Effects of 3D nanocomposite bioceramic scaffolds on the immune response

The interaction of new nanocomposite mesoporous glass/hydroxyapatite (MGHA) scaffolds with immune cells involved in both innate and acquired immunity has been studied in vitro as an essential aspect of their biocompatibility assessment. Since the immune response can be affected by the degradation products of bioresorbable scaffolds and scaffold surface changes, both processes have been evaluated. No alterations in proliferation and viability of RAW-264.7 macrophage-like cells were detected after culture on MGHA scaffolds which did not induce cell apoptosis. However, a slight cell size decrease and an intracellular calcium content increase were observed after contact of this cell line with MGHA scaffolds or their extracts. Although no changes in the percentages of RAW cells with low and high contents of reactive oxygen species (ROS) are observed by the treatment with 7 day extracts, this study has revealed modifications of these percentages after direct contact with scaffolds and by the treatment with 24 h extracts, related to the high reactivity/bioactivity of this MGHA nanocomposite at initial times. Furthermore, when normal fresh murine spleen cells were used as an experimental model closer to physiological conditions, no significant alterations in the activation of different immune cell subpopulations were detected in the presence of 24 h MGHA extract. MGHA scaffolds did not affect either the spontaneous apoptosis or intracellular cytokine expression (IL-2, IL-10, IFN-gamma, and TNF-alpha.) after 24 h treatment. The results obtained in the present study with murine immune cell subpopulations (macrophages, lymphocytes B, lymphocytes T and natural killer cells) support the biocompatibility of the MGHA material and suggest an adequate host tissue response to their scaffolds upon their implantation

Metabolomic response of osteosarcoma cells to nanographene oxide-mediated hyperthermia.

Nanographene oxide (nGO)-mediated hyperthermia has been increasingly investigated as a localized, minimally invasive anticancer therapeutic approach. Near InfraRed (NIR) light irradiation for inducing hyperthermia is particularly attractive, because biological systems mostly lack chromophores that absorb in this spectral window, facilitating the selective heating and destruction of cells which have internalized the NIR absorbing- nanomaterials. However, little is known about biological effects accompanying nGO-mediated hyperthermia at cellular and molecular levels. In this work, well-characterized pegylated nGO sheets with a hydrodynamic size of 300 nm were incubated with human Saos-2 osteosarcoma cells for 24 h and their internalization verified by flow cytometry and confocal microscopy. No effect on cell viability was observed after nGO uptake by Saos-2 cells. However, a proliferation delay was observed due to the presence of nGO sheets in the cytoplasm. 1H NMR metabolomics was employed to screen for changes in the metabolic profile of cells, as this could help to improve understanding of cellular responses to nanomaterials and provide new endpoint markers of effect. Cells inter- nalizing nGO sheets showed noticeable changes in several metabolites compared to control cells, including decreased levels of several amino acids, taurine and creatine and increased levels of phosphocholine and ur- idine/adenosine nucleotides. After NIR irradiation, cells showed decreases in glutamate and uridine nucleotides, together with increases in glycerophosphocholine and adenosine monophosphate. Overall, this study has shown that the cellular metabolome sensitively responded to nGO exposure and nGO-mediated hyperthermia and that NMR metabolomics is a powerful tool to investigate treatment responses

Mesoporous Silica Nanoparticles Decorated with Carbosilane Dendrons as New Non-viral Oligonucleotide Delivery Carriers

A novel nanosystem based on mesoporous silica nanoparticles covered with carbosilane dendrons grafted on the external surface of the nanoparticles is reported. This system is able to transport single-stranded oligonucleotide into cells, avoiding an electrostatic repulsion between the cell membrane and the negatively charged nucleic acids thanks to the cationic charge provided by the dendron coating under physiological conditions. Moreover, the presence of the highly ordered pore network inside the silica matrix would make possible to allocate other therapeutic agents within the mesopores with the aim of achieving a double delivery. First, carbosilane dendrons of second and third generation possessing ammonium or tertiary amine groups as peripheral functional groups were prepared. Hence, different strategies were tested in order to obtain their suitable grafting on the outer surface of the nanoparticles. As nucleic acid model, a single-stranded DNA oligonucleotide tagged with a fluorescent Cy3 moiety was used to evaluate the DNA adsorption capacity. The hybrid material functionalised with the third generation of a neutral dendron showed excellent DNA binding properties. Finally, the cytotoxicity as well as the capability to deliver DNA into cells, was tested in vitro by using a human osteoblast-like cell line, achieving good levels of internalisation of the vector DNA/carbosilane dendron-functionalised material without affecting the cellular viability

Multifunctional pH sensitive 3D scaffolds for treatment and prevention of bone infection

Multifunctional-therapeutic three-dimensional (3D) scaffolds have been prepared. These biomaterials are able to destroy the S. aureus bacterial biofilm and to allow bone regeneration at the same time. The present study is focused on the design of pH sensitive 3D hierarchical meso-macroporous 3D scaffolds based on MGHA nanocomposite formed by a mesostructured glassy network with embedded hydroxyapatite nanoparticles, whose mesopores have been loaded with levofloxacin (Levo) as antibacterial agent. These 3D platforms exhibit controlled and pH-dependent Levo release, sustained over time at physiological pH (7.4) and notably increased at infection pH (6.7 and 5.5), which is due to the different interaction rate between diverse Levo species and the silica matrix. These 3D systems are able to inhibit the S. aureus growth and to destroy the bacterial biofilm without cytotoxic effects on human osteoblasts and allowing an adequate colonization and differentiation of preosteoblastic cells on their surface. These findings suggest promising applications of these hierarchical MGHA nanocomposite 3D scaffolds for the treatment and prevention of bone infection. Statement of Significance Multifunctional 3D nanocomposite scaffolds with the ability for loading and sustained delivery of an antimicrobial agent, to eliminate and prevent bone infection and at the same time to contribute to bone regeneration process without cytotoxic effects on the surrounding tissue has been proposed. These 3D scaffolds exhibit a sustained levofloxacin delivery at physiological pH (pH 7.4), which increasing notably when pH decreases to characteristic values of bone infection process (pH 6.7 and pH 5.5). In vitro competitive assays between preosteoblastic and bacteria onto the 3D scaffold surface demonstrated an adequate osteoblast colonization in entire scaffold surface together with the ability to eliminate bacteria contamination

Characterization of M1 and M2 polarization phenotypes in peritoneal macrophages after treatment with graphene oxide nanosheets

Macrophages play a key role in nanoparticle removal and are primarily responsible for their uptake and trafficking in vivo. Due to their functional plasticity, macrophages display a spectrum of phenotypes between two extremes indentified as pro-inflammatory M1 and reparative M2 macrophages, characterized by the expression of specific cell surface markers and the secretion of different cytokines. The influence of graphene oxide (GO) nanosheets functionalized with poly(ethylene glycol-amine) and labelled with fluorescein isothiocyanate (FITC-PEG-GO) on polarization of murine peritoneal macrophages towards M1 and M2 phenotypes was evaluated in basal and stimulated conditions by flow cytometry and confocal microscopy through the expression of different cell markers: CD80 and iNOS as M1 markers, and CD206 and CD163 as M2 markers. Although FITC-PEG-GO did not induce M1 or M2 macrophage polarization after 24 and 48 h in basal conditions, this nanomaterial decreased the percentage of M2 reparative macrophages. We have also compared control macrophages with macrophages that have or have not taken up FITC-PEG-GO after treatment with these nanosheets (GO+ and GO− cells, respectively). The CD80 expression diminished in GO+ macrophages after 48 h of GO treatment but the CD206 expression in GO+ population showed higher values than in both GO- population and control macrophages. In the presence of pro-inflammatory stimuli (LPS and IFN-γ), a significant decrease of CD80+ cells was observed after treatment with GO. This nanomaterial also induced significant decreases of CD206+ and CD163+ cells in the presence of reparative stimulus (IL-4). The CD80, iNOS and CD206 expression was lower in both GO− and GO+ cells than in control macrophages. However, higher CD163 expression was obtained in both GO− and GO+ cells in comparison with control macrophages. All these facts suggest that FITC-PEG-GO uptake did not induce the macrophage polarization towards the M1 pro-inflammatory phenotype, promoting the control of the M1/M2 balance with a slight shift towards M2 reparative phenotype involved in tissue repair, ensuring an appropriate immune response to these nanosheets

Autonomous learning of the bioinorganic chemistry and biomaterials laboratory and characterisation techniques used through the use of e-learning tools. Enhancing the understanding of students with disabilities, hearing difficulties or foreigners

Concesión por parte de la Oficina para la Calidad de la prorroga para la entrega de la memoria del proyecto nº 52 hasta el 31 de diciembre de 2022El proyecto tiene como objetivo fundamental la elaboración de material audiovisual para la mejora de las prácticas de la asignatura de Química Bioinorgánica y Biomateriales de cuarto curso del grado en Farmacia para su satisfactorio desarrollo tanto en un escenario totalmente on-line como presencial. Para el buen desarrollo de las prácticas en el laboratorio es necesario que el alumno disponga de una información adecuada que le permita conocer los distintos aspectos del proceso experimental a realizar a través de la elaboración de una página web disponible en el campus virtual. Para ello, la práctica habitual es utilizar un manual de prácticas de laboratorio en el que se aporta información sobre conceptos básicos, material a utilizar, procedimiento experimental, seguridad, etc… de cada práctica. Hay tres aspectos de gran importancia desde el punto de vista formativo: 1) afianzar bien los conceptos básicos en los que se apoya el trabajo experimental posterior, 2) el manejo de los software que permitan el tratamiento de los resultados e interpretación de los datos utilizando herramientas e-learning y 3) facilitar la comprensión de los dos objetivos anteriores a personas con discapacidades auditivas y/o idioma incluyendo subtítulos en español e inglés en los videos elaborados. En dicha elaboración se tendrá muy en cuenta, la inclusión de los alumnos con diversidad presentando dicha información de manera visual, estructurada, secuencial y en pasos ordenados. Estas herramientas de aprendizaje estarán dirigidas inicialmente a los alumnos de cursos superiores del Grado en Farmacia, Química, Ingeniería de Materiales y Máster Universitario en Biomateriales por lo que para comprobar su impacto y eficacia seleccionaemos a estudiantes de los departamentos implicados antes de ponerlos a disposición de los alumnos. Este proyecto se basa en tres Competencias transversales instrumentales del Espacio Europeo de Educación Superior (EEES): Comunicación oral y escrita en la/s lengua/s materna/s, Comunicación en lengua extranjera, y Utilización de las Tecnologías de la información y la Comunicación (TIC) en el ámbito de estudio y contexto profesional. Estas competencias son clave para el desarrollo académico y profesional de los alumnos/as, especialmente para alumnos/as con algún tipo de discapacidad. Creemos que, mediante este proyecto, se realiza un esfuerzo para mejorar la inclusión de estos alumnos/as tanto en un contexto online como presencial en un laboratorio de Química. Por las razones anteriormente expuestas, un grupo de profesores de los Departamentos de Química en Ciencias Farmacéuticas y el de Bioquímica y Biología Molecular se propone elaborar un material didáctico que permita el autoaprendizaje del alumno en la parte experimental de la asignatura optativa de Química Bioinorgánica y Biomateriales del Grado en Farmacia, si bien, este material también será de utilidad para complementar otras asignaturas presentes en los Grados de Biología o de Ingeniería de Materiales e, incluso en el Máster en Biomateriales de la UCM.The main objective of the project is the development of audiovisual material to improve the practices of the Bioinorganic Chemistry and Biomaterials subject in the fourth year of the degree in Pharmacy for its satisfactory development both in a fully online and on-site scenario. For the good development of the laboratory practicals it is necessary that the student has adequate information that allows him/her to know the different aspects of the experimental process to be carried out through the development of a web page available on the virtual campus. To this end, the usual practice is to use a laboratory practice manual which provides information on basic concepts, materials to be used, experimental procedure, safety, etc., for each practice. There are three aspects of great importance from the training point of view: 1) to consolidate the basic concepts on which the subsequent experimental work is based, 2) the use of software that allows the processing of the results and interpretation of the data using e-learning tools and 3) to facilitate the understanding of the two previous objectives for people with hearing and/or language disabilities by including subtitles in Spanish and English in the videos produced. The inclusion of students with diversity will be taken into account in the elaboration of the videos by presenting the information in a visual, structured, sequential and orderly manner. These learning tools will initially be aimed at students in the upper years of the Bachelor's Degree in Pharmacy, Chemistry, Materials Engineering and the Master's Degree in Biomaterials, so in order to check their impact and effectiveness we will select students from the departments involved before making them available to students. This project is based on three transversal instrumental competences of the European Higher Education Area: Oral and written communication in the mother tongue(s), Communication in a foreign language, and use of information and communication technologies (ICT) in the field of study and professional context. These competences are key for the academic and professional development of students, especially for students with disabilities. We believe that, through this project, an effort is being made to improve the inclusion of these students both in an online and face-to-face context in a chemistry laboratory. For the above reasons, a group of lecturers from the Departments of Chemistry in Pharmaceutical Sciences and Biochemistry and Molecular Biology propose to develop a teaching material that allows students to learn by themselves in the experimental part of the optional subject of Bioinorganic Chemistry and Biomaterials of the Degree in Pharmacy, although this material will also be useful to complement other subjects in the Degrees in Biology or Materials Engineering and even in the Master's Degree in Biomaterials of the University of Valencia, where it will also be useful to complement other subjects in the Degrees in Biology or Materials Engineering and even in the Master's Degree in Biomaterials of the Universidad Complutense.Depto. de Química en Ciencias FarmacéuticasFac. de FarmaciaFALSEunpu

In-hospital and 6-month outcomes in patients with COVID-19 supported with extracorporeal membrane oxygenation (EuroECMO-COVID): a multicentre, prospective observational study

Extracorporeal membrane oxygenation (ECMO) has been widely used in patients with COVID-19, but uncertainty remains about the determinants of in-hospital mortality and data on post-discharge outcomes are scarce. The aims of this study were to investigate the variables associated with in-hospital outcomes in patients who received ECMO during the first wave of COVID-19 and to describe the status of patients 6 months after ECMO initiation