The Efficacy and Safety of an Intra-articular Dual-Acting Antibacterial Agent (TNP-2092) for Implant Infection-Associated Methicillin-Resistant Staphylococcus aureus.

Owing to the presence of microbial biofilm on the implant, the eradication of biofilm-associated infections poses a challenge for antibiotic therapies. The study aimed to investigate the efficacy and safety of the novel antibiotic agent TNP-2092 in the context of implant infections. In vivo, rats with periprosthetic joint infection (PJI) treated with antibiotics showed an increase in body weight and decrease in swelling, temperature, and width of knee, compared with the control group. Meanwhile, inflammatory markers in synovium and serum were decreased in the TNP-2092 group, consistent with the pathological results. Moreover, TNP-2092 was effective in eliminating bacteria and disruption biofilm formation, and further alleviated the abnormal bone absorption and reactive bone changes around the prosthesis. In conclusion, intra-articular injection of TNP-2092 is safe and effective in treating knee PJI in a rat model. The study provides a foundation for the future utilization of TNP-2092 in the management of implant-related infections.

Nanoemulsions of essential oils against multi-resistant microorganisms: An integrative review.

Microbial resistance to drugs continues to be a global public health issue that demands substantial investment in research and development of new antimicrobial agents. Essential oils (EO) have demonstrated satisfactory and safe antimicrobial action, being used in pharmaceutical, cosmetic, and food formulations. In order to improve solubility, availability, and biological action, EO have been converted into nanoemulsions (NE). This review identified scientific evidence corroborating the antimicrobial action of nanoemulsions of essential oils (NEEO) against antibiotic-resistant pathogens. Using integrative review methodology, eleven scientific articles evaluating the antibacterial or antifungal assessment of NEEO were selected. The synthesis of evidence indicates that NEEO are effective in combating multidrug-resistant microorganisms and in the formation of their biofilms. Factors such as NE droplet size, chemical composition of essential oils, and the association of NE with antibiotics are discussed. Furthermore, NEEO showed satisfactory results in vitro and in vivo evaluations against resistant clinical isolates, making them promising for the development of new antimicrobial and antivirulence drugs.

Assessing the efficiency and microbial diversity of H2S-removing biotrickling filters at various pH conditions.

This study aimed to investigate the operation of three parallel biotrickling filters (BTFs) in removing H2S at different pH conditions (haloalkaliphilic, neutrophilic, and acidophilic) and their associated microbial population in the biodesulfurization process. BTF columns were inoculated with enriched inoculum and experiments were performed by gradually reducing Empty Bed Retention Time (EBRT) and increasing inlet concentration in which the maximum removal efficiency and maximum elimination capacity in EBRT 60 s reached their maximum level in haloalkaline condition (91% and 179.5 g S-H2S m-3 h-1). For visualizing the attached microbial biofilms on pall rings, Scanning Electron Microscopy (SEM) was used and microbial community structure analysis by NGS showed that the most abundant phyla in haBTF, nBTF, and aBTF belong to Gammaproteobacteria, Betaproteobacteria, and Acidithiobacillia, respectively. Shannon and Simpson indexes evaluation showed a lower diversity of bacteria in the aBTF reactor than that of nBTF and haBTF and beta analysis indicated a different composition of bacteria in haBTF compared to the other two filters. These results indicated that the proper performance of BTF under haloalkaliphilic conditions is the most effective way for H2S removal from air pollutants of different industries.

Chemical composition, cytotoxicity, antimicrobial, antibiofilm, and anti-quorum sensing potential of Mentha Piperita essential oil against the oral pathogen Streptococcus mutans.

Dental caries is a highly prevalent oral disease affecting billions of individuals globally. The disease occurs chemically as a result of breakdown of the tooth surface attributed to metabolic activity in colonizing biofilm. Biofilms, composed of exopolysaccharides and proteins, protect bacteria like Streptococcus mutans, which is notable for its role in tooth decay due to its acid-producing abilities. While various antimicrobial agents may prevent biofilm formation, these drugs often produce side effects including enamel erosion and taste disturbances. This study aimed to examine utilization of the Mentha piperita essential oil as a potential antibiofilm activity agent against S. mutans. M. piperita oil significantly (1) reduced bacterial biofilm, (2) exhibited a synergistic effect when combined with chlorhexidine, and (3) did not induce cell toxicity. Chemical analysis identified the essential oil with 99.99% certainty, revealing menthol and menthone as the primary components, constituting approximately 42% and 26%, respectively. Further, M. piperita oil eradicated preformed biofilms and inhibited biofilm formation at sub-inhibitory concentrations. M. piperita oil also interfered with bacterial quorum sensing communication and did not produce any apparent cell toxicity in immortalized human keratinocytes (HaCaT). M. piperita represented an alternative substance for combating S. mutans and biofilm formation and a potential combination option with chlorhexidine to minimize side effects. An in-situ performance assessment requires further studies.

Characterization of Staphylococcus Species Isolates from Sheep Milk with Subclinical Mastitis: Antibiotic Resistance, Enterotoxins, and Biofilm Production.

Subclinical mastitis represents one of the most contagious diseases affecting animals involved in dairy production systems. Although coagulase-negative staphylococci (CoNSs) have been considered minor pathogens for many years, they have recently emerged as opportunistic pathogens in mastitis disorders. The objectives of this work were to assess the antimicrobial resistance profile and the ability to produce a biofilm in comparison with a reference strain and to search for genes related to biofilm production, antimicrobial resistance, and enterotoxins in 18 isolates of Staphylococcus species from the milk of sheep with subclinical mastitis, collected from different Sicilian farms. This knowledge is essential to provide basic information on the pathogenicity and virulence of staphylococcal species and their impact on animal health. All isolates were resistant to ampicillin, 88.8% to streptomycin, 77.7% to gentamicin, 44.4% to chloramphenicol, 27.7% to erythromycin, and 11.1% to tetracycline, and two isolates were strong biofilm producers. Antibiotic resistance gene profiling showed that 16.6% of isolates possess the blaZ gene, whereas the search of biofilm-associated genes revealed the occurrence of the sasC gene in 33.3% of isolates, the ica gene in 27.7%, and bap and agr (accessory gene regulator) genes in 16.6% of isolates. Altogether, the results of this study indicate that CoNSs can acquire virulence genes and could have a role as pathogens in subclinical mastitis.

Microbial granules on reactors performance during organic butyrate digestion: clean production.

This critical review for anaerobic degradation of complex organic compounds like butyrate using reactors has been enormously applied for biogas production. Biogas production rate has a great impact on: reactor granulation methanogenesis, nutrient content, shear velocity, organic loading and loss of nutrients taking place in the reactor continuously. Various technologies have been applied to closed anaerobic reactors to improve biogas production and treatment efficiency. Recent reviews showed that the application of closed anaerobic reactors can accelerate the degradation of organics like volatile fatty acid-butyrate and affect microbial biofilm formation by increasing the number of methanogens and increase methane production 16.5 L-1 CH4 L-1 POME-1. The closed anaerobic reactors with stable microbial biofilm and established organic load were responsible for the improvement of the reactor and methane production. The technology mentioned in this review can be used to monitor biogas concentration, which directly correlates to organic concentrations. This review attempts to evaluate interactions among the: degradation of organics, closed anaerobic reactors system, and microbial granules. This article provides a useful picture for the improvement of the degradation of organic butyrate for COD removal, biogas and methane production in an anaerobic closed reactor.

Recent reviews showed that the application of closed anaerobic reactors can accelerate the degradation of organic compounds, such as volatile fatty acid-butyrate, and affect microbial biofilm formation by increasing the number of methanogens, thus enhancing biogas production. The closed anaerobic reactors with stable microbial biofilm established the organic load and improved the performance of the reactor for methane production. The technology used involves monitoring biogas concentrations which correlates with organic concentrations. This review attempts to evaluate interactions among: the degradation of organics, closed anaerobic reactors system, and microbial granules. This review, therefore, provides a useful picture for the improvement of butyrate degradation for COD removal and methane production with the help of various anaerobic closed reactors. The performance of UASBR depends on granulation. The granulation process in UASB reactors can be divided into four steps: (1) Transport of cells to the surface of other cells; (2) Initial reversible adsorption by physicochemical forces; (3) Irreversible adhesion of the cells by microbial appendages and/or polymers; and (4) Multiplication of the cells and development of the granules. Any factor which can complement any one of the four steps will be able to accelerate the granulation process and shorten the startup time of UASB reactors.

Effects of two novel denture cleansers on multispecies microbial biofilms, stain removal and the denture surface: an in vitro study.

BACKGROUND: The continuously increasing demand for removable denture appliances and the importance of adequate denture cleaning have led to the development of various denture cleansing products. The aim of this study was to evaluate the efficacy of two novel denture cleansing agents (GE and TM) and three commonly available cleansers (0.5% sodium hypochlorite; NaClO, 0.12% chlorhexidine gluconate; CHX, and Polident®; POL) on multispecies microbial biofilm formation, stain removal and physical properties of dentures. METHODS: The antimicrobial activities of denture cleansing agents were determined against major oral opportunistic pathogens including Streptococcus mutans, Staphylococcus aureus, Escherichia coli and Candida albicans, using time-kill assays. Multispecies microbial biofilms grown on acrylic resins for 72 h were generated to determine the antibiofilm effects of cleansing agents by confocal laser scanning microscopy (CLSM). Evaluations of the tea and coffee stain removal properties and the alterations in the physical properties of dentures were also performed. The toxicity of cleanser residues released from denture acrylics to fibroblast cells was investigated using MTT assay. RESULTS: All denture cleansing agents tested could effectively kill oral bacteria and Candida albicans. Furthermore, after immersion for more than 3 h, the cleansers Polident®, GE and TM could efficiently penetrate and inhibit multispecies denture biofilms with effects similar to 10 min of immersion in 0.5% NaClO. However, immersion in 0.12% CHX for 20 min showed less antibiofilm activity. The NaClO solution had the highest efficacy for removing stains from the artificial teeth. Conversely, the CHX solution enhanced tea and coffee staining, and the teeth immersed in this solution showed clinically unacceptable colour changes (ΔE > 5.5). However, the colour differences of teeth stained and immersed in POL, GE and TM cleansers were in the clinically acceptable range. There was no significant difference among the POL, GE and TM cleansers in terms of stain removal efficacy. The cleansers GE and TM did not alter the surface roughness and colour of the materials, moreover the residues of both cleansers did not exhibit cytotoxicity. CONCLUSION: Two novel denture cleansing agents containing natural products, GE and TM exhibited effective antimicrobial activity, antibiofilm and stain removal capabilities without toxicity or disturbance of the physical properties of acrylics.

Bacterial biofilm prevalence in dental unit waterlines: a systematic review and meta-analysis.

BACKGROUNDS: Numerous studies have shown that dental unit water lines (DUWLs) are often contaminated by a wide range of micro-organisms (bacteria, fungi, protozoa) and various prevalence have been reported for it in previous studies. Therefore, this review study aims to describe the prevalence of bacterial biofilm contamination of DUWLs. METHODS: This is a systematic review and meta-analysis in which the related keywords in different international databases, including Medline (via PubMed) and Scopus were searched. The retrieved studies were screened and the required data were extracted from the included studies. Three standard methods including American Dental Association (ADA), The Center for Disease Control and Prevention (CDC) and contaminated > 100 CFU/ml(C-100) standards were used to assess the bacterial biofilm contamination of DUWLs. All studies that calculated the prevalence of bacterial biofilm contamination of DUWLs, and English full-text studies were included in the meta-analysis. Studies that did not have relevant data or used unusual laboratory methods were excluded. Methodological risk of bias was assessed by a related checklist and finally, the data were pooled by fixed or random-effect models. RESULTS: Seven hundred and thirty-six studies were identified and screened and 26 related studies were included in the meta-analysis. The oldest included study was published in 1976 and the most recent study was published in 2020. According to the ADA, CDC and C-100 standards, the prevalence of bacterial contamination was estimated to be 85.0% (95% confidence interval (CI): 66.0-94.0%), 77.0% (95%CI: 66.0-85.0%) and 69.0% (95%CI: 67.0-71.0%), respectively. The prevalence of Legionella Pneumophila and Pseudomonas Aeruginosa in DUWLs was estimated to be 12.0% (95%CI: 10.0-14.0%) and 8.0% (95%CI: 2.0-24.0%), respectively. CONCLUSION: The results of this review study suggested a high prevalence of bacterial biofilm in DUWLs; therefore, the use of appropriate disinfecting protocol is recommended to reduce the prevalence of contamination and reduce the probable cross-infection.

A Rapidly Responsive Sensor for Wireless Detection of Early and Mature Microbial Biofilms.

Biofilm-associated infections, which are able to resist antibiotics, pose a significant challenge in clinical treatments. Such infections have been linked to various medical conditions, including chronic wounds and implant-associated infections, making them a major public-health concern. Early-detection of biofilm formation offers significant advantages in mitigating adverse effects caused by biofilms. In this work, we aim to explore the feasibility of employing a novel wireless sensor for tracking both early-stage and matured-biofilms formed by the medically relevant bacteria Staphylococcus aureus and Pseudomonas aeruginosa. The sensor utilizes electrochemical reduction of an AgCl layer bridging two silver legs made by inkjet-printing, forming a part of near-field-communication tag antenna. The antenna is interfaced with a carbon cloth designed to promote the growth of microorganisms, thereby serving as an electron source for reduction of the resistive AgCl into a highly-conductive Ag bridge. The AgCl-Ag transformation significantly alters the impedance of the antenna, facilitating wireless identification of an endpoint caused by microbial growth. To the best of our knowledge, this study for the first time presents the evidence showcasing that electrons released through the actions of bacteria can be harnessed to convert AgCl to Ag, thus enabling the wireless, battery-less, and chip-less early-detection of biofilm formation.

[Evaluation of the antibacterial effect of passive nanoimpregnation of tooth root dentin by real-time PCR]. / Otsenka antibakterial'nogo effekta passivnoi nanoimpregnatsii dentina kornya zuba metodom polimeraznoi tsepnoi reaktsii v real'nom vremeni.

OBJECTIVE: Assessment of antibacterial properties of a new intracanal paste based on calcium hydroxocuprate (CHC) and silver nanoparticles hydrosol for passive root impregnation. MATERIAL AND METHODS: The study included 55 teeth with 69 root canals belonging patients with chronic apical periodontitis. The main group, including 44 root canals, was filled with a new paste based on CHC and silver nanoparticles for 7 days after preparation and irrigation. In the control group, 25 root canals were sealed with an aqueous paste of calcium hydroxide for 14 days. The presence of the endodontic microorganisms was evaluated by real-time PCR. RESULTS AND DISCUSSION: Further analysis showed that the amount of the DNA, common for P. gingivalis, T. forsythia and T. denticola, after treatment was less in the main group, where the new paste was applied. These results were significant at the p=0.05 level (p=0.005, p=0.006, p=0.003 according to each mentioned bacterial sample). No significant differences were found between the groups in the number of genome equivalents specific for P. intermedia and F. nucleatum (p=0.543, p=0.554). CONCLUSION: These findings suggest that the new method of passive root impregnation with the CHC and silver nanoparticles paste may be an effective method for the treatment of chronical apical periodontitis.

Boswellic Acids as Effective Antibacterial Antibiofilm Agents.

Boswellic acids are biologically active pentacyclic terpenoid compounds derived from Boswellia sp. plants. Extracts containing these acids have a number of positive effects on human health, especially in the treatment of inflammation, arthritis, or asthma. With increasing resistance to common antibiotics, boswellic acid-containing extracts could serve as an alternative or work in synergy with commonly available preparations. This study aims to determine the effect of boswellic acids on suspension cells and biofilms of Staphylococcus epidermidis, Enterococcus faecalis, and Escherichia coli. The antimicrobial and antibiofilm effect found was compared with commonly available antibiotics to control these undesirable microorganisms. The synergistic effect of boswellic acids and common antibiotics on the growth of these microorganisms was also determined. All tested microorganisms showed a positive additive effect of antibiotics and boswellic acid extract. The most significant effect was found in Enterococcus faecalis ATCC 29212 in a combination of 0.2 × MIC80 erythromycin (0.2 mg/L) and 0.8 × MIC80 boswellic acid extract (16 mg/L).

Effect of quorum quenchers on virulence factors production and quorum sensing signalling pathway of non-mucoid, mucoid, and heavily mucoid Pseudomonas aeruginosa.

Quorum quenching (QQ), a mechanism which inhibits, interferes or inactivates quorum sensing, has been investigated for control of biofilms instigated by quorum sensing process. Application of quorum quenchers (QQs) provides the possibility to investigate how different phenotypes of Pseudomonas aeruginosa (non-mucoid, mucoid, and heavily mucoid strains) modulate their gene expression to form biofilms, their quorum sensing (QS) mediated biofilm to be formed, and their virulence expressed. The mRNA expression of the AHL-mediated QS circuit and AHL-mediated virulence factors in P. aeruginosa was investigated in presence of QQs. qPCR analysis showed that farnesol and tyrosol actively reduce the expression of the synthase protein, LasI and RhlI, and prevent production of 3OC12-HSL and C4-HSL, respectively. Also, the use of farnesol and tyrosol significantly moderated gene expression for exo-proteins toxA, aprA, LasB, as well as rhlAB, which are responsible for rhamnolipid production. Our findings were promising, identifying several suppressive regulatory effects of furanone and Candida albicans QS signal molecules, tyrosol, and farnesol on the AHL-mediated P. aeruginosa QS network and related virulence factors.

Interactions Between Plastic, Microbial Biofilms and Gammarus pulex: An Initial Investigation.

There is increasing evidence that microbial biofilms which form on the surface of marine plastics can increase plastics palatability, making it more attractive to organisms. The same information, however, does not exist for freshwater systems. This study observed the response of the freshwater amphipod Gammarus pulex when exposed to 3 cm-diameter discs of biofilm-covered plastic, both alone and when presented alongside its natural food. G. pulex did not fragment or consume the plastic materials, and the presence of colonised plastic in the immediate environment did not alter the amount of time organisms spent interacting with their natural food. This study provides baseline information for virgin and microbially colonised low-density polyethylene and polylactic acid film. Further studies, with other types of plastic possessing different physical properties and with different microbial biofilm compositions are now required to build further understanding of interactions between plastic, microbial biofilms, and freshwater shredding invertebrates.

Synthesis and Bioactivity of Ancorinoside B, a Marine Diglycosyl Tetramic Acid.

The sponge metabolite ancorinoside B was prepared for the first time in 16 steps and 4% yield. It features a ß-d-galactopyranosyl-(1→4)-ß-d-glucuronic acid tethered to a d-aspartic acid-derived tetramic acid. Key steps were the synthesis of a fully protected d-lactose derived thioglycoside, its attachment to a C20-aldehyde spacer, functionalization of the latter with a terminal N-(ß-ketoacyl)-d-aspartate, and a basic Dieckmann cyclization to close the pyrrolidin-2,4-dione ring with concomitant global deprotection. Ancorinoside B exhibited multiple biological effects of medicinal interest. It inhibited the secretion of the cancer metastasis-relevant matrix metalloproteinases MMP-2 and MMP-9, and also the growth of Staphylococcus aureus biofilms by ca 87% when applied at concentrations as low as 0.5 µg/mL. This concentration is far below its MIC of ca 67 µg/mL and thus unlikely to induce bacterial resistance. It also led to a 67% dispersion of preformed S. aureus biofilms when applied at a concentration of ca 2 µg/mL. Ancorinoside B might thus be an interesting candidate for the control of the general hospital, catheter, or joint protheses infections.

Efficacy of Er:YAG laser irradiation for decontamination and its effect on biocompatibility of different titanium surfaces.

BACKGROUND: Erbium yttrium-aluminum-garnet (Er:YAG) laser have been shown to be suitable for decontamination of titanium surfaces at a wide range of energy settings, however, high intensity of laser irradiation destroy titanium surface and low intensity cannot remove enough microbial biofilm. The aim of this study was to investigate the optimal energy setting of Er:YAG laser for decontamination of sandblasted/acid-etched (SLA) and hydroxyapatite (HA) titanium surfaces. MATERIAL AND METHODS: After supragingival biofilm construction in vivo, SLA and HA titanium discs were divided into three groups: blank control (BC, clean discs), experimental control (EC, contaminated discs) and experimental groups (EP, contaminated discs irradiated by Er:YAG laser at 40, 70, and 100 mJ/pulse). Scanning electron microscopy (SEM), live/dead bacterial fluorescent detection, and colony counting assay were used to detect the efficacy of laser decontamination. To investigate the effect of laser decontamination on titanium surface biocompatibility, MC3T3-E1 cell adhesion and proliferation activity were examined by SEM and CCK-8 assay. RESULTS: Er:YAG laser irradiation at 100 mJ/pulse removed 84.1% of bacteria from SLA titanium surface; laser irradiation at 70 and 100 mJ/pulse removed 76.4% and 77.85% of bacteria from HA titanium surface respectively. Laser irradiation improved MC3T3-E1 cell adhesion on both titanium surfaces. For SLA titanium discs, 100 mJ/pulse group displayed excellent cellular proliferation activity higher than that in BC group (P < 0.01). For HA titanium discs, 70 mJ/pulse group showed the highest activity comparable to BC group (P > 0.05). CONCLUSIONS: With regards to efficient microbial biofilm decontamination and biocompatibility maintenance, Er:YAG laser at 100 mJ/pulse and 70 mJ/pulse are considered as the optimal energy settings for SLA titanium and HA titanium surface respectively. This study provides theoretical basis for the clinical application of Er:YAG laser in the treatment of peri-implantitis.

Microbiological findings in early and late implant loss: an observational clinical case-controlled study.

BACKGROUND: Implants are a predictable and well-established treatment method in dentistry. Nevertheless, looking at possible failures of dental implants, early and late loss have to be distinguished. The intent of the study was to report microbiological findings on the surface of implants with severe peri-implantitis, which had to be explanted. METHODS: 53 specimens of implants from 48 patients without severe general illnesses have been examined. The groups investigated were implants that had to be removed in the period of osseointegration (early loss, 13 patients with 14 implants) or after the healing period (late loss, 14 patients with 17 implants). The implant losses were compared with two control groups (implants with no bone loss directly after completed osseointegration, two to four months after implant placement (17 patients with 17 implants) and implants with no bone loss and prosthetic restoration for more than three years (5 patients with 5 implants)). Data about the bacteria located in the peri-implant sulcus was collected using amplification and high throughput sequencing of the 16S rRNA gene. RESULTS: The biofilm composition differed substantially between individuals. Both in early and late implant loss, Fusobacterium nucleatum and Porphyromonas gingivalis were found to be abundant. Late lost implants showed higher bacterial diversity and in addition higher abundances of Treponema, Fretibacterium, Pseudoramibacter and Desulfobulbus, while microbial communities of early loss implants were very heterogeneous and showed no significantly more abundant bacterial taxa. CONCLUSIONS: Specific peri-implant pathogens were found around implants that were lost after a primarily uneventful osseointegration. P. gingivalis and F. nucleatum frequently colonized the implant in early and late losses and could therefore be characteristic for implant loss in general. In general, early lost implants showed also lower microbial diversity than late losses. However, the microbial results were not indicative of the causes of early and late losses.

[Colonization activity of conditionally pathogenic microorganisms and features of biofilm formation on the surface of thermoplastic polymer material]. / Kolonizatsionnaya aktivnost' uslovno-patogennykh mikroorganizmov i osobennosti obrazovaniya bioplenki na poverkhnosti stomatologicheskogo termoplastichnogo polimernogo materiala.

THE PURPOSE OF THE STUDY: Was to assess the microbial colonization and biofilm-forming activity of conditionally pathogenic microorganisms in vitro to samples of acrylic-free thermoplastic polymer material and to assess the possibility of its use for the manufacture of removable tooth splinting structures by injection method. MATERIALS AND METHODS: The reference strains S. aureus, S. epidermidis, S. pyogenes, E. faecalis, E. coli, and C. albicans. Biofilm-forming activity was assessed by the level of ethanol extraction of a 0.1% aqueous solution of gentian violet by measuring on a PowerWave X microplate reader (USA). RESULTS: A low degree of severity of microbial colonization and sufficient colonization resistance to the formation of a microbial biofilm of E. coli (0.374±0.056 cu opt. Pl.), S. aureus (0.272±0.039 cu. Opt. Pl.), S. epidermdis (0.299±0.028 cu opt. Pl.), S. ryogenes (0.399±0.069 cu opt. Pl.), E. faecalis (0.401±0.089 cu opt. Pl.). Moreover, C. albicans strains form a more pronounced biofilm (0.425±0.104 cu opt. Pl.) in comparison with other strains studied in the experiment. CONCLUSION: Samples from a thermoplastic polymer based on polyoxymethylene have satisfactory biomedical characteristics and can be used at the stages of dental orthopedic treatment of patients without oral candidiasis.

Alternative method in Galleria mellonella larvae to study biofilm infection and treatment.

In vivo studies are crucial decision-maker step in order to translate in vitro data to an applied therapy. Considering this we describe a simple method that analyzes and quantifies biofilm formation inside the Galleria mellonella larvae. Toothbrush bristles were employed as an abiotic surface to mimic a medical device. A standardized inoculum of Staphylococcus aureus was systemically injected in the larvae together with the insertion of a bristle in the last proleg pair. After incubation adhered cells were detached from bristles and quantified by colony-forming units (CFU) counting using staphylococci-selective medium. About 3 × 106 CFU of S. aureus were recovered from bristles and scanning electron microscopy (SEM) images confirmed biofilm formation. Control group did not show adherent bacteria, as demonstrated by absence of CFU counting and SEM images, indicating that the insertion procedure is free of bacterial contamination. We present a feasible method to evaluate bacterial biofilm formation in vivo that in the near future can be used to evaluate antibiofilm compounds.

Usnic Acid: Potential Role in Management of Wound Infections.

Usnic acid (UA) is a secondary lichen metabolite extensively studied for the broad variety of biological features. The most interesting property of UA is its antimicrobial activity against Gram-positive bacteria growing either in planktonic or in biofilm mode. In this chapter, the most relevant studies assessing usnic acid activity against microbial biofilms have been summarized and the potential role of UA in the management of biofilm-based wound infections has been critically discussed. Additionally, an overview of the main strategies adopted so far to reduce drug toxicity and increase bioavailability is given in the perspective of a safe use of UA in the clinical management of infected wounds.

Synthesis, Characterization, and Bacterial Fouling-Resistance Properties of Polyethylene Glycol-Grafted Polyurethane Elastomers.

Despite advances in material sciences and clinical procedures for surgical hygiene, medical device implantation still exposes patients to the risk of developing local or systemic infections. The development of efficacious antimicrobial/antifouling materials may help with addressing such an issue. In this framework, polyethylene glycol (PEG)-grafted segmented polyurethanes were synthesized, physico-chemically characterized, and evaluated with respect to their bacterial fouling-resistance properties. PEG grafting significantly altered the polymer bulk and surface properties. Specifically, the PEG-grafted polyurethanes possessed a more pronounced hard/soft phase segregated microstructure, which contributed to improving the mechanical resistance of the polymers. The better flexibility of the soft phase in the PEG-functionalized polyurethanes compared to the pristine polyurethane (PU) was presumably also responsible for the higher ability of the polymer to uptake water. Additionally, dynamic contact angle measurements evidenced phenomena of surface reorganization of the PEG-functionalized polyurethanes, presumably involving the exposition of the polar PEG chains towards water. As a consequence, Staphylococcus epidermidis initial adhesion onto the surface of the PEG-functionalized PU was essentially inhibited. That was not true for the pristine PU. Biofilm formation was also strongly reduced.