Hydrogen sulfide (H2S) has been increasingly recognized as an important signaling molecule that regulates both blood pressure and neuronal activity. Attention has been drawn to its interactions with another gasotransmitter, nitric oxide (NO). Here, we provide evidence that the physiological effects observed upon the application of sodium nitroprusside (SNP) and H2S can be ascribed to the generation of nitroxyl (HNO), which is a direct product of the reaction between SNP and H2S, not a consequence of released NO subsequently reacting with H2S. Intracellular HNO formation has been confirmed, and the subsequent release of calcitonin gene-related peptide from a mouse heart has been demonstrated. Unlike with other thiols, SNP reacts with H2S in the same way as rhodanese, i.e., the cyanide transforms into a thiocyanate. These findings shed new light on how H2S is understood to interact with nitroprusside. Additionally, they offer a new and convenient pharmacological source of HNO for therapeutic purposes.

The spherical vaterite CaCO3 microcrystals are nowadays widely used as sacrificial templates for fabrication of various microcarriers made of biopolymers (e.g., proteins, nucleic acids, enzymes) due to porous structure and mild template elimination conditions. Here, we demonstrated for the first time that polymer microcarriers with tuned internal nanoarchitecture can be designed by employing the CaCO3 crystals of controlled porosity. The layer-by-layer deposition has been utilized to assemble shell-like (hollow) and matrix-like (filled) polymer capsules due to restricted and free polymer diffusion through the crystal pores, respectively. The crystal pore size in the range of few tens of nanometers can be adjusted without any additives by variation of the crystal preparation temperature in the range 7-45 °C. The temperature-mediated growth mechanism is explained by the Ostwald ripening of nanocrystallites forming the crystal secondary structure. Various techniques including SEM, AFM, CLSM, Raman microscopy, nitrogen adsorption-desorption, and XRD have been employed for crystal and microcapsule analysis. A three-dimensional model is introduced to describe the crystal internal structure and predict the pore cutoff and available surface for the pore diffusing molecules. Inherent biocompatibility of CaCO3 and a possibility to scale the porosity in the size range of typical biomacromolecules make the CaCO3 crystals extremely attractive tools for template assisted designing tailor-made biopolymer-based architectures in 2D to 3D targeted at drug delivery and other bioapplications.

In this study, the effect of temperature on the build-up of exponentially growing polyelectrolyte multilayer films was investigated. It aims at understanding the multilayer growth mechanism as crucially important for the fabrication of tailor-made multilayer films. Model poly(L-lysine)/hyaluronic acid (PLL/HA) multilayers were assembled in the temperature range of 25-85 °C by layer-by-layer deposition using a dipping method. The film growth switches from the exponential to the linear regime at the transition point as a result of limited polymer diffusion into the film. With the increase of the build-up temperature the film growth rate is enhanced in both regimes; the position of the transition point shifts to a higher number of deposition steps confirming the diffusion-mediated growth mechanism. Not only the faster polymer diffusion into the film but also more porous/permeable film structure are responsible for faster film growth at higher preparation temperature. The latter mechanism is assumed from analysis of the film growth rate upon switching of the preparation temperature during the film growth. Interestingly, the as-prepared films are equilibrated and remain intact (no swelling or shrinking) during temperature variation in the range of 25-45 °C. The average activation energy for complexation between PLL and HA in the multilayers calculated from the Arrhenius plot has been found to be about 0.3 kJ mol(-1) for monomers of PLL. Finally, the following processes known to be dependent on temperature are discussed with respect to the multilayer growth: (i) polymer diffusion, (ii) polymer conformational changes, and (iii) inter-polymer interactions.

signifi cantly precise control over the scaffold architecture. Well-defi ned internal structure (porosity) determines both cellular infi ltration into the scaffold and its material properties. Pore size and distribution ensuring diffusion of nutrients into the scaffold and removal of metabolic products is of high importance. [ 4,11 ] Ionically crosslinked polymer gels such as alginate hydrogels have been extensively developed as scaffolds for tissue engineering. [12][13][14][15] The adjustable kinetics of the alginate gel degradation at neutral pH [ 1,16,17 ] gives an option to use the gels for multiple applications as wound dressings, anti-adhesive, and repair materials. [ 14,[18][19][20][21] Characteristics of alginate gels (hydration, softness, porosity, swelling in water, etc.) can be adjusted using a certain fabrication approach and by variation of interactions between charged polyanionic groups and crosslinking counterions; [ 13,17,[22][23][24][25] these interactions are known to be strongly affected by ionic strength, molecule length and structure, solvent pH, salt concentration, and temperature conditions. [26][27][28][29][30][31] This allows the fabrication of alginate scaffolds possessing desired properties corresponding to a certain tissue, for instance cartilage, [ 12 ] or dura mater. [ 21 ] Porous alginate gels can provide space and mechanical support to seed biological cells for tissue formation, [ 1,13,32 ] also allowing a controlled release of gel-laden drugs (peptides and proteins) trapped within the alginate network. [ 14,24,25,33,34 ] To achieve loading of both biological cells and bioactive molecules, the internal structure of hydrogels has to be controlled on the scale of nano-and micrometers. Adjustment of the gel geometry has been demonstrated on substrate surfaces using different micropatterning techniques including a light-addressable electrolytic system, [ 14 ] electrodeposition, [ 15 ] electrochemical patterning, [ 35 ] or the benchtop method using Nylon mesh. [ 11 ] The internal structure of alginate gels has been patterned with regular tube-like pores, [ 16,36 ] interconnected ordered honeycomb pores, [ 12 ] and sponge-like isotropic pores. [37][38][39] However, most of the approaches for alginate gel fabrication lack a precise control over the microstructure and require special equipment. Encapsulation of molecules of interest with controlled spatial distribution is rather complicated or even impossible. There is a need to develop a simple approach for design of a scaffold with welldefi ned internal structure providing both a space to culture cells and cavities to host/release encapsulated (bio)active molecules.Fabrication of porous alginate hydrogels with a well-controlled architecture useful for tissue engineering is still a challenge. Here, CaCO 3 -based templating is utilized to design stable alginate gels with controlled pore dimensions in the range of 5-50 µm. The mechanism of pore formation is studied considering two factors affecting the pore size: i) osmotic pressure generat...

Polyelectrolyte multilayer films are nowadays very attractive for bioapplications due to their tunable properties and ability to control cellular response. Here we demonstrate that multilayers made of hyaluronic acid and poly-l-lysine act as high-capacity reservoirs for small charged molecules. Strong accumulation within the film is explained by electrostatically driven binding to free charges of polyelectrolytes. Binding and release mechanisms are discussed based on charge balance and polymer dynamics in the film. Our results show that transport of molecules through the film-solution interface limits the release rate. The multilayers might serve as an effective platform for drug delivery and tissue engineering due to high potential for drug loading and controlled release.