The demand for real-time monitoring of cell functions and cell conditions has dramatically increased with the emergence of organ-on-a-chip (OOC) systems. However, the incorporation of co-cultures and microfluidic channels in OOC systems increases their biological complexity and therefore makes the analysis and monitoring of analytical parameters inside the device more difficult. In this work, we present an approach to integrate multiple sensors in an extremely thin, porous and delicate membrane inside a liver-on-a-chip device. Specifically, three electrochemical dissolved oxygen (DO) sensors were inkjet-printed along the microfluidic channel allowing local online monitoring of oxygen concentrations. This approach demonstrates the existence of an oxygen gradient up to 17.5% for rat hepatocytes and 32.5% for human hepatocytes along the bottom channel. Such gradients are considered crucial for the appearance of zonation of the liver. Inkjet printing (IJP) was the selected technology as it allows drop on demand material deposition compatible with delicate substrates, as used in this study, which cannot withstand temperatures higher than 130 °C. For the deposition of uniform gold and silver conductive inks on the porous membrane, a primer layer using SU-8 dielectric material was used to seal the porosity of the membrane at defined areas, with the aim of building a uniform sensor device. As a proof-of-concept, experiments with cell cultures of primary human and rat hepatocytes were performed, and oxygen consumption rate was stimulated with carbonyl-cyanide-4-(trifluoromethoxy)phenylhydrazone (FCCP), accelerating the basal respiration of 0.23 ± 0.07 nmol s-1/106 cells up to 5.95 ± 0.67 nmol s-1/106 cells s for rat cells and the basal respiration of 0.17 ± 0.10 nmol s-1/106 cells by up to 10.62 ± 1.15 nmol s-1/106 cells for human cells, with higher oxygen consumption of the cells seeded at the outflow zone. These results demonstrate that the approach of printing sensors inside an OOC has tremendous potential because IJP is a feasible technique for the integration of different sensors for evaluating metabolic activity of cells, and overcomes one of the major challenges still remaining on how to tap the full potential of OOC systems.

A novel microelectrode array sensor was fabricated using MEMS technology on a needle, and then applied to real-time measurement of dissolved oxygen (DO) inside biofilms. The sensor consists of eleven gold disk microelectrodes and a rectangular auxiliary electrode along them and an external & internal reference electrode. Three kinds of sensors were designed and their responses were characterized and evaluated under various environmental conditions. The arrays exhibit a linear response to DO in the 0 -8 mg L -1 concentration range in water, high sensitivity, repeatability, and low limits of detection (< 0.11 mg L -1 ) and quantification (0.38 mg L -1 ). The sensors were then validated against a commercial Clark-type microelectrode and applied to profiling of DO in a heterotrophic biofilm cultivated in a flat-plate bioreactor. It is shown that the sensor array can provide a multipoint, simultaneous snapshot profile of DO inside a biofilm with high spatial resolution due to its micrometric dimensions. We conclude that this new sensor array represents a powerful tool for sensing of DO biofilms and in numerous bioprocesses.

Biodegradation process modeling is an essential tool for the optimization of biotechnologies related to gaseous pollutants 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.2

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The purpose of this work was to evaluate the technical and economical feasibility of converting three in-series chemical scrubbers to biotrickling filters for the simultaneous removal at neutral pH of 10000 m 3 /h containing H 2 S and volatile organic compounds (VOCs) at a wastewater treatment plant (WWTP). The conversion was based on previous conversion protocols and mainly required of replacing the original packing material by a structured packing and recycle pumps, besides modifying the controls and operation of the reactors. Complete removal of H 2 S and VOCs on a routine basis was reached in a longer period of time compared to previous conversions reported. Under the conditions at the WWTP, i.e., a gas contact time of about 1.4 seconds per reactor and pH controlled between 6.5 and 6.8, inlet average concentrations below 10 ppm v of H 2 S and below 5 ppm v for VOCs were often completely degraded in the reactors. The first and second reactors played a major role in H 2 S removal. Long-term operation of the biotrickling filters proved the ability of the biotrickling filter to handle progressive load variations of H 2 S and VOCs. However, fast, sudden load changes often lead to reduced removal efficciencies. Odor removal showed average efficiencies above 80%. Limited outlet odor concentration was attributed to limited removal of VOCs, which was assessed through gas chromatographymass spectrometry of selected gas samples. The conversion showed as economically viable taking into account the theoretical consumption of chemicals needed for the absorption and oxidation of both H 2 S and VOCs.

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A novel sensing device for simultaneous dissolved oxygen (DO) and pH monitoring specially designed for biofilm profiling is presented in this work. This device enabled the recording of instantaneous DO and pH dynamic profiles within biofilms, improving the tools available for the study and the characterization of biological systems. The microsensor consisted of two parallel arrays of microelectrodes. Microelectrodes used for DO sensing were bare gold electrodes, while microelectrodes used for pH sensing were platinum-based electrodes modified using electrodeposited iridium oxide. The device was fabricated with a polyimide (Kapton®) film of 127 µm as a substrate for minimizing the damage caused on the biofilm structure during its insertion. The electrodes were covered with a Nafion® layer to increase sensor stability and repeatability and to avoid electrode surface fouling. DO microelectrodes showed a linear response in the range 0–8 mg L−1, a detection limit of 0.05 mg L−1, and a sensitivity of 2.06 nA L mg−1. pH electrodes showed a linear super-Nernstian response (74.2 ± 0.7 mV/pH unit) in a wide pH range (pH 4−9). The multi-analyte sensor array was validated in a flat plate bioreactor where simultaneous and instantaneous pH and DO profiles within a sulfide oxidizing biofilm were recorded. The electrodes spatial resolution, the monitoring sensitivity, and the minimally invasive features exhibited by the proposed microsensor improved biofilm monitoring performance, enabling the quantification of mass transfer resistances and the assessment of biological activity.