Design of silicone interfaces with antibacterial properties.

Silicone implants are widely used for plastic or reconstruction medical applications. However, they can cause severe infections of inner tissues due to bacterial adhesion and biofilm growth on implant surfaces. The development of new antibacterial nanostructured surfaces can be considered as the most promising strategy to deal with this problem. In this article, we studied the influence of nanostructuring parameters on the antibacterial properties of silicone surfaces. Nanostructured silicone substrates with nanopillars of various dimensions were fabricated using a simple soft lithography technique. Upon testing of the obtained substrates, we identified the optimal parameters of silicone nanostructures to achieve the most pronounced antibacterial effect against the bacterial culture of Escherichia coli. It was demonstrated that up to 90% reduction in bacterial population compared to flat silicone substrates can be achieved. We also discussed possible underlying mechanisms behind the observed antibacterial effect, the understanding of which is essential for further progress in this field.

Constitutive chitosanase from Bacillus thuringiensis B-387 and its potential for preparation of antimicrobial chitooligomers.

The article proves the ability of the entomopathogenic strain B. thuringiensis var. dendrolimus B-387 to high the constitutive production (3-12.5 U/mL) of extracellular chitosanase, that was found for the first time. The enzyme was purified in 94-fold by ultrafiltration, affinity sorption and cation-exchange chromatography and characterized biochemically. The molecular mass of the chitosanase determined using SDS-PAGE is 40 kDa. Temperature and pH-optima of the enzyme are 55 °C and pH 6.5, respectively; the chitosanase was stable under 50-60 °C and pH 4-10.5. Purified chitosanase most rapidly (Vmax ~ 43 µM/mL × min, KM ~ 0.22 mg/mL, kcat ~ 4.79 × 104 s-1) hydrolyzed soluble chitosan of the deacetylation degree (DD) 85% by endo-mode, and did not degrade colloidal chitin, CM-cellulose and some other glucans. The main reaction products of the chitosan enzymolysis included chitobiose, chitotriose and chitotetraose. In addition to small chitooligosaccharides (CHOs), the studied chitosanase also generated low-molecular weight chitosan (LMWC) with average Mw in range 14-46 kDa and recovery 14-35%, depending on the enzyme/substrate ratio and incubation temperature. In some cases, the chitosan (DD 85 and 50%) oligomers prepared using crude chitosanase from B. thuringiensis B-387 indicated higher antifungal and antibacterial activities in vitro in comparison with the initial polysaccharides. The data obtained indicate the good prospect of chitosanase B-387 for the production of bioactive CHOs.