Effects of ZnO nanoparticles on aerobic denitrifying bacteria Enterobacter cloacae strain HNR.

The aerobic denitrification process is a promising and cost-effective alternative to the conventional nitrogen removal process. Widely used ZnO nanoparticles (NPs) will inevitably reach wastewater treatment plants, and cause adverse impacts on aerobic denitrification and nitrogen removal. Therefore, a full understanding of the responses and adaption of aerobic denitrifiers to ZnO NPs is essential to develop effective strategies to reduce adverse effects on wastewater treatment. In this study, the responses and adaption to ZnO NPs were investigated of a wild type strain (WT) and a resistant type strain (Re) of aerobic denitrifying bacteria Enterobacter cloacae strain HNR. When exposed to 0.75 mM ZnO NPs, the nitrate removal efficiency of Re was 11.2% higher than that of WT. To prevent ZnO NPs entering cells by adsorption, the production of extracellular polymeric substances (EPS) of WT and Re strains increased 13.2% and 43.9%, respectively. The upregulations of amino sugar and carbohydrate-related metabolism contributed to the increase of EPS production, and the increased nitrogen metabolism contributed to higher activities of nitrate and nitrite reductases. Interestingly, cationic antimicrobial peptide resistance contributed to resist Zn (II) released by ZnO NPs, and many antioxidative stress-related metabolism pathways were upregulated to resist the oxidative stress resulting from ZnO NPs. These findings will guide efforts to improve the aerobic denitrification process in an environment polluted by NPs, and promote the application of aerobic denitrification technologies.

Control of osteoblast cells adhesion and spreading by microcontact printing of extracellular matrix protein patterns.

In this study, we report a simple method for creating extracellular matrix (ECM) protein patterns to control osteoblast cell adhesion and spreading. The fibronectin patterns are directly produced on polystyrene (PS) surfaces by microcontact printing (µCP). Confocal laser scanning microscopy (CLSM) images show that protein patterns are successfully fabricated on PS surfaces. Newborn rat osteoblast cells are then seeded on these protein patterns and cultured for 4 days. The results demonstrate that osteoblast cells preferentially adhere and grow on the protein areas. The pattern dimensions have significant influences on cell behaviors, including cell adhesion, spreading, distribution, and growth direction. Therefore, it is possible to control the cell morphology and even cell function by carefully designing the pattern shapes and sizes. The present study suggests that the ECM protein patterns can be used to modify biomaterials' surfaces and spatially control the morphologies of osteoblast cells. We believe that our work could find applications for creating patterned bioactive surfaces to control cell adhesion, spreading and cell function. It may be helpful for the development of novel implantable biomaterials, such as artificial bone implants, where control of interfacial biological interactions between implants and cells would be preferable.

Enhancing the antibacterial activity of biomimetic HA coatings by incorporation of norvancomycin.

BACKGROUND: Bacterial infections associated with the use of biomaterials remain a great challenge for orthopedic surgery. The main purpose of the work discussed in this paper was to improve the antibacterial activity of a biomimetic calcium phosphate (CP) coating widely used in orthopedic biomaterials by incorporation of norvancomycin in the biomimetic process. METHODS: CP coating and CP coating containing norvancomycin were produced on a titanium alloy (Ti6Al4V) surface by a biomimetic process. The morphology, surface crystal structure, and concentrations of elements in the coatings were characterized by scanning electron microscopy (SEM), X-ray diffraction (XRD), and energy-dispersive X-ray spectroscopy (EDX), respectively. The amount of norvancomycin and its release were investigated by UV-visible spectroscopy. MTT was used to investigate cell behavior. The morphology of adhered bacteria was observed by SEM. Antibacterial activity was expressed as inhibition zone by using Staphylococcus aureus (ATCC 25923) as model bacteria. RESULTS: Results from SEM, EDX, and XRD revealed formation of a hydroxyapatite (HA) coating. The amount of antibiotic in the CP coating increased with increasing concentration of norvancomycin in the coating solution, followed by a plateau when the concentration of norvancomycin in the coating solution reached 600 mg/l. Approximately 2.16 µg norvancomycin per mg coating was co-precipitated with the CP layer onto titanium alloy discs when 600 mg/l norvancomycin coating solution was applied. The norvancomycin had a fast release profile followed by slow release. The MTT test of osteoblast cell cultures suggested that coatings containing norvancomycin did not cause any cytotoxicity compared with the CP coating and control titanium plate. The antibacterial activity test showed that the norvancomycin released from the coatings inhibited the growth of Staphylococcus aureus; more bacteria were found on the CP coating than on the norvancomycin-loaded coating. CONCLUSIONS: A norvancomycin-loaded HA-like coating was successfully obtained on titanium surfaces. The norvancomycin incorporated had no negative effects on osteoblast cell behavior. The released norvancomycin results in excellent antibacterial activity of Ca-P coatings. Therefore, incorporation of norvancomycin can enhance antibacterial activity and the norvancomycin-loaded CP coating can be used to inhibit post-surgical infections in orthopaedics.