ABSTRACT
We report development of a nanoparticle-based, X-ray-activated anticancer "nanodrug" composed of yttrium oxide (Y(2)O(3)) nanoscintillators, a fragment of the HIV-1 TAT peptide, and psoralen. In this formulation, X-ray radiation is absorbed by the Y(2)O(3) nanoscintillators, which then emit UVA light. Absorption of UVA photons by nanoparticle-tethered psoralen has the potential to cross-link adenine and thymine residues in DNA. UVA-induced cross-linking by free psoralen upon activation with UVA light has previously been shown to cause apoptosis in vitro and an immunogenic response in vivo. Studies using the PC-3 human prostate cancer cell line demonstrate that X-ray excitation of these psoralen-functionalized Y(2)O(3) nanoscintillators yields concentration-dependent reductions in cell number when compared to control cultures containing psoralen-free Y(2)O(3) nanoscintillators.
Subject(s)
Ficusin/pharmacology , Nanotechnology/methods , Prostatic Neoplasms/drug therapy , Prostatic Neoplasms/radiotherapy , Adenine/chemistry , Apoptosis , Cell Line, Tumor , DNA/chemistry , Drug Screening Assays, Antitumor/methods , Humans , Male , PUVA Therapy/methods , Thymine/chemistry , X-Rays , Yttrium/chemistry , tat Gene Products, Human Immunodeficiency Virus/chemistryABSTRACT
We describe the development and application of a co-functionalized nanoprobe and biodelivery platform combining a nuclear targeting peptide (NTP) for improved cellular uptake and intracellular targeting with p-mercaptobenzoic acid (pMBA) as a surface-enhanced Raman scattering (SERS) reporter for tracking and imaging. The nuclear targeting peptide, an HIV-1 protein-derived TAT sequence, has been previously shown to aid entry of cargo through the cell membrane via normal cellular processes, and furthermore, to localize small cargo to the nucleus of the cell. Previous work in our lab has verified cell uptake and distribution of the nanoprobes in clinically relevant mouse and human cell lines. In this work, two-dimensional SERS mapping was used to track the spatial and temporal progress of nanoparticle uptake in PC-3 human prostate cells and to characterize localization at various time points, demonstrating the potential for an intracellularly targeted multiplexed nanobiosensing system with excellent sensitivity and specificity. Silver nanoparticles co-functionalized with the TAT peptide showed greatly enhanced cellular uptake over the control nanoparticles lacking the targeting moiety. The ability to detect and monitor nanoprobe trafficking using SERS spectroscopy offers an improved alternative over previous tracking and detection methods such as light microscopy and fluorescence methods. The development of multifunctional nanoconstructs for intracellular delivery has potential clinical applications in early detection and selective treatment of disease in affected cells. Other applications include use in basic research aimed at understanding the inner workings of living cells and how they respond to chemical and biological stimuli.
Subject(s)
Cell Nucleus/metabolism , Metal Nanoparticles/chemistry , Signal Processing, Computer-Assisted , Spectrum Analysis, Raman/methods , Benzoates/chemistry , Benzoates/metabolism , Cell Line, Tumor , Cell Membrane/metabolism , Humans , Male , Peptide Fragments/chemistry , Peptide Fragments/metabolism , Prostatic Neoplasms/metabolism , Silver/chemistry , Silver/pharmacokinetics , Sulfhydryl Compounds/chemistry , Sulfhydryl Compounds/metabolism , tat Gene Products, Human Immunodeficiency Virus/chemistry , tat Gene Products, Human Immunodeficiency Virus/metabolismABSTRACT
We report the first demonstration of surface-enhanced Raman spectroscopy (SERS) detection of para-mercapto benzoic acid (pMBA) and surface-enhanced resonance Raman spectroscopy (SERRS) detection of brilliant cresyl blue (BCB) and cresyl violet perchlorate (CVP) with continuous-wave excitation from a stand-off distance of 15 meters. We further report the first stand-off SERRS detection of BCB and CVP at that same distance in the presence of ambient fluorescent and incandescent/blackbody background light. These preliminary results suggest that it is possible to detect sub-nanomole amounts of material at reasonable distances with eye-safe laser powers using stand-off SERRS and serve as proof-of-concept highlighting the potential extension of stand-off Raman spectroscopy to include SERS and SERRS for remote, eye-safe chemical detection, analysis, and imaging in the presence of ambient background light.
