Droplet Friction on Superhydrophobic Surfaces Scales With Liquid-Solid Contact Fraction.

It is generally assumed that contact angle hysteresis of superhydrophobic surfaces scales with liquid-solid contact fraction, however, its experimental verification has been problematic due to the limited accuracy of contact angle and sliding angle goniometry. Advances in cantilever-based friction probes enable accurate droplet friction measurements down to the nanonewton regime, thus suiting much better for characterizing the wetting of superhydrophobic surfaces than contact angle hysteresis measurements. This work quantifies the relationship between droplet friction and liquid-solid contact fraction, through theory and experimental validation. Well-defined micropillar and microcone structures are used as model surfaces to provide a wide range of different liquid-solid contact fractions. Micropillars are known to be able to hold the water on top of them, and a theoretical analysis together with confocal laser scanning microscopy shows that despite the spiky nature of the microcones droplets do not sink into the conical structure either, rendering a diminishingly small liquid-solid contact fraction. Droplet friction characterization with a micropipette force sensor technique reveals a strong dependence of the droplet friction on the contact fraction, and the dependency is described with a simple physical equation, despite the nearly three-orders-of-magnitude difference in liquid-solid contact fraction between the sparsest cone surface and the densest pillar surface.

Real-time selective detection of dopamine and serotonin at nanomolar concentration from complex in vitro systems.

Electrochemical sensors provide means for real-time monitoring of neurotransmitter release events, which is a relatively easy process in simple electrolytes. However, this does not apply to in vitro environments. In cell culture media, competitively adsorbing molecules are present at concentrations up to 350 000-fold excess compared to the neurotransmitter-of-interest. Because detection of dopamine and serotonin requires direct adsorption of the analyte to electrode surface, a significant loss of sensitivity occurs when recording is performed in the in vitro environment. Despite these challenges, our single-walled carbon nanotube (SWCNT) sensor was capable of selectively measuring dopamine and serotonin from cell culture medium at nanomolar concentration in real-time. A primary midbrain culture was used to prove excellent biocompatibility of our SWCNT electrodes, which is a necessity for brain-on-a-chip models. Most importantly, our sensor was able to electrochemically record spontaneous transient activity from dopaminergic cell culture without altering the culture conditions, which has not been possible earlier. Drug discovery and development requires high-throughput screening of in vitro models, being hindered by the challenges in non-invasive characterization of complex neuronal models such as organoids. Our neurotransmitter sensors could be used for real-time monitoring of complex neuronal models, providing an alternative tool for their characterization non-invasively.

Ascorbic acid does not necessarily interfere with the electrochemical detection of dopamine.

It is widely stated that ascorbic acid (AA) interferes with the electrochemical detection of neurotransmitters, especially dopamine, because of their overlapping oxidation potentials on typical electrode materials. As the concentration of AA is several orders of magnitude higher than the concentration of neurotransmitters, detection of neurotransmitters is difficult in the presence of AA and requires either highly stable AA concentration or highly selective neurotransmitter sensors. In contrast to the common opinion, we show that AA does not always interfere electrochemical detection of neurotransmitters. The decay of AA is rapid in cell culture medium, having a half-time of 2.1 hours, according to which the concentration decreases by 93% in 8 hours and by 99.75% in 18 hours. Thus, AA is eventually no longer detected by electrodes and the concentration of neurotransmitters can be effectively monitored. To validate this claim, we used unmodified single-wall carbon nanotube electrode to measure dopamine at physiologically relevant concentration range (25-1000 nM) from human midbrain organoid medium with highly linear response. Finally, AA is known to affect dopamine oxidation current through regeneration of dopamine, which complicates precise detection of small amounts of dopamine. By designing experiments as described here, this complication can be completely eliminated.

Nanoscale geometry determines mechanical biocompatibility of vertically aligned nanofibers.

Vertically aligned carbon nanofibers (VACNFs) are promising material candidates for neural biosensors due to their ability to detect neurotransmitters in physiological concentrations. However, the expected high rigidity of CNFs could induce mechanical mismatch with the brain tissue, eliciting formation of a glial scar around the electrode and thus loss of functionality. We have evaluated mechanical biocompatibility of VACNFs by growing nickel-catalyzed carbon nanofibers of different lengths and inter-fiber distances. Long nanofibers with large inter-fiber distance prevented maturation of focal adhesions, thus constraining cells from obtaining a highly spread morphology that is observed when astrocytes are being contacted with stiff materials commonly used in neural implants. A silicon nanopillar array with 500 nm inter-pillar distance was used to reveal that this inhibition of focal adhesion maturation occurs due to the surface nanoscale geometry, more precisely the inter-fiber distance. Live cell atomic force microscopy was used to confirm astrocytes being significantly softer on the long Ni-CNFs compared to other surfaces, including a soft gelatin hydrogel. We also observed hippocampal neurons to mature and form synaptic contacts when being cultured on both long and short carbon nanofibers, without having to use any adhesive proteins or a glial monoculture, indicating high cytocompatibility of the material also with neuronal population. In contrast, neurons cultured on a planar tetrahedral amorphous carbon sample showed immature neurites and indications of early-stage apoptosis. Our results demonstrate that mechanical biocompatibility of biomaterials is greatly affected by their nanoscale surface geometry, which provides means for controlling how the materials and their mechanical properties are perceived by the cells. STATEMENT OF SIGNIFICANCE: Our research article shows, how nanoscale surface geometry determines mechanical biocompatibility of apparently stiff materials. Specifically, astrocytes were prevented from obtaining highly spread morphology when their adhesion site maturation was inhibited, showing similar morphology on nominally stiff vertically aligned carbon fiber (VACNF) substrates as when being cultured on ultrasoft surfaces. Furthermore, hippocampal neurons matured well and formed synapses on these carbon nanofibers, indicating high biocompatibility of the materials. Interestingly, the same VACNF materials that were used in this study have earlier also been proven to be capable for electrophysiological recordings and sensing neurotransmitters at physiological concentrations with ultra-high sensitivity and selectivity, thus providing a platform for future neural probes or smart culturing surfaces with superior sensing performance and biocompatibility.