XV Jornadas Anuales CIBER-BBN

Abstract

Rosmarinic acid is an attractive candidate for wound healing purposes because of its antioxidant, anti-inflammatory, and photoprotective functions, however, its poor bioavailability hampers its therapeutic outcome. In this context, synthesis of polymer conjugates is an alternative to enlarge its applications. This work describes the synthesis of novel water-soluble chitosan – rosmarinic acid conjugates (CSRA) that have great potential for skin applications. Chitosan was functionalized with different contents of rosmarinic acid as confirmed by ATR-FTIR, 1H-NMR and UV spectroscopies. CSRA conjugates presented three-fold radical scavenger capacity compared to the free phenolic compound. Films were prepared by solvent-casting procedure and the biological activity of the lixiviates was studied in vitro. Results revealed that lixiviates reduced activation of inflamed macrophages, improved antibacterial capacity against E. coli with respect to native chitosan and free rosmarinic acid, and also attenuated UVB-induced cellular damage and reactive oxygen species production in fibroblasts and keratinocytes.

Abstract

My talk will be focused on the immunity against SARS-CoV-2. First, understanding the immune responses to natural infection and then summarizing current knowledge of immunity conferred by vaccines.

Several lessons can be extracted from these data, such as the surprising (or not) strength of hybrid immunity induced by both infection and vaccination, the differences between vaccines, and the need for administration of additional vaccine doses. Finally I’ll will speculate on the future balance between the virus and the immunity at the global level.

Abstract

Adoptive cell therapy is an emerging alternative to regular procedures in cancer treatment that consists of using immune cells as drugs, conferring to the patients, potential long-term protection. Usually, the therapy consists of extracting autologous T cells from patients, followed by a specific selection or genetic modification of these cells to increase their effectivity against cancer, such as the introduction of chimeric antigen receptors (CARs). Then, these specific T cells are largely expanded and finally, reinfused to the patients to act as living drugs. Therefore, these personalized treatments require laborious and expensive laboratory procedures that should be alleviated to enable their broad use in the clinics. With the objective to improve the ex vivo expansion of therapeutic phenotypes of human (CAR) T cells, we used different 3D biomaterials to mimic the extracellular matrix (ECM) of the lymph nodes, consisting of polystyrene scaffolds and hydrogels. Different improvements in T cell proliferation, differentiation, and function have been observed depending on the used matrix. For example, the T cell rich population of peripheral blood mononuclear cells (PBMCs) have shown increases in cell proliferation, whereas the same population modified with CARs was able to increase its cytotoxicity compared to cultures that did not include a 3D scaffold.

Abstract

Cancer is currently one of the deadliest diseases causing millions of deaths every year. According to the world health organisation, cancer is the second leading cause of death globally, accounting for an estimated 9.6 million deaths in 2018. Despite the established standard strategies (surgery, radiotherapy and chemotherapy) have reasonable success for certain cancers, resistant cancer cells, recurrence and metastases remain common.

Photodynamic Therapy (PDT) based treatments are promising to work as a tool against cancer and pre-cancerous diseases. To optimize this treatment, it is required to maximize the Integral of Overlap of the volumetric distributions of the photosensitizer, of the molecular oxygen and the photon distribution of the activating light on the diseased tissue. Then, only diseased cell death through oxidative damage is optimized by means of direct and indirect photodynamic effects. A major common challenge is the difficulty of delivering the activation light, deep into the tissue or to activate deep internal diseased tissue volumes.

As Bioluminescence (BL) based self-illuminated nano-sources offers a very attractive alternative to enable Deep PDT treatments, NaTBiL exploratory project was proposed to demonstrate the potential on innovative therapeutic nano-medicine activated by light to treat deep cancer diseases with higher specificity and effectiveness and, also, to be a first step to set the bases for a subsequent ambitious projects in the frontiers of knowledge.

In the talk, after the motivation of the proposal, works, results and impacts of the project will be presented. Obtained results proved the feasibility to associate the light-emitting protein RLuc8 (which is a mutant from parental renilla luciferase with an improved light output) to quantum dots containing a biocompatible polymer surface decorated with carboxylic groups in order to react with the lysines of the protein forming an amide bond. The amount of linked protein was obtained to produce self-activated nano-sources. It was also proved on in vitro and in vivo experiments. The former, by means of PDT with the verteporfin photosensitizer and in the later, self-illumination was observed upon intravenous injection to healthy mice for at least 24 h post-administration with both NPs.

Results have been presented in several national and international meetings and they have, also, enabled the presentation of several ambitious project proposals (one European Ongoing and, other in evaluation process). Summing up, NaTBiL seed project have broadly reached the established objectives.

Abstract

Three-dimensional (3D) bioprinted culture systems allow to accurately control microenvironment components and analyze their effects at cellular and tissue levels. The main objective of this project was to identify, quantify and localize the effects of physical-chemical communication signals between tumor cells and the surrounding biomaterial stiffness over time, defining how aggressiveness increases in SK-N-BE(2) neuroblastoma (NB) cell line.

GelMA-AlgMA composite was selected to develop neuroblastoma 3D bioprinted tumor models. To study the effect of ECM stiffness in neuroblastoma cluster behavior, GelMA was combined with 0%, 1% and 2% AlgMA. Compressive modulus results showed that stiffness increased with AlgMA concentration. In concordance with previous experiments with pristine GelMA, 1% AlgMA addition presented 50% increase in stiffness. Indeed, 2% AlgMA stiffness was two-fold higher than 1% AlgMA.

SK-N-BE(2) and SH-SY5Y neuroblastoma cell lines were encapsulated in GelMA-AlgMA prepolymers and cultured in 3D bioprinted square scaffolds. Human NB and 1% AlgMA hydrogels presented similar Young´s modulus mean, and orthotopic NB mice tumors were equally similar to 0% and 1% AlgMA hydrogels. Porosity increased over time; cell cluster density decreased over time and with stiffness, and cell cluster occupancy generally increased with time and decreased with stiffness.

Immunohistological analyzes of paraffined sections revealed increased expression of PTBP1 and Ki67 in the stiffest GelMA-AlgMA cultures in SK-N-BE(2). The % of Ki67 positive cells in soft materials was 70%, while 90% of cells were positive in stiff materials. In this line, 2% AlgMA showed increased expression of Bcl2 (50% positive cells) and Bax (20% positive cells) markers in comparison to 0% AlgMA, where there was no expression of Bax apoptotic marker. SH-SY5Y showed a lower % of Ki67 cells than SK-N-BE(2). In contrast to 2D control, both neuroblastoma clusters of SK-N-BE(2) and SH-SY5Y showed upregulated expression of vitronectin

In addition, cell proliferation, mRNA metabolism and antiapoptotic activity advanced over time and with stiffness. This study shows the potential of the 3D in vitro cell model described herein to infer how intercellular space stiffness patterns drive the clinical behavior associated with NB patients.

Abstract

The decoding of human’s brain activity using electroencephalography (EEG) has numerous applications, ranging from the understanding of how our central nervous system works, to the detection of intentions and emotional states. In recent years, deep-learning models gained attention for this complex task due to their excellent performance and ability to extract complex features from raw data. In particular, convolutional neural networks (CNN) showed promising results in brain-computer interfaces (BCI) aimed at improving the autonomy and quality of life of severely disabled people. However, there are still shortcomings that prevented the development of their full potential. In this study, we proposed a novel CNN, called EEG-Inception, that improved the accuracy and calibration time of assistive BCIs. To the best of our knowledge, EEG-Inception is the first model to integrate Inception modules for EEG pattern recognition, which combined efficiently with other structures in a light architecture, improved the performance of our approach. The model was validated in a population of 73 subjects, of which 31 present motor disabilities. Results showed that EEG-Inception outperformed 5 previous approaches, yielding significant improvements of the accuracy with respect to previous methods such as rLDA, xDAWN + Riemannian geometry, CNN-BLSTM, DeepConvNet and EEGNet. Moreover, EEG-Inception required very few calibration trials to achieve state-of-the-art performances taking advantage of a novel training strategy that combines cross-subject transfer learning and fine-tuning to increase the feasibility of this approach for practical use in assistive applications. In summary, this study could be considered a step forward towards efficient processing of EEG signals for BCI applications using deep learning.

Abstract

Achievement of spatiotemporal control of growth factors production remains a main goal in tissue engineering. In the present work, we combined inducible transgene expression and near infrared (NIR)-responsive hydrogels technologies to develop a therapeutic platform for bone regeneration. A heat-activated and dimerizer-dependent transgene expression system was incorporated into mesenchymal stem cells to conditionally control the production of bone morphogenetic protein 2 (BMP-2). Genetically engineered cells were entrapped in hydrogels based on fibrin and plasmonic gold nanoparticles that transduced incident energy of an NIR laser into heat. In the presence of dimerizer, photoinduced mild hyperthermia induced the release of bioactive BMP-2 from NIRresponsive cell constructs. A critical size bone defect, created in calvaria of immunocompetent mice, was filled with NIR-responsive hydrogels entrapping cells that expressed BMP-2 under the control of the heat-activated and dimerizer-dependent gene circuit. In animals that were treated with dimerizer, NIR irradiation of implants induced BMP-2 production in the bone lesion. Induction of NIR-responsive cell constructs conditionally expressing BMP-2 in bone defects resulted in the formation of new mineralized tissue, thus indicating the therapeutic potential of the technological platform.

Abstract

Recent achievements in the field of immunotherapy, such as the development of engineered T cells used in adoptive cell therapy, are introducing more efficient strategies to combat cancer. Nevertheless, T cells are challenging to manufacture, manipulate, and control. For example, there are limitations in producing the large amounts of T cells needed for these therapies in a short period of time and in an economically viable manner. In this study, three-dimensional (3D) poly(ethylene) glycol hydrogels (PEG) covalently combined with low molecular weight heparin were engineered to resemble the lymph nodes, where T cells reproduce. In these hydrogels, PEG provides the needed structural and mechanical properties, whereas heparin is used as an anchor for the cytokine CCL21, which is present in the lymph nodes, and can affect cell migration and proliferation. The 3D structure of the hydrogel in combination with its loading capacity result in an increased primary human CD4+ T cell proliferation compared to the state-of-the-art expansion systems consisting of artificial antigen presenting cells. Thus, we present a new tool for adoptive T cell therapy to help achieving the large numbers of cells required for therapy of selected phenotypes targeted against cancer cells, by mimicking the lymph nodes.

Abstract

Mesoporous carbon nanoparticles (MCNs) hold the potential to revolutionize the field of drug delivery. In this work, we have synthesized a series of MCNs that have been functionalized with a special class of pH-responsive polymer to achieve localized drug delivery. The physicochemical features of these nanocarriers have been extensively evaluated along with the pH- responsiveness, which has been validated in vial, in vitro, and in vivo. Finally, the MCNs have been loaded with a chemotherapeutic, demonstrating promising antitumoral features.

Abstract

Current treatments against HIV-1 infection are based in a combined therapy that can control replication and diseases progression. However, the appearance of resistances mainly in low- and middle-income countries and the fact that HIV-1 can infect a wide range of cells developing reservoirs, bring to light the need of new therapeutic approaches. Here, we present two potential therapeutic strategies using dendritic carbosilane systems. The first one consists in the use of dendrimers as nanocarriers for small RNAs, in particular microRNA, able to target HIV infection. The effective delivery of these miRNAs is a limiting factor in their therapeutic applications and for that they need to be protected and properly delivered. The second approach is based on the use of carbosilane dendrimers as antiviral systems per se, with the aim to obtain a topical vaginal microbicide that allow to reduce the risk of HIV -1 and other STIs. Therefore, studies in these two directions will be presented.

Abstract

Glioblastoma (GB) is the major aggressive primary brain tumour in adults, with poor survival and no cure. The participation of host immune system in cancer surveillance and successful response to therapy is widely recognized. However, there is a clear lack of early, non-invasive imaging biomarkers (IB) to evaluate such efficient immune response, allowing to halt unsuccessful treatment and providing evidence-based therapy-related decisions. Magnetic Resonance (MR)- derived data (MRI, MRSI, i.e. imaging, spectroscopic imaging) can be translated into IB of successful GB therapy through in vivo monitoring. We work towards a breakthrough innovative hashtag of multiparametric MR, combining MRI and MRSI data to achieve a surrogate IB of successful therapeutic outcome. Such approach definitely represents a step beyond in comparison with present follow-up therapy response strategies based in volume changes of the tumour mass. This is being first explored in preclinical subjects, allowing us for longitudinal analyses and molecular validation of MR-related findings. Our benchmarking results indicate that the MR- related findings may be closely associated to local host immune system changes triggered by therapy, provided immune respectful schedules are applied. The large amount of multiparametric data accumulated allow us to benefit from powerful state-of-the-art machine learning approaches. This, together with feature selection and interpretation methods help us in the extraction of MR- based significant IB. At present, there is no software approach allowing both fast, user-friendly post-processing of conventional MRI and MRSI, and encompassing machine learning developments. Accordingly, one of our ongoing goals is the development and implementation of a user-friendly, flexible software package able to combine machine learning analysis and 2D-3D data display. The potential of this intramural collaboration was recognized with an H2020 ATTRACT project for breakthrough ideas in 2019-2020.

Abstract

P. aeruginosa infections, like those caused by many other hospital bacteria, are becoming more difficult to treat because of the increasing development of antibiotic resistance processes and the fewer new antibiotics that are being developed. Currently the gold standard technology for diagnosis of infections is based on sample plate cultures, which have several limitations regarding sensitivity, especially in the case of previous antimicrobial therapy.

In this context, we have developed a highly sensitive, specific and reliable immunochemical assay to detect pyocyanin (PYO), the most important virulence factor of P. aeruginosa. The assay uses a high affinity monoclonal antibody raised against the precursor 1-hydroxyphenazine (1-OHphz) hapten derivatives. PYO is a redox-active phenazine responsible of the characteristic blue-green color of these bacteria that has been demonstrated to induce neutrophil apoptosis, interleukin release and weakness of the immune system. The microplate-based ELISA developed is able to achieve a LoD of 0.07 nM which is much below the concentrations reported to be found in clinical samples (130 M in sputa and 2.8 M in ear secretions.

To our knowledge does not exist in the market any rapid and specific diagnostic kit to detect this biomarker. Thus, the validation of this assay from a clinical perspective would render a faster, reliable diagnostic tool, opening the possibility to apply more appropriate specific treatment, reducing adverse effects and the development of antimicrobial resistance mechanisms. With this aim, the tasks included in this valorization project have included the assessment of PYO as biomarker of P. aeruginosa respiratory infections and the validation of the immunoassay for its detection. Moreover, the market and the need for an in vitro diagnostic (IVD) method have be evaluated, studying the possible commercialization of this assay implemented as a point of care device.

Abstract

Interest in the development of resorbable implants for bone regeneration focuses on the use of polymers and degradable metals. In this collaboration we have combined both types of materials to obtain composites with improved properties with respect to those of the individual materials. We have evaluated the behaviour of physical properties and their biological response to offer a description of these composites suitable for their subsequent implementation in implants for in vivo assessment.

Abstract

Physical activity has been shown to delay and even prevent the occurrence of pathologies and conditions associated with aging, including neurological disorders, some types of cancers and cardiovascular diseases. In this talk, we will focus our attention on the relationship between physical activity and cardiac function across a range of ages.

We will present results on the analysis of cardiac risk markers quantifying oscillations in the T wave of the electrocardiogram. We will illustrate how those markers vary as a function of age, body composition and fitness level.

Also, we will describe differences observed in measurements of heart rate variability, commonly used to assess autonomic nervous system activity, in individuals spanning from young adults to centenarians.

To better characterize cardiac activity in the oldest old and its relation to physical activity, we will describe a study on the benefits of resistance training in frail centenarians.

Abstract

Cancer cells and the stroma create dynamic pseudo-organs that contain a unique niche. During the progression of the disease, tumor cells typically rewire their metabolism, adapting to ever-changing environments. Such metabolic reprogramming has therefore profound implications in tumor proliferation, to an extent that monitoring such perturbations could harbor diagnostic and therapeutic relevance. In this context, surface-enhanced Raman scattering (SERS) can be used for the label-free detection of diverse molecules of interest among extracellular components. We present here the application of plasmonic superlattices comprising Au nanoparticles, to the precise SERS detection of selected metabolites and secreted immunomodulatory derivatives of the tumor metabolism. Finally, the effective plasmonic SERS substrate is combined with a hydrogel-based three-dimensional cancer model, which recreates the tumor microenvironment, for the real-time imaging of metabolite alterations and cytotoxic effects on tumor cells.

Abstract

In tissue engineering, biological, physical, and chemical inputs are combined to mimic cellular environments designed to fulfil different biomedical needs. Protein nanoparticles (pNPs) can simultaneously provide such physical and biochemical stimuli to cells when attached to surfaces and 3D scaffolds.[1] To provide a stable anchoring, a covalent binding of pNPs will be presented featuring a robust nanoscale topography with unprecedented mechanical stability and availability to influence cell morphology and orientation.[2]

Dynamic molecular interfaces that allow temporal control of cell behavior using an external stimulus, are very relevant for applications in biology, material sciences and medicine. Here we will present a cell adhesion study with spatio temporal control using stimuli-responsive self-assembled monolayers (SAMs) of an electroactive hydroquinone-benzoquinone (HQ-BQ) molecule that are used as a dynamic interface to immobilize pegitaled RGD functionalized peptides via interfacial reactions upon the application of a low electric potential.[3]

Density and spacing of RGD peptide at the nanoscale have already shown a significant influence on cell adhesion but its hierarchical nanostructuration influence is still rather unexplored. Here we present a versatile colloidal system based on fluid nanovesicles as a novel template for the hierarchic nanostructuration of RGD, anchoring it on the fluidic vesicle membrane. The engineered RGD-based nanovesicles are covalently anchored to surfaces. Such hierarchical substrates significantly enhance cell adhesion capabilities opening new pathways for the hierarchical immobilization of biomolecules on surfaces.

Abstract

Sleep disorders have a high prevalence. To investigate normal and disturbed sleep, we record biosignals both in the sleep laboratory and at home. Signals may be recorded directly, such as EEG, EOG, ECG, heart rate, respiration, pulse wave. Signals may be recorded with little contact or no contact systems such as actigraphy, body movement, bed sensors or bedside radiofrequency sensors. Some signal acquisitions are new in sleep research and require new technology and analysis concepts. We can identify wakefulness and sleep, we can derive details about sleep, such as light sleep, deep sleep, and REM sleep, arousals and sleep fragmentation. Not only classical methods in the time and frequency domain are used, but also more recent methods using statistical approaches are applied. Interaction between signals and causality analysis are important now.

Results allows recognizing normal and restorative sleep and identifying sleep disorders as well. Some sleep disorders have cardiovascular consequences and require personalized treatment. Sleep disordered breathing is the disorder with most cardiovascular consequences. Many diagnostic tools focus on this group of disorders. New diagnostic methods and perspectives are presented.

Abstract

The immune system contributes to maintaining the body's functional integrity through its two main functions: recognizing and destroying foreign external agents (invading microorganisms) and identifying and eliminating senescent cells, damaged or abnormal endogenous entities (such as cellular debris or misfolded/degraded proteins). Accordingly, the immune system can detect molecular and cellular structures with a spatial resolution down to 10 nm, which allows for the detection of molecular patterns expressed in a great variety of pathogens, including viral and bacterial proteins and nucleic acid sequences, and also expressed in abnormal cells. In this model, it is expected that nanostructured materials in the size range of proteins, protein aggregates, and viruses with different molecular coatings can engage in a sophisticated interaction with the immune system. As expected, it has been observed that nanoparticles can be recognized or passed undetected by the immune system. Once detected, they can be tolerated or induce defensive (inflammatory) or anti-inflammatory responses. This paper describes the observed mechanisms of the different modes of interaction between nanoparticles, especially inorganic nanoparticles, and the immune system and discusses their implications. This perspective should help to propose a set of selection rules for nanosafety-by-design and medical nanoparticles design.

Abstract

IMPaCT Genomics is a program of the IMPaCT infrastructure with the global objective of being the structural basis of a Precision Medicine strategy at the national level. Its specific objectives are:

• To align the network of Genomic Analysis Centers with the National Health System for the supply of highly complex genomic analysis services to the entire country.
• To demonstrate the functionality of the IMPaCT-Genomic infrastructure in use cases, linked to rare diseases, hereditary cancer and pharmacogenetics (including the identification of the causes of serious adverse reactions to drugs and vaccines).
• Population genomics: To genotype and sequence part of the cohort of the epidemiology program of IMPACT and to establish standards for PRS (Polygenic Risk Score) as well as mechanisms for the future incorporation of other use cases of Genomic Medicine within the IMPACT infrastructure.

Abstract

This talk will provide an overview of our work in the design of functionalised polymer nanoparticles and hybrid biomaterials for applications in healthcare. We engineer simple conceptually novel approaches to detect disease biomarkers, such as abnormally regulated enzymes, to extend the detection window for early disease diagnostics. We aim to design biosensing strategies that are simple, cost-effective and easy deploy to the point-of-care to democratise access to advanced diagnostic technology. I will discuss impactful biosensing applications for infectious and non-communicable diseases for example innovative smartphone enabled tests for epidemic surveillance in the field [2] and injectable nanoparticle-based sensing probes for in vivo detection of cancer that produce a colorimetric response in urine in under 1 hour [3]. We use advanced manufacture techniques to engineer complex 3D architectures that mimic anisotropic and multiscale tissue structures and produce spatially arranged bioinstructive materials to transform the regenerative medicine field [4]. To support the discovery of innovative biomaterials, we develop an array of characterisation techniques such as Raman spectroscopy characterisation for high-throughput tracking of surface functionalisation in single nanoparticles [4], visualisation of 3-dimensional structures for tissue engineering applications and FIB-SEM for investigating the cell-material interface. I will discuss how these versatile approaches can be applied to heal the body and detect diseases earlier.

Abstract

Nano-pharmaceuticals have the potential to drive the scientific and technological uplift, offering great clinical and socio-economic benefits to the society, industry, key stakeholders and patients. Nevertheless, affordable and advanced testing, manufacturing facilities and services are main prerequisites for successful implementation of these advances to further enhance the growth and innovation capacity. The establishment of current good manufacturing practice (cGMP) in nano-pharmaceutical production at large scale is the key step to successfully transfer nano-pharmaceuticals from lab to industrial scale. Due to the lack of resources to implement GMP manufacturing at-site, the scale-up and production of innovative nano-pharmaceuticals are still challenging to main players of the EU nanomedicine market, start-ups and SMEs. To allow successful implementation of the nano-pharmaceuticals in the nanomedicine field, there is an urgent need to establish a science- and regulatory-based Open Innovation Test Bed (OITB). PHOENIX aims to enable the seamless, timely and cost-friendly transfer of nano-pharmaceuticals from lab bench to clinical trials by providing the necessary advanced, affordable and easily accessible PHOENIX-OITB. PHOENIX-OITB offers a consolidated network of facilities, technologies, services and expertise for all the technology transfer aspects from characterisation, testing, verification up to scale up, GMP compliant manufacturing and regulatory guidance. The services and expertise provided by the PHOENIX-OITB will include production and characterisation under GMP conditions, safety evaluation, regulatory compliance and commercialisation boost.

Acknowledgement: This project has received funding from the European Union’s Horizon 2020 research and innovation programme under grant agreement No 953183.

Abstract

Innovative sectors can benefit from holistic support approaches at different stages of product development. Medtech is one of those sectors, facing also the challenges of the new European regulation for MDs and IVDs. To address the aforementioned challenges, the SAFE-N-MEDTECH consortium aims to bring a strong and competitive cooperation throughout the SAFE-N-MEDTECH OITB. This Consortium will offer a multidisciplinary and market-oriented innovation approach to academics, SMEs, Healthcare providers and Industries for the translation to the market of nano-enabled MTs, based on a deep understanding and knowledge of the material-nanoproperties, their advance use and applications in MTs and other aspects involved in MT safety. SAFE-N-MEDTECH will build an innovative open access platform to offer companies and reference laboratories, the capabilities, knowhow, networks and services required for the development, testing, assessment, upscaling and market exploitation of nanotechnology-based Medical and Diagnosis Devices. This across the whole Life Cycle of nano-enabled MTs.

Abstract

Antifolates are structural analogs of folates, essential one-carbon donors in the synthesis of DNA in mammalian cells, and they work as inhibitors of key enzymes in folate metabolism, such as dihydrofolate reductase and thymidylate synthetase. Methotrexate was one of the first agents of this class and is still extensively used in the treatment of a variety of tumors, including acute lymphocytic leukemia, breast cancer, osteosarcoma, primary central nervous system lymphoma, and head and neck cancer. Above all, it is also commonly used in certain autoimmune diseases, such as rheumatoid arthritis or psoriasis.1 However, the clinical efficacy of methotrexate is often limited and compromised by toxic dose-related side effects, which leads to morbidity, interruption of the treatment, and occasional mortality. A promising approach to tackle this problem is to activate the drug exclusively at its desired place of action. In fact, in those diseases that would benefit from a highly localized treatment, a precise spatiotemporal control over the activity of a chemotherapeutic agent would allow reducing the concentration of active compound outside the target tissue, improving the tolerability and hence the efficacy of the treatment.

Light is a powerful tool in this respect: it offers unparalleled opportunities as a non-invasive regulatory signal for pharmacological applications because it can be delivered with high precision regarding space, time, intensity and wavelength.

We developed Phototrexate, the first photoswitchable antifolate, by incorporation of a photochromic unit into the structure of methotrexate.

Phototrexate was designed to be constitutively inactive in its thermodynamically stable configuration (E isomer), while it can be activated with light (Z isomer) to locally provide the pharmacological effects of the parent drug, as confirmed in our earlier experiments in vitro and in zebrafish larvae.2 We have performed studies to assess safety and tolerability, pharmacokinetics, pharmacodynamics, and efficacy of our compound in vitro and in preclinical animal models. These results indicate that Phototrexate is a drug candidate with high potential for development as an innovative light-regulated antifolate for cancer and psoriasis.

Abstract

CANDI-EYE valorization project has intended to advance in the development of a new medical device for the detection of Candida albicans. The system validated within the project was previously developed by IQMA-IDM-UPV in collaboration with the accredited group of severe infection of IISLaFe. The device consists of a gated nanoporous alummina support loaded with a reporter (rodamine B) and capped with a specific oligonucleotide able to hybridize with Candida albicans genomic DNA. In the presence of Candida albicans, a selective release of the reporter is obtained. Within the project, the system has been validated in around 100 real samples (such as blood, cerebrospinal fluid, peritoneal fluid, pleural fluid) from patients infected with Candida albicans and has demonstrated its operativity. Thanks to the results of the project, the design and procedure, which were protected with a patent application (P201731069) have been transferred to Match Biosystems S.L. who will develop the final kit and will pursue its commercialization.

Abstract

As part of the IMPaCT infrastructure to implement precision medicine research in our country, CIBER presented a proposal to build a cohort of 200,000 people representing the entire population residing in Spain in collaboration with all Autonomous Communities and the autonomous cities (Ceuta and Melilla).

Coordinated by the area of Epidemiology and Public Health (CIBERESP), the participation of all CIBER thematic areas serves to speed-up a process that has taken four to five years in other countries, deciding the information and clinical data to be collected from participants and ensuring that it will be useful for multiple purposes, both for clinical and public health research. IMPacT cohort will serve as a platform to analyze health inequalities, monitor main health problems, formulate predictive models of disease, quantify the effect of different exposures and evaluate health policies. Innovative procedures for collecting biological samples and evaluating exposure variables based on a complete bio-psycho-social health models will be considered. Recruitment will be stablished in selected primary health care units all around the country. The IMPaCT cohort will integrate for the first time on a large scale in Spain clinical, epidemiological and biological data collected de novo in participants and their computerized medical records within the SNS. The IMPaCT cohort will respond to research questions about the effects on health of the lifestyles of residents in Spain. This initiative follows the model of the already existing national cohorts in countries like UK, USA, France, Germany and Sweden, something that facilitates the internationalization of the project, in its scientific and health components.

The cohort will be at the service of the Ministry of Health, as one more element of knowledge generation that helps decision-making in the field of public health and health management. The group of participating entities guarantees the integration of the IMPaCT cohort in the autonomous health services, becoming a research platform available to the entire scientific community.

During the lecture, I will comment how more than 100 researchers are already contributing to the design and protocols of this new infraestructure.

Abstract

BNN is a non-profit research organization owned by the BioNanoNet Association. We are responsible and devoted for pursuing the objectives of the Association, taking special care of the needs and requests of our members. The BioNanoNet Association aims to strengthen innovative research by promoting cooperation and creating synergies among people working in the fields of Health & Safety, Data & Sustainability and Enabling Technologies.

By connecting our members to national and international strategic stakeholders, together we establish and coordinate thematic platforms that lead initiatives in different fields. As a consequence, we jointly contribute to shape the R&D&I landscape in Europe. BNN’s core competences are distributed along different services that we implement not only to benefit the association members but also external customers and partnerships. BNN’s competences will be presented, as key factor of success of our on-going initiatives and projects with the aim to look for future cross-collaborations and/or initiatives.

Abstract

ANIMATE is a platform to promote behavioural changes and the acquisition of healthy lifestyles related to diet and physical exercise. It is based on an mHeath tool, that provides self-registration of diet, physical exercise, weigth, diet-related mood and health status, and generates personal notifications, taking into account the patient goals. It combines several technologies and uses the personal motivation as the key of the behavioural changes. The core of ANÍMATE is the set of algorithms that process and analyzes the data recorded and provides personalized notifications (alerts, recommendations and congratulations), weekly reports and a motivating progress visualization to empower the patient and to generate the self-confidence he need to achieve his goals. ANíMATE is running in Sant Pau Hospital where a clinical trial is going to start in November, with 35 overweigth patients, during 4 months.

Abstract

The standard treatment in glioblastoma patients includes maximal safe resection followed by concomitant treatment with chemoradiotherapy including temozolomide, and adjuvant temozolomide. The first MR follow-up study to evaluate treatment response is performed one month after concomitant treatment (P1M). Currently, it is not possible to confidently distinguish between tumour growth (true progression) and treatment-associated changes (pseudoprogression) with conventional MR, but MRSI could help distinguish among progression statuses. Magnetic Resonance Spectroscopy (MRS) is a non-invasive method of brain tumors, diagnosis and grading, surgical planning and assessment of response to therapy. Convex Non-negative matrix factorization(cNMF) is an unsupervised machine learning technique that is used for decomposition of signals. The purpose of this study was to train a classifier based on an additional P1M MRSI, that allows to early predict the behaviour of the suspicious contrast-enhancing regions (SCER) in the next MR exploration.

Abstract

Analyses of fongitudinal data allow studying trajectories of biomarkers in disease. A common method is the Linear Mixed Effects (LME) modelling, which can be implemented using a frequentist or a Bayesian framework. We studied these approaches using ADNI data and different dataset configurations. We analyzed the capability of both options to detect differences in the hippocampal volume across healthy controls (HC), mild cognitive impairment (MCI) and Alzheimer’s Disease (AD) patients. We used a random-removal permutation-based approach to calculate the minimum sample size. Both methods performed well in big-N datasets. We found that 115 subjects were needed to differentiate MCI converters vs non converters, and 147 to differentiate across groups. The Bayesian approach was more restrictive with low-N sets, but it allowed estimating the LME model in sets with high number of missing points, where the frequentist approach failed.

Abstract

Human induced pluripotent stem cell-derived cardiomyocytes (hiPSC-CMs) cultured on bio-printed scaffolds have shown promising results for cardiac function restoration. Nevertheless, pro-arrhythmicity favoured by reduced conduction velocity of the transplanted constructs has been poorly assessed. Here, we investigate the impact of the scaffold geometry on the electrical activation properties of hiPSC-CMs cultures. Electrophysiological models of hiPSC-CMs and the Finite Element Method were employed for computational simulation of hiPSC-CMs cultures. These models were calibrated to replicate experimentally measured activation time maps by adjusting parameters representative of fiber alignment and cell-to-cell coupling. Scaffolds with rectangular, auxetic and elongated hexagonal pore shapes were studied to determine the most biomimetic structure in terms of electrical propagation. Our results showed that elongated hexagons led to faster activation of hiPSC-CMs cultures by facilitating the alignment of cardiac fibers parallel to the depolarization wavefront.

Abstract

Early detection and grading of Diabetic Retinopathy (DR) are essential to determine an adequate treatment and prevent severe vision loss. However, the manual analysis of fundus images involves a work overload due to the increasing prevalence of DR and the limited resources in personnel and technology. We propose a novel end-to-end deep learning framework for automatic DR grading. Our approach is based on an attention mechanism which performs a separate attention of the dark and the bright structures of the retina. We used the largest retinal image dataset publicly available, containing over 88,000 images, and discarded those with poor quality. The proposed approach achieved 83.7% accuracy and a Quadratic Weighted Kappa of 0.78, which outperforms several state-of-the-art approaches. Our results suggest that our framework could be a diagnostic aid for the early detection and grading of DR.

Abstract

Conventional sinusoidal signals have been exclusively used in magnetic hyperthermia as the only alternating magnetic field waveform to excite magnetic nanoparticles. However, there are no theoretical nor experimental reasons that prevents the use of different waveforms. We aim to study the effect of various waveforms on the heat production effectiveness of magnetic nanoparticles, seeking to prove that signals with more significant slope values allow the nanoparticles to reach higher efficiency in heat generation. Also, we seek to point out the dependency of the nanoparticle power dissipation on the waveform's slope, and not only the frequency, magnetic field intensity and the nanoparticle size. The experimental results showed a remarkably higher heat production performance of the nanoparticles when exposed to trapezoidal and almost-square signals than conventional sinusoidal signals. The normalized power dissipation value was also calculated and prove its dependency on the slope. However, adjustments are necessary to the coil before proceeding with in vitro and in vivo studies.

Abstract

Alzheimer’s disease (AD) is a neurodegenerative disorder that, eventually, leads to dementia. In the last decades, multiple lines of research have analyzed numerous biological, clinical and sociodemographic factors, such as cerebrospinal fluid, brain activity, genetics, neurocognitive test, education level, and diet, among others, to better understand its pathogenesis. However, the study of the implications of each aspect involved with AD on their own, although relevant, has limitations. Since the brain is a hugely complex, interconnected system characterized by uncountable non-linear physiological relations, the study of the associations between factors even, at first, not directly related, is meaningful. In this regard, association networks provide us a powerful tool to integrate and summarize the complex relationships between the heterogeneous factors related with AD. This insight is crucial for gaining a further comprehension and obtaining the pathophysiological fingerprint of AD.

Abstract

Optical hyperthermia (OH) is a developed therapheutic strategy, which applies laser irradiation on near-infrared (NIR) photoabsorbers to generate heat for thermal ablation of cancer cells in a selective and safe way. Gold nanoparticles are the best candidate for this treatmenet due to their photothermal properties as Surface Plasmon Resonance (SPR), Biofunctionalization with differentes biomolecules as drugs or antibodies, NIR window used and Enhanced Permeability and Retention (ERP) effect. Some results are shown in the presentation. Temperature curves obtained show a great heat effectiveness conversion. No citotoxicity has been found in any nanoparticles or any concentration used. All nanoparticles are potencial candidates for this treatment. The synergic effect with laser irradiation and nanoparticles is enough to kill cancer cells avoiding damage in healthy tissue around.

Abstract

Pediatric obstructive sleep apnea is a frequent respiratory condition that has been linked to increased cardiovascular risk. After using adenotonsillectomy as treatment, there have been reported changes in classic heart rate variability (HRV) characteristics of the affected children. However, these changes could not be causally attributed to treatment outcomes. Furthermore, the scientific literature lacks of an evaluation of HRV approaches as a potential indicator of sleep apnea resolution. This research study shows that changes in specific HRV parameters that were statistically found are able to identify obstructive sleep apnea treatment outcomes. Besides, HRV activity in BW2, a specific apnea-related spectral band in the range 0.028-0.074 Hz, differentiates children achieving obstructive sleep apnea resolution when two different resolution definitions are used. Based on these results, we propose BW2 as a potential biomarker of pediatric obstructive sleep apnea resolution.

Abstract

Early detection and grading of Diabetic Retinopathy (DR) are essential to determine an adequate treatment and prevent severe vision loss. However, the manual analysis of fundus images involves a work overload due to the increasing prevalence of DR and the limited resources in personnel and technology. We propose a novel end-to-end deep learning framework for automatic DR grading. Our approach is based on an attention mechanism which performs a separate attention of the dark and the bright structures of the retina. We used the largest retinal image dataset publicly available, containing over 88,000 images, and discarded those with poor quality. The proposed approach achieved 83.7% accuracy and a Quadratic Weighted Kappa of 0.78, which outperforms several state-of-the-art approaches. Our results suggest that our framework could be a diagnostic aid for the early detection and grading of DR.

Abstract

Human life expectancy has increased worldwide, favoring the appearance of musculoskeletal diseases. Tissue engineering approaches based on the use of polymers that reproduce physiological characteristics of the tissue itself are of great interest for bone regeneration therapies. Our group has developed a series of smart cell culture supports which mimic bone’s piezoelectricity, a key factor in guiding mesenchymal stem cells (MSCs) osteogenic differentiation for their use in bone regeneration strategies. The combination of the piezoelectric polymer poly(vinylidene fluoride) (PVDF) with magnetostrictive nanoparticles (MNPs) allows to induce the piezoelectric effect generating an external magnetic field with a bioreactor. These cell culture platforms can be tailored in different shapes (porous membranes, films or microspheres); and functionalized with different biomolecules that resemble bone’s extracellular matrix. This offers a wide range of possibilities not only for bone, but for the regeneration of other electroactive tissues.

Abstract

Triple Negative Breast Cancer is characterized by a high degree of cancer cell heterogeneity. Among cancer cell subpopulations Cancer Stem Cells (CSCs) outstand due to their crucial role in tumor resistance, recurrence and progression. Using in vitro models fluorescently tagging CSCs from heterogenous cell lines we isolated extracellular vesicles (EVs) from CSCs and differentiated cancer cells (DCC) and assessed their impact on cancer associated fibroblasts. Interestingly, EV specific protein cargo depended on the producing cancer cell subset. When administered to CAF in vitro, these EVs exerted markedly distinct activities. Namely, EVsDCC promoted the activation of secretory iCAFs while EVsCSC activated a myofibroflastic CAF subset (myoCAFs). Further assessing in vivo of EV differential CAF activation on metastasis revealed a major contribution of EVCSC-activated myoCAFs on the macrometastatic burden. These results contribute to understand metastatic progression and provide new potential therapeutic targets.

Abstract

The most important challenges in cancer treatment are 1) avoid systemic side effects in non-desired organs and 2) develop alternative treatment strategies for metastatic and aggressive tumors. Nanomedicine was assumed to have answers to these issues, but delivering enough therapeutic nanoparticles (NPs) to tumors still remains an important challenge in nanomaterials-based treatments. Exosomes (nanosized extracellular vesicles) play a crucial role in cell communication processes and can be combined with NPs to improve their targeting capabilities. In this work, we leverage the ability of exosomes derived from stem cells to reach tumor areas successfully, being used as delivery vehicles for optical hyperthermia treatments. Once NIR-sensitive hollow gold NPs loaded exosomes reached primary subcutaneous solid tumors, they were irradiated with a NIR laser and almost complete tumor remission was obtained. More interestingly, those exosomal vehicles were also able to reach multinodular areas similar to those on advanced metastatic phases, eradicating most tumor growth regions in multiple cancerous nodules located in the pancreas region.

Abstract

We develop two targeted nanoparticles (NTs) against the CXCR4, which is overexpressed in metastatic stem cells of colorectal cancer (CRC). The catalytic domains of these NTs are based on toxins (Pseudomonas aeruginosa and Corynebacterium Diphteriae), and they have previously shown to produce pyroptosis in human CRC cell lines. Pyroptosis is a programmed inflammatory cell death that triggered the recruitment of immune cells through the release of proinflammatory interleukins. The pyroptosis happening in tumour would recruit proinflammatory immune cells to the tumour microenvironment, helping with the antitumour effect of the NTs. The objective is to describe the cell death mechanisms triggered by the treatment with NTs in mouse CRC cell lines and in CRC mouse models, and explain the changes in immune cell populations derived from the cell death mechanisms.

Abstract

The main objective of the ViSQCT project is to develop a device based on Quartz Crystal Resonators (QCR) for medical diagnosis, measuring the viscoelastic properties of human fluids. As a starting hypothesis, we propose that it is possible to design, develop and validate a medical device based on QCR, as well as the methodology of use of this, with which the analysis of the viscoelastic properties of some body fluids (synovial fluid and cerebrospinal fluid) is performed and whose results help in the diagnosis of pathologies associated with these fluids in an efficient, fast, economical and objective way. As first results, the viscosity of various phantoms that mimicked the viscosity of synovial and cerebrospinal fluids was measured for pathological and healthy cases. Currently, we are measuring real samples of synovial fluid, being able to differentiate between the viscosities of these samples, and whose ultimate goal will be to evaluate the efficiency and sensitivity of the sensor.

Abstract

Human Activity Recognition (HAR) plays an important role in behavior analysis, video surveillance, gestures recognition, gait analysis, and posture recognition. Given the recent progress of Artificial Intelligence (AI) applied to HAR, the inputs that are the data from wearable sensors can be treated as time-series from which movement events can be classified with high accuracy. In this study, a dataset of raw sensor data served as input to four different deep learning networks (DNN, CNN, LSTM, and CNN-LSTM). Differences in accuracy and learning time were then compared and evaluated for each model. An analysis of HAR was made based on an attempt to classify three activities: walking, sit-to-stand, and squatting. We also compared the performance of two different sensor data types: 3-axis linear acceleration measured from two inertial measurement units (IMUs) versus 3D acceleration of two retro-reflective markers from the high-end optoelectronic motion capture system (MOCAP). The dataset created from observations of ten subjects was preprocessed with labelling and sliding windows and then used as input to the four frameworks. The results indicate that, for HAR prediction, linear accelerations estimated using IMUs are as reliable as those measured using the MOCAP system. Also, the use of the hybrid CNN-LSTM framework for both methods resulted in higher accuracy (99%).

Abstract

Pediatric sleep apnea-hypopnea syndrome (SAHS) is associated with major cardiovascular illnesses. A successful development of the project will lead to a new tool for the diagnosis and screening of pediatric SAHS. This tool will have the ability to detect the children most severely affected by SAHS, but it also aims to identify those children at risk of suffering from cardiovascular issues (CVR) associated with SAHS. The screening tool is based on the overnight heart rate variability, and there is behind a strategy for industrial property protection to develop a clinical web application. While it is true that defining CVR indices can be straightforward −in fact, many CVR indices exist such as the metabolic syndrome or the Framingham index− this is not the case for the specific pediatric population. Here we present an approach, using standard clinical variables and corresponding estimative parameters for the definition of the CVR in children.

Abstract

This presentations is about a doctoral thesis which main objective is to develop a tool that will improve the current practice of the rehabilitation of Orientation and Mobility within visually impaired people. The tool, which is currently under development, will be a gamified system based in the analysis of inertial measurements unit sensors that measures the motion of the coverage of the long cane and the spatio-temporal gait parameters of the user. The presentation contains the process of the discovering of a gap in literature trough the description of a review paper written in the first stage of the doctoral advances and a second paper that describes the results of the motion analysis algorithms that have been developed so far. Also, a description of a further work in order to complete the gamified accessibility software of the system.

Abstract

The CiPA initiative from FDA has promoted the use of human models into the safety assessment workflow for early prediction of drug-induced cardiotoxicity. Yet, the effect of age, a primary risk factor for cardiac arrhythmias, is not regarded in these tools. The goal of this study is to create human in vitro and in silico models that contribute to predict drug cardiotoxic effects as a function of age. In vitro, an inducible progerin expression system has been knocked-in into human induced pluripotent stem cells (hiPSCs) so that its derived matured cardiomyocytes can be induced to age. In silico, experimentally-calibrated populations of adult and old human ventricular electrophysiological models have been generated by using the results of age-related transcriptomic data analysis. The potential for prediction of drug-induced cardiotoxic effects by combining the proposed in vitro and in silico models is presented.

Abstract

We have recently demonstrated that the transgenic TgF344-AD rat model of Alzheimer disease shows time-dependent alterations revealing dysfunction in several resting state networks. Moreover, the amplitude of the somatosensory network of the transgenic rats at 18 months of age was strongly correlated with the behavioral performance in a working memory task. In this study we present preliminary results regarding the effect of early behavioral training during 8 months on somatosensory networks in non-aged wild type and transgenic rats. Although no significant differences between genotypes were observed, there was a significant difference between non-trained and trained transgenic rats, pointing to an impact of cognitive training in the somatosensorial network of TgF344-AD rats, that could compensate for the initial pathology-related alterations in these animals.

Abstract

In this video we describe a new mHealth supported clinical pathway of care for people living with medically stable HIV. The EmERGE mHealth platform was co-designed with clinicians and community, developed using agile methodologies, integrated with hospital information systems and validated in a large prospective cohort study of 2251 participants. The evaluation was conducted using a tailored Health Technology Assessment: the Model for Assessment of Telemedicine Applications. Usability and acceptability were assessed through the System Usability Score and a Patient Reported Experience Measure. The platform was successfully deployed across diverse care settings in 5 European countries and used by 2251 patients and more than 20 clinicians for up to 30 months. Results have demonstrated that the EmERGE platform is feasible and acceptable; with a high level of usability and very positive patient reported experience. The EmERGE platform is a secure and GDPR compliant system with a complete set of functionalities that could be easily adapted to other clinical conditions, clinical sites and health systems thanks to its modular technical architecture.

Abstract

Cartilage damage is one of the major causes of disability. Unfortunately, it is one of the most difficult tissues to repair and there are few treatments available. Here we study the cartilage differentiation capacity of a new biomaterial composed by Chitosan (Ch), Hyaluronic Acid (HA) and Chondroitin Sulfate (CS) by mesenchymal stem cells. Expression analysis of skeletal differentiation markers determined that Ch-HA-SC promoted sox9 expression during initial differentiation stages, and an increase in the cartilage structural component aggrecan, both signs of a healthy cartilage. This biomaterial also presented increased expression of the bone marker osteocalcin, which suggests the biomaterial is able to sustain an endochondral ossification process. Histology staining of cartilage proteoglycans by Alcian Blue confirmed presence of mature chondrocytes and cartilage matrix. Therefore, these results support that the Ch-HA-SC biomaterial promotes chondrogenic differentiation and its potential for cartilage regeneration.

Abstract

Accumulation of misfolded a-synuclein (a-syn) is a hallmark of Parkinson’s disease (PD) thought to play important roles in the pathophysiology of the disease. Dendritic systems, able to modulate the folding of proteins, have emerged as promising new therapeutic strategies for PD treatment. Here, we set out to investigate the effects of dendrimers on endogenous a-syn accumulation in disease-relevant cell types from PD patients. For this purpose, we chose cationic carbosilane dendrimers of bow tie topology based on their performance at inhibiting a-syn aggregation in vitro. Dopamine neurons were differentiated from induced pluripotent stem cells generated from PD patients, which reportedly display abnormal accumulation of a-syn. Treatment of PD dopamine neurons with dendrimers was effective at preventing abnormal accumulation and aggregation of a-syn. Our results in a genuinely human experimental model of PD highlight the therapeutic potential of dendritic systems and open the way to developing safe and efficient therapies for delaying or even halting PD progression.

Abstract

Osteoarthritis treatments are based on non-steroidal anti-inflammatory drugs, COX-2 inhibitors and corticoids. Most of these drugs present cytotoxicity and low bioavailability in physiological conditions. Therefore, the goal of this work was to encapsulate three different hydrophobic anti-inflammatory drugs (celecoxib, tenoxicam and dexamethasone) into polymeric nanoparticles with potential applications in osteoarthritis. Nanoparticles presented hydrodynamic diameters of 120 nm and almost neutral surface charges. Encapsulation efficiencies were highly dependent of the drug water solubility, having the highest values for celecoxib. Nanoencapsulation reduced celecoxib and dexamethasone cytotoxicity in articular chondrocytes and macrophages. Besides, nanoparticles reduced the release of different inflammatory mediators by LPS-stimulated RAW264.7 and proved to be in vivo biocompatible. These findings suggest that these nanoparticles could be suitable candidates for the treatment of inflammatory processes associated to osteoarthritis due to the demonstrated in vitro activity as regulators of inflammatory mediators production.

Abstract

Multiple myeloma (MM) is a hematological neoplasia characterized by plasma cell abnormal proliferation and with bone marrow homing. Nowadays, drug resistance generation among patients is the main clinical problem. In the bone marrow, the microenviroment plays a key role in drug resistance generation. Different interactions such as cell-cell or cell-extracellular matrix participate of this process and are not represented in conventional cell culture models. This limits their use as in vitro testing models. In this context, our BIOMICROGEL project aims to develop a semi-solid cell culture media formed by biomimetic microspheres that attempts to redefine in vitro models for this disease including a better biomimicry of aspects such as extracellular matrix (ECM) mediated DR. In this presentation, the microgel concept will be presented as well as our results in microsphere’s fabrication and characterization and MM cell cultures in the system.

Abstract

HEPATOPRINT project proposes the novel design of more reliable strategies for prediction of hepatotoxicity in drug development and the early diagnose of drug-induced liver injury (DILI) as a personalized medicine application. It is based on biomimetic printable hydrogels that efficiently present growth factors from the solid-phase to control accurately and efficiently its delivery. Materials are selected to mimic the liver extracellular matrix using protein-polysaccharide combinations. These injectable hydrogels will be 3D bio-printed in culture for the automatic production of replicates to assess reproducible and high-throughput screenings. As a result, we will produce a new reliable platform for DILI prediction, able to make personalized studies to identify idiosyncratic toxicity reactions that could be apply in preclinical trials.

Abstract

Atraumatic mesh fixation approaches aim at reducing chronic pain and discomfort caused by classical suture fixation following hernia repair. This study compares the biological and mechanical response of a self-adhering (Adhesix), a self-gripping (ProGrip) and a conventional mesh (Surgipro) fixed with cyanoacrylate adhesive in a rabbit model of prosthetic hernia repair. Partial defects (6x4cm) were repaired with those meshes, and morphological and biomechanical performance of the implants were evaluated 14/90 days postoperatively (n=6 each). At both time-points, ProGrip and Surgipro exhibited better tissue incorporation and lower shrinkage than Adhesix. While ProGrip’s microhooks remained following 90 days, Adhesix’s hydrogel was resorbed by day 14. Macrophage response was higher in ProGrip and Surgipro implants. ProGrip showed the highest tensile strength and Adhesix the highest failure stretch. In this model, the self-gripping mesh displayed better biological and mechanical performance than the self-adhering one.

Abstract

Mesh infection following hernia repair is a devastating complication which increases morbidity and mortality rates in patients. This study aimed at assessing the effect on abdominal wall repair of coating meshes with two biopolymer carboxymethylcellulose-based antibacterial gels (antiseptic, antibiotic) in a preclinical model of Staphylococcus aureus infection. Partial abdominal wall defects (5x2cm) were created in New Zealand white rabbits. Four study groups (n=6) were established according to biopolymer gel coating and S. aureus infection. Fourteen days after surgery, histological studies and gene/protein expression of collagens type I and III were carried out. Compared to uncoated ones, antibacterial-coated meshes showed good integration within the host tissue, as well as higher collagen 1/3 mRNA ratio and collagen I protein expression. Our findings suggest that prophylactic antibacterial coating of meshes may help improving abdominal wall tissue repair in the presence of infection.

Abstract

Osteopenia is a condition characterized by a reduced bone mass which is present in a wide range of skeletal disorders. This affection leads in the end to frequent fractures and disability in many cases. In this presentation, we have introduced an overview of the work which is being carried out at the Group of Biomaterials, in collaboration with other organizations, to develop formulations that, by varying the loaded components, could be used as therapies at different osteopenic scenarios. Based on oxidized hyaluronic acid and carboxymethyl chitosan, injectable in situ gelling formulations with different biological/bioactive cargo were developed. Applications including controlled drug delivery, gene therapy and cell encapsulation are briefly explained.

Abstract

Osteoporosis is a generalized bone disease characterized by a decrease in mass and a deterioration of bone microarchitecture. The Smart Biomaterials Research Group (GIBI) has developed two different strategies to enface this pathology. The first one is the design macroporous scaffolds to boost bone regeneration. The second is the development of new treatments based in mesoporous silica nanoparticles. The macroporous scaffolds were made of mesoporous bioactive glasses, a highly bioactive bioceramics that promoted bone regeneration of high quality in an osteoporotic sheep model. On the other hand, mesoporous silica nanoparticles were loaded with SOST gene siRNA and osteostatine. These nanoparticles injected subcutaneously in ovacteromised micewere able to increase the expression of certain osteogenic genes, producing also an improvement in the bone microarchitecture.

Abstract

Biofilm-related infections associated to orthopaedic implants constitute one of most problematic chronic infection diseases with high morbidity, and high medical care expenses. In the Smart Biomaterials Research Group, we have developed of two different strategies for the design of new nanomaterials to manage bone infection: i) Prevention and ii) Healing. The properties of these materials must be adapted to achieve anti-infective performance and biocompatibility, which permit a good integration of the implant with the surrounding tissue. PREVENTION has been accomplished via the fabrication and developing of nanostructured surfaces onto metallic implants and the design of 3D scaffolds for bone regeneration incorporating antimicrobial agents as prophylactic treatment. Finally, HEALING has been tackled by the design of nanocarriers based on mesoporous silica nanoparticles as nanovehicles of different antimicrobial agents for local treatment of infection. Such strategies constitute a real breakthrough in the design of new nano-weapons for the management of bone infection.

Abstract

Cardiovascular diseases remain the main cause of morbidity and mortality worldwide. Despite the progress in the development of interventional and pharmacological agents to treat cardiovascular complications, the drug failure rate remains very high indicating the lack of appropriate assays for safety and efficiency in current drug screening methods. In this regard, organ-on-a-chip technology is gaining increasing attention by providing more efficient and predictive human-based platforms. Particularly, heart-on-a-chip, having the ability to recapitulate the important biological and physiological parameters of cardiac tissue, has recently emerged as a promising approach by providing a technological platform capable of accelerating cardiovascular drug development. Here, we will present a collaborative project aimed at developing a heart-on-a-chip device that combines the main cell types present in the myocardium, differentiated from human induced pluripotent stem cells, and provide proof-ofconcept evidence for its usefulness in cardiovascular drug development.

Abstract

In this work we have fabricated 3D printed plasmonic scaffolds with the aim to reproduce complex 3D cell models that resemble tumours and study their evolution. We propose the use of Surface Enhanced Raman Spectroscopy (SERS) to monitor the release of tumour metabolites from cancer cells that can grow within a 3D printed scaffold. Plasmonic inks were prepared by mixing plasmonic nanoparticles, such as gold nanorods, with a mixture of polymers with suitable rheological properties for printing. Once the scaffolds were prepared we performed the sensing of model molecules and proved their ability to detect them in a 3D fashion. SERS can be also used as an imaging tool after the labelling of both the scaffold and the cells with suitable Raman tags and study their behaviour inside the scaffold.

Abstract

Therapeutic nucleosides are still one of the most used drugs in cancer. Among them, 2’-deoxy-5-fluorouridine o floxuridine (FdU) is one of the most employed. One of the major problems described is cell resistance to the nucleoside antimetabolites in long treatments. Our strategy is based on the use of oligonucleotides carrying these FdU as prodrugs. We have prepared and evaluated conjugates of (FdU)5 with different delivery enhancers molecules (Cholesterol, Folic acid, GalNAc, Palmitic acid and PEG) in different cell lines. The results confirmed that the most active product was the Palmitic Acid conjugate but not the best internalized which was the Cholesterol conjugate. In the same way, we have prepared and evaluated (FdU)5 and (FdU)10 covalently attached to G-quadruplex structures. The results confirmed a higher uptake of the Gq structure and a higher toxicity of the (FdU)10 TG6T in resistant FdU cell lines.

Abstract

Over last decades, several nanoparticles have been approved as medicines for human use (1). Pharmacopeia recommends methods to control bacterial endotoxins to avoid attribution to the medicines of toxic effects that may be entirely due to these contaminants. Endotoxins levels 1EU/mL by LAL assay quantification are established for parenteral products (2).Endotoxins are negatively charged lipophilic molecules present in bacteria membrane, which can interfere with positively charged nanoparticles, giving misleading results in LAL assays (1). In our group, monodisperse nanoliposomes have been developed for the effective delivery of enzymes (3,4). Among different assays for endotoxin quantification, Gel-clot technique was selected to avoid interference from liposomal samples turbidity (5). We need to disrupt our liposomes through a treatment to avoid Low Endotoxin Recovery (LER) issues. Our methodology has shown to be effective to control the presence of endotoxins in liposomal products.

Abstract

Osteochondral defects are focal areas of damage within cartilage and subchondral bone tissues that cause loss of mobility and chronic pain. Osteochondral regeneration is particularly challenging due to the different physiological structures and functions of cartilage and bone. Recent studies demonstrated that large osteochondral defects are more efficiently healed when regeneration recapitulates bone development by a cartilage intermediate. During skeletogenesis, mesenchymal cells gather into cartilaginous nodules, setting the bases for both cartilage and bone formation. The existence of a common cartilage template suggests cell condensate’s structure could be paramount for an efficient osteochondral repair.

The NanoBuds project aims at engineering optimal osteochondral precursors, from a cell adhesion-guided cell condensation process, capable of recapitulating osteochondral development. The current presentation shows the most recent advances within the framework of the project.

Abstract

Our group is involved in engineering colloidal systems with potential applications in biomedicine. Currently, we are focused on preparing polymeric nanoparticles from nano-emulsion templating using low energy methods to be used in targeted breast cancer and the treatment of Alzheimer’s disease. In the latter case, our group is working in the context of an Early Stage and Intramural projects. These projects involve the encapsulation of natural products and functionalization of NPs’ surface with a series of targeting ligands like oligonucleotides, peptides and poly(ethylene glycol). The characterization of drug-loaded nanoparticles including DLS, z-potential and TEM microscopy as well as the evaluation of cumulative drug release and toxicity have been successfully accomplished. Further experiments involving the use of a BBB-model membrane and in vivo experiments will confirm the feasibility of these systems for the management of the Alzheimer’s disease.

Abstract

Our group is involved in engineering colloidal systems with potential applications in biomedicine. Currently, we are focused on preparing polymeric nanoparticles from nano-emulsion templating using low energy methods to be used in targeted breast cancer and the treatment of Alzheimer’s disease. In the latter case, our group is working in the context of an Early Stage and Intramural projects. These projects involve the encapsulation of natural products and functionalization of NPs’ surface with a series of targeting ligands like oligonucleotides, peptides and poly(ethylene glycol). The characterization of drug-loaded nanoparticles including DLS, z-potential and TEM microscopy as well as the evaluation of cumulative drug release and toxicity have been successfully accomplished. Further experiments involving the use of a BBB-model membrane and in vivo experiments will confirm the feasibility of these systems for the management of the Alzheimer’s disease.

Abstract

Although the mechanism of action of liposomes applied to the skin remains in debate, their penetration enhancement capability seems to depend on the deformability, which is unfortunately low in the case of conventional, stable liposomes. The demand for vesicular systems with enhanced properties has led to alternative nanovesicles like the Quatsomes (QS), homogeneous and stable upon long storage. As part of the FlexCAB project, we investigated the structure and mechanics of the QS membrane at the nano and mesoscale, by X-ray and atomic force microscopy/spectroscopy techniques. We show that ions present in the medium and the surfactant chain length, fine-tune the QS membrane thickness and lateral packing. These may translate into the flexibility of the QS vesicles, which could be easily tailored by adjusting the chemical composition and the surrounding medium, according to the specific need for each application.

Abstract

Chronic liver disease (CLD) has no effective treatments apart from reducing its biological complications. Simvastatin showed promising results as vasoprotective drug in experimental models of CLD, but also limiting adverse effects, mainly due to accumulation in muscles. Encapsulation of simvastatin in Pluronic® F127-micelles (PM127-simv) targeting liver sinusoidal endothelial cells (LSECs) is expected to improve the therapeutic window of simvastatin by alter its biodistribution. PM127-simv showed a high rate of cell internalization in rat LSECs and significantly lower toxicity than free simvastatin, improving cell phenotype. The in vivo biodistribution showed a high hepatic accumulation (50% of the injected PM). Remarkably, after one week of administration in a model of CLD, PM127-simv demonstrated superior effect than free simvastatin in reducing portal hypertension. Moreover, no signs of toxicity of PM127-simv were detected. Our results indicate PM-simv is a promising therapeutic approach to treat CLD.

Abstract

The use of Near-Infrared (NIR) irradiation in combination with photo-active materials is a promising cancer-treatment to achieve a selective and non-invasive therapy. However, conventional photosensitizers are organic molecules lack of stability in physiological media. Titanium dioxide (TiO2) as a classic heterogeneous catalyst promote ROS formation under ultraviolet-visible (UV-vis) radiation is proposed as a more robust promising alternative. To overcome UV-vis limitation, the combination of TiO2 with a NIR-sensible nanomaterial, Nitrogen Doped Carbon Dots (N-CDs) is presented. N-CDs can transform NIR light into UV-vis, suitable to perform the desired photocatalytic reactions and trigger tumor cell death. The focus of this work is set on the synthesis of highly monodisperse photoluminescent N-CDs by laser pyrolysis, with tunable optical properties and application as sensitizers for NIR-driven photocatalysis. This hybrid photocatalyst has shown promising results towards both the oxidation of key metabolites for tumor cell proliferation (i.e. glucose) and generation of toxic species (.OH) under NIR radiation.

Abstract

Cancer constitutes one of the main causes of death despite the wide range of existing therapies. Current efforts are being devoted to the optimization of alternative treatments (e.g. photodynamic therapy (PDT), starvation therapy (ST) or chemodynamic therapy (CDT)) to alter the tumor microenvironment (TME). Fast growth of tumors entails a dramatic increased blood vessel formation (angiogenesis) in order to maintain the accelerated metabolism of cancer cells, providing them with nutrients (especially glucose) and oxygen (O2). However, rapid growth provokes an irregular alignment of endothelial cells associated with the tumor, with two major consequences: (i) large spaces between cells are created, with enough margin for 20-200 nm-sized nanoparticles to penetrate into the interstitial space, known as Enhanced Permeability and Retention (EPR) effect and (ii) limited O2 levels supply inside tumour cells (hypoxia conditions). The present works aims at presenting a novel gold-platinum core-shell dendritic nanocatalyst (Figure 1 left) with enzyme-mimicking capabilities to convert high rates of glucose into gluconolactone (Figure 1 center) and simultaneously generate O2 from endogenous H2O2 to relief tumor hypoxia and re-establish the glucose oxidation cycle (Figure 1 center and right).

Abstract

Metastasis remains as the leading cause of death in cancer patients and therefore, smart nanocarriers for targeted drug delivery need to be developed. In this context, CXCR4 appears as a promising receptor for specific targeting, as its overexpression associates with tumor aggressiveness and metastatic initiating capacity. Our laboratory has developed a first generation of CXCR4-targeted multivalent nanoconjugates that selectively deliver genotoxic drugs to CXCR4+ metastatic stem cells. However, the currently used unspecific lysine-amine drug coupling method does permit control over the amount nor the specific location where the drug is being conjugated to. The production of a new generation of nanoconjugates using site-directed conjugation allows to significantly improve their therapeutic efficiency as well as the control, characterization, and homogeneity of the final nanoconjugate. Specifically, the Cysteine Coupling methodology implemented in THIOMABs has been used to upgrade our current drug coupling technology.

Abstract

A high percentage of the cancer patients present a mutation in a well-known gene called Kirsten rat sarcoma viral oncogene homolog (KRAS). When this mutation is present, there is an aberrant activation of the KRAS protein that will activate molecular cascades responsible for cell malignancy and tumor development. Unfortunately, until now any study as succeed to develop effective therapies specifically against the KRAS mutated cancers. We believe that the most efficient and specific way to achieve this objective is to deliver intracellularly a specific antibody against the KRAS isoform present in each tumor. The antibodies by itself do not have the capacity to internalize into the cells and required the help of carriers with a nanometric size. We will use polymeric micelles as carriers, since they present all the features of an ideal delivery system.

Abstract

A high percentage of the cancer patients present a mutation in a well-known gene called Kirsten rat sarcoma viral oncogene homolog (KRAS). When this mutation is present, there is an aberrant activation of the KRAS protein that will activate molecular cascades responsible for cell malignancy and tumor development. Unfortunately, until now any study as succeed to develop effective therapies specifically against the KRAS mutated cancers. We believe that the most efficient and specific way to achieve this objective is to deliver intracellularly a specific antibody against the KRAS isoform present in each tumor. The antibodies by itself do not have the capacity to internalize into the cells and required the help of carriers with a nanometric size. We will use polymeric micelles as carriers, since they present all the features of an ideal delivery system.

Abstract

The research line in 3D biopriting, regenerative medicine and tissue engineering carried out by the NanoBioCel Group is presented, firstly describing what is meant by advanced therapies and how this therapeutic approach can be carried out by 3D printing. In order to develop these lines of research, the Joint Research Laboratory (JRL) in advanced pharmaceutical therapies has been established between the nanobiocel group and the Tecnalia Corporation with the support of the Provincial Council of Alava. A presentation of the existing equipment in the Group with the different 3dbiopriting technologies is carried out, to finish with an overview of the projects in which the group is currently involved.

Abstract

The development of non-viral vectors for gene therapy is only a research line of NanoBioCel group, but has led to almost 40 scientific papers in 8 years of research in this exciting field in collaboration with other CIBER-BBN groups. Although the majority of current gene therapy drugs in the market are viral vectors, they show safety concerns and high cost of production. Hence, the development of non-viral vectors is in the spotlight of research community. The first step in their design, consist in verify that they fulfil suitable physicochemical parameters for gene therapy purposes, followed by in vitro experiments to assess their transfection efficiency and biocompatibility and, finally, the translation to in vivo administrations. Our short-term challenges include 1)developing 3D bioprinted tissues to test our non-viral vectors, 2)using microfluidics and 3)green technology to elaborate our non-viral vectors for gene delivery.

Abstract

Upon COVID-19 outbreak, extraordinary research efforts rapidly focused on developing accurate diagnostics and efficient vaccines against SARS-CoV-2 infection. However, there still is a concerning lack of specific anti-viral therapies able to prevent or reduce clinical complications in patients. Monoclonal Antibodies (mAbs) have been profiled as the most promising immunotherapy, capable of blocking SARS-CoV-2 infection at very early stages. Despite their great benefits, a major limitation hampers the implementation in public healthcare systems: a high costly and lengthy production process that results in excessively expensive treatments. We introduce plasmonic biosensors as a potential accelerator in the production of COVID-19 immunotherapies, simplifying the screening and evaluation of neutralizing mAbs through biomimetic label-free analysis. Our plasmonic sensors are biofunctionalized with an artificial cell membrane tethered with human ACE-2 receptors. The device can monitor and quantify the interaction with SARS-CoV-2 viruses and the blocking effect of neutralizing antibodies in just a few minutes and using a low amount of sample. The biosensor could be employed for the rapid screening of different mAb designs, dose-response studies, and efficacy tests towards new variants of SARS-CoV-2.

Abstract

Different mechanisms for the acquisition of optical activity in plasmonic systems are presented in the context of recent works developed in the Bionanoplasmonics lab. On one hand, these strategies include the colloidal synthesis of intrinsically chiral gold nanoparticles. In that regard, the group has reported a novel seed-mediated strategy able to produce chiral nanorods with tunable circular dichroism signals. On another hand, chiral self-assembly of achiral nanoparticles can promote collective chirality from plasmonic coupling between neighboring nanoparticles. Self-assembly of gold nanorods with amyloid fibers driven by electrostatic interactions proved to satisfy the requirements for plasmonic optical activity to arise. Such principle could be applied for the detection of amyloid aggregates in Parkinson's disease-affected brain samples.With such tools in mind, examples reported in literature are brought into consideration as a starting point for the formulation of new biosensing strategies.

Abstract

Pancreatic ductal adenocarcinoma (PDAC) is a dismal disease with a very bad prognosis, even when diagnosed early. Characterized by a dense desmoplastic stroma that inhibits drug penetration. In this sense, the combination of chemotherapy with hyperthermia (HT) has been proposed as a potential treatment for PDAC. In this work, the efficacy of nanoparticle-mediated hyperthermia in combination with gemcitabine and nab-Paclitaxel was in vivo studied. In vivo, biodistribution assays showed that nanoparticle remained mainly within the tumor after intratumoral administration, mainly concentrated around areas with high stromal component. Moreover, the combination of hyperthermia treatment with sub-optimal doses of gemcitabine and nab-Paclitaxel in BxPC-3 subcutaneous tumors, showed clear advantages over the use of chemotherapy treatment alone. Based in these studies, clinical investigational studies for Class III medical devices have already been approved for the nanoparticle and the magnetic field generator.

Abstract

Fabry disease (FD) is caused by a deficient activity of α-galactosidase A (GLA) enzyme that leads to the accumulation of glycosphingolipids, mainly globotriaosylceramide (Gb3). Fabry patients are mainly treated with the enzymatic replacement therapy (ERT) that comes with some limitations such the difficulty to reach some target tissues. In order to overcome these limitations, targeted nanoliposomes containing GLA were synthesized following the DELOS-SUSP methodology. We demonstrated that GLA-nanoliposomes (nanoGLA) reduced the Gb3 accumulation more efficiently than the free enzyme in cell cultures. Based in this good results we then tested the in vivo efficacy of nanoGLA in GLA KO Fabry mice after single dose or repeated dose administrations of 1 mg/Kg of GLA protein. After a single and repeated administration, nanoGLA reduced Gb3 levels of liver, spleen, kidneys, heart and brain more efficiently than the free enzyme.

Abstract

Amyotrophic lateral sclerosis (ALS) is a rapidly progressive degenerative motor neuron disease without effective treatment and without an accepted prognostic or diagnostic biomarker. Analysis of lipid metabolism has yielded many disease biomarkers with additional information on the underlying biological processes in a wide range of psychiatric and neurodegenerative disorders, including Alzheimer's and Parkinson's disease. Regarding ALS clinical and experimental studies have already shown that lipids also play an important role in the neurodegenerative process of ALS, and even blood cholesterol levels have been proposed to predict survival. Additionally, some lipid-lowering strategies have been tried in disease with early promising results. In a previous study in which we performed a comprehensive analysis of the blood lipidomic profile of ALS patients, we reported imbalances in long-chain fatty acids that suggested specific alterations in the use or production of such biomolecules. Subsequently, through the specific analysis of the relative amount of the different fatty acids in the lipid fraction in a follow-up cohort analyzed by complex computational methods, we were able to identify the participation of fatty acid elongation in the progression of ALS disease. A recent publication in the prestigious journal Nature has revealed a putative key role of very long chain saturated fatty acids in the toxicity exerted by the glia in neurodegenerative processes, postulating that the inhibition of elongation could be a first class therapy against ALS. In our hands we have observed that both genetic and pharmacological inhibition with a potent, selective, and orally bioavailable fatty acids elongation inhibitor, improves consistently neuromuscular phenotypes and life expectancy in Drosophila Melanogaster ALS models induced by loss of function of TDP-43.

Key references

Guttenplan et al. Neurotoxic reactive astrocytes induce cell death via saturated lipids. Nature 2021 Oct 6. doi: 10.1038/s41586-021-03960-y.

Abstract

IMPaCT-Data is one of the three pillars of IMPaCT funded by the ISCIII. The objective of IMPaCT-Data is to develop the first iteration of the Infrastructure and the necessary protocols to coordinate, integrate, manage and analyze clinical, medical imaging and genomic data, so that it is possible to implement Personalized Medicine in an effective and coordinated way.

The technical objectives should be understood as prototyping and testing of data management and analysis systems. These resources to the use cases derived from the activities of the other IMPaCT.

• First iteration of a federated data platform.
• Initial version of a cloud computing infrastructure.
• Implementation of data analysis, evaluation and FAIRification protocols.
• Initial systems for the extraction of information from health information systems.
• Extraction of quantitative information from medical images.
• Incorporation of genetic and genomic information.
• Portfolio of bioinformatics resources offered by Spain to ELIXIR.
• Initial systems for data integration.
• Implementation and evaluation of "demonstrators" of advanced interoperability functions.