Differential effects of early growth conditions on colour-producing nanostructures revealed through small angle X-ray scattering and electron microscopy.

The costs associated with the production and maintenance of colour patches is thought to maintain their honesty. Although considerable research on sexual selection has focused on structurally coloured plumage ornaments, the proximate mechanisms of their potential condition dependence, and thus their honesty, is rarely addressed, particularly in an experimental context. Blue tit (Cyanistes caeruleus) nestlings have ultraviolet (UV)-blue structurally coloured tail feathers, providing a unique opportunity for investigation of the causes of variation in their colour. Here, we examined the influence of early growing conditions on the reflectance and structural properties of UV-blue-coloured tail feathers of blue tit nestlings. We applied a two-stage brood size manipulation to determine which stage of development more strongly impacts the quality of tail feather colouration and microstructure. We used small-angle X-ray scattering (SAXS) and electron microscopy to characterise the nanoscale and microscale structure of tail feather barbs. Nestlings from the broods enlarged at a later stage of growth showed a sex-specific rectrix development delay, with males being more sensitive to this manipulation. Contrary to predictions, treatment affected neither the quality of the barbs' nanostructures nor the brightness and UV chroma of feathers. However, at the microscale, barbs' keratin characteristics were impaired in late-enlarged broods. Our results suggest that nanostructure quality, which determines the UV-blue colour in tail feathers, is not sensitive to early rearing conditions. Furthermore, availability of resources during feather growth seems to impact the quality of feather microstructure more than body condition, which is likely to be determined at an earlier stage of nestling growth.

Novel hybrid materials for preparation of bone tissue engineering scaffolds.

The organic-inorganic hybrid systems based on biopolymer hydrogels with dispersed silica nanoparticles were obtained and characterized in terms of their physicochemical properties, cytocompatibility and bioactivity. The hybrid materials were prepared in a form of collagen and collagen-chitosan sols to which the silica nanoparticles of two different sizes were incorporated. The ability of these materials to undergo in situ gelation under physiological temperature was assessed by microviscosity and gelation time determination based on steady-state fluorescence anisotropy measurements. The effect of silica nanoparticles addition on the physicochemical properties (surface wettability, swellability) of hybrid materials was analyzed and compared with those characteristic for pristine collagen and collagen-chitosan hydrogels. Biological studies indicate that surface wettability determined in terms of contact angle for all of the hybrids prepared is optimal and thus can provide satisfactory adhesion of fibroblasts. Cytotoxicity test results showed high metabolic activity of mouse as well as human fibroblast cell lines cultured on hybrid materials. The composition of hybrids was optimized in terms of concentration of silica nanoparticles. The effect of silica on the formation of bone-like mineral structures on exposition to simulated body fluid was determined. SEM images revealed mineral phase formation not only at the surfaces but also in the whole volumes of all hybrid materials developed suggesting their usefulness for bone tissue engineering. EDS and FTIR analyses indicated that these mineral phases consist of apatite-like structures.

Lysine-functionalized chondroitin sulfate improves the biological properties of collagen/chitosan-based injectable hydrogels.

Novel bioactive collagen/chitosan/lysine-functionalized chondroitin sulfate (CSmod) injectable hydrogels are presented. The modification of CS with amine groups introduced with lysine moieties (the degree of substitution about 21%) guarantees its covalent binding with the hydrogel network while genipin crosslinking. Both the physicochemical and biological features of developed hydrogels might be adjusted by playing with CSmod and crosslinking agent concentrations. It was revealed that materials became more hydrophobic with increased CSmod content, while crosslinking degree and enzymatic degradation studies established the influence of CSmod concentration and Ch:CSmod ratio on the crosslinking process. In situ rheological experiments verified the injectability of resulted systems. The biological in vitro evaluation demonstrated that all designed materials are biocompatible as they supported proliferation and adhesion of MG-63 cell line. In vitro biomineralization study employing simulated body fluid model revealed CSmod-content dependent bioactivity of obtained hydrogels. Importantly for pristine collagen/chitosan materials, the formation of apatite-like structures was not observed. Our findings demonstrate that developed injectable ColChCSmod hydrogels particularly system with the greatest CSmod concentration exhibits high bioactive potential, without the need of applying additional inducers what renders them promising materials within tissue engineering applications.

Sexual dichromatism, size dimorphism, and microscale anatomy of white wing stripe in blue tits.

Achromatic patches are a common element of plumage patterns in many bird species and there is growing body of evidence that in many avian taxa they can play a signaling role in mate choice. Although the blue tit Cyanistes caeruleus is a well-established model species in the studies on coloration, its white wing patch has never been examined in the context of sex-specific trait expression. In this exploratory study, we examined sexual size dimorphism and dichromatism of greater covert's dots creating white wing patch and analyzed its correlations with current body condition and crown coloration-a trait with established role in sexual selection. Further, we qualitatively analyzed microstructural barb morphology underlying covert's coloration. We found significant sexual dimorphism in the dot size independent of covert size and sexual dichromatism in both white dot and blue outer covert's vane spectral characteristics. Internal structure of covert barbs within the white dot was similar to the one found in barbs from the blue part that is, with a medullary area consisting of dead keratinocytes containing channel-type ß-keratin spongy nanostructure and centrally located air cavities. However, it lacked melanosomes which was the main observed difference. Importantly, UV chroma of covert's blue vane was positively correlated with crown UV chroma and current condition (the latter only in males), which should be a premise for further research on the signal function of the wing stripe.

Bioactive hydrogel scaffolds reinforced with alkaline-phosphatase containing halloysite nanotubes for bone repair applications.

Alkaline phosphatase (ALP), biomineralization promoting enzyme, was immobilized in halloysite (HAL) nanotubes and used as a bioactive component of the chitosan (CH) and chitosan-collagen (C-CH) hydrogel scaffolds for bone regeneration. The influence of HAL-ALP and collagen on the properties of the obtained materials was studied. 30 wt% of HAL-ALP increased significantly the swelling ratio of chitosan-based scaffolds. The presence of both: collagen and HAL-ALP had positive effect on the scaffolds' porosity, which was considerably higher for C-CH. The presence of HAL has improved the mechanical properties of both types of scaffolds, while the addition of 20% of collagen to the chitosan hydrogels have considerably decreased their storage modulus. Biomineralization tests showed that although mineral was formed in both CH and C-CH scaffolds with HAL-ALP, the process was more effective for collagen-containing hydrogels. Biological studies, performed using MG-63 osteoblast-like cell line showed that C-CH scaffolds, especially those after biomineralization, were a better material for cell adhesion and growth. Both types of scaffolds degraded slowly in physiological pH. C-CH scaffolds containing 30% of HAL-ALP have the highest potential as bioactive material for bone regeneration.

Genipin crosslinked bioactive collagen/chitosan/hyaluronic acid injectable hydrogels structurally amended via covalent attachment of surface-modified silica particles.

Collagen, chitosan and hyaluronic acid based multicomponent injectable and in situ gellating biomimetic hybrid materials for bone tissue engineering applications were prepared in one-step procedure. The bioactive phase in the form of surface-modified silica particles was introduced to the solutions of biopolymers and simultaneously crosslinked with genipin both the biopolymer matrix and dispersed particles at 37 °C. The novel approach presented here involved the use of silica particles which surfaces were priory functionalized with amino groups. That modification makes possible the covalent attachment of silica particles to the polymeric hydrogel network on crosslinking with genipin. That methodology is especially important as it makes possible to obtain the hybrid materials (biopolymer-silica particles) in which the problems related to the potential phase separation of mineral particles, hindering their in vivo application can be eliminated. The hybrids of various compositions were obtained and their physicochemical and biological properties were determined. The in vitro experiments performed under simulated body fluid conditions revealed that the amino-functionalized silica particles covalently attached to the biopolymeric network are still bioactive. Finally, the in vitro cell culture studies shown that the materials developed are biocompatible as they supported MG-63 cells adhesion, proliferation as well as Alkaline phosphatase (ALP) expression.

Alginate- and gelatin-based bioactive photocross-linkable hybrid materials for bone tissue engineering.

The paper presents the synthesis, the physico-chemical and the biological properties of novel hybrid materials prepared from photo-crosslinked gelatin/alginate-based hydrogels and silica particles exhibiting potential for the regeneration of bone tissue. Both alginate and gelatin were functionalized with methacrylate and methacrylamide moieties, respectively to render them photo-crosslinkable. Submicron silica particles of two sizes were dispersed within three types of polymeric sols including alginate, gelatin, and gelatin/alginate blends, which were subsequently photo-crosslinked. The swelling ratio, the gel fraction and the mechanical properties of the hybrid materials developed were examined and compared to these determined for reference hydrogel matrices. The in vitro cell culture studies have shown that the prepared materials exhibited biocompatibility as they supported both MEFs and MG-63 mitochondrial activity. Finally, the in vitro experiments performed under simulated body fluid conditions have revealed that due to inclusion of silica particles into the biopolymeric hydrogel matrices the mineralization was successfully induced.