Investigations on the impact of the introduction of the Aloe vera into the hydrogel matrix on cytotoxic and hydrophilic properties of these systems considered as potential wound dressings.

In the paper, synthesis of chitosan-based hydrogels modified with Aloe vera juice is presented. The novelty of the research was a combination of hydrogel materials with properties beneficial in viewpoint of their use as modern wound dressings and Aloe vera juice supporting the wound healing process. Hydrogels have been obtained via UV radiation. The impact of the amount of the crosslinking agent as well as the introduction of the Aloe vera juice into the hydrogel matrix has been determined. Performed measurements involved analysis of the swelling ability, characteristics of the surface roughness, determining the release profile of Aloe vera and the contact angles of hydrogels. Furthermore, the analysis of the dehydration process of the polymer membrane, investigations on the cytotoxicity of hydrogels via MTT reduction assay and the neutral red uptake assay as well as the studies on the pro-inflammatory activity have also been performed. It was proved that the addition of Aloe vera juice improves the hydrophilic properties of the materials (e.g. contact angle changed from 82.5° to 73.0°). Next, the use of 25% more of the crosslinker resulted even in the increase of the contact angle by 86%. Modified hydrogels showed higher swelling properties even by 15% than unmodified materials. Furthermore, obtained hydrogels show an ability to release Aloe vera - after 5 h approx. 80% of this additive has been released in an acidic environment. Tested materials do not exhibit cytotoxic properties, the addition of Aloe vera results in an improvement of the viability of L929 murine fibroblasts and, importantly, these materials show lower pro-inflammatory activity than the positive control. Performed investigations allow to state that obtained materials show a great application potential.

Social amoebae mating types do not invest unequally in sexual offspring.

Unequal investment by different sexes in their progeny is common and includes differential investment in the zygote and differential care of the young. The social amoeba Dictyostelium discoideum has a sexual stage in which isogamous cells of any two of the three mating types fuse to form a zygote which then attracts hundreds of other cells to the macrocyst. The latter cells are cannibalized and so make no genetic contribution to reproduction. Previous literature suggests that this sacrifice may be induced in cells of one mating type by cells of another, resulting in a higher than expected production of macrocysts when the inducing type is rare and giving a reproductive advantage to this social cheat. We tested this hypothesis in eight trios of field-collected clones of each of the three D. discoideum mating types by measuring macrocyst production at different pairwise frequencies. We found evidence that supported differential contribution in only two of the 24 clone pairs, so this pattern is rare and clone-specific. In general, we did not reject the hypothesis that the mating types contribute cells relative to their proportion in the population. We also found a significant quadratic relationship between partner frequency and macrocyst production, suggesting that when one clone is rare, macrocyst production is limited by partner availability. We were also unable to replicate previous findings that macrocyst production could be induced in the absence of a compatible mating partner. Overall, mating type-specific differential investment during sex is unlikely in microbial eukaryotes like D. discoideum.

Sex ratio and gamete size across eastern North America in Dictyostelium discoideum, a social amoeba with three sexes.

Theory indicates that numbers of mating types should tend towards infinity or remain at two. The social amoeba, Dictyostelium discoideum, however, has three mating types. It is therefore a mystery how this species has broken the threshold of two mating types, but has not increased towards a much higher number. Frequency-dependent selection on rare types in combination with isogamy, a form of reproduction involving gametes similar in size, could explain the evolution of multiple mating types in this system. Other factors, such as drift, may be preventing the evolution of more than three. We first looked for evidence of isogamy by measuring gamete size associated with each type. We found no evidence of size dissimilarities between gametes. We then looked for evidence of balancing selection, by examining mating type distributions in natural populations and comparing genetic differentiation at the mating type locus to that at more neutral loci. We found that mating type frequency varied among the three populations we examined, with only one of the three showing an even sex ratio, which does not support balancing selection. However, we found more population structure at neutral loci than the mating type locus, suggesting that the three mating types are indeed maintained at intermediate frequencies by balancing selection. Overall, the data are consistent with balancing selection acting on D. discoideum mating types, but with a sufficiently weak rare sex advantage to allow for drift, a potential explanation for why these amoebae have only three mating types.

Enzymatic mineralization of gellan gum hydrogel for bone tissue-engineering applications and its enhancement by polydopamine.

Interest is growing in the use of hydrogels as bone tissue-engineering (TE) scaffolds due to advantages such as injectability and ease of incorporation of active substances such as enzymes. Hydrogels consisting of gellan gum (GG), an inexpensive calcium-crosslinkable polysaccharide, have been applied in cartilage TE. To improve GG suitability as a material for bone TE, alkaline phosphatase (ALP), an enzyme involved in mineralization of bone by cleaving phosphate from organic phosphate, was incorporated into GG hydrogels to induce mineralization with calcium phosphate (CaP). Incorporated ALP induced formation of apatite-like material on the submicron scale within GG gels, as shown by FTIR, SEM, EDS, XRD, ICP-OES, TGA and von Kossa staining. Increasing ALP concentration increased amounts of CaP as well as stiffness. Mineralized GG was able to withstand sterilization by autoclaving, although stiffness decreased. In addition, mineralizability and stiffness of GG was enhanced by the incorporation of polydopamine (PDA). Furthermore, mineralization of GG led to enhanced attachment and vitality of cells in vitro while cytocompatibility of the mineralized gels was comparable to one of the most commonly used bone substitute materials. The results proved that ALP-mediated enzymatic mineralization of GG could be enhanced by functionalization with PDA.

Alginate-hydroxypropylcellulose hydrogel microbeads for alkaline phosphatase encapsulation.

There is a growing interest in using proteins as therapeutics agents. Unfortunately, they suffer from limited stability and bioavailability. We aimed to develop a new delivery system for proteins. ALP, a model protein, was successfully encapsulated in the physically cross-linked sodium alginate/hydroxypropylcellulose (ALG-HPC) hydrogel microparticles. The obtained objects had regular, spherical shape and a diameter of ∼4 µm, as confirmed by optical microscopy and SEM analysis. The properties of the obtained microbeads could be controlled by temperature and additional coating or crosslinking procedures. The slow, sustained release of ALP in its active form with no initial burst effect was observed for chitosan-coated microspheres at pH = 7.4 and 37 °C. Activity of ALP released from ALG/HPC microspheres was confirmed by the occurance of effectively induced mineralization. SEM and AFM images revealed formation of the interpenetrated three-dimensional network of mineral, originating from the microbeads' surfaces. FTIR and XRD analyses confirmed formation of hydroxyapatite.