Biosensors for bioengineering


The Biosensors for bioengineering group is a junior group under IBEC’s Tenure Track scheme.

 

Javier Ramón Azcón | Junior Group Leader ICREA
Francesco De Chiara | Postdoctoral Researcher
Juanma Fernandez Costa | Postdoctoral Researcher
Gerardo López Muñoz | Postdoctoral Researcher
Irene Marco Rius | Postdoctoral Researcher
Maria Alejandra Ortega Machuca | Postdoctoral Researcher
Júlia Rodríguez Comas | Postdoctoral Researcher
José Yeste Lozano | Postdoctoral Researcher
Marc Azagra Rodríguez | PhD Student
Laura Clua Ferré | PhD Student
Xiomara Gislen Fernández Garibay | PhD Student
Ferran Velasco Mallorquí | PhD Student
Natalia Kovaleva Dorozhkina | Masters Student
Jordina Balaguer Trias | Laboratory Assistant
Ainhoa Ferret Miñana | Laboratory Assistant
Rodrigo Alvarez Velasco | Visiting Researcher

 

About

Drug discovery pathway relies heavily on in vivo animal models and in vitro cell mediums. In the case of animal models we have not only some ethical problems but also the ability to extrapolate data to human conditions is limited and in vitro platforms often do not simulate the complex cell–cell and cell–matrix interactions crucial for regulating cell behaviour.  

Pancreas islet stained in blue for nuclei and red for actin inside a microporous microfibrillated cellulose-gelatin scaffold stained with green.

Biosensors for bioengineering group is focused in a new line of research that has become of extreme importance in the last years. The idea is to integrate biosensor technology and nanotechnology with stem cell research and with tissue engineering. Engineered tissues are integrated with biosensing technology to obtain microdevices for detecting cellular responses to external stimuli, monitoring the quality of the microenvironment (e.g., metabolites, nutrients), and supporting diverse cellular requirements. This research on 3D-functional engineered tissues is expected to develop knowledge of tissue construction and their functions and relation with some human diseases. Integration of fully functional tissues with microscale biosensor technology allowed us to obtain “organs-on-a-chip”. These chips could be used in pharmaceutical assays and could be a step toward the ultimate goal of producing in vitro drug testing systems crucial to the medicine and pharmaceutical industry. 

Engineered skeletal muscle from patients’ cells with myotonic dystrophy disease.

Human myotubes encapsulated in micropatterned hydrogel scaffold. Scale is 100 µm.

 

 

 

 

 

 

 

 

 

 

 

 

Projects

EU-funded projects
DAMOC · ‘Diabetes Approach by Multi-Organ-on-a-Chip’ (2017-2021) ERC Javier Ramón
BLOC · Benchtop NMR for Lab-on-Chip (2020-2022) European Comission FET-Open Javier Ramón
National Projects
INDUCT · Fabrication of a biomimetic in vitro model of the intestinal tube muscle wall: smooth muscle-on-a-chip (2018-2020) MINECO Javier Ramón
Privately funded projects
Tatami · Therapeutic targeting of MBNL microRNAs as innovative treatments for myotonic dystrophy Fundació bancaria “La Caixa” Javier Ramón

Publications


Velasco, Ferran, Fernandez-Costa, Juan M., Neves, Luisa, Ramon Azcon, Javier, (2020). Volumetric CNT-doped Gelatin-Cellulose scaffold for skeletal muscle tissue engineering Nanoscale Advances ahead,

Currently, the fabrication of scaffolds for engineered skeletal muscle tissue is unable to reach the millimeter size. The main drawbacks are the poor nutrients diffusion, lack of internal structure to align precursor cells as well as poor mechanical and electric properties. Herein, we present a combination of gelatin-carboxymethyl cellulose materials polymerised by a cryogelation process that allowed us to reach scaffold fabrication up to millimeters size and solve the main problems related with large size muscle tissue constructs. 1) By incorporating carbon nanotubes (CNT) we can improve the electrical properties of the scaffold, thereby enhancing tissue maturation when applying an electric pulse stimulus (EPS). 2) We have fabricated an anisotropic internal three-dimensional microarchitecture pore distribution with high aligned morphology to enhance cells alignment, cell fusion and myotubes formation. With this set up, we were able to generate a fully functional skeletal muscle tissue using a combination of EPS and our doped-biocomposite scaffold and obtain a mature tissue in a millimeter scale. We also characterized pore distribution, swelling, stiffness and conductivity of the scaffold. Moreover, we proved that the cells are viable and able to fuse in a three-dimensional (3D) functional myotubes throughout the scaffold. In conclusion, we fabricate a biocompatible and customizable scaffold for 3D cell culture suitable for a wide range of application such as organ-on-a-chip, drug screening, transplantation and disease modelling.


Ortega, María A., Fernández-Garibay, Xiomara, Castaño, Albert G., De Chiara, Francesco, Hernández-Albors, Alejandro, Balaguer-Trias, Jordina, Ramón-Azcón, Javier, (2019). Muscle-on-a-chip with an on-site multiplexed biosensing system for in situ monitoring of secreted IL-6 and TNF-α Lab on a Chip 19, 2568-2580

Despite the increasing number of organs-on-a-chip that have been developed in the past decade, limited efforts have been made to integrate a sensing system for in situ continual measurements of biomarkers from three-dimensional (3D) tissues. Here, we present a custom-made integrated platform for muscle cell stimulation under fluidic conditions connected with a multiplexed high-sensitivity electrochemical sensing system for in situ monitoring. To demonstrate this, we use our system to measure the release levels and release time of interleukin 6 and tumor necrosis factor alpha in vitro by 3D muscle microtissue under electrical and biological stimulations. Our experimental design has enabled us to perform multiple time point measurements using functionalized screen-printed gold electrodes with sensitivity in the ng mL−1 range. This affordable setup is uniquely suited for monitoring factors released by 3D single cell types upon external stimulation for metabolic studies.


de Goede, M., Dijkstra, M., Obregón, R., Ramón-Azcón, J., Martínez, Elena, Padilla, L., Mitjans, F., Garcia-Blanco, S. M., (2019). Al2O3 microring resonators for the detection of a cancer biomarker in undiluted urine Optics Express 27, (13), 18508-18521

Concentrations down to 3 nM of the rhS100A4 protein, associated with human tumor development, have been detected in undiluted urine using an integrated sensor based on microring resonators in the emerging Al2O3 photonic platform. The fabricated microrings were designed for operation in the C-band (λ = 1565 nm) and exhibited a high-quality factor in air of 3.2 × 105. The bulk refractive index sensitivity of the devices was ~100 nm/RIU (for TM polarization) with a limit of detection of ~10−6 RIU. A surface functionalization protocol was developed to allow for the selective binding of the monoclonal antibodies designed to capture the target biomarker to the surface of the Al2O3 microrings. The detection of rhS100A4 proteins at clinically relevant concentrations in urine is a big milestone towards the use of biosensors for the screening and early diagnosis of different cancers. Biosensors based on this microring technology can lead to portable, multiplexed and easy-to-use point of care devices.

Keywords: Distributed feedback lasers, Effective refractive index, Laser coupling, Polarization maintaining fibers, Refractive index, Scanning electron microscopy


De Chiara, F., Checcllo, C. U., Ramón-Azcón, J., (2019). High protein diet and metabolic plasticity in non-alcoholic fatty liver disease: Myths and truths Nutrients 11, (12), 2985

Non-alcoholic fatty liver disease (NAFLD) is characterized by lipid accumulation within the liver affecting 1 in 4 people worldwide. As the new silent killer of the twenty-first century, NAFLD impacts on both the request and the availability of new liver donors. The liver is the first line of defense against endogenous and exogenous metabolites and toxins. It also retains the ability to switch between different metabolic pathways according to food type and availability. This ability becomes a disadvantage in obesogenic societies where most people choose a diet based on fats and carbohydrates while ignoring vitamins and fiber. The chronic exposure to fats and carbohydrates induces dramatic changes in the liver zonation and triggers the development of insulin resistance. Common believes on NAFLD and different diets are based either on epidemiological studies, or meta-analysis, which are not controlled evidences; in most of the cases, they are biased on test-subject type and their lifestyles. The highest success in reverting NAFLD can be attributed to diets based on high protein instead of carbohydrates. In this review, we discuss the impact of NAFLD on body metabolic plasticity. We also present a detailed analysis of the most recent studies that evaluate high-protein diets in NAFLD with a special focus on the liver and the skeletal muscle protein metabolisms.

Keywords: High protein diet, Low carbohydrates, NAFLD, NASH, Physical activity


Hernández-Albors, Alejandro, Castaño, Albert G., Fernández-Garibay, Xiomara, Ortega, María Alejandra, Balaguer, Jordina, Ramón-Azcón, Javier, (2019). Microphysiological sensing platform for an in-situ detection of tissue-secreted cytokines Biosensors and Bioelectronics: X 2, 100025

Understanding the protein-secretion dynamics from single, specific tissues is critical toward the advancement of disease detection and treatments. However, such secretion dynamics remain difficult to measure in vivo due to the uncontrolled contributions from other tissue populations. Here, we describe an integrated platform designed for the reliable, near real-time measurements of cytokines secreted from an in vitro single-tissue model. In our setup, we grow 3D biomimetic tissues to discretize cytokine source, and we separate them from a magnetic microbead-based biosensing system using a Transwell insert. This design integrates physiochemically controlled biological activity, high-sensitivity protein detection (LOD < 20 pg mL−1), and rapid protein diffusion to enable non-invasive, near real-time measurements. To showcase the specificity and sensitivity of the system, we use our setup to probe the inflammatory process related to the protein Interleukine 6 (IL-6) and to the Tumor Necrosis Factor (TNF-α). We show that our setup can monitor the time-dependence profile of IL-6 and TNF-α secretion that results from the electrical and chemical stimulation of 3D skeletal muscle tissues. We demonstrate a novel and affordable methodology for discretizing the secretion kinetics of specific tissues for advancing metabolic-disorder studies and drug-screening applications.

Keywords: Microphysiological tissues, Tissue engineering, Electrochemical, biosensors, Magnetic particles, Skeletal muscle, Electric stimulation


García-Lizarribar, Andrea, Fernández-Garibay, Xiomara, Velasco-Mallorquí, Ferran, Castaño, Albert G., Samitier, Josep, Ramon-Azcon, Javier, (2018). Composite biomaterials as long-lasting scaffolds for 3D bioprinting of highly aligned muscle tissue Macromolecular Bioscience 18, (10), 1800167

Abstract New biocompatible materials have enabled the direct 3D printing of complex functional living tissues, such as skeletal and cardiac muscle. Gelatinmethacryloyl (GelMA) is a photopolymerizable hydrogel composed of natural gelatin functionalized with methacrylic anhydride. However, it is difficult to obtain a single hydrogel that meets all the desirable properties for tissue engineering. In particular, GelMA hydrogels lack versatility in their mechanical properties and lasting 3D structures. In this work, a library of composite biomaterials to obtain versatile, lasting, and mechanically tunable scaffolds are presented. Two polysaccharides, alginate and carboxymethyl cellulose chemically functionalized with methacrylic anhydride, and a synthetic material, such as poly(ethylene glycol) diacrylate are combined with GelMA to obtain photopolymerizable hydrogel blends. Physical properties of the obtained composite hydrogels are screened and optimized for the growth and development of skeletal muscle fibers from C2C12 murine cells, and compared with pristine GelMA. All these composites show high resistance to degradation maintaining the 3D structure with high fidelity over several weeks. Altogether, in this study a library of biocompatible novel and totally versatile composite biomaterials are developed and characterized, with tunable mechanical properties that give structure and support myotube formation and alignment.


Ino, Kosuke, Nashimoto, Yuji, Taira, Noriko, Ramón-Azcon, Javier, Shiku, Hitoshi, (2018). Intracellular electrochemical sensing Electroanalysis 30, (10), 2195-2209

Observing biochemical processes within living cell is imperative for biological and medical research. Fluoresce imaging is widely used for intracellular sensing of cell membranes, nuclei, lysosomes, and pH. Electrochemical assays have been proposed as an alternative to fluorescence-based assays because of excellent analytical features of electrochemical devices. Notably, thanks to the rapid progress of micro/nanotechnologies and electrochemical techniques, intracellular electrochemical sensing is making rapid progress, leading to a successful detection of intracellular components. Such insight can provide a deep understanding of cellular biological processes and, ultimately, define the human healthy and diseased states. In this review, we present an overview of recent research progress in intracellular electrochemical sensing. We focus on two main topics, electrochemical extraction of cytosolic contents from cells and intracellular electrochemical sensing in situ.

Keywords: Micro/nanoelectrode, Analytical electrochemistry, Intracellular sensing, Cell analysis


de Goede, M., Chang, L., Dijkstra, M., Obregón, R., Ramón-Azcon, J., Martínez, Elena, Padilla, L., Adan, J., Mitjans, F., García-Blanco, S.M., (2018). Al2O3 Microresonator based passive and active biosensors ICTON 2018 20th International Conference on Transparent Optical Networks , IEEE Computer Society (Bucharest, Romania) , 8473820

Al2O3 microresonators were realized for sensing applications of both passive and active devices. Passive microring resonators exhibited quality factors up to 3.2×105 in air. A bulk refractive index sensitivity of 100 nm/RIU was demonstrated together with a limit of detection of 10-6 RIU. Functionalizing their surface allowed for the label-free detection of the biomarker rhS100A4 from urine with a limit of detection of 3 nM. Furthermore, single-mode Al2O3:Yb3+ microdisk lasers were realized that could operate in an aqueous environment. Upon varying the bulk refractive index their lasing wavelength could be tuned with a sensitivity of 20 nm/RIU and a LOD of 3×10-6 RIU.


de Goede, M., Chang, L., Dijkstra, M., Obregón, R., Ramón-Azcon, J., Martínez, Elena, Padilla, L., Adan, J., Mitjans, F., García-Blanco, S.M., (2018). Al2O3 Mmicroresonators for passive and active sensing applications Sensors 2018 Optical Sensors , OSA - The Optical Society (Zurich, Switzerland) Part F110, 1-2

The Al2O3 waveguide technology was explored for sensing applications. Passive microring resonators with a quality factor in air of 3.2×105 were developed with a bulk refractive index sensitivity of ~100 nm/RIU and limit of detection of ~10-6 RIU. These were functionalized to detect the biomarker rhS100A4 from urine down to concentrations of 3 nM. Furthermore, Al2O3:Yb3+ microdisk lasers were realized that exhibited single mode lasing operation in water. Their lasing wavelength was tuned by varying the bulk refractive index and a bulk refractive index sensitivity of ~20 nm/RIU with a LOD of ~3×10-6 was achieved.


Mohammadi, M. H., Obregón, R., Ahadian, S., Ramón-Azcón, J., Radisic, M., (2017). Engineered muscle tissues for disease modeling and drug screening applications Current Pharmaceutical Design , 23, (20), 2991-3004

Animal models have been the main resources for drug discovery and prediction of drugs’ pharmacokinetic responses in the body. However, noticeable drawbacks associated with animal models include high cost, low reproducibility, low physiological similarity to humans, and ethical problems. Engineered tissue models have recently emerged as an alternative or substitute for animal models in drug discovery and testing and disease modeling. In this review, we focus on skeletal muscle and cardiac muscle tissues by first describing their characterization and physiology. Major fabrication technologies (i.e., electrospinning, bioprinting, dielectrophoresis, textile technology, and microfluidics) to make functional muscle tissues are then described. Finally, currently used muscle tissue models in drug screening are reviewed and discussed.

Keywords: Cardiac muscle, Drug screening, Engineering muscle, Human pharmacological response, Physiological similarity, Skeletal muscle


Obregón, R., Ramón-Azcón, J., Ahadian, S., (2017). Nanofiber composites in blood vessel tissue engineering Nanofiber Composites for Biomedical Applications (ed. Ramalingam, M., Ramakrishna, S.), Elsevier (Duxford, UK) Woodhead Publishing Series in Biomaterials, 483-506

Tissue engineering (TE) aims to restore function or replace damaged tissue through biological principles and engineering. Nanofibers are attractive substrates for tissue regeneration applications because they structurally mimic the native extracellular matrix. Composite nanofibers, which are hybrid nanofibers blended from natural and synthetic polymers, represent a major advancement in TE and regenerative medicine, since they take advantage of the physical properties of the synthetic polymer and the bioactivity of the natural polymer while minimizing the disadvantages of both. Although various nanofibrous matrices have been applied to almost all the areas of TE, in this chapter we will focus on nanofiber composites scaffolds for vascular TE.

Keywords: Blood vessels, Nanofiber composite, Tissue engineering, Vascularized tissue



(See full publication list in ORCID)


Equipment


Micro and nanofabrication techniques:

  • 3D microstructures on hydrogel materials
  • Mini-bioreactor for 3D cell culture
  • Microelectrodes fabrication
  • Synthesis and chemical modification of polymers and surfaces
  • Dielectrophoretic cells and micro particles manipulation

Characterization techniques:

  • Optical Microscopes (white light/epifluorescence)
  • Electrochemical techniques (Potentiometric/Amperometric/Impedance spectroscopy)
  • Immunosensing techniques (Fluorescence ELISA/Colorimetric ELISA/magneto ELISA)

Equipment:

  • Microfluidic systems (High precision syringe pumps/Peristaltic pumps/Micro valves)
  • Biological safety cabinet (class II)
  • Epifluorescence microscope for live-cell imaging

Access to the Nanotechnology Platform (IBEC Core Facilities): equipment for hot embossing lithography, polymer processing and photolithography, chemical wet etching, e-beam evaporation and surface characterization (TOF-SIMS)
Access to the Scientific and Technological Centers (University of Barcelona): equipment for surface analysis (XPS, AFM, XRD), organic structures characterization (NMR) and microscopy techniques (SEM, TEM, confocal)

Collaborations

  • Prof. Josep Samitier
    IBEC
  • Dr. Elena Martinez
    IBEC
  • Dr. Anna Novials
    Institut d’Investigacions Biomediques August Pi i Sunyer (IDIBAPS)
  • Dr. Ramon Gomís
    Institut d’Investigacions Biomediques August Pi i Sunyer (IDIBAPS)
  • Dr. Eduard Montanya
    The Bellvitge Biomedical Research Institute (IDIBELL)
  • Prof. Enric Bertran
    Physics and Engineering of Amorphous Materials and Nanostructures (FEMAN), Department of Applied Physics, University of Barcelona

 

News/Jobs

Javier Ramón, new ICREA professor at IBEC

Javier Ramón, Group Leader of the “Biosensors for Bioengineering” group at IBEC, has been appointed new Research Professor at the Catalan Institution for Research and Advanced Studies, ICREA. Currently IBEC hosts 8 ICREA Professors out of 22 group leaders.

Since last April 1, Dr. Javier Ramón, Group Leader at IBEC, has become part of the ICREA Research Professors community. ICREA is a foundation funded by the Catalan Government aimed at recruiting the most extraordinary and talented international scientific researchers. With this appointment, IBEC hosts 8 ICREA Professors and 2 ICREA Academia (UB affiliated professors), one of the highest numbers among all research centres in Catalonia.

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IBEC leads a European Project to evaluate drug response in organ-on-a-chip devices

A group of researchers from the Institute for Bioengineering of Catalonia (IBEC) leads the European project BLOC, an initiative led by researchers Javier Ramón and Irene Marco that seeks to evaluate the response to different drugs in metabolic diseases using organ-in-a- chip by using nuclear magnetic resonance (NMR). For this, the consortium will have a budget of almost 3 million euros, financed by the Horizon 2020 FET Open program.

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IBEC presents its capabilities in 3D bioprinting and in other industrial areas at the fourth INDUSTRY edition

During the 29th to 31st of October, IBEC participated in the fourth edition of “INDUSTRY: From Needs to Solutions”, the international meeting dedicated to 3D printing, and also the HELTHIO Days where Josep Samitier moderated the round table on 3D printing applications in healthcare.

In its fourth edition, IN(3D)USTRY becomes INDUSTRY to include more industrial sectors, the main goal is to cover the entire value chain and provide intelligent manufacturing solutions. The institute had a stand in the exhibition area, where the representatives of the IBEC Technology Transfer office welcomed the interested visitors to learn more about the 3D bioprinting capabilities of IBEC.

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Researchers at IBEC develop a bioengineering platform to detect pro-inflammatory molecules in muscular disorders

The Biosensors for bioengineering group led by Javier Ramón has developed a sensing platform for the in-situ detection of tissue-secreted pro-inflammatory molecules, the so-called cytokines. This new methodology opens a new door in the understanding of metabolic-disorders such those found in muscular diseases, as well as the development of drug-screening applications.

Although 40% of total body mass is skeletal muscle tissue, there is no specialized clinical doctor for the treatment of muscular diseases, according to the American Medical Association. The research group of Dr. Javier Ramón at IBEC works to fill this gap between muscle disorders and ad hoc therapies.

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IBEC researcher’s ERC project highlighted in Madrid exhibition

This weekend Javier Ramon’s European Research Council-funded project, DAMOC, was one of eight highlighted in a special exhibition in Madrid to mark the ERC’s tenth anniversary.

Alcobendas’ Museo Nacional de Ciencia y Tecnología (MUNCYT) displayed the most “outstanding” projects led by researchers in Spain as part of a full weekend of activities to celebrate the first decade of the prestigious funding body, which was launched in 2007 by the European Union and has funded nearly 7,000 researchers, among them six Nobel Prize winners.

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ERC funding for new diabetes approach at IBEC

javierramon2_tiIBEC’s Dr. Javier Ramón is one of just six researchers in Catalonia to have been awarded a 2016 Starting Grant by the European Research Council (ERC).

The senior researcher in the Biomimetic Systems for Cell Engineering group won funding for his project ‘Diabetes Approach by Multi-Organ-on-a-Chip’ (DAMOC) from Europe’s most prestigious funding body.

With the support, which will last for up to five years, Javier will start a new line to design a innovative new tool to test drugs for diabetes. As well as improving drug testing approaches, the multi-organ-on-a-chip device will provide new therapies to prevent the loss of beta cell mass and defects in the glucose uptake in skeletal muscle associated with type 2 diabetes.

“This project will give me the opportunity to have a multidisciplinary group of researchers working together from the beginning in a synchronized way, the most rewarding experience that a researcher can have,” he says.

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