UCNP-based Photoluminescent Nanomedicines for Targeted Imaging and Theranostics of Cancer.

Theranostic approach is currently among the fastest growing trends in cancer treatment. It implies the creation of multifunctional agents for simultaneous precise diagnosis and targeted impact on tumor cells. A new type of theranostic complexes was created based on NaYF4: Yb,Tm upconversion nanoparticles coated with polyethylene glycol and functionalized with the HER2-specific recombinant targeted toxin DARPin-LoPE. The obtained agents bind to HER2-overexpressing human breast adenocarcinoma cells and demonstrate selective cytotoxicity against this type of cancer cells. Using fluorescent human breast adenocarcinoma xenograft models, the possibility of intravital visualization of the UCNP-based complexes biodistribution and accumulation in tumor was demonstrated.

Neutrophil targeted nano-drug delivery system for chronic obstructive lung diseases.

The success of drug delivery to target airway cell(s) remains a significant challenge due to the limited ability of nanoparticle (NP) systems to circumvent protective airway-defense mechanisms. The size, density, surface and physical-chemical properties of nanoparticles are the key features that determine their ability to navigate across the airway-barrier. We evaluated here the efficacy of a PEGylated immuno-conjugated PLGA-nanoparticle (PINP) to overcome this challenge and selectively deliver drug to specific inflammatory cells (neutrophils). We first characterized the size, shape, surface-properties and neutrophil targeting using dynamic laser scattering, transmission electron microscopy and flow cytometry. Next, we assessed the efficacy of neutrophil-targeted PINPs in transporting through the airway followed by specific binding and release of drug to neutrophils. Finally, our results demonstrate the efficacy of PINP mediated non-steroidal anti-inflammatory drug-(ibuprofen) delivery to neutrophils in murine models of obstructive lung diseases, based on its ability to control neutrophilic-inflammation and resulting lung disease.

Bioconjugation of luminescent silicon quantum dots for selective uptake by cancer cells.

Conventional quantum dots have great potential in cancer-related imaging and diagnostic applications; however, these applications are limited by concerns about the inherent toxicity of their core materials (e.g., cadmium, lead). Virtually all imaging applications require conjugation of the imaging agent to a biologically active molecule to achieve selective uptake or binding. Here, we report a study of biocompatible silicon quantum dots covalently attached to biomolecules including lysine, folate, antimesothelin, and transferrin. The particles possess desirable physical properties, surface chemistry, and optical properties. Folate- and antimesothelin-conjugated silicon quantum dots show selective uptake into Panc-1 cells. This study contributes to the preclinical evaluation of silicon quantum dots and further demonstrates their potential as an imaging agent for cancer applications.

Multimodal imaging probes based on Gd-DOTA conjugated quantum dot nanomicelles.

Recently, multimodal nanoparticles integrating dual- or tri-imaging modalities into a single hybrid nanosystem have attracted plenty of attention in biomedical research. Here, we report the fabrication of two types of multimodal micelle-encapsulated nanoparticles, which were systematically characterized and thoroughly evaluated in terms of their imaging potential and biocompatibility. Optical and magnetic resonance (MR) imaging probes were integrated by conjugating DOTA-gadolinium (Gd) derivative to quantum dot based nanomicelles. Two amphiphilic block copolymer micelles, amine-terminated mPEG-phospholipid and amine-modified Pluronic F127, were chosen as the capping agents because of their excellent biocompatibility and ability to prevent opsonization and prolong circulation time in vivo. Owing to their different hydrophobic-hydrophilic structure, the micellar aggregates exhibited different sizes and protection of core QDs. This work revealed the differences between these nanomicelles in terms of the stability over a wide range of pH, along with their cytotoxicity and the capacity for chelating gadolinium, thus providing a useful guideline for tailor-making multimodal nanoparticles for specific biomedical applications.

Bioconjugated PLGA-4-arm-PEG branched polymeric nanoparticles as novel tumor targeting carriers.

In this study, we have developed a novel carrier, micelle-type bioconjugated PLGA-4-arm-PEG branched polymeric nanoparticles (NPs), for the detection and treatment of pancreatic cancer. These NPs contained 4-arm-PEG as corona, and PLGA as core, the particle surface was conjugated with cyclo(arginine-glycine-aspartate) (cRGD) as ligand for in vivo tumor targeting. The hydrodynamic size of the NPs was determined to be 150-180 nm and the critical micellar concentration (CMC) was estimated to be 10.5 mg l( - 1). Our in vitro study shows that these NPs by themselves had negligible cytotoxicity to human pancreatic cancer (Panc-1) and human glioblastoma (U87) cell lines. Near infrared (NIR) microscopy and flow cytometry demonstrated that the cRGD conjugated PLGA-4-arm-PEG polymeric NPs were taken up more efficiently by U87MG glioma cells, over-expressing the α(v)ß(3) integrin, when compared with the non-targeted NPs. Whole body imaging showed that the cRGD conjugated PLGA-4-arm-PEG branched polymeric NPs had the highest accumulation in the pancreatic tumor site of mice at 48 h post-injection. Physical, hematological, and pathological assays indicated low in vivo toxicity of this NP formulation. These studies on the ability of these bioconjugated PLGA-4-arm-PEG polymeric NPs suggest that the prepared polymeric NPs may serve as a promising platform for detection and targeted drug delivery for pancreatic cancer.

Biocompatible PEGylated gold nanorods as colored contrast agents for targeted in vivo cancer applications.

In this contribution, we report the use of a PEGylated gold nanorods formulation as a colored dye for tumor labeling in vivo. We have demonstrated that the nanorod-targeted tumor site can be easily differentiated from the background tissues by the 'naked eye' without the need of sophisticated imaging instruments. In addition to tumor labeling, we have also performed in vivo toxicity and biodistribution studies of PEGylated gold nanorods in vivo by using BALB/c mice as the model. In vivo toxicity studies indicated no mortality or adverse effects or weight changes in BALB/c mice treated with PEGylated gold nanorods. This finding will provide useful guidelines in the future development of diagnostic probes for cancer diagnosis, optically guided tumor surgery, and lymph node mapping applications.

[Characterization of biocompatible CdTe/znte quantum dots and its application in cell labeling].

Water-soluble CdTe/ZnTe core-shell quantum dots (QDs) coated with L-cysteine were synthesized in low-temperature aqueous-phase one-pot approach. The authors measured the spectral characteristics of QDs at different pH in various buffer solutions and under different excitation laser powers. The primary results show that the absorption spectra of QDs approximately overlap and the fluorescence spectra peaks have no shift in different pH solution. The fluorescence intensity increased linearly with increasing pH. With the incubation time in borate buffer solution, the fluorescence intensity decreased a little. Under strong power laser, the QDs were photobleached rapidly. However, QDs are strongly anti-photobleaching under appropriate laser power (< 100 microW). Thus, such QDs have good biological stability and optical stability. By conjugating the QDs with transferrin protein and constructing the targeted fluorescent nanoparticles, the authors labelled the HeLa cell successfully. Photobleaching experiments in vivo show that microenvironment inside cells affect the stability and accelerate the photobleaching of QDs.

Biomedical In Vivo Studies with ORMOSIL Nanoparticles Containing Active Agents.

Organically modified silica (ORMOSIL) nanoparticles have found many biomedical applications and emerged as biocompatible and efficient carriers of diagnostic and therapeutic agents, such as fluorophores, drugs, and DNA. Herein, we describe two major in vivo studies exemplifying the use of these nanoparticles as carriers of active agents. The first part of this report details a systemic administration and biodistribution of radiolabeled and fluorophore-incorporated ORMOSIL nanoparticles in mice. The second part of this report focuses on the use of ORMOSIL nanoparticles as carriers of plasmid DNA for nonviral gene delivery to the mouse brain. We provide detailed protocols describing preparation and characterization of ORMOSIL nanoparticles, methods used for loading the particles with active agents (e.g., radioimaging agents, plasmid DNA), and in vivo administration of the particles.

Robust dual modality antibacterial action using silver-Prussian blue nanoscale coordination polymer.

We report the synthesis of novel silver-doped Prussian blue nanoscale coordination polymers (SPB NCPs), for dual modality photothermal ablation and oxidative toxicity in bacterial cells. The comparison of SPB NCPs (having Fe-CN-Ag bonds) with the conventionally used Prussian blue nanoscale coordination polymers (PB NCPs, having Fe-CN-Fe bonds) was investigated in terms of their physical and therapeutic properties. It was observed that both PB and SPB NCPs have similar physical dimensions, crystalline phase and optical properties. Both these NCPs showed robust photothermal effect by heat generation (hyperthermia) upon exposure to red laser light. However, among the two, only SPB NCP showed oxidase-like activity by generating H2O2 in aqueous medium, presumably due to its silver content. In vitro antibacterial studies revealed that the SPB NCPs, but not PB NCPs, show inherent toxicity towards bacteria with an IC50 value close to 2.5 µg/ml. It can be inferred that this toxicity is oxidative in nature, as a result of the oxidase-like behaviour shown by SPB NCPs. Furthermore, light activation resulted in substantial additional antibacterial effect (photothermal toxicity) in bacterial cells treated with SPB NCPs. In comparison, marginal additional photothermal toxicity was observed in PB NCP-treated bacteria. Thus, we conclude that the combination of dual modality oxidative and photothermal toxicities demonstrated by SPB NCPs, but not by control PB NCPs, makes the former promising antibacterial agents at low dosages.

Biocompatible near-infrared quantum dots as ultrasensitive probes for long-term in vivo imaging applications.

A facile synthesis method to produce monodisperse, biocompatible, lysine crosslinked mercaptoundecanoic acid (MUA) CdSe(0.25)Te(0.75)/CdS near-infrared (NIR) quantum dots and use them as probes to study their long term in vivo distribution, clearance, and toxicity is presented. Large signal enhancements are demonstrated by these quantum dots, which enables their use as efficient and sensitive probes for live-animal imaging. An important finding is that mice intravenously injected with approximately 10.5 mg kg(-1) of NIR QDs survive for more than three months without any apparent adverse effect to their health. Furthermore, it is determined that there is a significant reduction in the number of the QDs in the liver and spleen three months post injection. In addition, histological analysis of heart, kidney, liver, spleen, and lung tissue indicates that there are no acute toxic effects from these lysine cross-linked MUA NIR QDs. This study suggests that these NIR QDs can be potentially used for long-term targeted imaging and therapy studies in vivo.

Gold Nanoparticle-Enhanced Photodynamic Therapy from Photosensitiser-Entrapped Ormosil Nanoparticles.

Herein, we investigate the effect of the surface attachment of gold nanoparticles (AuNPs) on photosensitiser-entrapped organically modified silica (ormosil) nanoparticles for enhanced photodynamic therapy (PDT). Photosensitiser methylene blue (MB)-entrapped ormosil nanoparticles of three different diameters (30/55/80 nm) were synthesised, and AuNPs were attached to their surface. Subsequent light irradiation with a 630 nm laser resulted in the production of singlet oxygen (1O2), with different amounts depending on the diameter of the ormosil nanoparticle. Enhancement in 1O2 production was observed upon the AuNP attachment on the ormosil surface. In vitro studies in pancreatic cancer (Panc-1) cells have demonstrated that cellular uptake and photodynamic cytotoxicity are also dependent on the size of ormosil nanoparticles and AuNP attachment. These studies pave the way for designing ormosil nanoparticles with suitable size and surface attachment for PDT applications.

Doxorubicin-loaded casein nanoparticles for drug delivery: Preparation, characterization and in vitro evaluation.

Casein, a milk protein that self-assembles to form micelles in aqueous solution, can bind to a wide range of drugs (hydrophilic and hydrophobic). Herein, a low cost and facile method was reported to prepare casein nanoparticles loaded with an anticancer drug, doxorubicin (DOX). The particles were fabricated by adding an excess of Ca2+ ions which brings the soluble casein present in the solution into the micellar framework to form dense nanoparticles. The binding between the drug and the macromolecule was confirmed using fluorescence studies. Circular Dichroism (CD) shows that upon addition of excess Ca2+ the protein chains rearrange. The nanoparticles were characterized by transmission electron microscopy (TEM), field emission scanning electron microscopy (FESEM), and dynamic light scattering (DLS). The release at pH 1 was higher than the physiological pH making this formulation potent for delivering the drug to the stomach via the oral route. The DOX attached with casein showed improved efficacy, i.e., better cytotoxicity against human pancreatic carcinoma cell line, PANC 1 cells as compared to the free drug of the same concentration, owing to higher cell uptake of the macromolecule.

Iron oxide-doped niosomes as drug carriers for magnetically targeted drug delivery.

Magnetism has wide applications in various fields, such as diagnostics, drug targeting, molecular biology, cell isolation, cell purification, hyperthermia, and radioimmunoassay. In this study, we synthesized niosomes doped with iron oxide nanoparticles and a fluorophore for potential applications in magnetically targeted drug delivery. Release kinetics of the fluorophore and cytotoxicity were assessed. The results demonstrate that niosomes doped with iron oxide nanoparticles can serve as proficient and effective drug carriers in magnetically targeted drug delivery.

Transfection of tyrosine kinase deleted FGF receptor-1 into rat brain substantia nigra reduces the number of tyrosine hydroxylase expressing neurons and decreases concentration levels of striatal dopamine.

The effects of HSV-1 amplicon and polyethyleneimine (PEI)-mediated transfection of dominant negative FGF receptor-1 mutant FGFR1(TK-) into the rat brain substantia nigra (SN) were examined in vivo to model the reduced FGF signaling documented to occur in Parkinson's disease. The number of SN neurons that expressed tyrosine hydroxylase (TH) was significantly reduced following HSV-1 FGFR1(TK-) intranigral delivery and similar changes were observed after PEI-mediated FGFR1(TK-) transfections. Further, we also observed a significantly lower striatal dopamine content following the PEI transfection of FGFR1(TK-). Thus, we conclude that reduced FGF signaling in the SN of Parkinsonian patients could play a role in the impaired dopaminergic transmission associated with the degenerative disease.

Assessment of viral and non-viral gene transfer into adult rat brains using HSV-1, calcium phosphate and PEI-based methods.

CNS gene transfer could provide new approaches to the modelling of neurodegenerative diseases and devising potential therapies. One such disorder is Parkinson's disease (PD), in which dysfunction of several different metabolic processes has been implicated. Here we review the literature on gene transfer systems based on herpes simplex virus type 1 (HSV-1) and non-viral polyethyleneimine (PEI) and calcium phosphate nanoparticle methods. We also assess the usefulness of various CNS gene delivery methods and present some of our own data to exemplify such usefulness. Our data result from vectors stereotaxically introduced to the substantia nigra (SN) of adult rats and evaluated 1 week and/or 1 month post injection using histochemical methods to assess recombinant ss-galactosidase enzyme activity. Gene transfer using PEI or calcium phosphate-mediated transfections was observed for both methods and PEI was comparable to that of HSV-1 amplicon. Our data show that the amplicon delivery was markedly increased when packaged with a helper virus and was similar to the expression profile achieved with a full-size replication-defective HSV-1 recombinant (8117/43). We also examine whether PEI or HSV-1 amplicon-mediated gene transfer could facilitate assessment of the biological effects induced by a dominant negative FGF receptor-1 mutant to model the reduced FGF signalling thought to occur in Parkinson's disease.

Synthesis of luminescent near-infrared AgInS2 nanocrystals as optical probes for in vivo applications.

Near infrared quantum dots have been receiving great attention as fluorescent optical probes for in vivo imaging applications. In this contribution, we report the synthesis and surface functionalization of cadmium free ternary AgInS2 nanocrystals emitting in the near infrared range for successful in vitro and in vivo bioimaging applications. The FDA approved triblock copolymer Pluronic F127 was used to encapsulate the nanocrystals and made them dispersible in aqueous solution. By employing a whole body small animal optical imaging setup, we were able to use the AgInS2 nanocrystals formulation for passive targeted delivery to the tumor site. The ultra-small crystal size, near-infrared emitting luminescence, and high quantum yield make the AgInS2 nanocrystals an attractive candidate as a biological contrast agent for cancer sensing and imaging.

Assessing clinical prospects of silicon quantum dots: studies in mice and monkeys.

Silicon nanocrystals can provide the outstanding imaging capabilities of toxic heavy-metal-based quantum dots without employing heavy metals and have potential for rapid progression to the clinic. Understanding the toxicity of silicon quantum dots (SiQDs) is essential to realizing this potential. However, existing studies of SiQD biocompatibility are limited, with no systematic progression from small-animal to large-animal studies that are more clinically relevant. Here, we test the response of both mice and monkeys to high intravenous doses of a nanoconstruct created using only SiQDs and FDA-approved materials. We show that (1) neither mice nor monkeys show overt signs of toxicity reflected in their behavior, body mass, or blood chemistry, even at a dose of 200 mg/kg. (2) This formulation did not biodegrade as expected. Elevated levels of silicon were present in the liver and spleen of mice three months post-treatment. (3) Histopathology three months after treatment showed adverse effects of the nanoformulation in the livers of mice, but showed no such effects in monkeys. This investigation reveals that the systemic reactions of the two animal models may have some differences and there are no signs of toxicity clearly attributable to silicon quantum dots.

Non-invasive tumor detection in small animals using novel functional Pluronic nanomicelles conjugated with anti-mesothelin antibody.

In this study QDs were encapsulated in carboxylated PluronicF127 (F127COOH) triblock polymeric micelles and conjugated with anti-mesothelin antibody for the purpose of alleviating potential toxicity, enhancing the stability and improving targeting efficiency of CdTe/ZnS quantum dots (QDs) in tumors. The amphiphilic triblock polymer of F127COOH contains hydrophilic carboxylated poly(ethylene oxide) (PEO) and hydrophobic poly(propylene oxide) (PPO) units. After encapsulating QDs into carboxylated F127 (F127COOH-QD) micelles, the particles were conjugated with anti-mesothelin antibodies to allow targeting of cancerous areas. The size of the monodispersed spherical QD-containing micelles was determined to be ∼120 nm by dynamic light scattering (DLS). The critical micelle concentration (CMC) was estimated to be 4.7 × 10(-7) M. In an in vitro study, the anti-methoselin antibody conjugated F127COOH (Me-F127COOH-QD) nanomicelles showed negligible cytotoxicity to pancreatic cancer cells (Panc-1). Confocal microscopy demonstrated that the Me-F127COOH-QD nanomicelles were taken up more efficiently by Panc-1 cells, due to antibody mediated targeting. An in vivo imaging study showed that Me-F127COOH-QD nanomicelles accumulated at the pancreatic tumor site 15 min after intravenous injection. In addition, the low in vivo toxicity of the nanomicellar formulation was evaluated by pathological assays. These results suggest that anti-mesothein antibody conjugated carboxylated F127 nanomicelles may serve as a promising nanoscale platform for early human pancreatic cancer detection and targeted drug delivery.

Near-infrared phosphorescent polymeric nanomicelles: efficient optical probes for tumor imaging and detection.

We report a formulation of near-infrared (near-IR) phosphorescent polymeric nanomicelles and their use for in vivo high-contrast optical imaging, targeting, and detection of tumors in small animals. Near-IR phosphorescent molecules of Pt(II)-tetraphenyltetranaphthoporphyrin (Pt(TPNP)) were found to maintain their near-IR phosphorescence properties when encapsulated into phospholipid nanomicelles. The prepared phosphorescent micelles are of approximately 100 nm size and are highly stable in aqueous suspensions. A large spectral separation between the Pt(TPNP) absorption, with a peak at approximately 700 nm, and its phosphorescence emission, with a peak at approximately 900 nm, allows a dramatic decrease in the level of background autofluorescence and scattered excitation light in the near-IR spectral range, where the signal from the phosphorescent probe is observed. In vivo animal imaging with subcutaneously xenografted tumor-bearing mice has resulted in high contrast optical images, indicating highly specific accumulation of the phosphorescent micelles into tumors. Using optical imaging with near-IR phosphorescent nanomicelles, detection of smaller, visually undetectable tumors has also been demonstrated.

Biocompatible luminescent silicon quantum dots for imaging of cancer cells.

Luminescent silicon quantum dots (Si QDs) have great potential for use in biological imaging and diagnostic applications. To exploit this potential, they must remain luminescent and stably dispersed in water and biological fluids over a wide range of pH and salt concentration. There have been many challenges in creating such stable water-dispersible Si QDs, including instability of photoluminescence due their fast oxidation in aqueous environments and the difficulty of attaching hydrophilic molecules to Si QD surfaces. In this paper, we report the preparation of highly stable aqueous suspensions of Si QDs using phospholipid micelles, in which the optical properties of Si nanocrystals are retained. These luminescent micelle-encapsulated Si QDs were used as luminescent labels for pancreatic cancer cells. This paves the way for silicon quantum dots to be a valuable optical probe in biomedical diagnostics.