Bacterial Biofilm Development Research Articles

Research Progress on the Combination of Quorum-Sensing Inhibitors and Antibiotics against Bacterial Resistance.

Bacterial virulence factors and biofilm development can be controlled by the quorum-sensing (QS) system, which is also intimately linked to antibiotic resistance in bacteria. In previous studies, many researchers found that quorum-sensing inhibitors (QSIs) can affect the development of bacterial biofilms and prevent the synthesis of many virulence factors. However, QSIs alone have a limited ability to suppress bacteria. Fortunately, when QSIs are combined with antibiotics, they have a better therapeutic effect, and it has even been demonstrated that the two together have a synergistic antibacterial effect, which not only ensures bactericidal efficiency but also avoids the resistance caused by excessive use of antibiotics. In addition, some progress has been made through in vivo studies on the combination of QSIs and antibiotics. This article mainly expounds on the specific effect of QSIs combined with antibiotics on bacteria and the combined antibacterial mechanism of some QSIs and antibiotics. These studies will provide new strategies and means for the clinical treatment of bacterial infections in the future.

Chestnut Honey Is Effective against Mixed Biofilms at Different Stages of Maturity.

The irresponsible overuse of antibiotics has increased the occurrence of resistant bacterial strains, which represents one of the biggest patient safety risks today. Due to antibiotic resistance and biofilm formation in bacteria, it is becoming increasingly difficult to suppress the bacterial strains responsible for various chronic infections. Honey was proven to inhibit bacterial growth and biofilm development, offering an alternative solution in the treatment of resistant infections and chronic wounds. Our studies included chestnut honey, valued for its high antibacterial activity, and the bacteria Pseudomonas aeruginosa, methicillin-resistant Staphylococcus aureus, and S. epidermidis, known to form multi-species biofilm communities. Minimum inhibitory concentrations (MIC) of chestnut honey were determined for each bacterial strain. Afterwards, the mixed bacterial biofilms were treated with chestnut honey at different stages of maturity (incubation times: 2, 4, 6, 12, 24 h). The extent of biofilm inhibition was measured with a crystal violet assay and demonstrated by scanning electron microscopy (SEM). As the incubation time increased and the biofilm became more mature, inhibition rates decreased gradually. The most sensitive biofilm was the combination MRSA-S. epidermidis, with a 93.5% inhibition rate after 2 h of incubation. Our results revealed that chestnut honey is suitable for suppressing the initial and moderately mature stages of mixed biofilms.

Alteration of stainless-steel surface potential by modifying topography inhibits the development of bacterial biofilm

Alteration of stainless-steel surface potential by modifying topography inhibits the development of bacterial biofilm

Nanostructured Coatings Based on Graphene Oxide for the Management of Periprosthetic Infections.

To modulate the bioactivity and boost the therapeutic outcome of implantable metallic devices, biodegradable coatings based on polylactide (PLA) and graphene oxide nanosheets (nGOs) loaded with Zinforo™ (Zin) have been proposed in this study as innovative alternatives for the local management of biofilm-associated periprosthetic infections. Using a modified Hummers protocol, high-purity and ultra-thin nGOs have been obtained, as evidenced by X-ray diffraction (XRD) and transmission electron microscopy (TEM) investigations. The matrix-assisted pulsed laser evaporation (MAPLE) technique has been successfully employed to obtain the PLA-nGO-Zin coatings. The stoichiometric and uniform transfer was revealed by infrared microscopy (IRM) and scanning electron microscopy (SEM) studies. In vitro evaluation, performed on fresh blood samples, has shown the excellent hemocompatibility of PLA-nGO-Zin-coated samples (with a hemolytic index of 1.15%), together with their anti-inflammatory ability. Moreover, the PLA-nGO-Zin coatings significantly inhibited the development of mature bacterial biofilms, inducing important anti-biofilm efficiency in the as-coated samples. The herein-reported results evidence the promising potential of PLA-nGO-Zin coatings to be used for the biocompatible and antimicrobial surface modification of metallic implants.

Anti-Bacterial Activity of Green Synthesised Silver and Zinc Oxide Nanoparticles against Propionibacterium acnes.

Propionibacterium acnes plays a critical role in the development of acne vulgaris. There has been a rise in the number of patients carrying P. acnes strains that are resistant to antibiotics. Thus, alternative anti-microbial agents are required. Zinc oxide (ZnO-NPs) and silver (Ag-NPs) nanoparticles can be used against several antibiotic-resistant bacteria. The impact of Ag-NPs and ZnO-NPs against two clinical strains of P. acnes, P1 and P2, and a reference strain, NCTC747, were investigated in this research. A chemical approach for the green synthesis of Ag-NPs and ZnO-NPs from Peganum harmala was employed. The microtiter plate method was used to examine the effects of NPs on bacterial growth, biofilm development, and biofilm eradication. A broth microdilution process was performed in order to determine minimal inhibitory (MIC) concentrations. Ag-NPs and ZnO-NPs had a spherical shape and average dimensions of 10 and 50 nm, respectively. MIC values for all P. acnes strains for Ag-NPs and ZnO-NPs were 125 µg/mL and 250 µg/mL, respectively. Ag-NP and ZnO-NP concentrations of 3.9- 62.5 µg/mL and 15-62.5 µg/mL significantly inhibited the growth and biofilm formation of all P. acnes strains, respectively. ZnO-NP concentrations of 15-62.5 μg/mL significantly inhibited the growth of NCTC747 and P2 strains. The growth of P1 was impacted by concentrations of 31.25 μg/mL and 62.5 μg/mL. Biofilm formation in the NCTC747 strain was diminished by a ZnO-NP concentration of 15 μg/mL. The clinical strains of P. acnes were only affected by ZnO-NP titres of more than 31.25 μg/mL. Established P. acne biofilm biomass was significantly reduced in all strains at a Ag-NP and ZnO-NP concentration of 62.5 µg/mL. The findings demonstrated that Ag-NPs and ZnO-NPs exert an anti-bacterial effect against P. acnes. Further research is required to determine their potential utility as a treatment option for acne.

Engineering Bacterial Biofilm Development and Structure via Regulation of Silver Nanoparticle Density in Graphene Oxide Composite Coating.

Graphene-based composites have shown significant potential in the treatment of biofilm infections in clinical settings due to their exceptional antimicrobial properties and specific mechanisms. Nevertheless, a comprehensive understanding of the influence exerted by nanoparticles embedded in the composites on the development and structure of biofilms is still lacking. Here, we fabricate different graphene oxide-silver nanoparticle (GAg) composite-modified substrates (GAgS) with varying densities of silver nanoparticles (AgNPs) and investigate their effects on planktonic bacterial adhesion, subsequent biofilm formation, and mature biofilm structure. Our findings indicate that the initial attachment of Pseudomonas aeruginosa cells during biofilm formation is determined by the density of AgNPs on the GAgS surface. In contrast, the subsequent transition from adherent bacteria to the biofilm is determined by GAgS's synergistic antimicrobial effect. There exists a threshold for the inhibitory performance of GAgS, where the 20 μg/cm2 GAg composite completely prevents biofilm formation; below this concentration, GAgS delays the development of the biofilm and causes structural changes in the mature biofilm with enhanced bacterial growth and increased production of extracellular polymeric substance. More importantly, GAgS have minimal impact on mammalian cell morphology and proliferation while not inducing hemolysis in red blood cells. These results suggest that GAg composites hold promise as a therapeutic approach for addressing medical devices and implant-associated biofilm infections.

The Influence of Hydroxyapatite, Titanium, and Yttria-Stabilized Zirconia Oxide on Diversity of Bacterial Cultures Grown from Human Dental Biofilm Specimens in an In Vitro Model

Background: Titanium (TI) and yttria stabilized-zirconia oxide (YSZ) are dental materials commonly utilized at the soft-tissue interface surrounding dental implants. The influence of these surfaces on bacterial adhesion and biofilm development may affect clinical performance and patient susceptibility to inflammatory peri-implant disease. The purpose of this study was to evaluate the influence of the substrate material on biofilm diversity. Methods: Biofilms were cultured on TI, YSZ, and hydroxyapatite (HA) surfaces (control) using plaque specimens obtained from three human donors. Duplicate cultures grew for one, two, three, six or nine days. Biofilm diversity was then analyzed using 16S rRNA sequencing. The Shannon Diversity Index (SDI) was calculated for each experimental group. Microbial profiles were intercompared in a pairwise fashion to establish dissimilarity scores, which were recorded in a distance dissimilarity matrix. Results: A total of 16 taxa were identified, and relative abundances of the predominant phyla and genera did not appear statistically different across experimental groups. Biofilms grown on HA surfaces exhibited significantly higher alpha diversity compared with those formed on TI or YSZ (p<0.0001), although biofilms cultured on TI and YSG surfaces exhibited comparable diversity. Statistically significant differences in beta diversity associated with substrate (p=0.012) and growth period (p=0.001) were detected. Conclusions: Under the conditions described, biofilms grown on TI or YSZ appeared significantly less complex than those formed on HA. Transmucosal implant abutment surface characteristics represent one modifiable factor potentially influencing risk of peri-implant disease. Among risk of peri-implant disease. Among multiple considerations in abutment design, biofilm diversity performance may represent a clinically relevant benchmark.

Biofilm formation and manipulation with optical tweezers.

Some bacterial species form biofilms in suboptimal growth and environmental conditions. Biofilm structures allow the cells not only to optimize growth with nutrient availability but also to defend each other against external stress, such as antibiotics. Medical and bioengineering implications of biofilms have led to an increased interest in the regulation of bacterial biofilm formation. Prior research has primarily focused on mechanical and chemical approaches for stimulating and controlling biofilm formation, yet optical techniques are still largely unexplored. In this paper, we investigate the biofilm formation of Bacillus subtilis in a minimum biofilm-promoting medium (MSgg media) and explore the potential of optical trapping in regulating bacterial aggregation and biofilm development. Specifically, we determine the most advantageous stage of bacterial biofilm formation for optical manipulation and investigate the impact of optical trapping at different wavelengths on the aggregation of bacterial cells and the formation of biofilm. The investigation of optically regulated biofilm formation with optical tweezers presents innovative methodologies for the stimulation and suppression of biofilm growth through the application of lasers.

The effect of 3D-printed bone tissue scaffolds geometrical designs on bacterial biofilm formation

Bone fractures are recognized as a global health problem. A common strategy to tackle this issue is to employ a tissue engineering scaffold to accelerate tissue healing. However, one of the main challenges that can result in delaying the recovery is the risk of bacterial infections. This study aims to assess the impact of the geometry and the porosity of tissue scaffolds on the Staphylococcus aureus biofilm formation. Three triply periodic minimal surface designs of Schwarz primitive (SP), gyroid (GY), and Schwarz diamond (SD) and re-entrant auxetic (RE) design were examined and compared to a reference design (RD) considering two different porosity levels of 75% and 45%. The amount of biofilm was quantified using crystal violet assay and was visualized using scanning electron microscopy. The SP scaffold, with low porosity, exhibited a significantly less amount of bacterial biofilm formation and was regarded as having the best design among the others, while the SD with low porosity showed the greatest amount of biofilm. The morphological analysis was also in line with the crystal violet assay results. On the other hand, the surface roughness was affected by the complexity, geometrical variations, and limitations of fused filament fabrication three-dimensional printing. For the RD, SP, GY, and SD designs, an increase in surface roughness was demonstrated to increase the production of bacterial biofilms. Without statistical significance, the RE design showed the opposite trend. Contrary to other designs, the increase in pore size of the SP and GY designs was associated with the development of bacterial biofilms. This study suggests that it is possible to minimize the likelihood of bacterial biofilm formation by optimizing the scaffold geometry and its manufacturing.

A versatile micromodel technology to explore biofilm development in porous media flows.

Bacterial biofilms that grow in porous media are critical to ecosystem processes and applications ranging from soil bioremediation to bioreactors for treating wastewater or producing value-added products. However, understanding and engineering the complex phenomena that drive the development of biofilms in such systems remains a challenge. Here we present a novel micromodel technology to explore bacterial biofilm development in porous media flows. The technology consists of a set of modules that can be combined as required for any given experiment and conveniently tuned for specific requirements. The core module is a 3D-printed micromodel where biofilm is grown into a perfusable porous substrate. High-precision additive manufacturing, in particular stereolithography, is used to fabricate porous scaffolds with precisely controlled architectures integrating flow channels with diameters down to several hundreds of micrometers. The system is instrumented with: ultraviolet-C light-emitting diodes; on-line measurements of oxygen consumption and pressure drop across the porous medium; camera and spectrophotometric cells for the detection of biofilm detachment events at the outlet. We demonstrate how this technology can be used to study the development of Pseudomonas aeruginosa biofilm for several days within a network of flow channels. We find complex dynamics whereby oxygen consumption reaches a steady-state but not the pressure drop, which instead features a permanent regime with large fluctuations. We further use X-ray computed microtomography to image the spatial distribution of biofilms and computational fluid dynamics to link biofilm development with local flow properties. By combining the advantages of additive manufacturing for the creation of reproducible 3D porous microarchitectures with the flow control and instrumentation accuracy of microfluidics, our system provides a platform to study the dynamics of biofilm development in 3D porous media and to rapidly test new concepts in process engineering.

Review on Anti-Biofilm Potential of CRISPR-Cas System on Various Pathogens

The development of bacterial biofilms entails regulatory as well as signalling mechanisms that regulate the shift from a mobile to astationary state of existence, the production of the extracellular polymeric matrix, and the progression of 3-dimensional biofilm formation withemphasis on how easily they may be created and how crucial they are in biological, ecological, and industrial settings, biofilms are the subject ofextensive research. Biofilms and a variety of pathogenic human disorders are frequently linked. Since bacteria in biofilms can resist antibiotics, theimmune system, and other treatments, biofilm infections are typically long-lasting. Many prokaryotes include CRISPR-associated proteins (Cas), a solidadaptable immunological system that may be programmed to damage the bacterial genomes and induce cell death. Short palindromic repeats that aregrouped and adequately spaced together make up CRISPR-Cas. In light of this, CRISPR-Cas can be seen as an exciting strategy to address andovercome antibiotic resistance. Furthermore, the CRISPR-Cas system can create "precise antimicrobials" that target bacterial infections according tospecific DNA sequences. This CRISPR-Cas technique is susceptible to drug-resistant microorganisms due to its selective targeting of the genesinvolved in biofilm formation, pathogenicity, and antibiotic resistance. However, this method requires potent vectors for the CRISPR-Cas system toaccess the bacterial genomes. As vectors, genetically engineered Phage, liposomes, and lipid-mediated nanoparticles are exciting options. Thistechnique has been used to prevent extracellular and intracellular from forming biofilms. The most current developments in creating innovations andpossible advantages of the various CRISPR-Cas delivery methods for the deliberate eradication of bacterial pathogens will be covered in this review,focussing mainly on the anti-biofilm potential, which is found to be one of the primary causes of the difficulty of irradiation of Multi-drug resistantbacteria. Additionally, each distribution system's positive aspects are highlighted, along with its challenges and potential for advancement in the future.

Ceftazidime-assisted synthesis of ultrasmall chitosan nanoparticles for biofilm penetration and eradication of Pseudomonas aeruginosa

Pseudomonas aeruginosa (P. aeruginosa) infections present a grave threat to immunocompromised individuals, particularly those with cystic fibrosis due to the development of bacterial biofilms. In this study, we engineered self-assembling chitosan-ceftazidime nanoparticles (CSCE) capable of effectively penetrating biofilms and eradicating P. aeruginosa. The CSCE nanoparticles were synthesized through ionic cross-linking, combining negatively charged ceftazidime with positively charged chitosan, resulting in uniform nanoparticles measuring approximately 40 nm in diameter, exhibiting high dispersity and excellent biocompatibility. Remarkably, these nanoparticles exhibited significant inhibition of P. aeruginosa growth, reduced pyocyanin production, and diminished biofilm formation, achieving a maximum inhibition rate of 22.44%. Furthermore, in vivo investigations demonstrated enhanced survival in mice with abdominal P. aeruginosa infection following treatment with CSCE nanoparticles, accompanied by reduced levels of inflammatory cytokines Interleukin-6 (125.79 ± 18.63 pg/mL), Interleukin-17 (125.67 ± 5.94 pg/mL), and Tumor Necrosis Factor-α (135.4 ± 11.77 pg/mL). Critically, mice treated with CSCE nanoparticles showed no presence of bacteria in the bloodstream following intraperitoneal P. aeruginosa infection. Collectively, our findings highlight the potential of these synthesized nanoparticles as effective agents against P. aeruginosa infections.

Microfluidics for Biofilm Studies

Biofilms are multicellular communities held together by a self-produced extracellular matrix and exhibit a set of properties that distinguish them from free-living bacteria. Biofilms are exposed to a variety of mechanical and chemical cues resulting from fluid motion and mass transport. Microfluidics provides the precise control of hydrodynamic and physicochemical microenvironments to study biofilms in general. In this review, we summarize the recent progress made in microfluidics-based biofilm research, including understanding the mechanism of bacterial adhesion and biofilm development, assessment of antifouling and antimicrobial properties, development of advanced in vitro infection models, and advancement in methods to characterize biofilms. Finally, we provide a perspective on the future direction of microfluidics-assisted biofilm research.

Chromatography-Free Synthesis of β-Carboline 1-Hydrazides and an Investigation of the Mechanism of Their Bioactivity: The Discovery of β-Carbolines as Promising Antifungal and Antibacterial Candidates.

Agrochemical science prioritizes the discovery and effective synthesis of innovative and promising lead compounds. Herein, we developed an efficient column chromatography-free synthesis for β-carboline 1-hydrazides via a mild CuBr2-catalyzed oxidation and investigated the antifungal and antibacterial activities and mechanisms for these compounds. In our study, compounds 4de (EC50 = 0.23 μg·mL-1) and 4dq (EC50 = 0.11 μg·mL-1) displayed the best efficacy, demonstrating enhancements in inhibitory activity of more than 20-fold against Ggt compared to silthiopham (EC50 = 2.39 μg·mL-1). Additionally, compound 4de (EC50 = 0.21 μg·mL-1) demonstrated outstanding in vitro antifungal activities as well as in vivo curative activities against Fg. According to preliminary mechanistic studies, β-carboline 1-hydrazides led to the accumulation of reactive oxygen species, destruction of cell membranes, and dysregulation of histone acetylation. Furthermore, several substances exhibited antibacterial activity against Psg and Cms by preventing the development of bacterial biofilms.

Preparation of antibacterial PHBV/GO nanocomposite membranes via electrospinning

Poly(3-hydroxybutyrate-co-3-hydroxypentanoate) (PHBV)/graphene oxide (GO) composite membranes were electrospun with an aim at fabricating antibacterial nanofibrous scaffolds for tissue engineering application. Randomly and aligned oriented PHBV/GO fibrous membrane were prepared by controlling the mixed solvent, GO concentration and process parameters, and their effect on the morphology and structure of the membranes was investigated. When 10 wt% GO nanosheets was incorporated into the composite spinning dope, PHBV/GO membrane presented a 18.7 MPa breaking strength and 5.51 × 102 MPa tensile modulus, much greater than 14.3 MPa and 1.76 × 102 MPa of PHBV membrane. Colonies forming units of E.coli and S.aureus reduced by 81.7% and 71.8% for randomly oriented PHBV/GO, and 84.2% and 73.4% for aligned oriented PHBV/GO when compared to PHBV membrane. The development of bacterial biofilm on PHBV/GO membrane reduced greatly when incorporating 5–10 wt% GO. The antibacterial PHBV/GO membranes with better mechanical properties showed great promise for further studies on bone tissue repair.

Fucoidan-Containing, Low-Adhesive Siloxane Coatings for Medical Applications: Inhibition of Bacterial Growth and Biofilm Development.

The deposition of low-adhesive siloxane coatings is a current trend for the non-toxic control of bacterial growth and biofilm formation. Total elimination of biofilm formation has not been reported so far. The aim of this investigation was to study the ability of a non-toxic, natural, biologically active substance, such as fucoidan, to inhibit bacterial growth on similar medical coatings. The fucoidan amount was varied, and its impact on the bioadhesion-influencing surface characteristics, as well as on bacterial cell growth, was investigated. The inclusion of up to 3-4 wt.% brown algae-derived fucoidan in the coatings increases their inhibitory effect, more significantly on the Gram-positive bacterium S. aureus than on the Gram-negative bacterium Escherichia coli. The biological activity of the studied siloxane coatings was ascribed to the formation of a low-adhesive, biologically active surface top layer consisting of siloxane oil and dispersed water-soluble fucoidan particles. This is the first report on the antibacterial activity of fucoidan-containing medical siloxane coatings. The experimental results give reason to expect that relevantly selected, natural biologically active substances can be efficient in the non-toxic control of bacterial growth on medical devices and, as a result, medical device-associated infections.

Synergistic effects of biofilm-producing PGPR strains on wheat plant colonization, growth and soil resilience under drought stress

Drought stress substantially impedes crop productivity throughout the world. Microbial based approaches have been considered a potential possibility and are under study. Based on our prior screening examination, two distinct and novel biofilm-forming PGPR strains namely Bacillus subtilis-FAB1 and Pseudomonas azotoformans-FAP3 are encompassed in this research. Bacterial biofilm development on glass surface, microtiter plate and seedling roots were assessed and characterized quantitatively and qualitatively by light and scanning electron microscopy. Above two isolates were further evaluated for their consistent performance by inoculating on wheat plants in a pot-soil system under water stresses. Bacterial moderate tolerance to ten-day drought was recorded on the application of individual strains with wheat plants; however, the FAB1 + FAP3 consortium expressively improved wheat survival during drought. The strains FAB1 and FAP3 displayed distinct and multifunctional plant growth stimulating attributes as well as effective roots and rhizosphere colonization in combination which could provide sustained wheat growth during drought. FAB1 and FAP3-induced alterations cooperatively conferred improved plant drought tolerance by controlling physiological traits (gs, Ci, E, iWUE and PN), stress indicators (SOD, CAT, GR, proline and MDA content) and also maintained physico-chemical attributes and hydrolytic enzymes including DHA, urease, ALP, protease, ACP and β glucosidase in the soil. Our findings could support future efforts to enhance plant drought tolerance by engineering the rhizobacterial biofilms and associated attributes which requires in-depth exploration and exploiting potential native strains for local agricultural application.

Main approaches of biofilm formation in composite resins: a concise systematic review

Introduction: Composite resins are widely used in clinical dental practice to restore the structure lost by caries and dental defects, achieving aesthetic and functional results. Failures in the photoactivation process can cause problems in the mechanical resistance to wear in regions of direct occlusal contact on the restorative material. This occurs due to the reduction in the degree of conversion of monomers into polymers, compromising the clinical performance of the composite resin. Polymerization shrinkage leads to microleakage of the restoration, leading to staining and the development of bacterial biofilms. Objective: It was to carry out a systematic review of the main scientific evidence of biofilm formation on composite resin veneers, identifying potential predictors, as well as proposals to mitigate this problem. Methods: The systematic review rules of the PRISMA Platform were followed. The search was carried out from December 2022 to March 2023 in the Scopus, PubMed, Science Direct, Scielo, and Google Scholar databases, using articles from 2015 to 2022. The quality of the studies was based on the GRADE instrument and the risk of bias was analyzed accordingly, according to the Cochrane instrument. Results and Conclusion: A total of 125 articles were found, 25 articles were evaluated and 20 were included and developed in this systematic review study. Considering the Cochrane tool for risk of bias, the overall assessment resulted in 10 studies with a high risk of bias and 70 studies that did not meet GRADE. It was concluded that the surface properties of resin-based composite materials, as well as surface treatments, can strongly affect bacterial adhesion and biofilm formation. In addition, scientific evidence highlights that cariogenic biofilm formation can alter the surface properties of materials, thus stimulating bacterial adhesion and biofilm formation. Photoactivation allows the curing and crosslinking of polymer chains, essential for the clinical longevity of the composite resin. Insufficient photoactivation culminates in marginal microleakage in restorations and biofilm accumulation on surfaces exposed to the oral environment.

Review of Antimicrobial Nanocoatings in Medicine and Dentistry: Mechanisms of Action, Biocompatibility Performance, Safety, and Benefits Compared to Antibiotics.

This review discusses topics relevant to the development of antimicrobial nanocoatings and nanoscale surface modifications for medical and dental applications. Nanomaterials have unique properties compared to their micro- and macro-scale counterparts and can be used to reduce or inhibit bacterial growth, surface colonization and biofilm development. Generally, nanocoatings exert their antimicrobial effects through biochemical reactions, production of reactive oxygen species or ionic release, while modified nanotopographies create a physically hostile surface for bacteria, killing cells via biomechanical damage. Nanocoatings may consist of metal nanoparticles including silver, copper, gold, zinc, titanium, and aluminum, while nonmetallic compounds used in nanocoatings may be carbon-based in the form of graphene or carbon nanotubes, or composed of silica or chitosan. Surface nanotopography can be modified by the inclusion of nanoprotrusions or black silicon. Two or more nanomaterials can be combined to form nanocomposites with distinct chemical or physical characteristics, allowing combination of different properties such as antimicrobial activity, biocompatibility, strength, and durability. Despite their wide range of applications in medical engineering, questions have been raised regarding potential toxicity and hazards. Current legal frameworks do not effectively regulate antimicrobial nanocoatings in matters of safety, with open questions remaining about risk analysis and occupational exposure limits not considering coating-based approaches. Bacterial resistance to nanomaterials is also a concern, especially where it may affect wider antimicrobial resistance. Nanocoatings have excellent potential for future use, but safe development of antimicrobials requires careful consideration of the "One Health" agenda, appropriate legislation, and risk assessment.

Staphylococcus aureus Behavior on Artificial Surfaces Mimicking Bone Environment

Infections, which interfere with bone regeneration, may be a critical issue to consider during the development of biomimetic material. Calcium phosphate (CaP) and type I collagen substrates, both suitable for bone-regeneration dedicated scaffolds, may favor bacterial adhesion. Staphylococcus aureus possesses adhesins that allow binding to CaP or collagen. After their adhesion, bacteria may develop structures highly tolerant to immune system attacks or antibiotic treatments: the biofilms. Thus, the choice of material used for scaffolds intended for bone sites is essential to provide devices with the ability to prevent bone and joint infections by limiting bacterial adhesion. In this study, we compared the adhesion of three different S. aureus strains (CIP 53.154, SH1000, and USA300) on collagen- and CaP-coating. Our objective was to evaluate the capacity of bacteria to adhere to these different bone-mimicking coated supports to better control the risk of infection. The three strains were able to adhere to CaP and collagen. The visible matrix components were more important on CaP- than on collagen-coating. However, this difference was not reflected in biofilm gene expression for which no change was observed between the two tested surfaces. Another objective was to evaluate these bone-mimicking coatings for the development of an in vitro model. Thus, CaP, collagen-coatings, and the titanium-mimicking prosthesis were simultaneously tested in the same bacterial culture. No significant differences were found compared to adhesion on surfaces independently tested. In conclusion, these coatings used as bone substitutes can easily be colonized by bacteria, especially CaP-coating, and must be used with an addition of antimicrobial molecules or strategies to avoid bacterial biofilm development.