Development of Radiosterilized Porcine Skin Electrosprayed with Silver Nanoparticles Prevents Infections in Deep Burns.

Extensive burns represent a significant challenge in biomedicine due to the multiple systemic and localized complications resulting from the major skin barrier loss. The functionalization of xenografts with nanostructured antibacterial agents proposes a fast and accessible application to restore barrier function and prevent localized bacterial contamination. Based on this, the objective of this work was to functionalize a xenograft by electrospray deposition with silver nanoparticles (AgNPs) and to evaluate its antibiofilm and cytotoxic effects on human fibroblasts. Initially, AgNPs were synthesized by a green microwave route with sizes of 2.1, 6.8, and 12.2 nm and concentrations of 0.055, 0.167, and 0.500 M, respectively. The AgNPs showed a size relationship directly proportional to the concentration of AgNO3, with a spherical and homogeneous distribution determined by high-resolution transmission electron microscopy. The surface functionalization of radiosterilized porcine skin (RPS) via electrospray deposition with the three AgNP concentrations (0.055, 0.167, and 0.500 M) in the epidermis and the dermis showed a uniform distribution on both surfaces by energy-dispersive X-ray spectroscopy. The antibiofilm assays of clinical multidrug-resistant Pseudomonas aeruginosa showed significant effects at the concentrations of 0.167 and 0.500 M, with a log reduction of 1.3 and 2.6, respectively. Additionally, viability experiments with human dermal fibroblasts (HDF) exposed to AgNPs released from functionalized porcine skin showed favorable tolerance, with retention of viability more significant than 90% for concentrations of 0.05 and 0.167 M after 24 h exposure. Antibacterial activity combined with excellent biocompatibility makes this biomaterial a candidate for antibacterial protection by inhibiting bacterial biofilms in deep burns during early stages of development.

Evaluation of anti-biofilm and cytotoxic effect of a gel formulation with Pluronic F-127 and silver nanoparticles as a potential treatment for skin wounds.

The skin wounds cause serious burden to healthcare systems. The lack of sterility of the innate barrier function of the skin facilitates the development of microbial communities within the wound environment especially in biofilm form. Since biofilm is difficult to eradicate, new treatments have been established, such as silver nanoparticles (AgNPs), which antimicrobial and anti-biofilm properties have been studied, nevertheless, their toxic effects are known too. Different concentrations of AgNPs stabilized with a biocompatible and thermo-reversible vehicle as hydrogel Pluronic F-127 were synthesized, those formulations presented interesting thermo-reversibility which could be used to apply on wounds. The formulations (Gel 62.5, 125, and 250 ppm of AgNPs) proposed in this study showed in vitro a total inhibition of clinical strains (Staphylococcus aureus and Pseudomonas aeruginosa) in planktonic form, as well as, anti-biofilm activity was archived with the formulation of Gel 250 ppm, a total inhibition of biofilm formation with mixed culture was registered in the first 30 min of biofilm growth; even more, the viability of human fibroblasts with all gels formulations was >95%, in contrast to silver sulfadiazine cream 1% which showed the highest cytotoxic effect. PF-127 gel with AgNPs could be a prophylactic treatment for skin wounds, because its activity in critical steps on biofilm formation.

Adhesion forces of biofilms developed in vitro from clinical strains of skin wounds.

A biofilm is a very complex consortium formed by a mix of different microorganisms, which have become an important health problem, because its formation is a resistance mechanism used by bacteria against antibiotics or the immune system. In this work, we show differences between some physicochemical properties of biofilms in mono- and multi-species, formed by bacteria from clinical samples of infected chronic wounds. Of the most prevalent bacteria in wounds, two mono- and one multi-species biofilms were developed in vitro by Drip Flow Reactor: one biofilm was developed by S. aureus, other by P. aeruginosa, and a third one by the mix of both strains. With these biofilms, we determined microbial growth by plate counting, and their physicochemical characterization by Atomic Force Microscopy, Raman Micro-Spectroscopy and Scanning Electron Microscopy. We found that the viability of S. aureus was less than P. aeruginosa in multi-species biofilm. However, the adhesion force of S. aureus is much higher than that of P. aeruginosa, but it decreased while that of P. aeruginosa increased in the multi-species biofilm. In addition, we found free pyrimidines functional groups in the P. aeruginosa biofilm and its mix with S. aureus. Surprisingly, each bacterium alone formed single layer biofilms, while the mix bacteria formed a multilayer biofilm at the same observation time. Our results show the necessity to evaluate biofilms from clinically isolated strains and have a better understanding of the adhesion forces of bacteria in biofilm multispecies, which could be of prime importance in developing more effective treatments against biofilm formation.