Ana Moya

Prototype of a Nanostructured Sensing Contact Lens for Noninvasive Intraocular Pressure Monitoring

PURPOSE To present the application of a new sensor based on a flexible, highly piezoresistive, nanocomposite, all-organic bilayer (BL) adapted to a contact lens (CL) for non-invasive monitoring intraocular pressure (IOP). METHODS A prototype of a sensing CL, adapted to a pig eyeball, was tested on different enucleated pig eyes. A rigid, gas-permeable CL was designed as a doughnut shape with a 3-mm hole, where the BL film-based sensor was incorporated. The sensor was a polycarbonate film coated with a polycrystalline layer of the highly piezoresistive molecular conductor β-(ET)₂I₃, which can detect deformations caused by pressure changes of 1 mm Hg. The pig eyeballs were subjected to controlled-pressure variations (low-pressure transducer) to register the electrical resistance response of the CL sensor to pressure changes. Similarly, a CL sensor was designed according to the anatomic characteristics of the eye of a volunteer on the research team. RESULTS A good correlation (r² = 0.99) was demonstrated between the sensing CL electrical response, and IOP (mm Hg) changes in pig eyes, with a sensitivity of 0.4 Ω/mm Hg. A human eye test also showed the high potential of this new sensor (IOP variations caused by eye massage, blinking, and eye movements were registered). CONCLUSIONS A new nanostructured sensing CL for continuous monitoring of IOP was validated in an in vitro model (porcine eyeball) and in a human eye. This prototype has adequate sensitivity to continuously monitor IOP. This device will be useful for glaucoma diagnosis and treatment.

Simultaneous monitoring of Staphylococcus aureus growth in a multi-parametric microfluidic platform using microscopy and impedance spectroscopy.

We describe the design, construction, and characterization of a scalable microfluidic platform that allows continuous monitoring of biofilm proliferation under shear stress conditions. Compared to other previous end-point assay studies, our platform offers the advantages of integration into multiple environments allowing simultaneous optical microscopy and impedance spectroscopy measurements. In this work we report a multi-parametric sensor that can monitor the growth and activity of a biofilm. This was possible by combining two interdigitated microelectrodes (IDuEs), and punctual electrodes to measure dissolved oxygen, K+, Na+ and pH. The IDuE has been optimized to permit sensitive and reliable impedance monitoring of Staphylococcus aureus V329 growth with two- and four-electrode measurements. We distinguished structural and morphological changes on intact cellular specimens using four-electrode data modeling. We also detected antibiotic mediated effects using impedance. Results were confirmed by scanning electrode microscopy and fluorescence microscopy after live/dead cell staining. The bacitracin mediated effects detected with impedance prove that the approach described can be used for guiding the development of novel anti-biofilm agents to better address bacterial infection.

Biofilm dynamics characterization using a novel DO-MEA sensor: mass transport and biokinetics

Biodegradation process modeling is an essential tool for the optimization of biotechnologies related to gaseous pollutant treatment. In these technologies, the predominant role of biofilm, particularly under conditions of no mass transfer limitations, results in a need to determine what processes are occurring within the same. By measuring the interior of the biofilms, an increased knowledge of mass transport and biodegradation processes may be attained. This information is useful in order to develop more reliable models that take biofilm heterogeneity into account. In this study, a new methodology, based on a novel dissolved oxygen (DO) and mass transport microelectronic array (MEA) sensor, is presented in order to characterize a biofilm. Utilizing the MEA sensor, designed to obtain DO and diffusivity profiles with a single measurement, it was possible to obtain distributions of oxygen diffusivity and biokinetic parameters along a biofilm grown in a flat plate bioreactor (FPB). The results obtained for oxygen diffusivity, estimated from oxygenation profiles and direct measurements, revealed that changes in its distribution were reduced when increasing the liquid flow rate. It was also possible to observe the effect of biofilm heterogeneity through biokinetic parameters, estimated using the DO profiles. Biokinetic parameters, including maximum specific growth rate, the Monod half-saturation coefficient of oxygen, and the maintenance coefficient for oxygen which showed a marked variation across the biofilm, suggest that a tool that considers the heterogeneity of biofilms is essential for the optimization of biotechnologies.

Resemblance of the human liver sinusoid in a fluidic device with biomedical and pharmaceutical applications: ORTEGA-RIBERA et al.

Maintenance of the complex phenotype of primary hepatocytes in vitro represents a limitation for developing liver support systems and reliable tools for biomedical research and drug screening. We herein aimed at developing a biosystem able to preserve human and rodent hepatocytes phenotype in vitro based on the main characteristics of the liver sinusoid: unique cellular architecture, endothelial biodynamic stimulation, and parenchymal zonation. Primary hepatocytes and liver sinusoidal endothelial cells (LSEC) were isolated from control and cirrhotic human or control rat livers and cultured in conventional in vitro platforms or within our liver‐resembling device. Hepatocytes phenotype, function, and response to hepatotoxic drugs were analyzed. Results evidenced that mimicking the in vivo sinusoidal environment within our biosystem, primary human and rat hepatocytes cocultured with functional LSEC maintained morphology and showed high albumin and urea production, enhanced cytochrome P450 family 3 subfamily A member 4 (CYP3A4) activity, and maintained expression of hepatocyte nuclear factor 4 alpha (hnf4α) and transporters, showing delayed hepatocyte dedifferentiation. In addition, differentiated hepatocytes cultured within this liver‐resembling device responded to acute treatment with known hepatotoxic drugs significantly different from those seen in conventional culture platforms. In conclusion, this study describes a new bioengineered device that mimics the human sinusoid in vitro, representing a novel method to study liver diseases and toxicology.

Inkjet-printed dissolved oxygen and pH sensors on flexible plastic substrates

There are a broad range of applications such as analytical sensors, biosensing and medical applications that require the monitoring of dissolved oxygen (DO) and pH using sensitive, stable, compact and low cost sensors. Here we develop full inkjet printing sensors to measure DO and pH. They have been fabricated using commercially available gold and platinum inks in plastic substrates. The inks are specially designed formulation which allows their sintering at temperatures as low as 150 and 190 °C for Au and Pt respectively. This is a key point in the development of low-cost sensors made on plastic and paper substrates. These sensors integrate in a single platform all the basic elements for pH and DO recording, allowing the measures without any external electrode. The DO is directly measured with a gold working electrode, and the pH sensors is achieved after electrodepositing iridium oxide film over platinum working electrode. The printed electrodes for DO sensing exhibits excellent linearity between 0 and 8 mg L _ 1 range, with correlation factors greater than 0.99, obtaining low limits of detection, 0.17 mgL _ 1 and a sensitivity of 0.06 A(mgL) _ 1. IrOx pH sensors exhibit a super-Nernstian response in sensitivity repeatedly and reversibly between 65 mV/pH in the pH range of 3 to 10. This work demonstrates that these sensors are suitable for the determination of DO and pH and provide a cost-effective solution for future electrochemical monitoring systems.

Inkjet-printed sulfide-selective electrode

Inkjet printing technology has emerged as an alternative manufacturing method for low-cost production of electrodes. Despite significant progress, there is still a lack in the production of ion-selective electrodes. Herein, the two-step fabrication of the first inkjet-printed sulfide-selective electrode (IPSSE) is described. The two-step fabrication consists of printing a silver electrode followed by an electrochemical deposition of sulfide to produce a second kind electrode (Ag/Ag2S). The performance of this novel device was tested using potentiometric measurements. Nernstian response (-29.4 ± 0.3 mV·decade-1) was obtained within concentrations of 0.03-50 mM with a response time of ∼3 s. Furthermore, river/sea-spiked environmental samples and samples from a bioreactor for sulfate reduction to sulfide were measured and compared against a commercial sensor giving no significant differences. The IPSSE described in this work showed good reproducibility and durability during daily measurements over 15 days without any special storage conditions. Considering all the current challenges in inkjet-printed ion-selective electrodes, this different fabrication approach opens a new perspective for mass production of all-solid state ion-selective electrodes.

Flexible microfluidic bio-lab-on-a-chip multi-sensor platform for electrochemical measurements

A novel polymeric microfluidic multi-sensor system has been developed using rapid prototyping techniques and low-cost materials which permits the dynamic online monitoring of several cell culture parameters in a wide range of biomedical applications. The multi-sensor is made of polymeric flexible substrate with integrated gold microband electrodes for electrochemical measurements of Dissolved Oxygen (DO) and Hydrogen (H+), Sodium (Na+) and Potassium (K+) cations. The platform integrates several working electrodes (WE), pseudoreferences (pRE) and a counter electrode (CE) for electrochemical detection. The bio-lab-on-a-chip allows the detection of changes in real time, with rapid response, allowing to measure small volumes and fast cell metabolisms changes. The reliable analytical performance of the platform in in-vitro conditions has been demonstrated. DO was detected with activated gold electrode, and H+, Na+ and K+ were detected with the developed ion-selective microelectrodes (μISE) using electropolymerized polypyrrole (PPy) film as a contact layer, allowing a good performance of the sensor.