A Superparamagnetic Composite Hydrogel Scaffold as In Vivo Dynamic Monitorable Theranostic Platform for Osteoarthritis Regeneration.

Osteoarthritis (OA) is a prevalent disease, characterized by subchondral fractures in its initial stages, which has no precise and specific treatment now. Here, a novel multifunctional scaffold is synthesized by photopolymerizing glycidyl methacrylate-modified hyaluronic acid (GMHA) as the matrix in the presence of hollow porous magnetic microspheres based on hydroxyapatite. In vivo subchondral bone repairing results demonstrate that the scaffold's meticulous design has most suitable properties for subchondral bone repair. The porous structure of inorganic particles within the scaffold facilitates efficient transport of loaded exogenous vascular endothelial growth factor (VEGF). The Fe3O4 nanoparticles assembled in microspheres promote the osteogenic differentiation of bone marrow mesenchymal stem cells and accelerate the new bone generation. These features enable the scaffold to exhibit favorable subchondral bone repair properties and attain high cartilage repair scores. The therapy results prove that the subchondral bone support considerably influences the upper cartilage repair process. Furthermore, magnetic resonance imaging monitoring demonstrates that Fe3O4 nanoparticles, which are gradually replaced by new bone during osteochondral defect repair, allow a noninvasive and radiation-free assessment to track the newborn bone during the OA repair process. The composite hydrogel scaffold (CHS) provides a versatile platform for biomedical applications in OA treatment.

Synthesis of controlled-size starch nanoparticles and superparamagnetic starch nanocomposites by microemulsion method.

In this study, a synthesis process based on the microemulsion method (ME) was developed with the aim to produce controlled-size starch nanoparticles (SNPs). Several formulations were tested for the preparation of the W/O microemulsions varying the organic/aqueous phase ratios and co-stabilizers concentrations. SNPs were characterized in terms of size, morphology, monodispersity and crystallinity. Spherical shape particles with mean sizes 30-40 nm were prepared. The method was then used to simultaneously synthesize SNPs and iron oxide nanoparticles with superparamagnetic properties. Starch-based nanocomposites with superparamagnetic properties and controlled size were obtained. Therefore, the microemulsion method developed could be considered an innovative technology for the design and development of novel functional nanomaterials. The starch-based nanocomposites were evaluated in terms of morphology and magnetic properties, and they are being considered as promising sustainable nanomaterials for different biomedical applications.

Field-Pulse-Induced Annealing of 2D Colloidal Polycrystals.

Two-dimensional colloidal crystals are of considerable fundamental and practical importance. However, their quality is often low due to the widespread presence of domain walls and defects. In this work, we explored the annealing process undergone by monolayers of superparamagnetic colloids adsorbed onto fluid interfaces in the presence of magnetic field pulses. These systems present the extraordinary peculiarity that both the extent and the character of interparticle interactions can be adjusted at will by simply varying the strength and orientation of the applied field so that the application of field pulses results in a sudden input of energy. Specifically, we have studied the effect of polycrystal size, pulse duration, slope and frequency on the efficiency of the annealing process and found that (i) this strategy is only effective when the polycrystal consists of less than approximately 10 domains; (ii) that the pulse duration should be of the order of magnitude of the time required for the outer particles to travel one diameter during the heating step; (iii) that the quality of larger polycrystals can be slightly improved by applying tilted pulses. The experimental results were corroborated by Brownian dynamics simulations.

Comparative Analysis of Postoperative Complications of Sentinel Node Identification Using the SentiMag® Method and the Use of a Radiotracer in Patients with Breast Cancer.

(1) Background: The purpose of the study was a retrospective, comparative assessment of complications of the surgical sentinel node biopsy (SNB) procedure in breast cancer using the radiotracer method and the SentiMag® method on groups of patients after 3.5 years of use. (2) Methods: The material was a group of 345 patients with primary surgical breast cancer who underwent the SNB procedure with the use of a radiotracer in combination with wide local excision (WLE), simple amputation (SA) with SNB and an independent SNB procedure in the period from May 2018 to January 2021 in the Department of Oncological Surgery. Of the patients who were monitored in the Hospital Outpatient Clinic, 300 were enrolled. The analyzed group was compared in terms of the occurrence of the same complications with the group of 303 patients also operated on in our center in the period from January 2014 to September 2017, in which SN identification was performed using the SentiMag® method. (3) Results: The most common complications found were sensation disorders in the arm, which occurred in 16 (14.1%) patients using the radiotracer method, SentiMag®-11 (9.9%). By comparing the complication rate between the methods with the radiotracer (n = 300) and SentiMag® (n = 303), no significant differences were found. (4) Conclusions: Sentinel node (SN) identification using the radiotracer method and the SentiMag® method are comparable diagnostic methods in breast cancer, with a low risk of complications.

Myocardial inflammation and energetics by cardiac MRI: a review of emerging techniques.

The role of inflammation in cardiovascular pathophysiology has gained a lot of research interest in recent years. Cardiovascular Magnetic Resonance has been a powerful tool in the non-invasive assessment of inflammation in several conditions. More recently, Ultrasmall superparamagnetic particles of iron oxide have been successfully used to evaluate macrophage activity and subsequently inflammation on a cellular level. Current evidence from research studies provides encouraging data and confirms that this evolving method can potentially have a huge impact on clinical practice as it can be used in the diagnosis and management of very common conditions such as coronary artery disease, ischaemic and non-ischaemic cardiomyopathy, myocarditis and atherosclerosis. Another important emerging concept is that of myocardial energetics. With the use of phosphorus magnetic resonance spectroscopy, myocardial energetic compromise has been proved to be an important feature in the pathophysiological process of several conditions including diabetic cardiomyopathy, inherited cardiomyopathies, valvular heart disease and cardiac transplant rejection. This unique tool is therefore being utilized to assess metabolic alterations in a wide range of cardiovascular diseases. This review systematically examines these state-of-the-art methods in detail and provides an insight into the mechanisms of action and the clinical implications of their use.

Magnetic field-enhanced agglutination as a readout for rapid serologic assays with human plasma.

Recent virus outbreaks have revealed a critical need for large scale serological assays. However, many available tests either require a cumbersome, costly apparatus or lack the availability of full automation. In order to address these limitations, we describe a homogeneous assay for antibody detection via measurement of superparamagnetic particles agglutination. Application of a magnetic field permits to overcome the limitations governed by Brownian translational diffusion in conventional assays and results in an important acceleration of the aggregation process as well as an improvement of the limit of detection. Furthermore, the use of protein-concentrated fluid such as 5 times-diluted human plasma does not impair the performances of the method. Screening of human plasma samples shows a strict discrimination between seropositive and seronegative samples in an assay duration as short as 14 s. The sensitivity of this method, combined with its quickness and simplicity, makes it a promising diagnostic tool.

Static and Dynamic Self-Assembly of Pearl-Like-Chains of Magnetic Colloids Confined at Fluid Interfaces.

Magnetic colloids adsorbed at a fluid interface are unique model systems to understand self-assembly in confined environments, both in equilibrium and out of equilibrium, with important potential applications. In this work the pearl-chain-like self-assembled structures of superparamagnetic colloids confined to a fluid-fluid interface under static and time-dependent actuations are investigated. On the one hand, it is found that the structures generated by static fields transform as the tilt angle of the field with the interface is increased, from 2D crystals to separated pearl-chains in a process that occurs through a controllable and reversible zip-like thermally activated mechanism. On the other hand, the actuation with precessing fields about the axis perpendicular to the interface induces dynamic self-assembled structures with no counterpart in non-confined systems, generated by the interplay of averaged magnetic interactions, interfacial forces, and hydrodynamics. Finally, how these dynamic structures can be used as remotely activated roller conveyors, able to transport passive colloidal cargos at fluid interfaces and generate parallel viscous flows is shown. The latter can be used in the mixture of adsorbed molecules and the acceleration of surface-chemical reactions, overcoming diffusion limitations.

Pro-osteogenesis and in vivo tracking investigation of a dental implantation system comprising novel mTi implant and HYH-Fe particles.

Insufficient early osteogenesis seriously affects the later stage osteogenic quality and osseointegration of dental implants. To promote early osteogenesis, we first designed a Ti dental implant with a built-in magnet (mTi) to produce a local static magnetic field (SMF). Then, a dental implantation system comprising the mTi implant and the superparamagnetic hydroxyapatite (HA:Yb/Ho-Fe, named HYH-Fe) particles was implanted into the alveolar bone of beagles. The results showed that the mTi + HYH-Fe group displayed better early osteogenesis and later stage osseointegration than the Ti + HA and mTi + HA groups. A combination of the local SMF (mTi) and superparamagnetic HYH-Fe particles had a positive effect on the pro-osteogenesis of Ti implants. The results also indicated that week 10 could be adopted as the key time point to evaluate the early osteogenic effect of the mTi + HYH-Fe implantation system, which would be a promising prospect for promotion of osteogenesis, in vivo tracking investigation of material-bone relationships, and clinical applications.

Diagnostic Applications of Ultrasmall Superparamagnetic Particles of Iron Oxide for Imaging Myocardial and Vascular Inflammation.

Cardiac magnetic resonance (CMR) is at the forefront of noninvasive methods for the assessment of myocardial anatomy, function, and most importantly tissue characterization. The role of CMR is becoming even more significant with an increasing recognition that inflammation plays a major role for various myocardial diseases such as myocardial infarction, myocarditis, and takotsubo cardiomyopathy. Ultrasmall superparamagnetic particles of iron oxide (USPIO) are nanoparticles that are taken up by monocytes and macrophages accumulating at sites of inflammation. In this context, USPIO-enhanced CMR can provide valuable additional information regarding the cellular inflammatory component of myocardial and vascular diseases. Here, we will review the recent diagnostic applications of USPIO in terms of imaging myocardial and vascular inflammation, and highlight some of their future potential.

Substrate for the Myocardial Inflammation-Heart Failure Hypothesis Identified Using Novel USPIO Methodology.

OBJECTIVES: The purpose of this study was to identify where ultrasmall superparamagnetic particles of iron oxide (USPIO) locate to in myocardium, develop a methodology that differentiates active macrophage uptake of USPIO from passive tissue distribution; and investigate myocardial inflammation in cardiovascular diseases. BACKGROUND: Myocardial inflammation is hypothesized to be a key pathophysiological mechanism of heart failure (HF), but human evidence is limited, partly because evaluation is challenging. USPIO-magnetic resonance imaging (MRI) potentially allows specific identification of myocardial inflammation but it remains unclear what the USPIO-MRI signal represents. METHODS: Histological validation was performed using a murine acute myocardial infarction (MI) model. A multiparametric, multi-time-point MRI methodology was developed, which was applied in patients with acute MI (n = 12), chronic ischemic cardiomyopathy (n = 7), myocarditis (n = 6), dilated cardiomyopathy (n = 5), and chronic sarcoidosis (n = 5). RESULTS: USPIO were identified in myocardial macrophages and myocardial interstitium. R1 time-course reflected passive interstitial distribution whereas multi-time-point R2* was also sensitive to active macrophage uptake. R2*/R1 ratio provided a quantitative measurement of myocardial macrophage infiltration. R2* behavior and R2*/R1 ratio were higher in infarcted (p = 0.001) and remote (p = 0.033) myocardium in acute MI and in chronic ischemic cardiomyopathy (infarct: p = 0.008; remote p = 0.010), and were borderline higher in DCM (p = 0.096), in comparison to healthy controls, but were no different in myocarditis or sarcoidosis. An R2*/R1 threshold of 25 had a sensitivity and specificity of 90% and 83%, respectively, for detecting active USPIO uptake. CONCLUSIONS: USPIO are phagocytized by cardiac macrophages but are also passively present in myocardial interstitium. A multiparametric multi-time-point MRI methodology specifically identifies active myocardial macrophage infiltration. Persistent active macrophage infiltration is present in infarcted and remote myocardium in chronic ischemic cardiomyopathy, providing a substrate for HF.

Development of an LDL Receptor-Targeted Peptide Susceptible to Facilitate the Brain Access of Diagnostic or Therapeutic Agents.

Blood-brain barrier (BBB) crossing and brain penetration are really challenging for the delivery of therapeutic agents and imaging probes. The development of new crossing strategies is needed, and a wide range of approaches (invasive or not) have been proposed so far. The receptor-mediated transcytosis is an attractive mechanism, allowing the non-invasive penetration of the BBB. Among available targets, the low-density lipoprotein (LDL) receptor (LDLR) shows favorable characteristics mainly because of the lysosome-bypassed pathway of LDL delivery to the brain, allowing an intact discharge of the carried ligand to the brain targets. The phage display technology was employed to identify a dodecapeptide targeted to the extracellular domain of LDLR (ED-LDLR). This peptide was able to bind the ED-LDLR in the presence of natural ligands and dissociated at acidic pH and in the absence of calcium, in a similar manner as the LDL. In vitro, our peptide was endocytosed by endothelial cells through the caveolae-dependent pathway, proper to the LDLR route in BBB, suggesting the prevention of its lysosomal degradation. The in vivo studies performed by magnetic resonance imaging and fluorescent lifetime imaging suggested the brain penetration of this ED-LDLR-targeted peptide.

Molecular Imaging of Galectin-1 Expression as a Biomarker of Papillary Thyroid Cancer by Using Peptide-Functionalized Imaging Probes.

Thyroid cancers are the most frequent endocrine cancers and their incidence is increasing worldwide. Thyroid nodules occur in over 19-68% of the population, but only 7-15% of them are diagnosed as malignant. Diagnosis relies on a fine needle aspiration biopsy, which is often inconclusive and about 90% of thyroidectomies are performed for benign lesions. Galectin-1 has been proposed as a confident biomarker for the discrimination of malignant from benign nodules. We previously identified by phage display two peptides (P1 and P7) targeting galectin-1, with the goal of developing imaging probes for non-invasive diagnosis of thyroid cancer. The peptides were coupled to ultra-small superparamagnetic particles of iron oxide (USPIO) or to a near-infrared dye (CF770) for non-invasive detection of galectin-1 expression in a mouse model of papillary thyroid cancer (PTC, as the most frequent one) by magnetic resonance imaging and fluorescence lifetime imaging. The imaging probes functionalized with the two peptides presented comparable image enhancement characteristics. However, those coupled to P7 were more favorable, and showed decreased retention by the liver and spleen (known for their galectin-1 expression) and high sensitivity (75%) and specificity (100%) of PTC detection, which confirm the aptitude of this peptide to discriminate human malignant from benign nodules (80% sensitivity, 100% specificity) previously observed by immunohistochemistry.

Specific biological responses following dextran-coated ultra-small superparamagnetic particles of iron oxides administration.

Aim: The potential bio-related risks of dextran-coated ultra-small superparamagnetic particles of iron oxides (D-USPIO) were assessed. Materials & methods: Metabolic responses of D-USPIO in BALB/C mice were obtained using 1H-NMR-based metabolomic strategy combined with the traditional biochemical assay. Results: The metabolomic analyses of biological fluids (plasma and urine) and organs (liver, kidney and spleen) indicated that the disturbance, impairment and recovery of the physiological functions were related to the metabolic response to D-USPIO. The correlations between the biofluids and tissue metabolomes described the specific metabolic information of D-USPIO on their in vivo transportation, absorption, biodistribution and excretion. Conclusion: Metabolomic analysis provides preliminary validation for the use of D-USPIO in clinical medicine, and the results help to understand the potential adverse effects of the similar bio-nanomaterials further serve to their synthesis optimization and biocompatibility improvement.

Particle-Based Microfluidic Quartz Crystal Microbalance (QCM) Biosensing Utilizing Mass Amplification and Magnetic Bead Convection.

Microfluidic quartz crystal microbalances (QCM) can be used as powerful biosensors that not only allow quantifying a target analyte, but also provide kinetic information about the surface processes of binding and release. Nevertheless, their practical use as point-of-care devices is restricted by a limit of detection (LoD) of some ng/cm². It prohibits the measurement of small molecules in low concentrations within the initial sample. Here, two concepts based on superparamagnetic particles are presented that allow enhancing the LoD of a QCM. First, a particle-enhanced C-reactive protein (CRP) measurement on a QCM is shown. The signal response could be increased by a factor of up to five by utilizing the particles for mass amplification. Further, a scheme for sample pre-preparation utilizing convective up-concentration involving magnetic bead manipulation is investigated. These experiments are carried out with a glass device that is fabricated by utilizing a femtosecond laser. Operation regimes for the magnetic manipulation of particles within the microfluidic channel with integrated pole pieces that are activated by external permanent magnets are described. Finally, the potential combination of the concepts of mass amplification and up-concentration within an integrated lab-on-a chip device is discussed.

Ultra-small superparamagnetic iron oxide contrast agents for lymph node staging of high-risk prostate cancer.

Ultrasmall superparamagnetic particles of iron oxide (USPIOs) imaged with magnetic resonance imaging (MRI) have been proposed as an experimental method for visualizing lymph node (LN) metastases. The method does not require ionizing radiation, yet can detect small nodes that are involved with metastases. USPIOs are naturally taken up by macrophages that deposit in the normal LN creating a low signal region in normal areas; areas within the node that do not show this loss of signal are likely involved by tumor although there can be other causes (fibrosis or inflammation). However, the lack of approved USPIOs that are clinically available hinders adoption and larger studies. The proposed indications for USPIO MRI, including specific compounds and imaging methods are discussed.

Rapid detection of unconjugated estriol in the serum via superparamagnetic lateral flow immunochromatographic assay.

Unconjugated estriol (uE3) is one of the main naturally occurring estrogens that plays an important role in growth and development of the fetus. Usually, the level of uE3 is very low in men and non-pregnant women, but in pregnant women, the level of estriol has been found to be quite high. Therefore, the combination of uE3, AFP, and hCG is now widely used for Down Syndrome screening as a triple marker. Here, we developed a superparamagnetic lateral flow immunochromatographic assay to quantitatively detect uE3. The detection limit of this assay was 0.86 nmol/L and the linear range for the determination of uE3 was from 1 to 100 nmol/L. The detection time was 15 min and the assay had very low cross-reactivity with estrone (E1), estradiol (E2), and progesterone. The coefficient of variation (CV) of intra- and inter-assay ranged from 5% to 13%. The magnetic signals were stable under 37 °C within 7 d. Moreover, the concentrations of uE3 measured by lateral flow immunochromatographic assay in 230 serum samples collected from pregnant women at the Chinese People's Liberation Army General Hospital had a good correlation with those measured by time-resolved fluorescence immunoassay (R = 0.946).

Toward a new and noninvasive diagnostic method of papillary thyroid cancer by using peptide vectorized contrast agents targeted to galectin-1.

The incidence of papillary thyroid cancer has increased these last decades due to a better detection. High prevalence of nodules combined with the low incidence of thyroid cancers constitutes an important diagnostic challenge. We propose to develop an alternative diagnostic method to reduce the number of useless and painful thyroidectomies using a vectorized contrast agent for magnetic resonance imaging. Galectin-1 (gal-1), a protein overexpressed in well-differentiated thyroid cancer, has been targeted with a randomized linear 12-mer peptide library using the phage display technique. Selected peptides have been conjugated to ultrasmall superparamagnetic particles of iron oxide (USPIO). Peptides and their corresponding contrast agents have been tested in vitro for their specific binding and toxicity. Two peptides (P1 and P7) were selected according to their affinity toward gal-1. Their binding has been revealed by immunohistochemistry on human thyroid cancer biopsies, and they were co-localized with gal-1 by immunofluorescence on TPC-1 cell line. Both peptides induce a decrease in TPC-1 cells' adhesion to gal-1 immobilized on culture plates. After coupling to USPIO, the peptides preserved their affinity toward gal-1. Their specific binding has been corroborated by co-localization with gal-1 expressed by TPC-1 cells and by their ability to compete with anti-gal-1 antibody. The peptides and their USPIO derivatives produce no toxicity in HepaRG cells as determined by MTT assay. The vectorized contrast agents are potential imaging probes for thyroid cancer diagnosis. Moreover, the two gal-1-targeted peptides prevent cancer cell adhesion by interacting with the carbohydrate-recognition domain of gal-1.

An Explanatory Case on the Limitations of Lymph Node Staging in Recurrent Prostate Cancer.

We describe the case of a 54-year-old man with nodal recurrence prostate cancer after radical prostatectomy, salvage external-beam radiotherapy and salvage lymph node dissection. The patient was evaluated with a lymphotropic ultrasmall superparamagnetic particles of iron oxide (USPIO)-MRI and a 68Ga radiolabelled prostate specific membrane antigen (Ga68-PSMA) PET-CT scan which enhanced persistent localized nodal disease. The patient was then considered for a second robot-assisted extended S-LND. Differently from preoperative imaging, pathology report revealed a wide nodal involvement mirroring a metastatic disease. The current manuscript is an explanatory case on the limitations of lymph node imaging in prostate cancer recurrence.

Sentinel lymph node (sln) in breast cancer. Review of current identification methods.

The subject of the paper is the review of the current methods for identifying the sentinel lymph node in breast cancer treatment, taking into account the latest techniques and based on the up-to-date analyses of many years of experience in using this method.

MRI detection of endothelial cell inflammation using targeted superparamagnetic particles of iron oxide (SPIO).

BACKGROUND: There is currently no clinical imaging technique available to assess the degree of inflammation associated with atherosclerotic plaques. This study aims to develop targeted superparamagnetic particles of iron oxide (SPIO) as a magnetic resonance imaging (MRI) probe for detecting inflamed endothelial cells. METHODS: The in vitro study consists of the characterisation and detection of inflammatory markers on activated endothelial cells by immunocytochemistry and MRI using biotinylated anti-P-selectin and anti-VCAM-1 (vascular cell adhesion molecule 1) antibody and streptavidin conjugated SPIO. RESULTS: Established an in vitro cellular model of endothelial inflammation induced with TNF-α (tumor necrosis factor alpha). Inflammation of endothelial cells was confirmed with both immunocytochemistry and MRI. These results revealed both a temporal and dose dependent expression of the inflammatory markers, P-selectin and VCAM-1, on exposure to TNF-α. CONCLUSION: This study has demonstrated the development of an in vitro model to characterise and detect inflamed endothelial cells by immunocytochemistry and MRI. This will allow the future development of contrast agents and protocols for imaging vascular inflammation in atherosclerosis. This work may form the basis for a translational study to provide clinicians with a novel tool for the in vivo assessment of atherosclerosis.