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The simultaneous detection and quantification of several iron-containing species in biological matrices is a challenging issue. Especially in the frame of studies using magnetic nanoparticles for biomedical applications, no gold-standard technique has been described yet and combinations of different techniques are generally used. In this work, AC magnetic susceptibility measurements are used to analyze different organs from an animal model that received a single intratumor administration of magnetic nanoparticles. The protocol used for the quantification of iron associated with the magnetic nanoparticles is carefully described, including the description of the preparation of several calibration standard samples of nanoparticle suspensions with different degrees of dipolar interactions. The details for the quantitative analysis of other endogenous iron-containing species such as ferritin or hemoglobin are also described. Among the advantages of this technique are that tissue sample preparation is minimal and that large amounts of tissue can be characterized each time (up to hundreds of milligrams). In addition, the very high specificity of the magnetic measurements allows for tracking of the nanoparticle transformations. Furthermore, the high sensitivity of the instrumentation results in very low limits of detection for some of the iron-containing species. Therefore, the presented technique is an extremely valuable tool to track iron oxide magnetic nanoparticles in samples of biological origin., This work was funded by the Ministerio de Ciencia, Innovación y Universidades (MCIU), the Agencia Estatal de Investigación (AEI), and Fondo Europeo de Desarrollo Regional (FEDER) through the MACBETH project (PGC2018-096016-B-I00 to R.M.F and L.G.). Funding was also received from the European Commission through the TBMED project (DT-NMBP-02-2018, ID 814439, to R.M.F.) and the MagicCellGene Project (M-ERA.NET COFUND call 2016, funded by Ministerio de Economía y Competitividad, MINECO, Spain, in the framework of the PCIN-2017-060 project, to V.G.). Authors also acknowledge support from Gobierno de Aragón and Fondos FEDER for funding the Bionanosurf (E15_17R) research group. MINECO and FSE/Agencia Estatal de Investigación are also acknowledged for the Ramón y Cajal subprogram grants RYC-2015-17640 to R.M.F., RYC-2014-15512 to L.G., and RYC2019-026860-I to M.M.. Y.F-A. thanks the Santander-Universidad Zaragoza Fellowship program for her PhD position.

Trabajo presentado a la Conference on Advanced Materials and Devices for Nanomed (AMA4MED), celebrada en Valencia (España) del 3 al 4 de mayo de 2022.

Resumen del trabajo presentado a la 5th Spanish Conference on Biomedical Applications of Nanomaterials (SBAN), celebrada on-line del 3 al 10 de septiembre de 2022.

Trabajo presentado a la 6th International Nanotech and Nanoscience Conference and Exhibition, celebrada virtualmente del 23 al 25 de junio de 2021.

Trabajo presentado a la 4th Spanish Conference on Biomedical Applications of Nanomaterials, celebrada online del 2 al 4 de junio de 2021.

Trabajo presentado a la V Reunión de Jóvenes Investigadores en Coloides e Interfases, celebrada en la Universidad de Zaragoza del 2 al 4 de marzo de 2020.

Trabajo presentado al III Workshop de NanoOncología, celebrado en la Universidad de Zaragoza del 14 al 15 de enero de 2020.

Póster presentado a la 2nd Spanish Conference on Biomedical Applications of Nanomaterials (SBAN), celebrada en Madrid del 6 al 7 de junio de 2019., Multiple Sclerosis (MS) is an inflammatory autoimmune disease of the central nervous system that can lead to severe disability. The wide variety of symptoms characteristic for MS such as numbness, weakness, paresis, visual problems, severe fatigue and depression and also intellectual and cognitive impairments are caused by by brain and spinal cord's inflammation that damages the myelin sheath. During the last years, significant progress has been made in understanding how pathogenic leukocytes migrate from the periphery to the CNS, and the critical roles played by the α4ß1and α4ß7 integrins have been studied extensively in animal models, and in patients in clinical trials. Thus, the counter ligands of these integrins (VCAM1 and MAdCAM1) are a perfect target to prevent the inflammatory process. Gold nanoparticles (AuNPs) show low toxicity and excellent optical properties, they represent a very powerful tool in imaging and therapy applications. In this work, we propose the functionalization of AuNPs with five different integrins ligand-like peptidomimetics to target the pathogenic leukocytes and create a challenging device for imaging and therapy of MS. Preliminary outcomes in in vitro experiments will be presented in order to lay the foundations for future study., This project has received funding from MCINN (Spain) (project "SAF2014-54763-C2-1-R"), from Gobierno de Aragón (predoctoral contract of Rafael Ramírez Jiménez) and from Fondo Social of Gobierno de Aragón (Spain).

Resumen del trabajo presentado a la 2nd Spanish Conference on Biomedical Applications of Nanomaterials (SBAN), celebrada en Madrid del 6 al 7 de junio de 2019., In recent years, there has been significant research focusing on the use of magnetic nanoparticles for biomedical applications. To date, most of this research has focused on the development of the particles, their surface functionalization and the experiments leading to the proof of concept of the different biomedical applications. However, before these particles reach the clinical practice, several aspects regarding their biological fate, biodistribution, and toxicity, must be assessed. Previous research on the biodistribution and degradation of magnetic nanoparticles after systemic administration has shown that nanoparticles accumulate mainly in the spleen and liver of animals and that the degradation kinetics of magnetic nanoparticles may be very slow. However, there is little information regarding the biodistribution of magnetic nanoparticles when different administration routes are used. In this study, we have evaluated the degradation and biodistribution of magnetic nanoparticles after intratumoral injection. We have used a human pancreatic heterotopic-xenograft tumour model in Balb/c nude mice. Animals received a single intratumoral administration of magnetic nanoparticles. The migration of the particles to other organs was evaluated in two different scenarios: one in which no further treatment was applied to the animals and other in with an alternating magnetic field for hyperthermia treatment was applied three times along a week. The biodistribution and degradation over time of the particles has been followed by AC magnetic susceptibility. These experiments have allowed us to evaluate the effect of the magnetic hyperthermia treatment on the nanoparticles biodistribution.

Resumen del trabajo presentado a la IV Reunión de Jóvenes Investigadores en Coloides e Interfases, celebrada en Córdoba del 7 al 9 de febrero de 2018.

Resumen del trabajo presentado a la 12th International Conference on the Scientific and Clinical Applications of Magnetic Carriers, celebrada en Copenhague (Dinamarca) del 22 al 26 de mayo de 2018., Magnetic hyperthermia is a promising therapy for the localized treatment of cancer. Under the exposure to an external alternating magnetic field, magnetic nanoparticles (MNPs) act as heating agents inducing cell death in the treated region. Understanding the molecular mechanisms involved in the cellular damage generated by this treatment is crucial for the successful application of this therapy. In this study, 12 nm spherical MNPs coated with PMAO (poly (maleic anhydride-alt-1-octadecene) and functionalized with glucose were prepared by thermal decomposition. In order to evaluate the influence of the nanoparticle location in the treatment efficacy, two different 3D cell culture models, based on collagen gels, were prepared using a macrophage cell line, RAW264.7, (Figure 1). The first model (Model 1) kept all the particles inside the cells while the second model (Model 2) had particles both inside and outside the cells. The first model mimics a scenario where MNPs are administered intravenously with an active targeting, and the second one mimics intratumoral administration. The MNPs uptake and cell death mechanisms induced after the hyperthermia treatment (377 kHz, 13kA/m and 30 minutes, DM100-DM3 nB nanoscale Biomagentics,) were evaluated by flow cytometry. Interestingly, the cell death pathway was different depending on the MNPs location. Necrosis was observed 24h after magnetic hyperthermia application in the model where the nanoparticles are located just inside the cells. In contrast, apoptosis was detected in the model where the particles are inside and outside the cells. In addition, in the second model, our results evidenced an enhancement of the nanoparticles uptake after the exposure to the alternating magnetic field. This observation could justify the repetition of the treatment to obtain a better antitumor effect.Our results demonstrate the potential efficacy of magnetic hyperthermia in the treatment of malignant tumours. In addition, the use of 3D cell culture models for the optimization of hyperthermia treatments (type and dose of MNPs, repetition cycles, field amplitude and frequency, etc) presents several advantages, allowing the obtention of information in a more realistic way than monolayer cell cultures and reducing the number of animals required for preclinical tests.

Trabajo presentado a la 3rd International Conference of Polyol Mediated Synthesis (IC-PMS), celebrada en el Instituto de Ciencia de Materiales de Madrid (CSIC) del 25 al 27 de junio de 2018.

Trabajo presentado al NanoSafety Cluster Scientific Meeting, celebrado en Atenas (Grecia) del 22 al 23 de marzo de 2018.

Resumen del póster presentado a la 8ª Jornada de Jóvenes Investigadores (Química y Física) de Aragón, celebrada en Zaragoza el dia 22 de noviembre de 2018., La esclerosis múltiple es una enfermedad crónica autoinmune que se caracteriza por la desmielinización del sistema nervioso central lo que daña severamente su funcionalidad. Se relaciona con procesos de inflamación causados por linfocitos T autorreactivos que sobreexpresan las integrinas α4β1 y α4β7, gracias a las cuales llegan al cerebro cruzando la barrera hematoencefálica. Así, los ligandos de estas integrinas (VCAM1 and MAdCAM1) pueden ser una diana perfecta para prevenir estos procesos de inflamación. Las nanopartículas de oro presentan baja toxicidad y propiedades que las hacen idóneas para aplicaciones en terapia y diagnóstico. En este trabajo proponemos la funcionalización con seis peptidomiméticos de los ligandos VCAM1 and MAdCAM1 para que interaccionen con las integrinas presentes en células T autorreactivas y crear un instrumento de imagen y terapia de la esclerosis múltiple. Los peptidomiméticos se acoplaron sobre nanoesferas de oro de 14 nm recubiertas con ácido ω-mercaptopolietilenglicol-α-carboxílico. La funcionalización se llevó a cabo a través de formación de un enlace amida para lo que se estudiaron diferentes condiciones de reacción. Se realizó la caracterización físico-química de las nanopartículas funcionalizadas y se estudió su internalización en células Jurkat (línea inmortalizada de linfocitos T humanos) encontrando una mayor internalización de las nanopartículas funcionalizadas.

Resumen del trabajo presentado a la 12th International Conference on the Scientific and Clinical Applications of Magnetic Carriers, celebrada en Copenhague (Dinamarca) del 22 al 26 de mayo de 2018., Due to their size-tunable physical and chemical properties, magnetic nanoparticles (MNPs) have demonstrated a wide range of applications in healthcare and life science. In particular, the well-known property of remotely inducing local heat when an alternating magnetic field (AMF) is applied has opened a new area of research called Magnetic Hyperthermia. Traditionally, the activation of these MNPs as nanoheaters has been used seeking the destruction of tumoral cells. In this work, however, we want to show that it is possible to use the heat generated by AMF for a different purpose: to locally reach an optimal temperature (45ºC) that enhances the conversion of a prodrug (indole-3-acetic acid) into a drug (peroxylated radicals) by a therapeutic enzyme (horseradish peroxidase, HRP). has been optimized. Different biohybrids (biosilica+MNP+HRP) were prepared using polyethylenimines of different molecular weight (1300 and 60000 daltons) as amino catalysts for the formation of the silica nanoparticles. Besides, the combination of enzyme crosslinking and the addition of polyols during encapsulation, provided a 250 times increase in the halflife time of the enzyme at 50°C. After carrying out and extensive physicochemical characterization (SEM, TEM, DLS, SQUID, ICP-OES...), it was demonstrated that by applying AMF it is possible to reach local temperatures higher than 45°C within the enzymes microenvironment without an increase in the overall temperature of the reaction medium. These results show that it is possible to think about developing a novel >on/off> switch approach for the remote conversion of pro-drugs in cancer therapy by enzymes of thermal origin that show little or no activity at 37ºC. This would allow a precise remote control of the therapy that solves a limitation of the current strategies that could not avoid its activation outside the target site.

Resumen del trabajo presentado a la 1st Spanish Conference on Biomedical Applications of Nanomaterials (SBAN), celebrada en Madrid del 7 al 8 de junio de 2018., Magnetic hyperthermia is a promising therapy for the localized treatment of cancer. Under the exposure to an external alternating magnetic field, magnetic nanoparticles (MNPs) act as heating agents inducing cell death in the treated region. Understanding the molecular mechanisms involved in the cellular damage generated by this treatment is crucial for the successful application of this therapy. In this study, 12 nm MNPs coated with PMAO (poly(maleic anhydride-alt-1-octadecene) and functionalized with glucose were prepared by thermal decomposition. In order to evaluate the influence of the nanoparticle location in the treatment efficacy, two different 3D cell culture models, based on collagen gels, were prepared using a macrophage cell line, RAW264.7 (Figure 1). The first model (Model 1) kept all the particles inside the cells while the second model (Model 2) had particles both inside and outside the cells. The first model mimics a scenario where MNPs are administered intravenously with an active targeting, and the second one mimics intratumoral administration. The MNPs uptake and cell death mechanisms induced after the hyperthermia treatment (377.5 kHz, 13 kA/m and 30 minutes, DM100-DM3 nB nanoscale Biomagnetics,) were evaluated by flow cytometry. Interestingly, the cell death pathway was different depending on the MNP location. Necrosis was observed 24 h after magnetic hyperthermia application in the model where the nanoparticles are located just inside the cells. In contrast, apoptosis was detected in the model where the particles are inside and outside the cells. In addition, in the second model, our results evidenced an enhancement of the nanoparticles uptake after the exposure to the alternating magnetic field. This observation could justify the repetition of the treatment to obtain a better antitumor effect. Our results demonstrate the potential efficacy of magnetic hyperthermia in the treatment of malignant tumours. In addition, the use of 3D cell culture models presents several advantages, allowing to obtention information in a more realistic way than monolayer cell cultures and reducing the number of animals required for preclinical tests.

Trabajo presentado al III Congreso de Materiales para Retos de la Sociedad, celebrado en Jaca (Huesca) del 5 al 7 de febrero de 2018.

Trabajo presentado al Workshop en NanoOncología, celebrado en la Universidad de Zaragoza el 16 de enero de 2018.

Resumen del trabajo presentado a la I Jornada de jóvenes investigadores de cerámica y vidrio en el ICMA, celebrada el 20 de marzo de 2018 en la Universidad de Zaragoza., Los materiales cerámicos con aplicaciones médicas o biocerámicas tienen un amplio abanico de aplicaciones, entre todas ellas destaca el uso de nanopartículas (NPs) de óxidos de hierro para el tratamiento del cáncer. Estas NPs poseen propiedades magnéticas que permiten su uso en el tratamiento antitumoral mediante el uso de hipertermia magnética. Esta aproximación consiste en la utilización de campos magnéticos alternos para producir calor utilizando las nanopartículas magnéticas (MNPs) como agentes de calentamiento. El aumento de temperatura en la zona tumoral conlleva cambios en la estructura del tejido, mejorando la accesibilidad de otros agentes quimioterápicos, así como la muerte celular por el efecto térmico local. En los ensayos realizados previamente en nuestro grupo de investigación se han utilizado MNPs preparadas mediante descomposición térmica, con un núcleo de óxido de hierro, recubierto de PMAO (poli (anhídrido maleico-alt-1-octadeceno)) y marcadas con TAMRA (tetrametilrodamina). Para mejorar la internalización de las nanopartículas dentro de las células, éstas son funcionalizadas con glucosa antes de empezar los estudios in vitro. Ensayos in vitro mediante el uso de modelos de gel 3D de colágeno y la línea celular de macrófagos (ATCC® TIB-71™), han demostrado una buena internalización de las nanoparticulas así como un prometedor efecto antitumoral. Para optimizar el tratamiento por hipertermia se va a estudiar una serie de MNPs con distinto tamaño de núcleo. Se evaluará su valor del SAR (Specific Absorption Rate), un parámetro clave para valorar su capacidad para el tratamiento mediante hipertermia magnética. Posteriormente, se realizarán estudios en células tumorales, evaluando la internalización y el efecto del tratamiento por hipertermia en geles 3D de células de cáncer colorectal humano (ATCC® CCL-247™), mediante el uso de técnicas de biología molecular y bioquímica (técnicas de medida de viabilidad celular, citometría de flujo, microscopía confocal y análisis de proteínas). Con todo ello se confirmará la reproducibilidad de los resultados anteriormente obtenidos en macrófagos, permitiendo además discernir que características de las MNPs producen mejores resultados. Por último, aquellas nanoparticulas que hayan mostrado mejores resultados pasarán a los ensayos in vivo, en los cuales se inocularán células tumorales, para después proceder al tratamiento con MNPs e hipertermia magnética., L.B agradece financiación de la beca Santander-Universidad de Zaragoza. L.G agradece al Ministerio de Economía y Competitividad el contrato Ramón y Cajal (RYC-2014-15512).

Trabajo presentado a la I Jornada de jóvenes investigadores de cerámica y vidrio en el ICMA, celebrada el 20 de marzo de 2018 en la Universidad de Zaragoza.

Resumen del trabajo presentado a la 12th International Conference on the Scientific and Clinical Applications of Magnetic Carriers, celebrada en Copenhague (Dinamarca) del 22 al 26 de mayo de 2018., [Introduction]: Magnetic Hyperthermia (MH) has been proved as an efficient treatment against tumours but still presents some drawbacks such as the limitations of the alternating magnetic field (AMF) generation sources or the need to achieve high concentrations of Magnetic Nanoparticles (MNPs) in the target tissue. Here, we present two different approaches of in vivo MH treatment with the same tumour model but different heating generation materials, combining one of them with a chemotherapeutic agent release such as doxorubicin. The results show, in both of them, an inhibition in the tumour growth. [Materials and Methods]: The animal model used consisted of an allograft and heterotopic tumour model, where EL4 murine lymphoma cell line from the strain C57BL/6N was injected subcutaneously in the right flank of immunocompetent C57BL/6 mice. One week after the cell injection the MNPs were administered intratumorally and the same day and two consecutive days MH was performed. Each exposure time was 30 min at 105 kHz and 18 kA·m-1. The first material that was used consisted of iron oxide nanoparticles (IONP) of 12nm synthetized by thermal decomposition, coated with an amphiphilic polymer (PMAO) and functionalize with Glucose to provide stability in biological fluids and to promote cell internalization. The second type of material consisted of a silica mesoporous matrix that contains IONP and doxorubicin. The silica particles were coated with a thermosensitive polymer with a Lower Critical Solution Temperature (LCST) of 42ºC2. Tumour dimensions (length, with and height) and mice weight were daily measured with a Vernier. After the last AMF exposure mice were maintained 3 days or until tumours started to ulcerate. MH treatments were made in group of 5 mice each, and all the needed control groups to validate the treatment were performed. [Results]: In both cases, 48h after the last magnetic hyperthermia treatment an inhibitory effect in the tumour growth was observed. Confocal microscopy images of the tumour revealed a good penetration of the material in both approaches. In the case of silica mesoporous material, it is noteworthy that none of the two control, where the effect of the free doxorubicin and the effect of the heat released by the MNPs due to the AMF exposition were analysed as single treatments, showed antitumor effect. However, a synergistic effect of the combination of both treatments was achieved.

Trabajo presentado a la Annual Action Conference and MC meeting (RADIOMAG), celebrada en Bilbao (España) del 27 al 28 de abril de 2017.

Póster presntado a la 5th International Conference on Multifunctional, Hybrid and Nanomaterials, celebrada en Lisboa (Portugal) del 6 al 10 de marzo de 2017.

Hoy en día se habla de la importancia e influencia de la literatura en el desarrollo académico, cultural y personal de los ciudadanos. Así la formación de lectores literarios competentes es una de las prioridades de la educación. En esta línea, este trabajo de fin de grado se centra en la necesidad de profundizar en la forma de aproximar el discurso literario a las aulas de educación infantil, en la creencia de que el modo en que el mediador incorpora la lectura en el aula resulta fundamental en la formación del lector. Con este fin, se propone el estudio del enfoque Dime de A. Chambers, a través de lo que él denomina conversaciones literarias. El análisis de las respuestas lectoras de un grupo de Primero de Educación Infantil, partiendo del álbum como propuesta, confirma la adecuación del citado enfoque a la formación de lectores literarios desde los primeros años.

Resumen del trabajo presentado al XIV Congreso de la Sociedad Española de Biología Molecular celebrado en Malaga del 12 al 15 de diciembre de 2011 y posteriormente al INTERMAG 2012 celebrado en Vancouver (Canadá) del 7 al 11 de mayo., This contribution is focused on the increasing interest for the application of magnetic nanoparticles in life sciences. There exist a broad scientific scenario concerning how new inorganic functional nanostructured materials interact with elemental biological units, in particulars with cells. In the field of medicine new therapy and diagnosis agents are being investigated with the goals of a selective drug delivery and an early diagnosis. Although incipient and promising applications in the clinic are encouraging the new field of nanobiomedicine, this discipline still is in the infancy but the scientific interest is rapidly increasing and nowadays it is considered as the main candidate for effective treatment of cancer and neurodegenerative diseases. [1,2] Other emerging activity in this field concerns with the application of nanoparticles in plants [3,4]. One can image the great impact in agriculture, ecology and health food. There exists an overwhelming lack of information concerning the interaction of functional nanoparticles with plants, which is a topic of major interest in environment, phytosanitary and food industries. In this contribution a broad overview on the interdisciplinary investigations carried out in collaboration with several research groups is given. The reported results deal with iron nanoparticles encapsulated in a carbon matrix (Fe@C). The magnetic behavior and the biocompatibility of these nanostructures provide an excellent frame of work in order to visualize, drive, heat, change the magnetic neighborhood of cells, tissues and organs. The magnetic behavior of the nanoparticles has a strong influence on the local internal fields of the tissues modifying the protons resonance of H atom in water; this is relevant for enhancing the MRI contrast. Location of magnetic nanoparticles can be favored by steady magnetic fields and local heating (magnetic hyperthermia) by application of radiofrequency electromagnetic fields. Cell separation or labeling is other important scenario for application of nanoparticles in cell biology. Internalization of magnetic nanoparticles favors gene therapy by application of magnetic fields. The scope of this talk is to establish a parallelism between application of nanotechnology in animal and plant kingdoms, a clear example is the selective drug or chemical release with clear application in therapy or in agriculture.