Nanoparticle-DNA-polymer composites for hepatocellular carcinoma cell labeling, sensing, and magnetic resonance imaging.

This paper describes comparative studies and protocols in (1) self-assembling of ultrasmall superparamagnetic iron oxide nanoparticle (NP), circular plasmid DNA, and branched polyethylenimine (PEI) composites; (2) magnetofection; (3) gene delivery, (4) magnetic resonance imaging (MRI), and (5) cytotoxicity of the composites toward hepatocellular carcinoma HepG2 cells.

Gold and iron oxide hybrid nanocomposite materials.

This critical review provides an overview of current research activities that focused on the synthesis and application of multi-functional gold and iron oxide (Au-Fe(x)O(y)) hybrid nanoparticles and nanocomposites. An introduction of synthetic strategies that have been developed for generating Au-Fe(x)O(y) nanocomposites with different nanostructures is presented. Surface functionalisation and bioconjugation of these hybrid nanoparticles and nanocomposites are also reviewed. A variety of applications such as theranostics, gene delivery, biosensing, cell sorting, bio-separation, and catalysis is discussed and highlighted. Finally, future trends and perspectives of these sophisticated nanocomposites are outlined. Underpinning the fundamental requirements for effectively forming Au-Fe(x)O(y) hybrid nanocomposite materials would shed light on future development of nanotheranostics, nanomedicines, and chemical technologies. It would be interesting to investigate such multi-component composite nanomaterials with different novel morphologies in the near future to advance chemistry, biology, medicine, and engineering multi-disciplinary research (120 references).

Durable mesenchymal stem cell labelling by using polyhedral superparamagnetic iron oxide nanoparticles.

Small polyhedral superparamagnetic iron oxide (SPIO) nanoparticles (<10 nm) coated with a thin layer of silica were prepared (SPIO@SiO(2) and SPIO@SiO(2)-NH(2)). Surface modification of the small polyhedral silica-coated SPIO nanoparticles with amines led to substantially higher mesenchymal stem cell (MSC) labelling efficiency without the use of additional transfecting agents. Therefore, amine surface-modified nanoparticles (SPIO@ SiO(2)-NH(2)) appeared to be the preferred candidate for MSC labelling. In vitro studies demonstrated that controlled labelling of SPIO@SiO(2) and SPIO@SiO(2)-NH(2) did not cause MSC death or proliferation inhibition. MSCs labelled with SPIO@SiO(2)-NH(2) nanoparticles retained differentiation potential and showed osteogenic, adipogenic and chondrogenic differentiations. The noncytotoxic polyhedral SPIO@SiO(2)-NH(2) nanoparticle-labelled MSCs were successfully implanted in rabbit brain and erector spinae muscle, and demonstrated long-lasting, durable MRI labelling efficacy after 8-12 weeks.

Evaluation of biocompatible alginate- and deferoxamine-coated ternary composites for magnetic resonance imaging and gene delivery into glioblastoma cells.

BACKGROUND: This paper describes comparative studies in cytotoxicities, magnetic resonance imaging (MRI), and gene delivery into glioblastoma U87MG or U138MG cells with ternary composites that are consist of superparamagnetic iron oxide (SPIO) nanoparticles (NPs) (size: 8-10 nm) with different surface coatings, circular plasmid DNA (pDNA) (~4 kb) equipped with fluorescent/luminescent probe, and branched polyethylenimine (25 kDa, PDI 2.5). METHODS: Three types of SPIO-NPs were used, including: (I) naked iron oxide NPs with Fe-OH surface group (Bare-NP); (II) iron oxide NPs with a coating of alginate (Alg-NPs); and (III) iron oxide NPs with a coating of deferoxamine (Def-NPs). By tuning the polyethylenimine (PEI)/NP ratios and with a fixed DNA amount, different ternary composites were employed for NP/gene transfection into glioblastoma U87MG or U138MG cells, which were then characterized by Prussian blue staining, in vitro MRI, green fluorescence protein (GFP) fluorescence and luciferase assay. RESULTS: Among the composites prepared, 0.2 ng PEI/0.5 µg DNA/1.0 µg Bare-NP ternary composite possessed the best cellular uptake efficiency of NP to the cytoplasm, following the trend Bare-NP > Alg-NP > Def-NP. This observation was consistent to the MRI assessments with in vitro T 2 relaxivity (r 2) values of 46.0, 35.5, and 23.7 s(-1)·µM(-1)·Fe, respectively. For cellular uptake efficiency of the pDNA, all variations of PEI/NP ratios of the composites did not yield significant differences. However, cellular uptake efficiencies of pDNA in the ternary composites in U138MG cells were generally higher than that of U87MG cells by an order of magnitude. Exceptionally, the ternary composite 0.2 ng PEI/0.5 µg DNA/1.0 µg Bare-NP possessed a lowered luciferase activity RLU for gene expression in U138MG cells. A total of 0.2 ng PEI/0.5 µg DNA/0.1 µg Bare-NP would be uptaken to the cell nucleus with the highest luciferase activity. A working concentration range of PEI with at least 15% higher cell viabilities than lipofectamine was 0.1 to 0.2 ng/well. The cytotoxicities became significant when 0.5 ng/well PEI was present in the ternary composites. CONCLUSIONS: The as-prepared composites offer potential biomedical applications in simultaneous gene delivery, imaging contrast enhancement, and metabolism study.

Enhanced cellular uptake and gene delivery of glioblastoma with deferoxamine-coated nanoparticle/plasmid DNA/branched polyethylenimine composites.

Ternary composite nanomaterials based on deferoxamine-coated superparamagnetic iron oxide nanoparticles (8-10 nm), circular plasmid DNA (~4 kb) with fluorescent/luminescent reporter group, and branched polyethylenimine (25 kDa, PDI = 2.5) were prepared and compared in terms of their efficiencies in transfecting brain tumor cells at low concentration.

Type III-B rotaxane dendrimers.

Type III-B first generation [3]rotaxane and second generation [4]rotaxane dendrimers have been synthesized via (1) a modified copper-catalyzed alkyne-azide cycloaddition (CuAAC), (2) Glaser-Hay's acetylenic oxidative homo-coupling, and (3) amide formation. The dendron does not reveal obvious cytotoxicities in L929 fibroblast cells. The rotaxane dendrimers can capture ammonia and are switchable both in solution and on surfaces.

Ternary hybrid nanocomposites for gene delivery and magnetic resonance imaging of hepatocellular carcinoma cells.

This paper describes comparative studies in magnetic resonance imaging (MRI) and gene deliveries toward hepatocellular carcinoma (HCC) HepG2 cells with ternary composites that consist of superparamagnetic iron oxide (SPIO) nanoparticles (NPs) (8-10 nm) with deferoxamine coating, circular plasmid DNA (~4 kb) equipped with green fluorescent probe, and branched polyethylenimine (PEI) (25 kDa, PDI 2.5). The packaging of the ternary complexes has been characterized by agarose gel retardation assay. By tuning the PEI/NP ratios and with a fixed DNA amount, different ternary composites have been employed for NP/gene transfection towards HepG2 cells, which have been characterized by in vitro MRI and green fluorescence protein (GFP) fluorescence.

Efficacy and Durability in Direct Labeling of Mesenchymal Stem Cells Using Ultrasmall Superparamagnetic Iron Oxide Nanoparticles with Organosilica, Dextran, and PEG Coatings.

We herein report a comparative study of mesenchymal stem cell (MSC) labeling using spherical superparamagnetic iron oxide (SPIO) nanoparticles containing different coatings, namely, organosilica, dextran, and poly(ethylene glycol) (PEG). These nanomaterials possess a similar SPIO core size of 6-7 nm. Together with their coatings, the overall sizes are 10-15 nm for all SPIO@SiO2, SPIO@dextran, and SPIO@PEG nanoparticles. These nanoparticles were investigated for their efficacies to be uptaken by rabbit bone marrow-derived MSCs without any transfecting agent. Experimentally, both SPIO@SiO2 and SPIO@PEG nanoparticles could be successfully uptaken by MSCs while the SPIO@dextran nanoparticles demonstrated limited labeling efficiency. The labeling durability of SPIO@SiO2 and SPIO@PEG nanoparticles in MSCs after three weeks of culture were compared by Prussian blue staining tests. SPIO@SiO2 nanoparticles demonstrated more blue staining than SPIO@PEG nanoparticles, rendering them better materials for MSCs labeling by direct uptake when durable intracellullar retention of SPIO is desired.

pH-controllable supramolecular systems.

This Focus Review surveys representative examples of pH-controllable supramolecular systems with interesting features and state-of-the-art applications such as 1) conformational changes within individual molecules; 2) folding/unfolding of polymers; 3) simultaneous binding of cations and anions; 4) logic function; 5) ON-OFF switchable colorimetric sensing; 6) translocation of macrocycle-in-rotaxane molecules; 7) large-scale movement within molecules; and 8) regulation of the substrate flow in nanocontainers. In particular, systems will be discussed that involve: pH-induced conformational changes of a resorcinarene cavitand and a bis(iron porphyrin) complex; pH control in assembly and disassembly of supramolecular systems stabilized with different major noncovalent interactions; pH-driven movements of interlocked molecules involving rotaxanes, molecular elevators, and molecular muscles; and, finally, multicomponent supramolecular systems immobilized on solid supports as pH-responsive nanovalves for the controlled release of specific substrates. Recent advances in the understanding of pH-controllable supramolecular systems have led to the construction of meaningful molecular machines for electronic and biological applications that are amenable to control by simple perturbation with acids and bases.

Biological and magnetic contrast evaluation of shape-selective Mn-Fe nanowires.

One-dimensional composite Mn-Fe oxide nanostructures of different sizes (nanoneedles, nanorods, and nanowires) were prepared by a linker-induced organization of manganese-doped iron oxide nanoparticles. The nanostructures were obtained by the treatment of MnFe(2)O(4) nanoparticles in the presence of cystamine. The average lengths of nanoneedle, nanorod, and nanowire are approximately 400, 800, and 1000 nm, respectively. High-resolution transmission electron microscopy (HR-TEM), energy-dispersive X-ray (EDX) spectroscopy, and inductively coupled plasma-optical emission spectroscopy (ICP-OES) were employed to characterize the morphologies and the elemental contents of the nanostructures. As an example of their potential applications, these nanostructures were explored as the cell-labeling agents for magnetic resonance imaging (MRI). The magnetic contrast properties of the nanostructures were characterized by a 1.5 T (Tesla) whole body MR system. 10 microg/mL of the nanostructures caused substantial negative contrast. After in vitro incubation, the nanostructures could be effectively incorporated into the cells of a monocyte/macrophage cell line (RAW264.7). These cells' viability and proliferation potential were not affected when the labeling concentration was less than 50 microg/mL.

Discrete functional gold nanoparticles: hydrogen bond-assisted synthesis, magnetic purification, supramolecular dimer and trimer formation.

Amine monofunctional gold nanoparticles (1-AuNPs) were synthesized by employing a solid-supported technique and pH-switchable pseudorotaxane formation. Purification was repeatedly facilitated using crown ether peripherally coated superparamagnetic iron oxide microspheres to yield the monofunctional gold nanoparticles in excellent yield. The product and its related intermediate superstructures were characterized by IR and X-ray photoelectron spectroscopies. Novel supramolecular dimers and trimers were prepared by titrating the 1-AuNPs with bisDB24C8 and trisDB24C8 at different ratios. UV/visible absorption spectroscopic analyses of the supramolecular dimer and trimer solutions, which were formed by mixing their separate components in different ratios, indicated the gradual appearance of two distinct plasmonic resonance bands at 620 and approximately 700 nm. Furthermore, TEM images of the dimers revealed a significant amount of dimer pairs on the surface, while the TEM images of the trimers demonstrated the presence of both dimers and trimers. The trimers appeared as triangular or near-linear shapes.