Deciphering the Parameters to Produce Highly Reproducible and Scalable Iron Oxide Nanoparticles

Nanomedicine has been long hailed as a game changer for treating several ailments, but its translation from bench to bedside is facing some hurdles. Over the past few decades, there have been a plethora of reports regarding the synthesis of nanomaterials and, in particular, of iron oxide nanoparticles. However, very few reports discuss the role of stirring speed, reproducibility, and scalability. This work attempts to comprehensively revisit the most widely used existing protocols and discuss how the particle size or shape varies when certain parameters are altered and different precursors and solvents are used. It also discusses the probability of reproducing and scaling up the reactions while deciphering the effect of the ramp rate on size and shape. Lastly, it upgrades the existing methods and suggests a modification to produce highly reproducible and scalable nanoparticles of ∼4 nm, which can be further tuned to ∼2 nm by merely modifying the stirring speed.Ministerio de Ciencia e Innovación ID2020-118448RBC21, PID2020-118448RBC22Ministerio de Economia, Industria y Competitividad P18-RT-1663/PAIDI20Junta de Andalucía RH-0040-2021, P20_00727/PAIDI202

Synthesis and Characterization of Elongated-Shaped Silver Nanoparticles as a Biocompatible Anisotropic SERS Probe for Intracellular Imaging: Theoretical Modeling and Experimental Verification

Progress in the field of biocompatible SERS nanoparticles has promising prospects for biomedical applications. In this work, we have developed a biocompatible Raman probe by combining anisotropic silver nanoparticles with the dye rhodamine 6G followed by subsequent coating with bovine serum albumin. This nanosystem presents strong SERS capabilities in the near infrared (NIR) with a very high (2.7 × 107) analytical enhancement factor. Theoretical calculations reveal the effects of the electromagnetic and chemical mechanisms in the observed SERS effect for this nanosystem. Finite element method (FEM) calculations showed a considerable near field enhancement in NIR. Using density functional quantum chemical calculations, the chemical enhancement mechanism of rhodamine 6G by interaction with the nanoparticles was probed, allowing us to calculate spectra that closely reproduce the experimental results. The nanosystem was tested in cell culture experiments, showing cell internalization and also proving to be completely biocompatible, as no cell death was observed. Using a NIR laser, SERS signals could be detected even from inside cells, proving the applicability of this nanosystem as a biocompatible SERS probe.España, Regional Ministry of Economy, Junta de Andalucía, P07-FQM-02595 (to CC), P10-FQM-06615 (to JMOM), P10-CTS-6928 (to DP) and PAIDI2020 Program (FQM319 to RFM and CTS677 to DP)Junta de Andalucía, PI-0070/2008 (to PZ) and PI-0068/2008 (to DP

Highly water-stable rare ternary Ag-Au-Se nanocomposites as long blood term X-rays computed tomography contrast agents

X-ray computed tomography (CT) is a powerful and widely used medical non-invasive technique that requires intravenously administration of contrast agents to enhance the sensitivity and visualization of soft tissues. In this work, we have developed a novel CT contrast agent based on ternary Ag-Au-Se chalcogenide nanoparticles. A facile and gentle ligand exchange by using a 3 kDa PEGylated ligand with a dithiol dihydrolipoic as an anchor resulted in highly water-soluble and monodisperse nanoparticles. Moreover, the injected PEGylated ternary NPs presented excellent characteristics as a CT contrast agent with high bioavailability, low cytotoxicity and long blood circulation times with slow uptake by the mononuclear phagocyte system, thus being ideal for in vivo imaging

Passive targeting of high-grade gliomas via the EPR effect: a closed path for metallic nanoparticles?

Passive tumor targeting via the enhanced permeability and retention (EPR) effect has long been considered the most effective mechanism for the accumulation of nanoparticles inside solid tumors. However, several studies have demonstrated that the EPR effect is largely dependent on the tumor type and location. Particularly complex is the situation in brain tumors, where the presence of the blood–brain tumor barrier (BBTB) adds an extra limiting factor in reaching the tumor interstitium. However, it remains unclear whether these restraints imposed by the BBTB prevent the EPR effect from acting as an efficient tumor targeting mechanism for metallic nanoparticles. In this work, we have studied the EPR effect of metallic magnetic nanoparticles (MMNPs) in a glioblastoma (GBM) model by parametric MRI. Our results showed that only MMNPs ≤50 nm could reach the tumor interstitium, whereas larger MMNPs were unable to cross the BBTB. Furthermore, even for MMNPs around 30–50 nm, the amount of them found within the tumor was scarce and restricted to the vicinity of large tumor vessels, indicating that the BBTB strongly limits the passive accumulation of metallic nanoparticles in brain tumors. Therefore, active targeting becomes the most reasonable strategy to target metallic nanoparticles to GBMs.Ministerio de Economía, Industria y Competitividad de España - CTQ2017-86655-RMinisterio de Ciencia e Innovación de España - PID2020- 118448RB-C21 y PID2020-118448RB-C22Consejería de Salud de la Junta de Andalucía - OH-0026-201

Bi-Magnetic Core-Shell CoFe2O4@MnFe2O4 Nanoparticles for In Vivo Theranostics

In this work, we report the synthesis and characterization of three magnetic nanosystems, CoFe2O4, CoFe2O4@ZnFe2O4, and CoFe2O4@MnFe2O4, which were developed as potential theranostic agents for magnetic hyperthermia and magnetic resonance imaging (MRI). These nanosystems have been thoroughly characterized by X-ray Diffraction (XRD), Transmission Electron Miscroscopy (TEM), Dark Field-TEM (DF-TEM), Vibrating Sample Magnetometry (VSM), and inductive heating, in order to elucidate their structure, morphology, and magnetic properties. The bi-magnetic CoFe2O4@ZnFe2O4 and CoFe2O4@MnFe2O4 nanoparticles (NPs) exhibited a core-shell structure with a mean average particle size of 11.2 ± 1.4 nm and 14.4 ± 2.4 nm, respectively. The CoFe2O4@MnFe2O4 NPs showed the highest specific absorption rate (SAR) values (210-320 W/g) upon exposure to an external magnetic field, along with the highest saturation magnetization (Ms). Therefore, they were selected for functionalization with the PEGylated ligand to make them stable in aqueous media. After the functionalization process, the NPs showed high magnetic relaxivity values and very low cytotoxicity, demonstrating that CoFe2O4@MnFe2O4 is a good candidate for in vivo applications. Finally, in vivo MRI experiments showed that PEGylated CoFe2O4@MnFe2O4 NPs produce high T2 contrast and exhibit very good stealth properties, leading to the efficient evasion of the mononuclear phagocyte system. Thus, these bi-magnetic core-shell NPs show great potential as theranostic agents for in vivo applications, combining magnetic hyperthermia capabilities with high MRI contrast.España, MINECO CTQ2017-86655-

Comprehensive Toxicity Assessment of PEGylated Magnetic Nanoparticles for in vivo applications

Magnetic nanoparticles (MNPs) represent one of the greatest promises for the development of a new generation of diagnostic agents for magnetic resonance imaging, with improved specificity and safety. Indeed, during the last decade the number of studies published in this field has grown exponentially. However, the clinical translation achieved so far has been very limited. This situation is likely related to the fact that most studies are focused on the in vitro characterization of these new nanomaterials, and very few provide an exhaustive in vivo characterization, where key aspects, such as pharmacokinetics, bioavailability, and, most importantly, toxicity, are properly evaluated. In this work, we propose a protocol for the comprehensive assessment of the toxicity of MNPs, based on the use of zebrafish embryos as an intermediate screening step between cell culture assays and studies in rodents. MNPs with different cores, ferrite and manganese ferrite oxide, and sizes between 3 and 20 nm, were evaluated. Cell viability at a concentration of 50 μg/mL of PEGylated MNPs was above 90 % in all cases. However, the exposure of zebrafish embryos to manganese based MNPs at concentrations above 100 μg/ mL showed a low survival rate (< 50 %). In contrast, no mortality (survival rate ∼100 %) and normal hatching rate were obtained for the iron oxide MNPs. Based on these results, together with the physicochemical and magnetic properties (r2 = 153.6 mM-1·s-1), the PEGylated 20 nm cubic shape iron oxide MNPs were selected and tested in mice, showing very good MRI contrast and, as expected, absence of toxicity.Consejería de Andalucía Sanidad de la Junta de Andalucía - PI2013-0559Ministerio español de Economía y Competitividad - CTQ2017-86655-RUniversidad de Sevilla - V Plan Propi

Scaling Up Magnetic Nanobead Synthesis with Improved Stability for Biomedical Applications

The growing interest in multifunctional nano-objects based on polymers and magnetic nanoparticles for biomedical applications motivated us to develop a scale-up protocol to increase the yield of polymeric magnetic nanobeads while aiming at keeping the structural features at optimal conditions. The protocol was applied to two different types of magnetic ferrite nanoparticles: the Mn-ferrite selected for their properties as contrast agents in magnetic resonance imaging and iron oxide nanostar shaped nanoparticles chosen for their heat performance in magnetic hyperthermia. At the same time, some experiments on surface functionalization of nanobeads with amino modified polyethyelene glycol (PEG) molecules have provided further insight into the formation mechanism of magnetic nanobeads and the need to cross-link the polymer shell to improve the stability of the beads, making them more suitable for further manipulation and use. The present work summarizes the most important parameters required to be controlled for the upscaling of nanobead synthesis in a bench protocol and proposes an alternative cross-linking strategy based on prefunctionalization of the polymer prior to the nanobead formation as a key parameter to improve the nanobead structural stability in solutions at different pHs and during surface functionalization

PEGylated Terbium-Based Nanorods as Multimodal Bioimaging Contrast Agents

Diagnostic imaging strongly relies on the use of contrast agents (CAs). In general terms, current CAs present undesirable side effects that encourage researchers and pharmaceutical companies to continually search for safer and more versatile alternatives. Here, we describe the synthesis and characterization of terbium-based nanorods (TbNRs) as a potential alternative to traditional CAs for magnetic resonance imaging (MRI) and X-ray computed tomography (CT). The paramagnetism and high atomic number of Tb provide TbNRs with both magnetic relaxivity and X-ray attenuation capabilities. After surface functionalization with a polyethylene glycol (PEG)-derived ligand, TbNRs showed high colloidal stability in physiological media. Additionally, toxicity studies conducted in cell cultures and zebrafish embryos demonstrated the safety of the as-synthesized TbNRs, thus supporting their potential use as CAs. Lastly, in vivo imaging experiments in mice demonstrated that TbNRs produce remarkable contrast enhancement on both MRI and CT.The authors want to express special thanks to Alejandro Domínguez for his invaluable help with the in vivo toxicity experiments. The authors also thank Dr. Juan F. López for his support with the TEM experiments, Dr. Iñaki Orue for VSM measurement, Dr. John Pearson and Luisa Macías for assistance with the cell experiments and helpful discussion, Reyes Molina for assistance with animal experiments, and Maria Somoza for helping with the MRI experiments. Authors thank BIONAND’s Nanoimaging Unit. Optical microscopy, TEM, CT, and MRI experiments have been performed in the ICTS “NANBIOSIS”, more specifically in the U28 Unit at BIONAND.Funding for open access charge: Universidad de Málaga / CBUA. The authors want to express special thanks to Alejandro Domínguez for his invaluable help with the in vivo toxicity experiments. The authors also thank Dr. Juan F. López for his support with the TEM experiments, Dr. Iñaki Orue for VSM measurement, Dr. John Pearson and Luisa Macías for assistance with the cell experiments and helpful discussion, Reyes Molina for assistance with animal experiments, and Maria Somoza for helping with the MRI experiments. Authors thank BIONAND’s Nanoimaging Unit. Optical microscopy, TEM, CT, and MRI experiments have been performed in the ICTS “NANBIOSIS”, more specifically in the U28 Unit at BIONAND

Biological Evaluation of Carbohydrate-based Aprepitant Analogs for Neuroblastoma Treatment

Different studies using Aprepitant, a NK1R antagonist currently used as a clinical drug for treating chemotherapy-related nausea and vomiting, have demonstrated that pharmacological inhibition of NK1R effectively reduces the growth of several tumor types such as neuroblastoma (NB). In a previous work, we demonstrated that a series of carbohydrate-based Aprepitant analogs, derived from either D-galactose or L-arabinose, have shown high affinity and NK1R antagonistic activity with a broad-spectrum anticancer activity and an important selectivity. In this new study, we explore the selective cytotoxic effects of these derivatives for the treatment of NB. Furthermore, we describe the design and stereoselective synthesis of a new generation of D-glucose derivatives as Aprepitant analogs, supported by docking studies. This approach showed that most of our carbohydrate-based analogs are significantly more selective than Aprepitant. The galactosyl derivative 2α, has demonstrated a marked in vitro selective cytotoxic activity against NB, with IC50 values in the same range as those of Aprepitant and its prodrug Fosaprepitant. Interestingly, the derivative 2α has shown similar apoptotic effect to that of Aprepitant. Moreover, we can select the glucosyl amino derivative 10α as an interesting hit exhibiting higher in vitro cytotoxic activity against NB than Aprepitant, being 1.2 times more selective.Ministerio de Ciencia, Innovación y Universidades PID2019-104767RB-I0

"Nanohybrids" based on pH-responsive hydrogels and inorganic nanoparticles for drug delivery and sensor applications.

Allyl-PEG capped inorganic NPs, including magnetic iron oxide (IONPs), fluorescent CdSe/ZnS quantum dots (QDs), and metallic gold (AuNPs of 5 and 10 nm) both individually and in combination, were covalently attached to pH-responsive poly(2-vinylpyridine-co-divinylbenzene) nanogels via a facile and robust one-step surfactant-free emulsion polymerization procedure. Control of the NPs associated to the nanogels was achieved by the late injection of the NPs to the polymerization solution at a stage when just polymeric radicals were present. Remarkably, by varying the total amount of NPs injected, the swelling behavior could be affected. Furthermore, the magnetic response as well as the optical features of the nanogels containing either IONPs or QDs could be modified. In addition, a radical quenching in case of gold nanoparticles was observed, thus affecting the final nanogel geometry