Oncostatin M triggers brain inflammation by compromising blood-brain barrier integrity.

Oncostatin M (OSM) is an IL-6 family member which exerts neuroprotective and remyelination-promoting effects after damage to the central nervous system (CNS). However, the role of OSM in neuro-inflammation is poorly understood. Here, we investigated OSM's role in pathological events important for the neuro-inflammatory disorder multiple sclerosis (MS). We show that OSM receptor (OSMRß) expression is increased on circulating lymphocytes of MS patients, indicating their elevated responsiveness to OSM signalling. In addition, OSM production by activated myeloid cells and astrocytes is increased in MS brain lesions. In experimental autoimmune encephalomyelitis (EAE), a preclinical model of MS, OSMRß-deficient mice exhibit milder clinical symptoms, accompanied by diminished T helper 17 (Th17) cell infiltration into the CNS and reduced BBB leakage. In vitro, OSM reduces BBB integrity by downregulating the junctional molecules claudin-5 and VE-cadherin, while promoting secretion of the Th17-attracting chemokine CCL20 by inflamed BBB-endothelial cells and reactive astrocytes. Using flow cytometric fluorescence resonance energy transfer (FRET) quantification, we found that OSM-induced endothelial CCL20 promotes activation of lymphocyte function-associated antigen 1 (LFA-1) on Th17 cells. Moreover, CCL20 enhances Th17 cell adhesion to OSM-treated inflamed endothelial cells, which is at least in part ICAM-1 mediated. Together, these data identify an OSM-CCL20 axis, in which OSM contributes significantly to BBB impairment during neuro-inflammation by inducing permeability while recruiting Th17 cells via enhanced endothelial CCL20 secretion and integrin activation. Therefore, care should be taken when considering OSM as a therapeutic agent for treatment of neuro-inflammatory diseases such as MS.

Integrating Thermal Sensors in a Microplate Format: Simultaneous Real-Time Quantification of Cell Number and Metabolic Activity.

Microplates have become a standard tool in the pharmaceutical industry and academia for a broad range of screening assays. One of the most commonly performed assays is the cell proliferation assay, which is often used for the purpose of drug discovery. Microplate readers play a crucial role in this field, as they enable high-throughput testing of large sample numbers. Common drawbacks of the most popular plate reader technologies are that they are end-point-based and most often require the use of detection reagents. As a solution, with this work, we aim to expand the possibilities of real-time and label-free monitoring of cell proliferation inside a microplate format by introducing a novel thermal-based sensing approach. For this purpose, we have developed thin-film sensors that can easily be integrated into the bottom of standard 96-well plates. First, the accuracy and precision of the sensors for measuring temperature and thermal effusivity are assessed via characterization experiments. These experiments highlight the fast response of the sensors to changes in temperature and thermal effusivity, as well as the excellent reproducibility between different sensors. Later, proof-of-principle measurements were performed on the proliferation of Saccharomyces cerevisiae. The proliferation measurements show that the thermal sensors were able to simultaneously detect relative changes in cell number as well as changes in metabolic activity. This dual functionality makes the presented sensor technology a promising candidate for monitoring microplate assays.

Pulsed Thermal Method for Monitoring Cell Proliferation in Real-Time.

The study of cell proliferation is of great importance for medical and biological research, as well as for industrial applications. To render the proliferation process accurately over time, real-time cell proliferation assay methods are required. This work presents a novel real-time and label-free approach for monitoring cell proliferation by continuously measuring changes in thermal properties that occur at the sensor interface during the process. The sensor consists of a single planar resistive structure deposited on a thin foil substrate, integrated at the bottom of a cell culture reservoir. During measurement, the structure is excited with square wave current pulses. Meanwhile, the temperature-induced voltage change measured over the structure is used to derive variations in the number of cells at the interface. This principle is demonstrated first by performing cell sedimentation measurements to quantify the presence of cells at the sensor interface in the absence of cell growth. Later, cell proliferation experiments were performed, whereby parameters such as the available nutrient content and the cell starting concentration were modified. Results from these experiments show that the thermal-based sensor is able to accurately measure variations in the number of cells at the interface. Moreover, the influence of the modified parameters could be observed in the obtained proliferation curves. These findings highlight the potential for the presented thermal method to be incorporated in a standardized well plate format for high-throughput monitoring of cell proliferation.

Monitoring Body Fluids in Textiles: Combining Impedance and Thermal Principles in a Printed, Wearable, and Washable Sensor.

This work explores the feasibility of coupling two different techniques, the impedance and the transient plane source (TPS) principle, to quantify the moisture content and its compositional parameters simultaneously. The sensor is realized directly on textiles with the use of printing and coating technology. Impedance measurements use the fluid's electrical properties, while the TPS measurements are based on the thermal effusivity of the liquid. Impedance and TPS measurements show equal competency in measuring the fluid volume with a lowest measurable quantity of 0.5 µL, enabling ultralow volume passive measurements for sweat analysis. Both sensor principles were tested by monitoring the drying of a wet cloth and the measurements show perfect repeatability and accuracy. Nevertheless, when the biofluid property changes, the TPS sensor does not reflect this information on its readings, whereas, on the other hand, impedance can provide information on compositional changes. However, since the volume of the fluid changes simultaneously, one cannot differentiate between a volume change and a compositional change from impedance measurements alone. Therefore, we show in this work that we can apply impedance to measure the compositional properties; meanwhile, the TPS measurements accurately carry out volume measurements irrespective of the interferences from its compositional variations. To prove this, both of these techniques are applied for the quantification and composition monitoring of sweat, showing the capability to measure moisture content and compositional parameters simultaneously. TPS measurements can also be an indicator of the local temperature of the medium confined by the sensor, and it does not influence the fluid parameters. Compiling both impedance and thermal sensors in a single platform triggers smart wearable prospects of metering the liquid volume and simultaneously analyzing other compositional changes and body temperature. Finally, the repeatability and stability of the sensor readings and the washability of the device are tested. This device could be a potential sensing tool in real-life applications, such as wound monitoring and sweat analysis, and could be a promising addition toward future smart wearable sensors.