Biosensors for bioengineering

The Biosensors for bioengineering group is a junior group under IBEC’s Tenure Track scheme.
Javier Ramón Azcón | Group Leader / ICREA Research Professor
Irene Marco Rius | Senior Researcher
Francesco De Chiara | Postdoctoral Researcher
Juanma Fernandez Costa | Postdoctoral Researcher
Gerardo López Muñoz | 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
Alba Herrero Gómez | Laboratory Technician
Eduard Martin Lasierra | Masters Student
Ainoa Tejedera Villafranca | Masters Student
Jordina Balaguer Trias | Laboratory Assistant
Ainhoa Ferret Miñana | Laboratory Assistant
Rodrigo Alvarez Velasco | Visiting Researcher


Formation of 3D ESC aggregates in GelMA hydrogel using dielectrophoresis (DEP). The stem cells in the GelMA prepolymer were introduced into the 100 µm height chamber and localized by DEP forces to the low electric field regions within the microelectrodes. The GelMA prepolymer was then exposed to UV light, embedding the cells in a stable microscale organization. Scale bar shows 100 µm.

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.

The 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.


Right: Picture of aligned C2C12 muscle cells within hydrogel as obtained by the dielectrophoresis (DEP) technique using 50 µm electrode 50 µm gap device (A). Phase contrast images of the aligned C2C12 muscle cells within hydrogel at different culture times (B and C). Scale bar shows 0.25 cm, 400 μm, and 50 μm in A, B, and C, respectively.


Myotubes differentiated in a groove-ridge topography GelMA-CNTs composite loaded with 0.3 mg/mL CNTs. Immunostaining of cell nuclei/myosin heavy chain showing the highly aligned C2C12 myotubes. Z-lines were also observed for the myotubes indicating high maturation of muscle myofibers. Scale bar show 20 µm.

Myotubes differentiated in a groove-ridge topography GelMA-CNTs composite loaded with 0.3 mg/mL CNTs. Immunostaining of cell nuclei/myosin heavy chain showing the highly aligned C2C12 myotubes. Z-lines were also observed for the myotubes indicating high maturation of muscle myofibers. Scale bar show 20 µm.



IBEC to launch Faster Future, a new fundraising initiative, on Giving Tuesday

Next week IBEC will launch Faster Future, a new crowdfunding initiative that aims to help accelerate research projects that are close to tackling major challenges in health.

Finding out about diabetes approaches at IBEC’s Setmana de la Ciencia event

Yesterday IBEC group leader Javer Ramon presented his research on developing organ-on-a-chip to study diabetes in IBEC’s public event forming part of this year’s Setmana de la Ciència.

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.

ERC funding for new diabetes approach at IBEC

IBEC’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).


EU-funded projects
‘Diabetes Approach by Multi-Organ-on-a-Chip’ (DAMOC) ERC Javier Ramón


Gómez-Domínguez, D., Epifano, C., Miguel, F., Castaño, A. G., Vilaplana-Martí, B., Martín, A., Amarilla-Quintana, S., Bertrand, A. T., Bonne, G., Ramón-Azcón, J., Rodríguez-Milla, M. A., Pérez de Castro, I., (2020). Consequences of Lmna exon 4 mutations in myoblast function Cells 9, (5), 1286

Laminopathies are causally associated with mutations on the Lamin A/C gene (LMNA). To date, more than 400 mutations in LMNA have been reported in patients. These mutations are widely distributed throughout the entire gene and are associated with a wide range of phenotypes. Unfortunately, little is known about the mechanisms underlying the effect of the majority of these mutations. This is the case of more than 40 mutations that are located at exon 4. Using CRISPR/Cas9 technology, we generated a collection of Lmna exon 4 mutants in mouse C2C12 myoblasts. These cell models included different types of exon 4 deletions and the presence of R249W mutation, one of the human variants associated with a severe type of laminopathy, LMNA-associated congenital muscular dystrophy (L-CMD). We characterized these clones by measuring their nuclear circularity, myogenic differentiation capacity in 2D and 3D conditions, DNA damage, and levels of p-ERK and p-AKT (phosphorylated Mitogen-Activated Protein Kinase 1/3 and AKT serine/threonine kinase 1). Our results indicated that Lmna exon 4 mutants showed abnormal nuclear morphology. In addition, levels and/or subcellular localization of different members of the lamin and LINC (LInker of Nucleoskeleton and Cytoskeleton) complex were altered in all these mutants. Whereas no significant differences were observed for ERK and AKT activities, the accumulation of DNA damage was associated to the Lmna p.R249W mutant myoblasts. Finally, significant myogenic differentiation defects were detected in the Lmna exon 4 mutants. These results have key implications in the development of future therapeutic strategies for the treatment of laminopathies.

Keywords: CRISPR, Laminopathy, LMNA, Nuclear envelope

Trueba-Santiso, A., Fernández-Verdejo, D., Marco-Rius, I., Soder-Walz, J. M., Casabella, O., Vicent, T., Marco-Urrea, E., (2020). Interspecies interaction and effect of co-contaminants in an anaerobic dichloromethane-degrading culture Chemosphere 240, 124877

An anaerobic stable mixed culture dominated by bacteria belonging to the genera Dehalobacterium, Acetobacterium, Desulfovibrio, and Wolinella was used as a model to study the microbial interactions during DCM degradation. Physiological studies indicated that DCM was degraded in this mixed culture at least in a three-step process: i) fermentation of DCM to acetate and formate, ii) formate oxidation to CO2 and H2, and iii) H2/CO2 reductive acetogenesis. The 16S rRNA gene sequencing of cultures enriched with formate or H2 showed that Desulfovibrio was the dominant population followed by Acetobacterium, but sequences representing Dehalobacterium were only present in cultures amended with DCM. Nuclear magnetic resonance analyses confirmed that acetate produced from 13C-labelled DCM was marked at the methyl ([2–13C]acetate), carboxyl ([1–13C]acetate), and both ([1,2–13C]acetate) positions, which is in accordance to acetate formed by both direct DCM fermentation and H2/CO2 acetogenesis. The inhibitory effect of ten different co-contaminants frequently detected in groundwaters on DCM degradation was also investigated. Complete inhibition of DCM degradation was observed when chloroform, perfluorooctanesulfonic acid, and diuron were added at 838, 400, and 107 μM, respectively. However, the inhibited cultures recovered the DCM degradation capability when transferred to fresh medium without co-contaminants. Findings derived from this work are of significant relevance to provide a better understanding of the synergistic interactions among bacteria to accomplish DCM degradation as well as to predict the effect of co-contaminants during anaerobic DCM bioremediation in groundwater. © 2019 Elsevier Ltd

Keywords: Bioremediation, Co-contaminants, Dehalobacterium, Dichloromethane, Inhibition

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 2, 2885-2896

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.

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

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 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

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

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)


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)


  • 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)


  • Prof. Josep Samitier
  • Dr. Elena Martinez
  • 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