The aim of this study was to evaluate the effect of a moxifloxacin-loaded organicinorganic sol-gel with different antibiotic concentration in the in vitro biofilm development and treatment against Staphylococcus aureus, S. epidermidis, and Escherichia coli, cytotoxicity and cell proliferation of MC3T3-E1 osteoblasts; and its efficacy in preventing the prosthetic joint infection (PJI) caused by clinical strains of S. aureus and E. coli using an in vivo murine model. Three bacterial strains, S. epidermidis ATCC 35984, S. aureus 15981, and, E. coli ATCC 25922, were used for microbiological studies. Biofilm formation was induced using tryptic-soy supplemented with glucose for 24 h, and then, adhered and planktonic bacteria were estimated using drop plate method and absorbance, respectively. A 24-h-mature biofilm of each species growth in a 96well plate was treated for 24 h using a MBECTM biofilm Incubator lid with pegs coated with the different types of sol-gel, after incubation, biofilm viability was estimated using alamrBlue. MC3T3-E1 cellular cytotoxicity and proliferation were evaluated using CytoTox 96 Non-Radioactive Cytotoxicity Assay and alamarBlue, respectively. The microbiological studies showed that sol-gel coatings inhibited the biofilm development and treated to a mature biofilm of three evaluated bacterial species. The cell studies showed that the sol-gel both with and without moxifloxacin were non-cytotoxic and that cell proliferation was inversely proportional to the antibiotic concentration containing by sol-gel. In the in vivo study, mice weight increased over time, except in the E. coliinfected group without coating. The most frequent symptoms associated with infection were limping and piloerection; these symptoms were more frequent in infected groups
Candida auris is a multiresistant pathogenic yeast commonly isolated from bloodstream infections in immunocompromised patients. In this work, we infected G. mellonella larvae with 105 CFU of a reference strains and two clinical isolates of C. albicans and C. auris and we compared the outcomes of infection between both species. Larvae were evaluated every 24 h for a total of 120 h following the G. mellonella Health Index Scoring System, and survival, activity, melanization and cocoon formation were monitored. Our results showed that clinical isolates were significantly more pathogenic than reference strains independently of the tested species, producing lower survival and activity scores and higher melanization scores, and being C. albicans strains more virulent than C. auris strains. We did not find differences in mortality between aggregative and non-aggregative C. auris strains, although non-aggregative strains produced significantly lower activity scores and higher melanization scores than aggregative ones. Survival assays using Galleria mellonella have been previously employed to examine and classify strains of this and other microbial species based on their virulence before scaling the experiments to a mammal model. Taken together, these results show how a more complete evaluation of the model can improve the study of C. auris isolates.
A simple approach for the fabrication of functional nanopatterned protein materials using protein engineering and soft-nanolithography and its implementation in optical devices based on distributed feedback (DFB) laser phenomena.
The precise synthesis of materials and devices with tailored complex structures and properties is a requisite for the development of the next generation of products based on nanotechnology. Nowadays, the technology for the generation of this type of devices lacks the precision to determine their properties and is accomplished mostly by 'trial and error' experimental approaches. The use of bottom-up approaches that rely on highly specific biomolecular interactions of small and simple components is an attractive approach for the templating of nanoscale elements. In nature, protein assemblies define complex structures and functions. Engineering novel bio-inspired assemblies by exploiting the same rules and interactions that encode the natural diversity is an emerging field that opens the door to create nanostructures with numerous potential applications in synthetic biology and nanotechnology. Self-assembly of biological molecules into defined functional structures has a tremendous potential in nano-patterning and the design of novel materials and functional devices. Molecular self-assembly is a process by which complex 3D structures with specified functions are constructed from simple molecular building blocks. Here we discuss the basis of biomolecular templating, the great potential of repeat proteins as building blocks for biomolecular templating and nano-patterning. In particular, we focus on the designed consensus tetratricopeptide repeats (CTPRs), the control on the assembly of these proteins into higher order structures and their potential as building blocks in order to generate functional nanostructures and materials.
Ti-doped ZnO thin films were obtained
with the aim of tailoring
ZnO film bioadhesiveness and making the optoelectronic properties
of ZnO materials transferable to biological environments. The films
were prepared on silicon substrates by sol–gel spin-coating
and subsequent annealing. A Ti–O segregation limits the ZnO
crystallite growth and creates a buffer out-layer. Consequently, the
Ti-doped ZnO presents slightly increased resistivity, which remains
in the order of 10
–3
Ω·cm. The strong
biochemical interference of Zn
2+
ions released from pure
ZnO surfaces was evidenced by culturing
Staphylococcus
epidermidis
with and without the Zn
2+
coupling
agent clioquinol. The Ti-doped ZnO surfaces showed a considerable
increase of bacterial viability with respect to pure ZnO. Cell adhesion
was assayed with human mesenchymal stem cells (hMSCs). Although hMSCs
find difficulties to adhere to the pure ZnO surface, they progressively
expand on the surface of ZnO when the Ti doping is increased. A preliminary
microdevice has been built on the Si substrate with a ZnO film doped
with 5% Ti. A one-dimensional micropattern with a zigzag structure
shows the preference of hMSCs for adhesion on Ti-doped ZnO with respect
to Si. The induced contrast of surface tension further induces a cell
polarization effect on hMSCs. It is suggested that the presence of
Ti–O covalent bonding on the doped surfaces provides a much
more stable ground for bioadhesion. Such fouling behavior suggests
an influence of Ti doping on film bioadhesiveness and sets the starting
point for the selection of optimal materials for implantable optoelectronic
devices.
We report the fabrication of a conductive biomaterial based on engineered proteins and patterned gold nanoparticles to overcome the challenge of charge transport on macroscopic protein-based materials. This approach has great value for bioelectronics.
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