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Cell microencapsulation is an attractive strategy for cell-based therapies that allows the implantation of genetically engineered cells and the continuous delivery of de novo produced therapeutic products. However, the establishment of a way to retrieve the implanted encapsulated cells in case the treatment needs to be halted or when cells need to be renewed is still a big challenge. The combination of micro and macroencapsulation approaches could provide the requirements to achieve a proper immunoisolation, while maintaining the cells localized into the body. We present the development and characterization of a porous implantable macrocapsule device for the loading of microencapsulated cells. The device was fabricated in polyamide by selective laser sintering (SLS), with controlled porosity defined by the design and the sintering conditions. Two types of microencapsulated cells were tested in order to evaluate the suitability of this device; erythropoietin (EPO) producing C2C12 myoblasts and Vascular Endothelial Growth Factor (VEGF) producing BHK fibroblasts. Results showed that, even if the metabolic activity of these cells decreased over time, the levels of therapeutic protein that were produced and, importantly, released to the media were stable. This work was done under the BIOPAN project (CIBER-BBN). Authors wish to thank the intellectual and technical assistance from the ICTS "NANBIOSIS", more specifically by the Drug Formulation Unit (U10) and the Micro-Nano Technology Unit (U8) of the CIBER in Bioengineering, Biomaterials & Nanomedicine (CIBERBBN). Also, they thank the support to research on cell microencapsulation from the University of the Basque Country UPV/EHU (EHUA 16/06) and the Basque Country Government (Grupos Consolidados, No ref: IT907-16). The authors acknowledge the financial support from the Ministerio de Economia y Competitividad (MINECO) (Spain) through Ramon y Cajal program (RYC-2013-14479). This work has made use of the Spanish ICTS Network MICRONANOFABS partially supported by MINECO.

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&reg, ) film of 127 &micro, m as a substrate for minimizing the damage caused on the biofilm structure during its insertion. The electrodes were covered with a Nafion&reg, layer to increase sensor stability and repeatability and to avoid electrode surface fouling. DO microelectrodes showed a linear response in the range 0&ndash, 8 mg L&minus, 1, a detection limit of 0.05 mg L&minus, 1, and a sensitivity of 2.06 nA L mg&minus, 1. pH electrodes showed a linear super-Nernstian response (74.2 &plusmn, 0.7 mV/pH unit) in a wide pH range (pH 4&minus, 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.

Epithelia and endothelia delineate tissue compartments and control their environments by regulating the passage of ions and solutes. This barrier function is essential for the development and maintenance of multicellular organisms, and its dysfunction is associated with numerous human diseases. Recent advances in biomaterials and microfabrication technologies have evolved in vitro approaches for modelling biological barriers. Current microphysiological systems have become more efficient and reliable in mimicking the cell microenvironment. Additionally, methods for the quantification of barrier permeability have long provided significant insight into their underlying mechanisms. In this review, we outline the current techniques to quantify the barrier function of engineered tissues, and we also give an overview of recent microphysiological systems of biological barriers that emulate the microenvironment and microarchitecture of native tissues., This work has been supported by grants from the Ministerio de Economía y Competitividad (MINECO) (SAF2014–62114-EXP, DPI2015–65401-C3–3-R, and RYC-2013-14479) and from CIBER in Bioengineering, Biomaterials & Nanomedicine (CIBER-BBN).

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.

The interconnection of different tissue-tissue interfaces may extend organ-on-chips to a new generation of sophisticated models capable of recapitulating more complex organ-level functions. Single interfaces are largely recreated in organ-on-chips by culturing the cells on opposite sides of a porous membrane that splits a chamber in two or by connecting the cells of two adjacent compartments through microchannels. However, it is difficult to interconnect more than one interface using these approaches. To address this challenge, we present a novel microfluidic device where cells are arranged in parallel compartments and are highly interconnected through a grid of microgrooves, which facilitates paracrine signaling and heterotypic cell-cell contact between multiple tissues. In addition, the device includes electrodes on the substrate for the measurement of transepithelial electrical resistance (TEER). Unlike conventional methods for measuring the TEER where electrodes are on each side of the cell barrier, a method with only electrodes on the substrate has been validated. As a proof-of-concept, we have used the device to mimic the structure of the blood-retinal barrier by co-culturing primary human retinal endothelial cells (HREC), a human neuroblastoma cell line (SH-SY5Y), and a human retinal pigment epithelial cell line (ARPE-19). Cell barrier formations were assessed by a permeability assay, TEER measurements, and ZO-1 expression. These results validate the proposed microfluidic device with microgrooves as a promising in vitro tool for the compartmentalization and monitoring of barrier tissues., This work is part of the requirements to achieve the PhD degree in Electrical and Telecommunication Engineering at the Universitat Autònoma de Barcelona, and it was supported by grants from Ministerio de Economía y Competitividad (MINECO) (SAF2014-62114-EXP and SAF2016-77784-R) and Fundació La Marató de TV3 (Exp. 201629-10). This work has made use of the Spanish ICTS Network MICRONANOFABS partially supported by MINECO and the ICTS 'NANBIOSIS', more specifically by the Micro-Nano Technology Unit of the CIBER in Bioengineering, Biomaterials & Nanomedicne (CIBER-BBN) at the IMB-CNM.

Transepithelial electrical measurements in the renal tubule have provided a better understanding of how kidney regulates electrolyte and water homeostasis through the reabsorption of molecules and ions (e.g., H2O and NaCl). While experiments and measurement techniques using native tissue are difficult to prepare and to reproduce, cell cultures conducted largely with the Ussing chamber lack the effect of fluid shear stress which is a key physiological stimulus in the renal tubule. To overcome these limitations, we present a modular perfusion chamber for long-term culture of renal epithelial cells under flow that allows the continuous and simultaneous monitoring of both transepithelial electrical parameters and transepithelial NaCl transport. The latter is obtained from electrical conductivity measurements since Na+ and Cl- are the ions that contribute most to the electrical conductivity of a standard physiological solution. The system was validated with epithelial monolayers of raTAL and NRK-52E cells that were characterized electrophysiologically for 5 days under different flow conditions (i.e., apical perfusion, basal, or both). In addition, apical to basal chemical gradients of NaCl (140/70 and 70/140 mM) were imposed in order to demonstrate the feasibility of this methodology for quantifying and monitoring in real time the transepithelial reabsorption of NaCl, which is a primary function of the renal tubule., This work is part of the requirements to achieve the PhD degree in Electrical and Telecommunication Engineering at the Universitat Autònoma de Barcelona, and it was supported by grants from Ministerio de Economía y Competitividad (MINECO) (SAF2014-62114-EXP, DPI2015-65401-C3-3-R, and DPI2011-28262-C04-02). This work has made use of the Spanish ICTS Network MICRONANOFABS partially supported by MINECO and the ICTS 'NANBIOSIS', more specifically by the Micro-Nano Technology Unit of the CIBER in Bioengineering, Biomaterials & Nanomedicne (CIBER-BBN) at the IMB-CNM.

An experimental study of a micromachined non-porous pillar array column performance under non-retentive conditions is presented. The same pillar structure has been fabricated in cyclo-olefin polymer (COP) chips with three different depths via hot embossing and pressure-assisted thermal bonding. The influence of the depth on the band broadening along with the already known contribution arising from the top and bottom cover plates has been studied. The experimental results exhibit reduced plate heights as low as 0.2, which are in agreement with the previous experimental work. Moreover, the constant values of the reduced Van Deemter expression are also in accordance with the previous studies. A more exhaustive study of the C-term band broadening is also presented, showing that comparing the space between the pillars with different open tubular rectangular channels offers a good estimation of the C-term band broadening that is obtained experimentally. These experimental results, hence, confirm that micromachined pillar array columns fabricated in COP can achieve the same performance as the ones fabricated in silicon for the presently studied pillar channel design.

In this work, we present the use of interdigitated electrodes (IDEs) for performing electrical impedance spectroscopy (EIS) measurements to monitor a microfluidic blood brain barrier model. In particular, an electrode configuration which would not impair the optical visualization of the cell culture is proposed. Numerical studies have been performed to evaluate the electrical impedance sensitivity of the proposed tetrapolar configuration along the cell barrier in a given microfluidic chamber geometry. The system has been validated using a home-made cyclo olefin polymer (COP) bioreactor and perforated poly (methyl methacrylate) (PMMA) sheets with different pore densities in order to simulate different cell barrier impedances.

WO 3 –Cr, SnO 2 –Pd and TiO 2 –Cr nanoparticles have been prepared following a sol–gel route. The additive-modified nanostructured metal oxides have been deposited on hotplate platforms by microdropping. Functional gas test of the sensitive layers to ammonia has been performed in stationary and pulsed mode of operation. The pulsed mode fastened the regeneration of the surface yielding an enlargement of the sensor response and a decrease of the transient time. The improved sensing response to ammonia was particularly marked for the WO 3 –Cr based gas sensor.

Experimental measurements to study the retention capacity and band broadening under retentive conditions using micromachined non-porous pillar array columns fabricated in cyclo olefin polymer are presented. In particular, three columns with different depths but with the same pillar structure have been fabricated via hot embossing and pressure-assisted thermal bonding. Separations of a mixture of four coumarins using varying mobile phase compositions have been monitored to study the relation between the retention factor and the ratio of organic solvent in the aqueous mobile phase. Moreover, the linear relation between the retention and the surface/volume ratio predicted in theory has been observed, achieving retention factors up to k=2.5. Under the same retentive conditions, minimal reduced plate height values of h(min)=0.4 have been obtained at retention factors of k=1.2. These experimental results are compared with the case of non-porous and porous silicon pillars. Similar results for the plate heights are achieved while retention factors are higher than the non-porous silicon column and considerably smaller than the porous pillar column, given the non-porous nature of the used cyclo olefin polymer. The feasibility of using this polymer column as an alternative to the pillar array silicon columns is corroborated.

Mesoporous silica with KIT-6 structure was investigated as a preconcentrating material in chromatographic systems for ammonia and trimethylamine. Its adsorption capacity was compared to that of existing commercial materials, showing its increased adsorption power. In addition, KIT-6 mesoporous silica efficiently adsorbs both gases, while none of the employed commercial adsorbents did. This means that KIT-6 Mesoporous silica may be a good choice for integrated chromatography/gas sensing micro-devices.

The current paper describes the development and characterization of a pillar array chip that is constructed out of a sandwich of cyclo olefin polymer (COP) sheets. The silicon master of a 5 cm long pillar array was embossed into the COP, yielding 4.3 microm deep pillars of 15.3 microm diameter with an external porosity of 43 % and a well designed sidewall region to avoid side wall induced band broadening. A closed channel configuration was obtained by pressure assisted thermal bonding to a non-processed COP lid. Injection of coumarin dye plugs and detection with a fluorescence microscope showed very close agreement of this channel configuration to theoretical expectations in terms of band broadening. This agreement is due to the low taper, the optimized sidewall region and the excellent bonding quality between the two polymer sheets, even at the pillar area. Under non-retained conditions (pure methanol as mobile phase), plate heights as low as 4 microm were obtained. Under retained conditions, using the native hydrophobic properties of the COP channel (in 70/30 v/v water/methanol mixture as mobile phase), a minimum plate height of 6 microm was obtained. A 4 component separation was successfully achieved, demonstrating that COP is a cheap and efficient alternative for silicon and silica based liquid chromatography formats.

The development of an integrated gas chromatographic system using micro and nanotechnologies is presented in this paper. For this purpose, the different components of the chromatographic system, namely the preconcentrator, the chromatographic column and the gas sensors are being investigated and developed, and the actual state of this investigation is presented. The proposed target application comes from the agrofood industry, in particular the determination of the fish freshness. The structure of the preconcentrator has been fabricated using deep reactive ion etching (DRIE). The same fabrication technique has been employed for the patterning of the silicon microcolumns, which have been sealed with Pyrex glass. Inlet and outlets have been connected and initial experiments of functionalization have been performed. Gas sensors have been obtained by microdeposition of doped WO 3 or SnO 2 nanomaterials on microhotplates and their responses to the gases of interest have been measured, proving that the target gas concentrations can be detected.

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.