19F Magnetic Resonance Research Articles

Fluorinated PLGA-PEG-Mannose Nanoparticles for Tumor-Associated Macrophage Detection by Optical Imaging and MRI.

Tumor-associated macrophages (TAMs) promote cancer growth and metastasis, but their role in tumor development needs to be fully understood due to the dynamic changes of tumor microenvironment (TME). Here, we report an approach to visualize TAMs by optical imaging and by Fluorine-19 (19F) magnetic resonance imaging (MRI) that is largely applied to track immune cells in vivo. TAMs are targeted with PLGA-PEG-mannose nanoparticles (NPs) encapsulating perfluoro-15-crown-5-ether (PFCE) as MRI contrast agent. These particles are preferentially recognized and phagocytized by TAMs that overexpress the mannose receptor (MRC1/CD206). The PLGA-PEG-mannose NPs are not toxic and they were up-taken by macrophages as confirmed by in vitro confocal microscopy. At 48 h after intravenous injection of PLGA-PEG-mannose NPs, 4T1 xenograft mice were imaged and fluorine-19 nuclear magnetic resonance confirmed nanoparticle retention at the tumor site. Because of the lack of 19F background in the body, observed 19F signals are robust and exhibit an excellent degree of specificity. In vivo imaging of TAMs in the TME by 19F MRI opens the possibility for detection of cancer at earlier stage and for prompt therapeutic interventions in solid tumors.

Multi‐ and dual‐tuned microstripline‐based transmit/receive switch for 7‐Tesla magnetic resonance imaging

AbstractThis article introduces two new designs of transmit/receive switches for 7‐Tesla magnetic resonance imaging. Both designs based on a microstripline coupler technology. In the first design, a different single frequency signals can be handled to/from a radio frequency coil using smart tuning network. In the second design, a dual‐tuned 1H/23Na transmit/receive switch is designed to handle a dual resonant signal to a dual resonant 1H/23Na coil, simultaneously and without tuning. In transmit and receive, the first design achieved good matching (‐20 dB), and low insertion loss (0.78 dB) for the 1H, 19F, 31P, 13C, and 23Na magnetic resonance signals with high isolation (61 dB). Similarly, the dual‐tuned 1H/23Na switch achieved good matching (<−11.5 dB) and low insertion loss (1.0 dB) with high isolation (67 dB), whereas for 1H/31P switch, good matching has been achieved (<−12 dB) and low insertion loss (0.96 dB) with high isolation (62 dB). The first proposed switch enables the use of the same switch with different coils resonating at Larmor frequencies of different atomic nuclei. The second switch can handle a dual resonant signal to a single dual resonant coil. The first design is promising in reducing the costs by using the same switch with different coils. The second design is promising in the integration with multichannel dual resonant coil.

Characterization of Ion Association and Solvation in NaPF6 Carbonate Electrolytes

In this work we aimed to determine the ions association and solvation in electrolytes comprised of NaPF6 in binary EC:PC, and ternary 5EC:3PC:2DEC carbonate solvents. We studied their local and bulk transport properties through multi-Nuclear (1H, 19F and 23Na) Magnetic Resonance spin-lattice relaxation time (T1 ) and self-diffusion coefficient (D) measurements. These studies were complemented with ionic conductivity and solution viscosity measurements. Results show several interesting features. Firstly, the Walden Product – which relates the molar conductivity and viscosity through a temperature-dependent constant – showed a dependence on temperature (see Figure 1). This suggested the dissociation of NaPF6 is dependent on temperature and is lower for the less polar electrolytes. Secondly, the sodium ions are preferentially solvated by PC molecules over EC. Thirdly, significant cation-anion association was observed for the less polar electrolytes. Lastly, this association was independent of temperature indicating that the temperature dependence of the Walden Product is primarily due to that of the viscosity. These results can aid in the improvement of ‘beyond lithium ion’ electrolytes for energy storage applications. Figure 1

Development of Off-On Switching 19F MRI Probes for Cathepsin K Activity Detection

Abstract Cathepsin K is a protease expressed in osteoclasts that degrades bone tissue, such as type I collagen fibers. Overexpression of cathepsin K is involved in osteoporosis, rheumatoid arthritis, and bone metastasis. Therefore, detecting cathepsin K activity is important for understanding the mechanism of these diseases and developing new drugs. However, current chemical probes cannot be employed for the detection of cathepsin K activity in animal deep-tissue. In this study, we developed novel 19F magnetic resonance imaging (MRI) probes (FLAME-(Gd-X), X = Acp, Deg, Deg2) to detect cathepsin K. In FLAME-(Gd-X), the Gd3+ complex was modified on the surface of perfluorocarbon-encapsulated silica nanoparticles through cathepsin K substrate and three different hydrophobic/hydrophilic linkers. The 19F NMR signal intensities of these probes were suppressed by the paramagnetic relaxation enhancement (PRE) effect of the Gd3+ complexes. The 19F MRI signal intensities of FLAME-Gd-Acp and FLAME-Gd-Deg specifically increased with the substrate cleavage by cathepsin K. The 19F MRI probes based on the PRE effect can be applied to the in vivo detection of cathepsin K activity.

Glyconanofluorides as Immunotracers with a Tunable Core Composition for Sensitive Hotspot Magnetic Resonance Imaging of Inflammatory Activity.

Nature-inspired nanosized formulations based on an imageable, small-sized inorganic core scaffold, on which biomolecules are assembled to form nanobiomimetics, hold great promise for both early diagnostics and developed therapeutics. Nevertheless, the fabrication of nanobiomimetics that allow noninvasive background-free mapping of pathological events with improved sensitivity, enhanced specificity, and multiplexed capabilities remains a major challenge. Here, we introduce paramagnetic glyconanofluorides as small-sized (<10 nm) glycomimetics for immunotargeting and sensitive noninvasive in vivo19F magnetic resonance imaging (MRI) mapping of inflammation. A very short T1 relaxation time (70 ms) of the fluorides was achieved by doping the nanofluorides’ solid crystal core with paramagnetic Sm3+, resulting in a significant 8-fold enhancement in their 19F MRI sensitivity, allowing faster acquisition and improved detectability levels. The fabricated nanosized glycomimetics exhibit significantly enhanced uptake within activated immune cells, providing background-free in vivo mapping of inflammatory activity, demonstrated in both locally induced inflammation and clinically related neuropathology animal models. Fabricating two types of nanofluorides, each with a distinct chemical shift, allowed us to exploit the color-like features of 19F MRI to map, in real time, immune specificity and preferred targetability of the paramagnetic glyconanofluorides, demonstrating the approach’s potential extension to noninvasive multitarget imaging scenarios that are not yet applicable for nanobiomimetics based on other nanocrystal cores.

Imaging of Inflammation in Spinal Cord Injury: Novel Insights on the Usage of PFC-Based Contrast Agents.

Labeling of macrophages with perfluorocarbon (PFC)-based compounds allows the visualization of inflammatory processes by 19F-magnetic resonance imaging (19F-MRI), due to the absence of endogenous background. Even if PFC-labeling of monocytes/macrophages has been largely investigated and used, information is lacking about the impact of these agents over the polarization towards one of their cell subsets and on the best way to image them. In the present work, a PFC-based nanoemulsion was developed to monitor the course of inflammation in a model of spinal cord injury (SCI), a pathology in which the understanding of immunological events is of utmost importance to select the optimal therapeutic strategies. The effects of PFC over macrophage polarization were studied in vitro, on cultured macrophages, and in vivo, in a mouse SCI model, by testing and comparing various cell tracking protocols, including single and multiple administrations, the use of MRI or Point Resolved Spectroscopy (PRESS), and application of pre-saturation of Kupffer cells. The blood half-life of nanoemulsion was also investigated by 19F Magnetic Resonance Spectroscopy (MRS). In vitro and in vivo results indicate the occurrence of a switch towards the M2 (anti-inflammatory) phenotype, suggesting a possible theranostic function of these nanoparticles. The comparative work presented here allows the reader to select the most appropriate protocol according to the research objectives (quantitative data acquisition, visual monitoring of macrophage recruitment, theranostic purpose, rapid MRI acquisition, etc.). Finally, the method developed here to determine the blood half-life of the PFC nanoemulsion can be extended to other fluorinated compounds.

Fluorine Labeling of Nanoparticles and In Vivo 19F Magnetic Resonance Imaging.

Fluorinated nanoparticles have increasing applications, but they are still challenging to prepare, especially in the case of water-soluble fluorinated nanoparticles. Herein, a fluorine labeling strategy is presented that is based on the conjugation of custom-made small fluorinated building blocks, obtained by simple synthetic transformations, with carboxylated gold nanoparticles through a convenient phase-transfer process. The synthesis of four fluorinated building blocks with different chemical shifts in 19F nuclear magnetic resonance and varied functionalities is reported, along with their conjugation onto nanoparticles. Fluorinated nanoparticles of small core size obtained by this conjugation methodology and by direct synthesis presented high transverse relaxation times (T2) ranging from 518 to 1030 ms, and a large number of equivalent fluorine atoms per nanoparticle (340-1260 fluorine atoms), which made them potential candidates for 19F magnetic resonance related applications. Finally, nontargeted fluorinated nanoparticles were probed by performing in vivo 19F magnetic resonance spectroscopy (19F MRS) in mice. Nanoparticles were detected at both 1 and 2 h after being injected. 19F MRI images were also acquired after either intravenous or subcutaneous injection. Their fate was studied by analyzing the gold content in tissues by ICP-MS. Thus, the present work provides a general fluorination strategy for nanoparticles and shows the potential use of small fluorinated nanoparticles in magnetic-resonance-related applications.

Fluorine-19 Magnetic Resonance Imaging for Detection of Amyloid β Oligomers Using a Keto Form of Curcumin Derivative in a Mouse Model of Alzheimer's Disease.

Recent evidence suggests that the formation of soluble amyloid β (Aβ) aggregates with high toxicity, such as oligomers and protofibrils, is a key event that causes Alzheimer’s disease (AD). However, understanding the pathophysiological role of such soluble Aβ aggregates in the brain in vivo could be difficult due to the lack of a clinically available method to detect, visualize, and quantify soluble Aβ aggregates in the brain. We had synthesized a novel fluorinated curcumin derivative with a fixed keto form, named as Shiga-Y51, which exhibited high selectivity to Aβ oligomers in vitro. In this study, we investigated the in vivo detection of Aβ oligomers by fluorine-19 (19F) magnetic resonance imaging (MRI) using Shiga-Y51 in an APP/PS1 double transgenic mouse model of AD. Significantly high levels of 19F signals were detected in the upper forebrain region of APP/PS1 mice compared with wild-type mice. Moreover, the highest levels of Aβ oligomers were detected in the upper forebrain region of APP/PS1 mice in enzyme-linked immunosorbent assay. These findings suggested that 19F-MRI using Shiga-Y51 detected Aβ oligomers in the in vivo brain. Therefore, 19F-MRI using Shiga-Y51 with a 7 T MR scanner could be a powerful tool for imaging Aβ oligomers in the brain.

Photoresponsive host-guest chemistry and relaxation time of fluorinated cyclodextrin and arylazopyrazole-functionalized DOTA metal complexes.

Light-responsive modulation of the longitudinal (T1) and transversal relaxation times of a fluorinated cyclodextrin has been achieved by host-guest complexation with arylazopyrazole-modified metal complexes in aqueous solution. This supramolecular concept can potentially be applied to the development of contrast agents for 19F magnetic resonance imaging (MRI).

Delicately Designed Cancer Cell Membrane-Camouflaged Nanoparticles for Targeted 19F MR/PA/FL Imaging-Guided Photothermal Therapy.

Our exploration of multimodal nanoprobes aims to combine photoacoustic (PA) imaging, 19F magnetic resonance (MR), and fluorescence (FL) imaging, which offers complementary advantages such as high spatial resolution, unlimited penetration, and high sensitivity to enable more refined images for accurate tumor diagnoses. In this research, perfluorocarbons (PFCs) and indocyanine green (ICG) are encapsulated by poly(lactic-co-glycolic acid) (PLGA) for intravital 19F MR/FL/PA tri-modal imaging-guided photothermal therapy. Then, it is coated with an A549 cancer cell membrane (AM) to fabricate versatile theranostic nanoprobes (AM-PP@ICGNPs). After systemic administration, FLI reveals time-dependent tumor homing of NPs with high sensitivity, 19F MRI provides tumor localization of NPs without background signal interference, and PAI illustrates the detailed distribution of NPs inside the tumor with high spatial resolution. What is more, AM-PP@ICGNPs accumulated in the tumor area exhibit a prominent photothermal effect (48.4 °C) under near infrared (NIR) laser irradiation and realize an enhanced antitumor response in vivo. These benefits, in combination with the excellent biocompatibility, make AM-PP@ICGNPs a potential theranostic nanoagent for accurate tumor localization and ultimately achieve superior cancer therapy.

Intratumoral Glutathione Activatable Nanoprobes for Fluorescence and 19F Magnetic Resonance Turn-On Imaging.

Tumor microenvironment turn-on nanoprobes that could specifically detect the occurrence of diseases possess great potential in early diagnosis. Here, a GSH activated nanoprobe was designed for fluorescence and 19F magnetic resonance (MR) dual-modal turn-on imaging of tumors. Specifically, fluorescence AgInS2 quantum dots (QDs for fluorescence imaging) were co-encapsulated with perfluoro-15-crown-5-ether (P19FCE for19F MRI) by amphiphilic polymers and further coated with in situ formed manganese dioxide (MnO2) nanoshells, which served as efficient fluorescence and 19F MR quenchers due to energy transfer and paramagnetic relaxation effects, respectively. The over-expressed GSH in tumors would decompose the MnO2 nanoshells, resulting in remarkable enhancement of both fluorescence and 19F MRI signals of the nanoprobes, accordingly lighting up the tumor site.

Effective Detection of Nafion®-Based Theranostic Nanocapsules through 19F Ultra-Short Echo Time MRI.

The application of the Three-Dimensional Ultra-Short Echo Time (3D UTE)pulse sequence at a high magnetic field for visualization of the distribution of 19F loaded theranostic core-shell nanocapsules with Nafion® (1,1,2,2-tetrafluoroethene; 1,1,2,2-tetrafluoro-2- [1,1,1,2,3,3-hexafluoro-3-(1,2,2-trifluoroethenoxy)propan-2-yl] oxyethanesulfonic acid) incorporated into the shell is presented. The nanocarriers were formed via the layer-by-layer technique with biodegradable polyelectrolytes: PLL (Poly-L-lysine), and with Nafion®: polymer with high 19F content. Before imaging, an MR (magnetic resonance) spectroscopy and T1 and T2 measurements were performed, resulting in values of T2 between 1.3 ms and 3.0 ms, depending on the spectral line. To overcome limitations due to such short T2, the 3D UTE pulse sequence was applied for 19F MR imaging. First Nafion® solutions of various concentrations were measured to check the detection limit of our system for the investigated molecule. Next, the imaging of a phantom containing core-shell nanocapsules was performed to assess the possibility of visualizing their distribution in the samples. Images of Nafion® containing samples with SNR ≥ 5 with acquisition time below 30 min for 19F concentration as low as 1.53 × 10−2 mmol 19F/g of sample, were obtained. This is comparable with the results obtained for molecules, which exhibit more preferable MR characteristics.

Hypoxia theranostics of a human prostate cancer xenograft and the resulting effects on the tumor microenvironment

Hypoxia is frequently observed in human prostate cancer, and is associated with chemoresistance, radioresistance, metastasis, and castrate-resistance. Our purpose in these studies was to perform hypoxia theranostics by combining in vivo hypoxia imaging and hypoxic cancer cell targeting in a human prostate cancer xenograft. This was achieved by engineering PC3 human prostate cancer cells to express luciferase as well as a prodrug enzyme, yeast cytosine deaminase, under control of hypoxic response elements (HREs). Cancer cells display an adaptive response to hypoxia through the activation of several genes mediated by the binding of hypoxia inducible factors (HIFs) to HRE in the promoter region of target gene that results in their increased transcription. HIFs promote key steps in tumorigenesis, including angiogenesis, metabolism, proliferation, metastasis, and differentiation. HRE-driven luciferase expression allowed us to detect hypoxia in vivo to time the administration of the nontoxic prodrug 5-fluorocytosine that was converted by yeast cytosine deaminase, expressed under HRE regulation, to the chemotherapy agent 5-fluorouracil to target hypoxic cells. Conversion of 5-fluorocytosine to 5-fluorouracil was detected in vivo by 19F magnetic resonance spectroscopy. Morphological and immunohistochemical staining and molecular analyses were performed to characterize tumor microenvironment changes in cancer-associated fibroblasts, cell viability, collagen 1 fiber patterns, and HIF-1α. These studies expand our understanding of the effects of eliminating hypoxic cancer cells on the tumor microenvironment and in reducing stromal cell populations such as cancer-associated fibroblasts.

Tumor micro-environment sensitive 19F-magnetic resonance imaging in vivo

19F MRI has been paid great attention because of its higher signal-to-noise ratio and larger chemical shift than 1H MRI. However, the low 19F content and sensitivity of environment limit its development in clinic. Herein, considering the large surface area and abundant metal active sites of metal organic framework (MOF) nanoparticles, we fabricated Zr-based MOF nanoparticles (UiO-66-F NPs) with an organic ligand of tetrafluoro-terephthalic acid (F4PTA) as a pH-sensitive 19F MRI contrast agent, which remained stable in a neutral condition, but could release free F4PTA in an acidic condition to enhance 19F MRI signal. As a proof of concept, UiO-66-F NPs-based contrast agent could turn on the 19F MRI signal in an acidic tumor micro-environment. The results provide an alternative strategy for the design of tumor micro-environment responsive 19F MRI contrast agents.

Perfluorocarbons-Based 19F Magnetic Resonance Imaging in Biomedicine.

Fluorine-19 (19F) magnetic resonance (MR) molecular imaging is a promising noninvasive and quantitative molecular imaging approach with intensive research due to the high sensitivity and low endogenous background signal of the 19F atom in vivo. Perfluorocarbons (PFCs) have been used as blood substitutes since 1970s. More recently, a variety of PFC nanoparticles have been designed for the detection and imaging of physiological and pathological changes. These molecular imaging probes have been developed to label cells, target specific epitopes in tumors, monitor the prognosis and therapy efficacy and quantitate characterization of tumors and changes in tumor microenvironment noninvasively, therefore, significantly improving the prognosis and therapy efficacy. Herein, we discuss the recent development and applications of 19F MR techniques with PFC nanoparticles in biomedicine, with particular emphasis on ligand-targeted and quantitative 19F MR imaging approaches for tumor detection, oxygenation measurement, smart stimulus response and therapy efficacy monitoring, et al.

Versatile Nickel(II) Scaffolds as Coordination‐Induced Spin‐State Switches for 19F Magnetic Resonance‐Based Detection

Abstract19F magnetic resonance (MR) based detection coupled with well‐designed inorganic systems shows promise in biological investigations. Two proof‐of‐concept inorganic probes that exploit a novel mechanism for 19F MR sensing based on converting from low‐spin (S=0) to high‐spin (S=1) Ni2+ are reported. Activation of diamagnetic NiL1 and NiL2 by light or β‐galactosidase, respectively, converts them into paramagnetic NiL0, which displays a single 19F NMR peak shifted by &gt;35 ppm with accelerated relaxation rates. This spin‐state switch is effective for sensing light or enzyme expression in live cells using 19F MR spectroscopy and imaging that differentiate signals based on chemical shift and relaxation times. This general inorganic scaffold has potential for developing agents that can sense analytes ranging from ions to enzymes, opening up diverse possibilities for 19F MR based biosensing.

Design of Thermoresponsive Polyamine Cross-Linked Perfluoropolyether Hydrogels for Imaging and Delivery Applications

Perfluorocarbons are versatile compounds with applications in 19F magnetic resonance imaging (MRI) and chemical conjugation to drugs and pH sensors. We present a novel thermoresponsive perfluorocarbon emulsion hydrogel that can be detected by 19F MRI. The developed hydrogel contains perfluoro(polyethylene glycol dimethyl ether) (PFPE) emulsion droplets that are stabilized through ionic cross-linking with polyethylenimine (PEI). Specifically, PFPE ester undergoes hydrolysis upon contact with aqueous PEI solution, resulting in an ionic bond between the PFPE acid and charged PEI amino groups. Due to the ionic nature of the PFPE/PEI bond, potassium buffer is required to preserve the hydrogel's pH and rheological and emulsion droplet stability. The presence of the surface cross-linked PFPE droplets does not affect the hydrogel's rheological behavior, drug loading, or drug release, and the hydrogel is nontoxic. We propose that the presented hydrogel can be adapted to a broad range of biomedical imaging and delivery applications.

Multiresponsive Nanoprobes for Turn-On Fluorescence/19F MRI Dual-Modal Imaging.

Multiresponsive nanoprobes are highly desirable for low background and highly sensitive imaging in biomedical applications. Herein, we design a glutathione (GSH)/pH dual-responsive nanoprobe capable of both fluorescence imaging in cells and 19F magnetic resonance imaging (19F MRI) in deep tissue, by encapsulating manganese oleate (Mn(OA)2) on the surface of fluorinated fluorescent quantum dots (F-ZnS:Mn2+). In this approach, Mn(OA)2 serves as an efficient quencher of both fluorescence and 19F MRI signal. Both the fluorescence and 19F MRI signal can be turned on by introducing glutathione (GSH) that breaks up the Mn-O bonds within Mn(OA)2 under weak acidity conditions (e.g., pH 6.0). The imaging results in cells and mice suggest that this novel strategy can offer a promising nanoprobe for turn-on fluorescence/19F MRI dual-modal tumor imaging.

Fluorinated Water-Soluble Poly(2-oxazoline)s as Highly Sensitive 19F MRI Contrast Agents

Recently, 19F magnetic resonance imaging (MRI) emerged as a powerful noninvasive diagnostic tool in modern medicine. Fluorinated polymer materials represent an attractive class of MRI contrast agents (CAs) due to their structural variability and tunable properties. Herein, we describe for the first time the 19F MRI of CAs based on fluorinated water-soluble poly(2-oxazoline)s (PAOx), a polymer class with increasing popularity in biomedical sciences. A series of fluorinated PAOx with increasing fluorine content were synthesized by controlled side-chain hydrolysis of poly(2-methyl-2-oxazoline) followed by reacylation of its ethylenimine units by difluoroacetic anhydride. As the increasing fluorine content leads to the copolymer hydrophobization, their composition was optimized for maximal 19F MRI performance while retaining good solubility in water. The magnetic properties of the water-soluble polymers were studied in vitro by 19F NMR and MRI, revealing their outstanding relaxation properties and imaging sensitivity. All CAs were found to be noncytotoxic for HeLa cells in vitro. Finally, the diagnostic potential of the new CAs was demonstrated by a successful in vivo 19F MRI visualization of the selected fluorinated polymer in rats.

Synthesis and 19F NMR parameters of a perfluoro-tert-butoxy tagged L-DOPA analogue

The robust multistep synthesis of a perfluoro-tert-butoxy (PFTB) tagged analogue of the non-proteinogenic amino acid 3,4-dihydroxy-L-phenylalanine (L-DOPA) via diastereoselective benzylation of Oppolzer’s sultam glycinate was developed. The new compound is characterized by a flexible alkoxy linker connecting PFTB to the biochemically and pharmacologically relevant L-DOPA scaffold, which facilitates the free motion of the fluorinated moiety. Therefore, the nine chemically equivalent fluorine atoms give rise to an intense and sharp 19F NMR singlet signal that is easily detected in an aqueous environment. In addition, the kinetics of 19F NMR relaxation processes in blood solution fit the requirements for the potential use of the new compound as a probe in 19F magnetic resonance imaging applications in translational clinical field.