Cellular studies of magnetic nanoparticles for biomedical applications

Tesis doctoral inédita leída en la Universidad Autónoma de Madrid, Facultad de Ciencias, Departamento de Biología. Fecha de lectura: 11-12-2015Proyectos de Investigación que han subvencionado esta tesis: CM S0505/MAT-000194 (Nanoestructuras magnéticas: fabricación, propiedades y aplicaciones biomédicas y tecnológicas) CM S2009/MAT-1726 (Fundamentos y aplicaciones de moléculas, nanopartículas y nanoestructuras magnéticas: de la espintrónica a la biomedicina) MICINN CTQ2010-20870-C03-03 (Nuevas estrategias para terapia fotodinámica dirigida. 2. Biología) Unión Europea, VII Programa Marco contrato nº 262943 (Multifunctional Nanotechnology for selective detection and treatment of cancer) MINECO CTQ2013-48767-C3-3-R (Desarrollo de dispositivos nanoestructurados como plataformas multimodales fototerapéuticas. 2. Biología) CM S2013/MIT-2850 (Nuevas fronteras del nanomagnetismo fundamental y aplicado

Multifunctional albumin-stabilized gold nanoclusters for the reduction of cancer stem cells

Controlled delivery of multiple chemotherapeutics can improve the effectiveness of treatments and reduce side effects and relapses. Here in, we used albumin-stabilized gold nanoclusters modified with doxorubicin and SN38 (AuNCs-DS) as combined therapy for cancer. The chemotherapeutics are conjugated to the nanostructures using linkers that release them when exposed to different internal stimuli (Glutathione and pH). This system has shown potent antitumor activity against breast and pancreatic cancer cells. Our studies indicate that the antineoplastic activity observed may be related to the reinforced DNA damage generated by the combination of the drugs. Moreover, this system presented antineoplastic activity against mammospheres, a culturing model for cancer stem cells, leading to an efficient reduction of the number of oncospheres and their size. In summary, the nanostructures reported here are promising carriers for combination therapy against cancer and particularly to cancer stem cells.This research was funded by the Spanish Ministry of Economy and Competitiveness (CTQ2016-78454-C2-2-R, SAF2014-56763-R, and SAF2017-87305-R), Comunidad de Madrid (S2013/MIT-2850), Asociación Española Contra el Cáncer, and IMDEA Nanociencia IMDEA Nanociencia acknowledges support from the ‘Severo Ochoa’ Programme for Centres of Excellence in R&D (MINECO, Grant SEV-2016-0686

Optical control of spatially localized red blood cell activity by holographic tweezing

Red blood cells possess unique biomechanical ability to squeeze through capillaries smaller than their size to enable gas and ion exchange. A key signature of their active biomechanics is the out-of-equilibrium fluctuation of the plasma membrane, also known as flickering motion. This active flickering is driven by motor proteins that connect the forces between the spectrin skeleton and the lipid bilayer. However, studying flickering motions in living red blood cells is challenging without altering their physical properties. Here, we implemented a holographic optical tweezer that sculpted a laser beam to create a force field distributed directly along the membrane equatorial contour. We show heterogeneous membrane flickering activity driven by membrane kickers in free-standing cells. Then we inhibited the active kickers by optical forces under minimal invasion, thus benchmarking the active motion against thermal fluctuations. Our work paves the way for optical control of biophysical forces, providing touchless strategies for mechanotransduction in living cells.Comment: 6 figure

Iron oxide nanosized clusters embedded in porous nanorods: A new colloidal design to enhance capabilities of MRI contrast agents

“This document is the Accepted Manuscript version of a Published Work that appeared in final form in ACS Nano, © 2010 American Chemical Society after peer review and technical editing by the publisher. To acces final work see “Iron oxide nanosized clusters embedded in porous nanorods: A new colloidal design to enhance capabilities of MRI contrast agents”, ACS Nano 4.4 (2010): 2095-2103, https://doi.org/10.1021/nn9013388"Development of nanosized materials to enhance the image contrast between the normal and diseased tissue and/or to indicate the status of organ functions or blood flow is essential in nuclear magnetic resonance imaging (MRI). Here we describe a contrast agent based on a new iron oxide design (superparamagnetic iron oxide clusters embedded in antiferromagnetic iron oxide porous nanorods). We show as a proof-of-concept that aqueous colloidal suspensions containing these particles show enhanced-proton relaxivities (i.e., enhanced MRI contrast capabilities). A remarkable feature of this new design is that large scale production is possible since aqueous-based routes are used, and porosity and iron oxide superparamagnetic clusters are directly developed from a single phase. We have also proved with the help of a simple model that the physical basis behind the increase in relaxivities lies on both the increase of dipolar field (interactions within iron oxide clusters) and the decrease of proton-cluster distance (porosity favors the close contact between protons and clusters). Finally, a list of possible steps to follow to enhance capabilities of this contrast agent is also included (partial coating with noble metals to add extra sensing capacity and chemical functionality, to increase the amount of doping while simultaneously carrying out cytotoxicity studies, or to find conditions to further decrease the size of the nanorods and to enhance their stability)We acknowledge financial support from the Spanish Ministerio de Ciencia e innovación trough MAT2008-03224/NAN and from the Comunidad Autónoma de Madrid under Project S-0505/MAT/019

Tailored functionalized magnetic nanoparticles to target breast cancer cells including cancer stem-like cells

Nanotechnology-based approaches hold substantial potential to avoid chemoresistance and minimize side effects. In this work, we have used biocompatible iron oxide magnetic nanoparticles (MNPs) called MF66 and functionalized with the antineoplastic drug doxorubicin (DOX) against MDA-MB-231 cells. Electrostatically functionalized MNPs showed effective uptake and DOX linked to MNPs was more efficiently retained inside the cells than free DOX, leading to cell inactivation by mitotic catastrophe, senescence and apoptosis. Both effects, uptake and cytotoxicity, were demonstrated by different assays and videomicroscopy techniques. Likewise, covalently functionalized MNPs using three different linkers—disulfide (DOX-S-S-Pyr, called MF66-S-S-DOX), imine (DOX-I-Mal, called MF66-I-DOX) or both (DOX-I-S-S-Pyr, called MF66-S-S-I-DOX)—were also analysed. The highest cell death was detected using a linker sensitive to both pH and reducing environment (DOX-I-S-S-Pyr). The greatest success of this study was to detect also their activity against breast cancer stem-like cells (CSC) from MDA-MB-231 and primary breast cancer cells derived from a patient with a similar genetic profile (triple-negative breast cancer). In summary, these nanoformulations are promising tools as therapeutic agent vehicles, due to their ability to produce efficient internalization, drug delivery, and cancer cell inactivation, even in cancer stem-like cells (CSCs) from patientsThis research was funded by the European Seventh Framework Program (grant agreement number 262943); the European Union’s Horizon 2020 research and innovation programme (grant agreement number 685795); Ministerio de Economía y Competitividad, Spain (grants CTQ2016-78454-C2-2-R, BIO2016-77367-C2-1-R and SAF2017-87305-R); Basque Government Elkartek KK- 2017/00008; Comunidad de Madrid (IND2017/IND-7809; S2017/BMD-3867 RENIM-CM and S2018/NMT-4321 NANOMAGCOST-CM); NIHR Manchester Biomedical Research Centre (IS-BRC-1215-20007) and Breast Cancer Now (MAN-Q2); co-financed by European Structural Cancers 2020, 12, 1397 17 of 19 and Investment Fund, Asociación Española Contra el Cáncer (Singulares 2014) and IMDEA Nanociencia. CIC biomaGUNE acknowledges Maria de Maeztu Units of Excellence Program from the Spanish State Research Agency (Grant MDM-2017-0720). IMDEA Nanociencia acknowledges support from the ‘Severo Ochoa’ Programme for Centres of Excellence in R&D (MINECO, Grant SEV-2016-0686

Characterization of interaction of magnetic nanoparticles with breast cancer cells

Background: Different superparamagnetic iron oxide nanoparticles have been tested for their potential use in cancer treatment, as they enter into cells with high effectiveness, do not induce cytotoxicity, and are retained for relatively long periods of time inside the cells. We have analyzed the interaction, internalization and biocompatibility of dimercaptosuccinic acid-coated superparamagnetic iron oxide nanoparticles with an average diameter of 15 nm and negative surface charge in MCF-7 breast cancer cells. Results: Cells were incubated with dimercaptosuccinic acid-coated superparamagnetic iron oxide nanoparticles for different time intervals, ranging from 0.5 to 72 h. These nanoparticles showed efficient internalization and relatively slow clearance. Time-dependent uptake studies demonstrated the maximum accumulation of dimercaptosuccinic acid-coated superparamagnetic iron oxide nanoparticles after 24 h of incubation, and afterwards they were slowly removed from cells. Superparamagnetic iron oxide nanoparticles were internalized by energy dependent endocytosis and localized in endosomes. Transmission electron microscopy studies showed macropinocytosis uptake and clathrin-mediated internalization depending on the nanoparticles aggregate size. MCF-7 cells accumulated these nanoparticles without any significant effect on cell morphology, cytoskeleton organization, cell cycle distribution, reactive oxygen species generation and cell viability, showing a similar behavior to untreated control cells. Conclusions: All these findings indicate that dimercaptosuccinic acid-coated superparamagnetic iron oxide nanoparticles have excellent properties in terms of efficiency and biocompatibility for application to target breast cancer cellsThe research leading to these results have received partial funding from the European Seventh Framework Programme (FP7/2007-2013) under the project MULTIFUN grant agreement no. 262943, and the project Nanofrontmag-CM (S2013/MIT-2850) from the Comunidad de Madrid. Additional grants were obtained from BFU 2011–29038 and CTQ2013-48767-C3-3-R from the Ministerio de Economia y Competitividad and S2009/Mat 1507 from the Comunidad de Madrid (to JLC), from EU FP7 project NAMDIATREAM (ref 246479) and from “la Caixa” / CNB International PhD Programme Fellowship

Efficient treatment of breast cancer xenografts with multifunctionalized iron oxide nanoparticles combining magnetic hyperthermia and anti-cancer drug delivery

Introduction: Tumor cells can effectively be killed by heat, e.g. by using magnetic hyperthermia. The main challenge in the field, however, is the generation of therapeutic temperatures selectively in the whole tumor region. We aimed to improve magnetic hyperthermia of breast cancer by using innovative nanoparticles which display a high heating potential and are functionalized with a cell internalization and a chemotherapeutic agent to increase cell death. Methods: The superparamagnetic iron oxide nanoparticles (MF66) were electrostatically functionalized with either Nucant multivalent pseudopeptide (N6L; MF66-N6L), doxorubicin (DOX; MF66-DOX) or both (MF66-N6LDOX). Their cytotoxic potential was assessed in a breast adenocarcinoma cell line MDA-MB-231. Therapeutic efficacy was analyzed on subcutaneous MDA-MB-231 tumor bearing female athymic nude mice. Results: All nanoparticle variants showed an excellent heating potential around 500 W/g Fe in the alternating magnetic field (AMF, conditions: H = 15.4 kA/m, f = 435 kHz). We could show a gradual inter- and intracellular release of the ligands, and nanoparticle uptake in cells was increased by the N6L functionalization. MF66-DOX and MF66-N6LDOX in combination with hyperthermia were more cytotoxic to breast cancer cells than the respective free ligands. We observed a substantial tumor growth inhibition (to 40% of the initial tumor volume, complete tumor regression in many cases) after intratumoral injection of the nanoparticles in vivo. The proliferative activity of the remaining tumor tissue was distinctly reduced. Conclusion: The therapeutic effects of breast cancer magnetic hyperthermia could be strongly enhanced by the combination of MF66 functionalized with N6L and DOX and magnetic hyperthermia. Our approach combines two ways of tumor cell killing (magnetic hyperthermia and chemotherapy) and represents a straightforward strategy for translation into the clinical practice when injecting nanoparticles intratumorallyThe described work was carried out within the project, Multifunctional Nanoparticles for the Selective Detection and Treatment of Cancer (Multifun), which is funded by the European Seventh Framework Program (FP7/2007-2013) under grant agreement number 262943. We thank Dr Vijay Patel and Liquids Research Ltd (Mentec, Deiniol Road, Bangor, Gwynedd, North Wales, UK,) for the supply of MF66 MNP. We gratefully acknowledge Julia Göring and Susann Burgold for technical assistance in carrying out in vivo experiments and Yvonne Ozegowski for animal handling. We thank Francisco J. Teran (Unidad Asociada de Nanobiotecnología CNB-CSIC & IMDEA Nanociencia, Madrid) for helpful discussions. AS and ALC acknowledge financial support from Ministerio de Economia y Competitividad (grants: SAF-15440 and BIO2012-34835) and IMDEA Nanociencia. This work was partially founded by the Comunidad de Madrid NANOFRONTMAG-CM project (S2013/MIT-2850) (IMDEA-Nanociencia)

Bienestar Adolescente: ¿Cuál es su relación con Habilidades Emocionales y Cognitivas?

Objetivo: Analizar la relación entre variables emocionales (IE-autopercibida y diferenciación emocional) y habilidades cognitivas (vocabulario y memoria de trabajo) con el bienestar-psicológico adolescente. Método: a una muestra de 221 adolescentes escolarizados se le administraron las escalas TMMS-21 y BIEPS-J, la prueba de Ordenamiento Dígito-Letra de la BIMET, un cuestionario de Vocabulario Emocional y la prueba BAIRES-A. Se realizaron correlaciones y se puso a prueba un modelo por medio de ecuaciones estructurales. Resultados: Muestran que la IE-autopercibida impacta de manera positiva sobre el bienestar y que este último se relaciona de forma positiva con los factores claridad y recuperación. La memoria de trabajo mostró tener un efecto indirecto, a través de la IE-autopercibida, sobre el bienestar-psicológico y una relación positiva con el factor recuperación. La diferenciación emocional, evaluada por medio del vocabulario emocional, y el vocabulario general no mostraron tener relación más que entre ambos constructos. Conclusión: Se destaca la importancia del efecto de la memoria de trabajo sobre la IE-autopercibida y la importancia de ésta última para el bienestar psicológico.Objetive: Analyze the relationship between emotional variables (Self-perceived Emotional Intelligence -self-perceived EI- and emotional differentiation), cognitive skills (vocabulary and working memory) and adolescent psychological well-being. Method: A sample of 221 adolescents were assessed with the scales TMMS-21 and BIEPS-J, the BIMET’s Digit-Letter Ordering subtest, an Emotional Vocabulary questionnaire and the BAIRES-A test. We carried out correlation analyses and we tested a model through structural equations. Results: showed that self-perceived EI has a positive impact on well-being and the latter, a positive relationship with clarity and retrieval of emotions. Working memory showed an indirect effect, through self-perceived EI, on psychological well-being and a positive relationship with the retrieval factor. Emotional differentiation, assessed by means of emotional vocabulary, and general vocabulary were only associated with each other. Conclusion: We emphasized the importance of the effect of working memory on self-perceived EI and the latter on psychological well-being.Fil: Calero, Alejandra Daniela. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina. Universidad de Buenos Aires. Facultad de Psicología. Instituto de Investigaciones; ArgentinaFil: Barreyro, Juan Pablo. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina. Universidad de Buenos Aires. Facultad de Psicología. Instituto de Investigaciones; ArgentinaFil: Formoso, Jesica. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Saavedra 15. Centro Interdisciplinario de Investigaciones en Psicología Matemática y Experimental Dr. Horacio J. A. Rimoldi; ArgentinaFil: Burin, Debora Ines. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Saavedra 15. Centro Interdisciplinario de Investigaciones en Psicología Matemática y Experimental Dr. Horacio J. A. Rimoldi; ArgentinaFil: Puebla, Macarena. No especifíca;Fil: Murphy, Micaela. Universidad de Buenos Aires. Facultad de Psicología. Instituto de Investigaciones; ArgentinaFil: Retta, Agustina. Universidad Austral. Facultad de Ciencias Biomédicas; ArgentinaFil: Romano, Sol. Universidad Austral. Facultad de Ciencias Biomédicas; Argentin

Bienestar Adolescente: ¿Cual es su relacion con Habilidades Emocionales y Cognitivas?

Objetivo: Analizar la relación entre variables emocionales (IE-autopercibida y diferenciación emocional) y habilidades cognitivas (vocabulario y memoria de trabajo) con el bienestar-psicológico adolescente. Método: a una muestra de 221 adolescentes escolarizados se le administraron las escalas TMMS-21 y BIEPS-J, la prueba de Ordenamiento Dígito-Letra de la BIMET, un cuestionario de Vocabulario Emocional y la prueba BAIRES-A. Se realizaron correlaciones y se puso a prueba un modelo por medio de ecuaciones estructurales. Resultados: Muestran que la IE-autopercibida impacta de manera positiva sobre el bienestar y que este último se relaciona de forma positiva con los factores claridad y recuperación. La memoria de trabajo mostró tener un efecto indirecto, a través de la IE-autopercibida, sobre el bienestar-psicológico y una relación positiva con el factor recuperación. La diferenciación emocional, evaluada por medio del vocabulario emocional, y el vocabulario general no mostraron tener relación más que entre ambos constructos. Conclusión: Se destaca la importancia del efecto de la memoria de trabajo sobre la IE-autopercibida y la importancia de ésta última para el bienestar psicológico.Objetive: Analyze the relationship between emotional variables (Self-perceived Emotional Intelligence -self-perceived EI- and emotional differentiation), cognitive skills (vocabulary and working memory) and adolescent psychological well-being. Method: A sample of 221 adolescents were assessed with the scales TMMS-21 and BIEPS-J, the BIMET’s Digit-Letter Ordering subtest, an Emotional Vocabulary questionnaire and the BAIRES-A test. We carried out correlation analyses and we tested a model through structural equations. Results: showed that self-perceived EI has a positive impact on well-being and the latter, a positive relationship with clarity and retrieval of emotions. Working memory showed an indirect effect, through self-perceived EI, on psychological well-being and a positive relationship with the retrieval factor. Emotional differentiation, assessed by means of emotional vocabulary, and general vocabulary were only associated with each other. Conclusion: We emphasized the importance of the effect of working memory on self-perceived EI and the latter on psychological well-being.Fil: Calero, Alejandra Daniela. Universidad de Buenos Aires. Facultad de Psicología. Instituto de Investigaciones; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; ArgentinaFil: Barreyro, Juan Pablo. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Saavedra 15. Centro Interdisciplinario de Investigaciones en Psicología Matemática y Experimental Dr. Horacio J. A. Rimoldi; ArgentinaFil: Formoso, Jesica. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Saavedra 15. Centro Interdisciplinario de Investigaciones en Psicología Matemática y Experimental Dr. Horacio J. A. Rimoldi; ArgentinaFil: Murphy, Micaela. Universidad Austral. Facultad de Ciencias Biomédicas; ArgentinaFil: Puebla, Valentina Macarena. Universidad Austral. Facultad de Ciencias Biomédicas; ArgentinaFil: Retta, Agustina María. Universidad Austral. Facultad de Ciencias Biomédicas; ArgentinaFil: Romano Sánchez, Sol Micaela. Universidad Austral. Facultad de Ciencias Biomédicas; ArgentinaFil: Burin, Debora Ines. Universidad de Buenos Aires. Facultad de Psicología. Instituto de Investigaciones; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentin

Prepandemic viral community-acquired pneumonia: Diagnostic sensitivity and specificity of nasopharyngeal swabs and performance of clinical severity scores

© 2022 The Authors. Journal of Medical Virology published by Wiley Periodicals LLC. This is an open access article under the terms of the Creative Commons Attribution-NonCommercial-NoDerivs License, which permits use and distribution in any medium, provided the original work is properly cited, the use is non-commercial and no modifications or adaptations are made.The objectives of this work were to assess the diagnostic sensitivity and specificity of nasopharyngeal (NP) swabs for viral community-acquired pneumonia (CAP) and the performance of pneumonia severity index (PSI) and CURB-65 severity scores in the viral CAP in adults. A prospective observational cohort study of consecutive 341 hospitalized adults with CAP was performed between January 2018 and March 2020. Demographics, comorbidities, symptoms/signs, analytical data, severity scores, antimicrobials, and outcomes were recorded. Blood, NP swabs, sputum, and urine samples were collected at admission and assayed by multiplex real time-PCR, bacterial cultures, and Streptococcus pneumoniae and Legionella pneumophila antigens detection, to determine the etiologies and quantify the viral load. The etiology was identified in 174 (51.0%) patients, and in 85 (24.9%) it was viral, the most frequent rhinovirus and influenza virus. The sensitivity of viral detection in sputum (50.7%) was higher than in NP swabs (20.9%). Compared with sputum, the positive predictive value and specificity of NP swabs for viral diagnosis were 95.8% and 96.9%, respectively. Performance of PSI and CURB-65 scores in all CAP with etiologic diagnosis were as expected, with mortality associated with higher values, but they were not associated with mortality in patients with viral pneumonia. NP swabs have lower sensitivity but high specificity for the diagnosis of viral CAP in adults compared with sputum, reinforcing the use NP swabs for the diagnostic etiology work-up. The PSI and CURB-65 scores did not predict mortality in the viral CAP, suggesting that they need to be updated scores based on the identification of the etiological agent.This work was supported by National Plan R + D + I 2013–2016 and Instituto de Salud Carlos III, Subdirección General de Redes y Centros de Investigación Cooperativa, Ministry of Economy, Industry, and Competitiveness, Spanish Network for Research in Infectious Diseases [REIPI RD16/0016/0009]; cofinanced by European Development Regional Fund “A way to achieve Europe”, Operative program Intelligent Growth 2014–2020; and supported by the grant PI17/01055 from the Instituto de Salud Carlos III. MAG, RAM and JSC [grant number CB21/13/00006] also received support from the CIBER de Enfermedades Infecciosas (CIBERINFEC), Instituto de Salud Carlos III, Ministerio de Ciencia e Innovación, cofinanced by the European Development Regional Fund. J.S.C. is a researcher belonging to the program “Nicolás Monardes” (C-0059-2018), Servicio Andaluz de Salud, Junta de Andalucía, Spain.Peer reviewe