensions confirm the consistency of the physical model mimicking the structure of a SWNT