Nanoshell magnetic resonance imaging contrast agents

Nanocontrast agents have great potential in magnetic resonance (MR) molecular imaging applications for clinical diagnosis. We synthesized Au(3)Cu(1) (gold and copper) nanoshells that showed a promising MR contrast effect. For in vitro MR images, the large proton r1 relaxivities brightened T(1)-weighted images. As for the proton-dephasing effect in T(2), Au(3)Cu(1) lightened MR images at the low concentration of 0.125 mg mL(-1) (3.84 x 10(-7) mM), and then the signal continuously decreased as the concentration increased. For in vivo MR imaging, Au(3)Cu(1) nanocontrast agents enhanced the contrast of blood vessels and suggested their potential use in MR angiography as blood-pool agents. We propose that (1) the cooperativity originating from the form of the nanoparticles and (2) the large surface area coordinated to water from their porous hollow morphology are important for efficient relaxivity. In a cytotoxicity and animal survival assay, Au(3)Cu(1) nanocontrast agents showed a dose-dependent toxic effect: the viability rate of experimental mice reached 83% at a dose of 20 mg kg(-1) and as much as 100% at 2 mg kg(-1).     

Hot-injection synthesis of iron/iron oxide core/shell nanoparticles for T2 contrast enhancement in magnetic resonance imaging

Here we report a new, bench-top synthesis for iron/iron oxide core/shell nanoparticles via the thermal decomposition of Fe(η(5)-C(6)H(3)Me(4))(2). The iron/iron oxide core/shell nanoparticles are superparamagnetic at room temperature and show improved negative contrast in T(2)-weighted MR imaging compared to pure iron oxides nanoparticles, and have a transverse relaxivity (r(2)) of 332 mM(-1) s(-1).     

Antibiofouling polymer-coated superparamagnetic iron oxide nanoparticles as potential magnetic resonance contrast agents for in vivo cancer imaging

We report the fabrication and characterization of antifouling polymer-coated magnetic nanoparticles as nanoprobes for magnetic resonance (MR) contrast agents. Magnetite superparamagnetic iron oxide nanoparticles (SPION) were coated with the protein- or cell-resistant polymer, poly(TMSMA-r-PEGMA), to generate stable, protein-resistant MR probes. Coated magnetic nanoparticles synthesized using two different preparation methods (in situ and stepwise, respectively) were both well dispersed in PBS buffer at a variety of pH conditions (pH 1-10). In addition, dynamic light scattering data revealed that their sizes were not altered even after 24 h of incubation in 10% serum containing cell culture medium, indicative of a lack of protein adsorption on their surfaces. When the antibiofouling polymer-coated SPION were incubated with macrophage cells, uptake was significantly lower in comparison to that of the popular contrast agent, Feridex I.V., suggesting that the polymer-coated SPION can be long-circulated in plasma by escaping from uptake by the reticular endothelial system (RES) such as macrophages. Indeed, when the coated SPION were administered to tumor xenograft mice by intravenous injection, the tumor could be detected in T2-weighted MR images within 1 h as a result of the accumulation of the nanomagnets within the tumor site. Although the poly(TMSMA-r-PEGMA)-coated SPION do not have any targeting ligands on their surface, they are potentially useful for cancer diagnosis in vivo.     

Drug-loaded and superparamagnetic iron oxide nanoparticle surface-embedded amphiphilic block copolymer micelles for integrated chemotherapeutic drug delivery and MR imaging

We report on the fabrication of organic/inorganic hybrid micelles of amphiphilic block copolymers physically encapsulated with hydrophobic drugs within micellar cores and stably embedded with superparamagnetic iron oxide (SPIO) nanoparticles within hydrophilic coronas, which possess integrated functions of chemotherapeutic drug delivery and magnetic resonance (MR) imaging contrast enhancement. Poly(ε-caprolactone)-b-poly(glycerol monomethacrylate), PCL-b-PGMA, and PCL-b-P(OEGMA-co-FA) amphiphilic block copolymers were synthesized at first by combining ring-opening polymerization (ROP), atom transfer radical polymerization (ATRP), and post- modification techniques, where OEGMA and FA are oligo(ethylene glycol) monomethyl ether methacrylate and folic acid-bearing moieties, respectively. A model hydrophobic anticancer drug, paclitaxel (PTX), and 4 nm SPIO nanoparticles were then loaded into micellar cores and hydrophilic coronas, respectively, of mixed micelles fabricated from PCL-b-PGMA and PCL-b-P(OEGMA-co-FA) diblock copolymers by taking advantage of the hydrophobicity of micellar cores and strong affinity between 1,2-diol moieties in PGMA and Fe atoms at the surface of SPIO nanoparticles. The controlled and sustained release of PTX from hybrid micelles was achieved, exhibiting a cumulative release of ~61% encapsulated drugs (loading content, 8.5 w/w%) over ~130 h. Compared to that of surfactant-stabilized single SPIO nanoparticles (r(2) = 28.3 s(-1) mM(-1) Fe), the clustering of SPIO nanoparticles within micellar coronas led to considerably enhanced T(2) relaxivity (r(2) = 121.1 s(-1) mM(-1) Fe), suggesting that hybrid micelles can serve as a T(2)-weighted MR imaging contrast enhancer with improved performance. Moreover, preliminary experiments of in vivo MR imaging were also conducted. These results indicate that amphiphilic block copolymer micelles surface embedded with SPIO nanoparticles at the hydrophilic corona can act as a new generation of nanoplatform integrating targeted drug delivery, controlled release, and disease diagnostic functions.     

Innovative synthesis of citrate-coated superparamagnetic Fe3O4 nanoparticles and its preliminary applications

In this study, we describe the development of a facile and effective route for the synthesis of Fe(3)O(4)-based T(1) contrast agent, which can be useful for in vivo magnetic resonance (MR) imaging. Citrate-coated Fe(3)O(4) nanoparticles (6 nm) with a narrow size distribution were synthesized by "one-pot green chemistry route" in diethylene glycol (DEG) solvent. The synthesized nanoparticles were characterized by different analytical techniques including XRD, TEM, HRTEM, and FTIR. At room temperature, nanoparticles exhibited superparamagnetic nature with high saturation magnetization. The longitudinal (r(1)) and transverse (r(2)) relaxivities were found to be 35.45 and 51.81 mM(-1)s(-1), respectively. Contrast agent developed by this method showed a relatively higher longitudinal relaxivity (r(1)) and the lowest relaxivity ratio (r(2)/r(1)=1.46) at 3T MR field. The anionic nature of citric acid facilitated non-specific internalization without impairment of cell viability and functionality. The in vitro studies showed both phagocitic and non-phagocytic uptake of these NPs. In vivo MR imaging of swine showed both T(1) and T(2) contrast effect.     

Ultrasmall water-soluble metal-iron oxide nanoparticles as T1-weighted contrast agents for magnetic resonance imaging

Using an improved hydrolysis method of inorganic salts assisted with water-bath incubation, ultrasmall water-soluble metal-iron oxide nanoparticles (including Fe(3)O(4), ZnFe(2)O(4) and NiFe(2)O(4) nanoparticles) were synthesized in aqueous solutions, which were used as T(1)-weighted contrast agents for magnetic resonance imaging (MRI). The morphology, structure, MRI relaxation properties and cytotoxicity of the as-prepared metal-iron oxide nanoparticles were characterized, respectively. The results showed that the average sizes of nanoparticles were about 4 nm, 4 nm and 5 nm for Fe(3)O(4), ZnFe(2)O(4) and NiFe(2)O(4) nanoparticles, respectively. Moreover, the nanoparticles have good water dispersibility and low cytotoxicity. The MRI test showed the strong T(1)-weighted, but the weak T(2)-weighted MRI performance of metal-iron oxide nanoparticles. The high T(1)-weighted MRI performance can be attributed to the ultrasmall size of metal-iron oxide nanoparticles. Therefore, the as-prepared metal-iron oxide nanoparticles with good water dispersibility and ultrasmall size can have potential applications as T(1)-weighted contrast agent materials for MRI.     

Multifunctional yolk-shell nanoparticles: a potential MRI contrast and anticancer agent

We report a new type of multifunctional nanomaterials, FePt@Fe2O3 yolk-shell nanoparticles, that exhibit high cytotoxicity originated from the FePt yolks and strong MR contrast enhancement resulting from the Fe2O3 shells. Encouraged by the recently observed high cytotoxicity of FePt@CoS2 yolk-shell nanoparticles, we used Fe2O3 to replace CoS2 as the shells to further explore the applications of the yolk-shell nanostructures. The ultralow IC50 value (238 +/- 9 ng of Pt/mL) of FePt@Fe2O3 yolk-shell nanoparticles likely originates from the fact that the slow oxidation and release of FePt yolks increases the cytotoxicity. Moreover, compared with two commercial magnetic resonance imaging (MRI) contrast agents, MION and Sinerem, the FePt@Fe2O3 yolk-shell nanoparticle showed stronger contrast enhancement according to their apparent transverse relaxivity values (r2* = 3.462 (microg/mL)(-1) s(-1)). The bifunctional FePt@Fe2O3 yolk-shell nanoparticles may serve both as an MRI contrast agent and as a potent anticancer drug. This work indicates that these unique yolk-shell nanoparticles may ultimately lead to new designs of multifunctional nanostructures for nanomedicine.     

Synthesis, characterization and examination of Gd[DO3A-hexylamine]-functionalized silica nanoparticles as contrast agent for MRI-applications

Spherical, nonporous and monodisperse silica nanoparticles (NPs) with a diameter of about 100 nm were synthesized and covalently functionalized with lanthanoid(III) (Ln=Gd or Y) chelate complexes, which serve as contrast agents (CAs) for magnetic resonance imaging (MRI). The materials were fully characterized after each synthetic step by different analytical methods, such as dynamic light scattering, scanning electron microscopy, DRIFT and NMR spectroscopy, thermogravimetry and elemental analysis, as well as zetapotential measurements. High surface concentrations of Gd(III) complexes (up to 50 μmol g(-1)) were determined by ICP-AES and T(1)-measurements, respectively. MRI experiments show the typical concentration-dependent increase of the longitudinal relaxation rate. T(1)-weighted images of samples with more than 25 μg NPs per 100 μL agar display a clear contrast enhancement in the agar layer. The transverse relaxivities r(2) of the materials are significantly higher than r(2) of the corresponding free Gd(III) complexes in water and medium, whereas the longitudinal relaxivities r(1) are slightly increased. Due to the high loading of Gd(III) complexes, the relaxivities per particle are remarkably high (up to 2.78×10(5) mM(-1) s(-1) for r(1)). Thus, new hybrid materials, based on nonporous silica NPs with high local relaxivity values were synthesized, which can serve as very effective CAs for MRI.     

Generation of superparamagnetic liposomes revealed as highly efficient MRI contrast agents for in vivo imaging

Maghemite (gamma-Fe2O3) nanocrystals stable at neutral pH and in isotonic aqueous media were synthesized and encapsulated within large unilamellar vesicles of egg phosphatidylcholine (EPC) and distearoyl-SN-glycero-3-phosphoethanolamine-N-[methoxy(poly(ethylene glycol))-2000] (DSPE-PEG(2000), 5 mol %), formed by film hydration coupled with sequential extrusion. The nonentrapped particles were removed by flash gel exclusion chromatography. The magnetic-fluid-loaded liposomes (MFLs) were homogeneous in size (195 +/- 33 hydrodynamic diameters from quasi-elastic light scattering). Iron loading was varied from 35 up to 167 Fe(III)/lipid mol %. Physical and superparamagnetic characteristics of the iron oxide particles were preserved after liposome encapsulation as shown by cryogenic transmission electron microscopy and magnetization curve recording. In biological media, MFLs were highly stable and avoided ferrofluid flocculation while being nontoxic toward the J774 macrophage cell line. Moreover, steric stabilization ensured by PEG-surface-grafting significantly reduced liposome association with the macrophages. The ratios of the transversal (r2) and longitudinal (r1) magnetic resonance (MR) relaxivities of water protons in MFL dispersions (6 < r2/r1 < 18) ranked them among the best T2 contrast agents, the higher iron loading the better the T2 contrast enhancement. Magnetophoresis demonstrated the possible guidance of MFLs by applying a magnetic field gradient. Mouse MR imaging assessed MFLs efficiency as contrast agents in vivo: MR angiography performed 24 h after intravenous injection of the contrast agent provided the first direct evidence of the stealthiness of PEG-ylated magnetic-fluid-loaded liposomes.     

Core/shell quantum dots with high relaxivity and photoluminescence for multimodality imaging

A series of core/shell CdSe/Zn1-xMnxS nanoparticles were synthesized for use in dual-mode optical and magnetic resonance (MR) imaging techniques. Mn2+ content was in the range of 0.6-6.2% and varies with the thickness of the shell or amount of Mn2+ introduced to the reaction. These materials showed high quantum yield (QY), reaching 60% in organic solvent. Water-soluble nanoparticles were obtained by capping the core/shell particles with amphiphilic polymer, and the QY values in water reached 21%. These materials also demonstrated high relaxivity with r1 values in the range of 11-18 mM-1 s-1 (at room temperature, 7 T). Both optical and MR imaging were performed on nanoparticles in aqueous solution and applied to cells in culture. The results showed that the QY and manganese concentration in the particles was sufficient to produce contrast for both modalities at relatively low concentrations of nanoparticles.     

Facile preparation of holmium(Ⅲ)-doped carbon nanodots for fluorescence/magnetic resonance dual-modal bioimaging

In this work, we report the synthesis of holmium(III)-doped carbon nanodots(Ho BCDs) as fluorescence/magnetic resonance(FL/MR) dual-modal imaging probes via a facile hydrothermal process using citrate acid(CA), branched-polyethylenimine(BPEI) and diethylenetriamine pentaacetic acid hydrate holmium(III) dihydrogen salt(Ho-DTPA) as carbon source, passivating reagent and holmium source, respectively.The thus prepared Ho BCDs exhibited ultra-small particle size(~4 nm), high water solubility and bright fluorescence with an absolute quantum yield of 8%. Additionally, grey-scaled T_1-weighted images of Ho BCDs solution appeared to be apparently brighter than that of deionized water and un-doped blue carbon nanodots(BCDs) solution. In addition, in vitro toxicity assay validated superior biocompatibility of Ho BCDs. Using He La cells as models, Ho BCDs-treated cells were observed to emit blue fluorescence located both in plasma and nucleus, and presented positive contrast enhancement in T_1-weighted images, suggesting their potentials for practical biomedical applications.     

Large-scale synthesis of uniform and extremely small-sized iron oxide nanoparticles for high-resolution T1 magnetic resonance imaging contrast agents

Uniform and extremely small-sized iron oxide nanoparticles (ESIONs) of < 4 nm were synthesized via the thermal decomposition of iron-oleate complex in the presence of oleyl alcohol. Oleyl alcohol lowered the reaction temperature by reducing iron-oleate complex, resulting in the production of small-sized nanoparticles. XRD pattern of 3 nm-sized nanoparticles revealed maghemite crystal structure. These nanoparticles exhibited very low magnetization derived from the spin-canting effect. The hydrophobic nanoparticles can be easily transformed to water-dispersible and biocompatible nanoparticles by capping with the poly(ethylene glycol)-derivatized phosphine oxide (PO-PEG) ligands. Toxic response was not observed with Fe concentration up to 100 μg/mL in MTT cell proliferation assay of POPEG-capped 3 nm-sized iron oxide nanoparticles. The 3 nm-sized nanoparticles exhibited a high r(1) relaxivity of 4.78 mM(-1) s(-1) and low r(2)/r(1) ratio of 6.12, demonstrating that ESIONs can be efficient T(1) contrast agents. The high r(1) relaxivities of ESIONs can be attributed to the large number of surface Fe(3+) ions with 5 unpaired valence electrons. In the in vivo T(1)-weighted magnetic resonance imaging (MRI), ESIONs showed longer circulation time than the clinically used gadolinium complex-based contrast agent, enabling high-resolution imaging. High-resolution blood pool MR imaging using ESIONs enabled clear observation of various blood vessels with sizes down to 0.2 mm. These results demonstrate the potential of ESIONs as T(1) MRI contrast agents in clinical settings.     

Facile preparation of holmium(III)-doped carbon nanodots for fluorescence/magnetic resonance dual-modal bioimaging

Holmium(III)-doped carbon nanodots (HoBCDs) were synthesized for the first time via hydrothermal treatment of citrate acid (CA), branched-polyethylenimine (BPEI) and diethylenetriamine pentaacetic acid hydrate holmium(III) dihydrogen salt (Ho-DTPA), and their dual-modality bioimaging applications were demonstrated by the use of HeLa cells. The as-prepared nanoprobes exhibited bright fluorescence with an absolute quantum yield of 8% and apparent contrast enhancement in T₁-weighted images.     

Manganese-Zinc Ferrites: Safe and Efficient Nanolabels for Cell Imaging and Tracking In Vivo

Manganese-zinc ferrite nanoparticles were synthesized by using a hydrothermal treatment, coated with silica, and then tested as efficient cellular labels for cell tracking, using magnetic resonance imaging (MRI) in vivo. A toxicity study was performed on rat mesenchymal stem cells and C6 glioblastoma cells. Adverse effects on viability and cell proliferation were observed at the highest concentration (0.55 mM) only; cell viability was not compromised at lower concentrations. Nanoparticle internalization was confirmed by transmission electron microscopy. The particles were found in membranous vesicles inside the cytoplasm. Although the metal content (0.42 pg Fe/cell) was lower compared to commercially available iron oxide nanoparticles, labeled cells reached a comparable relaxation rate R₂, owing to higher nanoparticle relaxivity. Cells from transgenic luciferase-positive rats were used for in vivo experiments. Labeled cells were transplanted into the muscles of non-bioluminescent rats and visualized by MRI. The cells produced a distinct hypointense signal in T₂- or T₂*-weighted MR images in vivo. Cell viability in vivo was verified by bioluminescence.     

Enhancement of relaxivity rates of Gd-DTPA complexes by intercalation into layered double hydroxide nanoparticles

In this paper we report the preparation and characterization of [Gd(dtpa)](2-) intercalated layered double hydroxide (LDH) nanomaterials. [Gd(dtpa)](2-) (gadolinium(III) diethylene triamine pentaacetate) was transferred into LDH by anionic exchange. The intercalation of [Gd(dtpa)](2-) into LDH was confirmed by X-ray diffraction for the new phase with the interlayer spacing of 3.5-4.0 nm and by FTIR for the characteristic vibration peaks of [Gd(dtpa)](2-). The morphology of the nanoparticles was influenced by the extent of [Gd(dtpa)](2-) loading, in which the poly-dispersity quality decreased as the [Gd(dtpa)](2-) loading was increased. Compared with the morphology of the original Mg(2)Al-Cl-LDH nanoparticles (hexagonal plate-like sheets of 50-200 nm), the modified LDH-Gd(dtpa) nanoparticles are bar-like with a width of 30-60 nm and a length of 50-150 nm. LDH-Gd(dtpa) was expected to have an increased water proton magnetic resonance relaxivity due to the intercalation of [Gd(dtpa)](2-) into the LDH interlayer that led to slower molecular anisotropic tumbling compared with free [Gd(dtpa)](2-) in solution. Indeed, LDH-nanoparticle suspension containing approximately 1.6 mM [Gd(dtpa)](2-) exhibits a longitudinal proton relaxivity r(1) of approximately 16 mM(-1) s(-1) and a transverse proton relaxivity r(2) of approximately 50 mM(-1) s(-1) at room temperature and a magnetic field of 190 MHz, which represents an enhancement four times (r(1)) and 12 times (r(2)) that of free [Gd(dtpa)](2-) in solution under the same reaction conditions. We have thus tailored LDH-nanoparticles into a novel contrast agent with strong relaxivity, promising for great potential applications in magnetic resonance imaging.     

Simple and generalized synthesis of oxide-metal heterostructured nanoparticles and their applications in multimodal biomedical probes

Heterostructured nanoparticles composed of metals and Fe3O4 or MnO were synthesized by thermal decomposition of mixtures of metal-oleate complexes (for the oxide component) and metal-oleylamine complexes (for the metal component). The products included flowerlike-shaped nanoparticles of Pt-Fe3O4 and Ni-Fe3O4 and snowmanlike-shaped nanoparticles of Ag-MnO and Au-MnO. Powder X-ray diffraction patterns showed that these nanoparticles were composed of face-centered cubic (fcc)-structured Fe3O4 or MnO and fcc-structured metals. The relaxivity values of the Au-MnO and Au-Fe3O4 nanoparticles were similar to those of the MnO and Fe3O4 nanoparticles, respectively. Au-Fe3O4 heterostructured nanoparticles conjugated with two kinds of 12-base oligonucleotide sequences were able to sense a complementary 24-mer sequence, causing nanoparticle aggregation. This hybridization-mediated aggregation was detected by the overall size increase indicated by dynamic light scattering data, the red shift of the surface plasmon band of the Au component, and the enhancement of the signal intensity of the Fe3O4 component in T2-weighted magnetic resonance imaging.     

A Gd3Al tetranuclear complex as a potential bimodal MRI/optical imaging agent

A new self-assembled gadolinium(III)-aluminum(III) complex (Gd(3)Al) was synthesized and characterized. The efficacy of this Gd(3)Al complex as a potential bimodal magnetic resonance imaging (MRI)/optical imaging agent has been evaluated. Relaxivity studies showed that the Gd(3)Al complex has higher relaxation efficiency (7.18 mM(-1) s(-1)) compared with the clinically used complex gadolinium-diethylenetriaminepentaacetic acid (Gd-DTPA, 3.9 mM(-1) s(-1)) at 400 MHz and 25 °C. In vitro T(1)-MR images on a 0.5 T magnetic field exhibited a remarkable enhancement of signal contrast for Gd(3)Al compared to Gd-DTPA. Furthermore, the Gd(3)Al complex exhibits bright-green luminescence with the emission spectrum centred at 510 nm. Live-cell fluorescence imaging reveals that the Gd(3)Al complex is permeable to cells and localizes to the cytoplasm. In view of the relaxometric and luminescent properties, this Gd(3)Al complex could serve as a potential bimodal MRI/optical imaging agent.     

Magnetic nanoparticles--templated assembly of protein subunits: a new platform for carbohydrate-based MRI nanoprobes

A new approach for the preparation of carbohydrate-coated magnetic nanoparticles is reported. In a first step, we show that the pH-driven assembly-disassembly natural process that occurs in apoferritin protein is effective for the encapsulation of maghemite nanoparticles of different sizes: 4 and 6 nm. In a second step, we demonstrate that the presence of functional amine groups in the outer shell of apoferritin allows functionalization with two carbohydrates, N-acetyl-D-glucosamine and d-mannose. High-resolution electron microscopy (HREM), high angle annular dark field scanning electron microscopy (HAADF-STEM), electron energy loss spectroscopy (EELS), X-ray diffraction (XRD), and SQUID technique have been used to characterize the magnetic samples, termed herein Apomaghemites. The in vivo magnetic resonance imaging (MRI) studies showed the efficiency in contrasting images for these samples; that is, the r(2) NMR relaxivities are comparable with Endorem (a commercial superparamagnetic MRI contrast agent). The r(2) relaxivity values as well as the pre-contrast and post-contrast T(2)*-weighted images suggested that our systems could be used as perspective superparamagnetic contrast agents for magnetic resonance imaging (MRI). The carbohydrate-functionalized Apomaghemite nanoparticles retained their recognition abilities, as demonstrated by the strong affinity with their corresponding carbohydrate-binding lectins.     

Superparamagnetic blocking and superspin-glass freezing in ultra small δ-(Fe(0.67)Mn(0.33))OOH particles

The magnetic properties of ultra-small (~2 nm) δ-(Fe(0.67)Mn(0.33))OOH nanoparticles prepared by a microemulsion technique have been investigated by magnetization and ac susceptibility measurements at variable frequency. The results provide evidence of two different magnetic regimes whose onset is identified by two maxima in the zero-field-cooled susceptibility: a large one, centered at ~150 K (T(mh)), and a narrow one at ~30 K (T(ml)). The two temperatures exhibit a different frequency dependence: T(mh) follows a Vogel-Fulcher law τ = τ(0)exp[(E(a)/k(B))/(T-T(0))], indicating a blocking of weakly interacting nanoparticle moments, whereas T(ml) follows a power law τ = τ(0)(T(g)/T(mν)-T(g))(α), suggesting a collective freezing of nanoparticle moments (superspin-glass state). This picture is coherent with the field dependence of T(ml) and T(mh) and with the temperature dependence of the coercivity, strongly increasing below 30 K.     

Surface modification of gadolinium oxide thin films and nanoparticles using poly(ethylene glycol)-phosphate

The performance of nanomaterials for biomedical applications is highly dependent on the nature and the quality of surface coatings. In particular, the development of functionalized nanoparticles for magnetic resonance imaging (MRI) requires the grafting of hydrophilic, nonimmunogenic, and biocompatible polymers such as poly(ethylene glycol) (PEG). Attached at the surface of nanoparticles, this polymer enhances the steric repulsion and therefore the stability of the colloids. In this study, phosphate molecules were used as an alternative to silanes or carboxylic acids, to graft PEG at the surface of ultrasmall gadolinium oxide nanoparticles (US-Gd(2)O(3), 2-3 nm diameter). This emerging, high-sensitivity "positive" contrast agent is used for signal enhancement in T(1)-weighted molecular and cellular MRI. Comparative grafting assays were performed on Gd(2)O(3) thin films, which demonstrated the strong reaction of phosphate with Gd(2)O(3) compared to silane and carboxyl groups. Therefore, PEG-phosphate was preferentially used to coat US-Gd(2)O(3) nanoparticles. The grafting of this polymer on the particles was confirmed by XPS and FTIR. These analyses also demonstrated the strong attachment of PEG-phosphate at the surface of Gd(2)O(3), forming a protective layer on the nanoparticles. The stability in aqueous solution, the relaxometric properties, and the MRI signal of PEG-phosphate-covered Gd(2)O(3) particles were also better than those from non-PEGylated nanoparticles. As a result, reacting PEG-phosphate with Gd(2)O(3) particles is a promising, rapid, one-step procedure to PEGylate US-Gd(2)O(3) nanoparticles, an emerging "positive" contrast agent for preclinical molecular and cellular applications.