A novel, rapid and simple method for the preparation of emulsion-templated monoliths in microfluidic channels based on thiol-ene chemistry is presented. The method allows monolith synthesis and anchoring inside thiol-ene microchannels in a single photoinitiated step. Characterization by scanning electron microscopy showed that the methanol-based emulsion templating process resulted in a network of highly interconnected and regular thiol-ene beads anchored solidly inside thiol-ene microchannels. Surface area measurements indicate that the monoliths are macroporous, with no or little micro- or mesopores. As a demonstration, galactose oxidase and peptide-N-glycosidase F (PNGase F) were immobilized at the surface of the synthesized thiol-ene monoliths via two different mechanisms. First, cysteine groups on the protein surface were used for reversible covalent linkage to free thiol functional groups on the monoliths. Second, covalent linkage was achieved via free primary amino groups on the protein surface by means of thiol-ene click chemistry and l-ascorbic acid linkage. Thus prepared galactose oxidase and PNGase F microreactors demonstrated good enzymatic activity in a galactose assay and the deglycosilation of ribonuclease B, respectively.
Stratum corneum ceramides are major determinants of skin barrier function. Although their physiological and pathological role has been widely investigated, to date no structure-activity relationships have been established. In this study, a series of short-chain ceramide analogues with polar head structure identical to ceramide NS, a sphingosine length of 12 carbons and an acyl chain length of 2–12 carbons was synthesized. Their effect on skin permeability was evaluated using porcine skin and two model drugs, theophylline and indomethacin, and compared to that of a physiological ceramide NS. The results showed that the ceramide chain length was crucial for their barrier properties. Ceramides with a 4- to 8-carbon acyl chain were able to increase skin permeability for both drugs up to 10.8 times with maximum effect at a 6-carbon acyl chain. No increase in permeability was found for ceramide analogues with 2- and 12-carbon acyl chains and ceramide NS. The same relationships were obtained for skin concentrations of the model drugs. The relationship between ceramide acyl chain length and its ability to perturb skin barrier showed striking similarity to the behavior of short-chain ceramides in sphingomyelin/phospholipid membranes and confirmed that short-chain ceramides do not act as natural ceramides and their use as experimental tools should be cautious.
Fluid manipulation on the micro-scale (microfluidics) is bringing new potential applications in a number of fields, including chemistry, biology and medicine. At sub-millimeter channel scale, some phenomena, unimportant at the macroscale, become an important force to consider when designing a microfluidics system. For example, the decrease in fluid mass causes the effects of viscosity to overcome the influence of inertia. Turbulent flow cannot be achieved at any realistic fluid velocity, making mixing a challenging task. The only phenomenon capable of blending liquids at microscale is diffusion and liquid streams can be flowed side-by-side for tens of minutes before they completely fuse together. The decrease in the channel size also leads to an increased surface-to-volume ratio, which increases the importance of surface effects, including adsorption, capillary action and surface wetting and/or electric double layer formation with related electrokinetic phenomena. While rivers cannot flow uphill, a stream of liquid can easily flow up against gravity inside a capillary. Similarly, the formation of electric double layer near the charged surface of a micro-channel or capillary can be applied for electrokinetic actuating. This review summarizes selected physical phenomena related to liquid-based (water solutions) microfluidics as described recently.
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