Antibiofilm Testing Research Articles

Microspheres based on galactomannan and Spondias purpurea L. extract to increase antifungal and antibiofilm efficacy against Candida spp.

The ongoing problem of an increasing resistance of Candida spp. to available antifungals, has made it necessary the search for new therapeutic alternatives. The aim of this work was to develop a microsphere based on Caesalpinia ferrea galactomannan and Spondias purpurea L. stem bark extract using the spray drying technique and evaluate its antimicrobial effect on biofilm formation and planktonic cells of Candida spp. Differential scanning calorimetry (DSC), infrared analysis (IR) and scanning electron microscopy (SEM) were used to characterize the microsphere, in addition to the encapsulation efficiency by HPLC to quantify the extract in the microsphere. In microbiological analyses, broth microdilution and antibiofilm tests were carried out. The results of the minimum inhibitory concentration (MIC) for the stem bark extract (SBE) were within 0.5-2 μg mL-1 and the galactomannan microsphere (GMB) 1-8 μg mL-1. As for the biofilm, the microsphere compared to the extract showed a statistically significant improvement at 8xMIC for C. albicans 1, while for C. auris, it was at 4xMIC and 8xMIC. Furthermore, SBE and GMB did not present toxicity. The study revealed that the synthesized microspheres have the potential to be used as an antifungal agent.

Novel biocompatible magnetron-sputtered silver coating for enhanced antibacterial properties and osteogenesis in vitro

Peri-implant infection is a serious complication in orthopedic surgery. This study aimed to reduce the incidence of peri-implant infection by developing a durable and safe antibacterial silver coating. We compared the antibacterial properties and process controllability of various coating techniques to identify the most effective method for silver coating. We refined substrate treatment techniques and coating thicknesses through antibacterial and scratch tests. Scanning electron microscopy (SEM) and energy-dispersive X-ray spectroscopy (EDS) were used to analyze the coating’s morphology and composition. Micron-sized magnetron sputtering silver coating samples underwent in vitro antibacterial testing, cytotoxicity testing, silver ion release testing, and osteogenic testing using membrane contact culture, CCK-8 assay, inductively coupled plasma (ICP) detection, and alkaline phosphatase (ALP) activity/osteogenic gene PCR. Magnetron sputtering demonstrated superior antibacterial properties, uniformity, and process controllability compared to alternative techniques. The optimal adhesion strength was achieved with a 0.5 μm coating thickness and a 400 mesh sandpaper pretreatment process, without compromising antibacterial efficacy. The coating showed near-perfect antiseptic results in antibacterial and anti-biofilm tests. Fibroblasts cultured in silver ion precipitation medium exhibited growth rates of 89% on day 30 and 88% on day 90, compared to 95% in the control group. The osteogenic test indicated that the magnetron sputtering silver coating promotes osteogenesis effectively. Our study demonstrated that micron-sized magnetron sputtering silver coating has potential for clinical use to prevent peri-implant infections in the future.

Choline and geranate ionic liquid for subgingival biofilm control

Periodontitis is a chronic inflammatory disease that causes destruction of the periodontium and eventual tooth loss. The priority in the periodontal treatment is to remove the subgingival biofilm. Chemical removal of biofilms using antimicrobial agents has been applied in clinical practice. However, their clinical effect is still limited because the agents must overcome biofilm’s significant drug tolerance, which is primarily caused by the extracellular matrix, a physical barrier that attenuates drug diffusion. This study aimed to study the use of ionic liquids (ILs), a new class of biocompatible materials, for controlling subgingival biofilms because of their excellent permeability. Choline and geranate (CAGE) IL was tested for its highly potent antiseptic behavior and permeability. Antibacterial tests revealed that the significant efficacy of CAGE against periodontopathic microorganisms was derived from their ability to destroy cell membrane, as demonstrated by membrane permeability assay and transmission electron microscopy imaging. Antibiofilm tests using two pathogenic biofilm models revealed that CAGE exerted efficacy against the biofilm-embedded bacteria, conspicuously neutralized the biofilms, and eventually destroyed the biofilm structure. Furthermore, the penetration of CAGE into the biofilm was visually confirmed using confocal laser scanning microscopy. This study highlighted the potential of CAGE as a powerful antibiofilm therapeutic.

Efficacy of ozonated olive oil against peri-implant microbes isolated from peri-implantitis.

This study aimed to investigate the antibacterial and antimicrobial activity of ozone gel against oral biofilms grown on titanium dental implant discs. The experiment used medical grade five titanium discs on which peri-implant isolated biofilms were grown. The experimental groups were control, Streptococcus mutans (S. mutans) and Granulicatella adiacens (G. adiacens), (n = 6). The oral microbes grown on titanium discs were exposed to ozone gel for 3 minutes and the antibacterial activity was assessed by turbidity test and adherence test for the antibiofilm activity test. Bacterial morphology and confluence were investigated by scanning electron microscopy (SEM), (n=3). Two bacterial species were identified from the peri-implant sample, S. mutans and G. adiacens. The results showed that adding ozone to the bacterial biofilm on titanium dental implants did not exhibit significant antibacterial activity against S. mutans. Moreover, there was no significant difference in antibiofilm activity between control and treatment groups. However, significant antibacterial and antibiofilm effect was exhibited by ozone gel against G. adiacens. Ozonated olive oil can be considered as a potential antimicrobial agent for disinfecting dental implant surfaces and treating peri-implantitis.

Inhibition of Clinical MRSA Isolates by Coagulase Negative Staphylococci of Human Origin.

Staphylococcus aureus is frequently highlighted as a priority for novel drug research due to its pathogenicity and ability to develop antibiotic resistance. Coagulase-negative staphylococci (CoNS) are resident flora of the skin and nares. Previous studies have confirmed their ability to kill and prevent colonization by S. aureus through the production of bioactive substances. This study screened a bank of 37 CoNS for their ability to inhibit the growth of methicillin-resistant S. aureus (MRSA). Deferred antagonism assays, growth curves, and antibiofilm testing performed with the cell-free supernatant derived from overnight CoNS cultures indicated antimicrobial and antibiofilm effects against MRSA indicators. Whole genome sequencing and BAGEL4 analysis of 11 CoNS isolates shortlisted for the inhibitory effects they displayed against MRSA led to the identification of two strains possessing complete putative bacteriocin operons. The operons were predicted to encode a nukacin variant and a novel epilancin variant. From this point, strains Staphylococcus hominis C14 and Staphylococcus epidermidis C33 became the focus of the investigation. Through HPLC, a peptide identical to previously characterized nukacin KQU-131 and a novel epilancin variant were isolated from cultures of C14 and C33, respectively. Mass spectrometry confirmed the presence of each peptide in the active fractions. Spot-on-lawn assays demonstrated both bacteriocins could inhibit the growth of an MRSA indicator. The identification of natural products with clinically relevant activity is important in today's climate of escalating antimicrobial resistance and a depleting antibiotic pipeline. These findings also highlight the prospective role CoNS may play as a source of bioactive substances with activity against critical pathogens.

The Potential of Parijoto Fruit Extract Fraction (Medinilla speciosa Blume) as Antibiofilm against Staphylococcus aureus Bacteria

Parijoto fruit extract (Medinilla speciosa Blume) contains flavonoids, saponins, tannins, and terpenoids and has antibacterial potential in inhibiting the growth of Staphylococcus aureus bacteria. S. aureus bacteria are bacteria that are capable of forming biofilms. This research aims to determine the potential of extracts and fractions (water, ethyl acetate, and n-hexane) in inhibiting the formation and degradation of S. aureus biofilms. Parijoto fruit was extracted using the maceration method with 96% ethanol solvent. The extract was fractionated using a liquid-liquid partition technique with water, ethyl acetate, and n-hexane as solvents. The antibiofilm activity test against the S. aureus bacteria ATCC 25923 was carried out by treatment inhibiting biofilm formation and biofilm degradation. The method is to read absorbance using a microplate reader with a wavelength of 595 nm. Optical density results to calculate IC50 and EC50 values. The results showed that the extract and fractions (water, ethyl acetate, and n-hexane) had the potential to inhibit the formation and degradation of S. aureus bacterial biofilms. The IC50 of biofilm inhibition in the extract, water fraction, ethyl acetate, and n-hexane against S. aureus bacteria was 2.51; 1.03; 1.10; 1.52 mg/ml. The EC50 value of the biofilm degradation test on the extract, water fraction, ethyl acetate, and n-hexane against S. aureus bacteria was 1.03; 1.17; 1.37; 1.88 mg/ml. The most active fraction of parijoto fruit (Medinilla specosa Blume) in inhibiting and degrading S. aureus bacterial biofilms is the water fraction.

Nanoemulsion Mouthwash Formulation of Bajakah Tampala (Spatholobus littoralis Hassk.) Skin Extract Against Candida albicans

Candida albicans can cause two infections in humans: superficial and systemic. The ability of C. albicans to infect the host is influenced by virulence factors and character changes so that it can fool the immune system. From the character change factor, C. albicans can form a biofilm. This study aims to determine the good concentration in inhibiting and determine the antifungal and antibiofilm activity of nanoemulsion mouthwash formulation of bajakah tampala (Spatholobus littoralis Hassk) skin extract against C. albicans. This research was conducted with an experimental method. The formulation used a spontaneous magnetic stirrer technique to make nanoemulsion preparations. Antifungal and antibiofilm tests were carried out by dilution method using a 96-well plate and a microplate reader with a wavelength of 620 nm to determine the percentage inhibition against C. albicans and determine MIC50 and MBIC50. The results showed that the nanoemulsion mouthwash formulation of S. littoralis inhibited the planktonic and biofilm of C. albicans. The concentration of 1% is expressed as MIC50 and MBIC50. Therefore, the nanoemulsion formulation of S. littoralis extract could inhibit the growth of C. albicans in the oral cavity.

A Three-Dimensional Model of Bacterial Biofilms and Its Use in Antimicrobial Susceptibility Testing.

(1) Background: The discrepant antimicrobial susceptibility between planktonic and biofilm bacterial modes poses a problem for clinical microbiology laboratories and necessitates a relevant 3D experimental model allowing bacteria to grow in biofilm mode, in vitro, for use in anti-biofilm susceptibility testing. (2) Methods: This work develops a 3D biofilm model consisting of alginate beads containing S. aureus biofilm and encased within two thick layers of alginate matrix. The constructed model was placed on a thin Boyden chamber insert suspended on a 24-well culture plate containing the culture medium. The antibacterial activity of bacitracin and chlorhexidine digluconate (CD), either combined or separately, against 2D S. aureus culture was compared to that in the 3D biofilm model. Quantitative analysis and imaging analysis were performed by assessing the bacterial load within the matrix as well as measuring the optical density of the culture medium nourishing the matrix. (3) Results: The 3D biofilm model represented the typical complex characteristics of biofilm with greater insusceptibility to the tested antimicrobials than the 2D culture. Only bacitracin and CD in combination at 100× the concentration found to be successful against 2D culture were able to completely eliminate the 3D biofilm matrix. (4) Conclusions: The 3D biofilm model, designed to be more clinically relevant, exhibits higher antimicrobial insusceptibility than the 2D culture, demonstrating that the model might be useful for testing and discovering new antimicrobial therapies. The data also support the view that combination therapy might be the optimal approach to combat biofilm infections.

Efficacy of disinfectant and bacteriophage mixture against planktonic and biofilm state of Listeria monocytogenes to control in the food industry

Fresh produce and animal-based products contaminated with Listeria monocytogenes have been the main cause of listeriosis outbreaks for many years. The present investigation explored the potential of combination treatment of disinfectants with a bacteriophage cocktail to control L. monocytogenes contamination in the food industry. A mixture of 1 minimal inhibitory concentration (MIC) of disinfectants (sodium hypochlorite [NaOCl], hydrogen peroxide [H2O2], and lactic acid [LA]) and multiplicity of infection (MOI) 100 of phage cocktail was applied to both planktonic cells in vitro and already-formed biofilm cells on food contact materials (FCMs; polyethylene, polypropylene, and stainless steel) and foods (celery and chicken meat). All the combinations significantly lowered the population, biofilm-forming ability, and the expression of flaA, motB, hlyA, prfA, actA, and sigB genes of L. monocytogenes. Additionally, in the antibiofilm test, approximately 4 log CFU/cm2 was eradicated by 6 h treatment on FCMs, and 3 log CFU/g was eradicated within 3 days on celery. However, <2 log CFU/g was eradicated in chicken meat, and regrowth of L. monocytogenes was observed on foods after 5 days. The biofilm eradication efficacy of the combination treatment was proven through visualization using scanning electron microscopy (SEM) and confocal microscopy. In the SEM images, the unusual behavior of L. monocytogenes invading from the surface to the inside was observed after treating celery with NaOCl+P or H2O2 + P. These results suggested that combination of disinfectants (NaOCl, H2O2, and LA) with Listeria-specific phage cocktail can be employed in the food industry as a novel antimicrobial and antibiofilm approach, and further research of L. monocytogenes behavior after disinfection is needed.

Freshwater algae Cladophora glomerata and Vaucheria sp. from Serbia as sources of bioactive compounds: Chemical analysis and biological activities

We examined potential biological activities of two taxa of freshwater algae, Cladophora glomerata and Vaucheria sp., from Serbia. The total phenolic and pigment contents, antioxidant potential, antimicrobial, antibiofilm activities, and cytotoxicity of the ethanol and acetone extracts were evaluated. The extracts were also subjected to Fourier transform infrared spectroscopy (FTIR) analysis. The levels of total phenolic compounds, chlorophylls a and b, and carotenoids varied based on both the algal taxa and the type of extracts. FTIR analysis showed the presence of lipids, unsaturated fatty acids, protein, carbohydrates, and phenols in the algal extracts. The extracts had moderate DPPH radical scavenging activity and lower reducing power compared with ascorbic acid. The antimicrobial activity expressed as minimum inhibitory concentrations ranged from 0.31 mg/mL to 10 mg/mL. The strains of Staphylococcus aureus and Bacillus cereus isolated from food samples, as well as S. aureus ATCC 25923, were the most sensitive. For the first time, the antibiofilm activity test revealed 98.7% inhibition of S. aureus biofilm formation. The extracts exhibited cytotoxic effects on choriocarcinoma JAR cells but without selectivity on normal fetal lung fibroblast MRC-5 cells. This is the first report on the biological activities of freshwater macroalgae from Serbia.

The potency of Actinomycetes InaCC A758 against dual-species biofilms: Candida albicans and Staphylococcus aureus

Candida albicans and Staphylococcus aureus can coexist to form a biofilm, leading to infections associated with biofilms. Actinomycetes produce secondary metabolites known as antibiotics, antifungals, antibiofilm, anticancer, and antimalarials. This study was aimed to explore the antibiofilm activity of secondary metabolites of Actinomycetes InaCC A758 extracts (InaCC A758) against dual-species biofilms, i.e., C. albicans and S. aureus. Ethyl acetate and chloroform were used as solvents in a maceration extraction technique to isolate the compound designated InaCC A758. The fractionation of the InaCC A758 extracts was carried out using semi-preparative HPLC. Identification of the compounds from the InaCC A758 extracts was performed using gas chromatography-mass spectrometry analysis. Antimicrobial and antibiofilm testing of the InaCC A758 extracts was done utilizing the micro broth dilution method. The morphology of the biofilms following treatment with the InaCC A758 extracts was visualized using scanning electron microscopy (SEM). The minimum inhibitory concentration of InaCC A758 against dual-species biofilms was 400 μg/ml. The concentrations of InaCC A758 required to inhibit 50% and 80% of dual-species biofilm formation (BIC50 and BIC80) ranged from 3.57–6.66 and 29.25–30.50 μg/ml, respectively. The concentrations needed to reduce formed dual-species biofilms by 50% and 80% (BRC50 and BRC80) ranged from 100.62–131.85 and 596.4–849.6 μg/ml, respectively. SEM observation showed a reduced number of cells and disruption of the cell membranes when exposed to the InaCC A758 extracts. The InaCC A758 contains seven compounds, i.e., dodecanoic acid 3-hydroxy-; 1,3,5-pentanetriol,3-methyl-; 1,3-dioxolane-4-methanol, 2-ethyl-; 2-cyclopropylcarbonyloxytridecane; 2-hepten-1- ol,(E); methyl 6-methyl heptanoate; and 11-octadecenoic acid, methyl ester. The InaCC A758 extract altered cell morphology and exhibited potential as an anti-biofilm agent, particularly against dual-species biofilms.

Liposomal and Liposomes-Film Systems as Carriers for Bioactives from Paeonia tenuifolia L. Petals: Physicochemical Characterization and Biological Potential.

Paeonia tenuifolia L. (steppe peony) petal extract was proficiently encapsulated into liposomes and biopolymer films in the current work, both times utilizing a single-step procedure. The encapsulation efficiency, size of the particles, and index of polydispersity (PDI), as well as the ζ potential of the obtained liposomes were determined, whereas in the case of films, the test included moisture content and mechanical property assessment. Fourier transform infrared spectroscopy (FT-IR) was used to evaluate the chemical composition and existence of numerous interactions in the systems. All the obtained encapsulates were subjected to antibacterial, antifungal and antibiofilm activity testing of the pathogens associated with human skin. The results indicated that the liposomes prepared using Phospholipon had the highest encapsulation efficiency (72.04%), making them the most favorable ones in the release study as well. The biological assays also revealed that Phospholipon was the most beneficial phospholipid mixture for the preparation of liposomes, whereas the film containing these liposomes did not have the ability to inhibit pathogen growth, making the double encapsulation of P. tenuifolia L. petal extract needless. These findings may be a first step toward the potential use of steppe peony extract-loaded films and liposomes in pharmaceutical and cosmetical industries.

Exploring the antibacterial mechanisms of chemically characterized essential oils from leaves and buds of Syzygium aromaticum (L.) Merr. et Perry against Staphylococcus aureus and Pseudomonas aeruginosa

Syzygium aromaticum (L.) Merr. et Perry is a medicinal plant extensively used in international pharmacopeia to treat different ailments, including infectious diseases. In this study, we designed to determine the chemical constituents of S. aromaticum essential oils extracted from leaves (CEOL) and floral buds (CEOB) and their antibacterial mechanism of action against S. aureus and P. aeruginosa. The volatile components from CEOL and CEOB were detected using gas chromatography coupled to a mass spectrometer (GC/MS). The results indicated that eugenol was the major component for both CEOL (65.75%) and CEOB (74.03%). The antibacterial findings demonstrated the potent antibacterial activity of CEOL and CEOB. These oils exhibited significant zones of inhibition against all tested bacteria, including S. aureus ATCC 29213, K. aerogenes ATCC 13048, P. aeruginosa ATCC 27853, and E. coli ATCC 25922, surpassing the effects of antibiotics. Additionally, they displayed low minimum inhibitory concentrations (MICs) and minimum bactericidal concentrations (MBCs), indicating their bactericidal nature (MBC/MIC < 4.0). Time-kill kinetics revealed that CEOB was more effective than CEOL at sub-MIC doses, eradicating S. aureus within 12 h and P. aeruginosa within 12–24 h. The cell membrane permeability test demonstrated that CEOL and CEOB treatment affected the relative conductivity of S. aureus and P. aeruginosa at both MIC and sub-MIC concentrations, indicating disruption of the bacterial membrane and influencing their growth and viability. Furthermore, the cell membrane integrity test showed that the addition of CEOL and CEOB to S. aureus and P. aeruginosa cultures led to significant protein release from the bacterial cells. The results of the anti-biofilm activity test confirmed the efficacy of S. aromaticum EOs, particularly CEOB, as potent anti-biofilm agents, suggesting its potential effect on inhibiting quorum sensing. Scanning electron microscope (SEM) microscopy further revealed significant alterations in bacterial cells treated with CEOB essential oil, including damage to the cell wall and distorted cell morphology, highlighting its impact on bacterial cell structure. Presented information suggests the use of clove EOs as natural alternatives for sustainable application in food and pharmaceutical industries.

Unlocking the Power of Onion Peel Extracts: Antimicrobial and Anti-Inflammatory Effects Improve Wound Healing through Repressing Notch-1/NLRP3/Caspase-1 Signaling.

Onion peels are often discarded, representing an unlimited amount of food by-products; however, they are a valuable source of bioactive phenolics. Thus, we utilized UPLC-MS/MS to analyze the metabolomic profiles of red (RO) and yellow (YO) onion peel extracts. The cytotoxic (SRB assay), anti-inflammatory (Griess assay), and antimicrobial (sensitivity test, MIC, antibiofilm, and SP-SDS tests) properties were assessed in vitro. Additionally, histological analysis, immunohistochemistry, and ELISA tests were conducted to investigate the healing potential in excisional skin wound injury and Candida albicans infection in vivo. RO extract demonstrated antibacterial activity, limited skin infection with C. albicans, and improved the skin's appearance due to the abundance of quercetin and anthocyanin derivatives. Both extracts reduced lipopolysaccharide-induced nitric oxide release in vitro and showed a negligible cytotoxic effect on MCF-7 and HT29 cells. When extracts were tested in vivo for their ability to promote tissue regeneration, it was found that YO peel extract had the greatest impact. Further biochemical analysis revealed that YO extract suppressed NLRP3/caspase-1 signaling and decreased inflammatory cytokines. Furthermore, YO extract decreased Notch-1 levels and boosted VEGF-mediated angiogenesis. Our findings imply that onion peel extract can effectively treat wounds by reducing microbial infection, reducing inflammation, and promoting tissue regeneration.

Molecular Docking Study of the C-10 Massoia Lactone Compound as an Antimicrobial and Antibiofilm Agent against Candida tropicalis.

Antimicrobial resistance is now considered a global health problem because it reduces the effectiveness of antimicrobial drugs. According to the World Health Organization (WHO), the highest mortality rate is associated with infections caused by multidrug-resistant microorganisms, with approximately 700,000 deaths worldwide each year. The aim of this study was to determine the potential of C-10 massoia lactone to inhibit the growth of fungi and C. tropicalis biofilm, and molecular docking studies were performed to determine the nature of the inhibition. The study was conducted using the microdilution method for antifungal and antibiofilm testing and designed with a molecular docking approach. Furthermore, an analysis using the scanning electron microscope (SEM) was performed to evaluate the mechanism of effect. The results obtained showed that C-10 massoia lactone can inhibit the growth of fungi by 84.21% w/v. Meanwhile, the growth of C. tropicalis biofilm in the intermediate phase was 80.23% w/v and in the mature phase was 74.23% w/v. SEM results showed that C-10 massoia lactone damaged the EPS matrix of C. tropicalis so that hyphal formation was hindered due to damage to fungal cells, resulting in a decrease in attachment, density, and lysis of C. tropicalis fungal cells. Based on molecular docking tests, C-10 massoia lactone was able to inhibit biofilm formation without affecting microbial growth, while docking C-10 massoia lactone showed a significant binding and has the potential as an antifungal agent. In conclusion, the C-10 massoia lactone compound has the potential as an antibiofilm against C. tropicalis, so it can become a new antibiofilm agent.

Biological Activity of Natural Formulation with Propolis, Lavender and Thyme Oil on Candida Species

Objective: Candida strains have been frequently associated with nosocomial infections in recent years. On the other hand, public prefer natural resources rather than synthetic materials. Therefore, it is important to research combine effects of natural products. This study aimed to investigate the antifungal and antibiofilm effectiveness of essential oils and propolis containing samples on Candida species and the toxicity of samples.&#x0D; Material-Method: Three different samples were prepared with propolis, lavender oil and thyme oil. Gas Chromatography-mass Spectrometry (GC/MS) GC/MS was used for the determination of organic compounds in samples. The antifungal effects of a natural product mix were tested against C. albicans, C. krusei and C. parapsilosis by disk diffusion method. MIC and MFC tests of the mixture were performed against a standard such as Nystatin. Antibiofilm tests were performed with the microplate system, taking into account the detected concentrations. Finally, in the cytotoxicity test investigated in the L929 cell line with the WST-1 kit.&#x0D; Results: C. albicans was determined as the most sensitive species (MIC50: 90 ppm, MIC90: 750 ppm MFC: 3120 ppm). In the antibiofilm tests, it was determined that the sample was more effective on C. albicans and it inhibited biofilm formation. Also it was determined the IC50 value was 5052 ppm.&#x0D; Conclusion: The natural product mixture, which has scientifically proven antifungal effect, has a feature that has high added value and contributes to the economy. But of course, it has to investigate by further studies. The findings of this study may lead to new therapeutic approaches with standardized natural products in the future.

Antibiotic-Loaded Boron Nitride Nanoconjugate with Strong Performance against Planktonic Bacteria and Biofilms.

Protecting surfaces from biofilm formation presents a significant challenge in the biomedical field. The utilization of antimicrobial component-conjugated nanoparticles is becoming an attractive strategy against infectious biofilms. Boron nitride (BN) nanomaterials have a unique biomedical application value due to their excellent biocompatibility. Here, we developed antibiotic-loaded BN nanoconjugates to combat bacterial biofilms. Antibiofilm testing included two types of pathogens, Staphylococcus aureus and Escherichia coli. Gentamicin was loaded on polydopamine-modified BN nanoparticles (GPBN) to construct a nanoconjugate, which was very effective in killing E. coli and S. aureus planktonic cells. GPBN exhibited equally strong capacity for biofilm destruction, tested on preformed biofilms. A 24 h treatment with the nanoconjugate reduced cell viability by more than 90%. Our results suggest that GPBN adheres to the surface of the biofilm, penetrates inside the biofilm matrix, and finally deactivates the cells. Interestingly, the GPBN coatings also strongly inhibited the formation of bacterial biofilms. Based on these results, we suggest that GPBN could serve as an effective means for treating biofilm-associated infections and as coatings for biofilm prevention.

Ex vivo comparison of V.A.C.® Granufoam Silver™ and V.A.C.® Granufoam™ loaded with a first-in-class bis-dialkylnorspermidine-terphenyl antibiofilm agent

Implementation of negative pressure wound therapy (NPWT) as a standard of care has proven efficacious in reducing both the healing time and likelihood of nosocomial infection among pressure ulcers and traumatic, combat-related injuries. However, current formulations may not target or dramatically reduce bacterial biofilm burden following therapy. The purpose of this study was to determine the antibiofilm efficacy of an open-cell polyurethane (PU) foam (V.A.C.® Granufoam™) loaded with a first-in-class compound (CZ-01179) as the active release agent integrated via lyophilized hydrogel scaffolding. An ex vivo porcine excision wound model was designed to perform antibiofilm efficacy testing in the presence of NPWT. PU foam samples loaded with a 10.0% w/w formulation of CZ-01179 and 0.5% hyaluronic acid were prepared and tested against current standards of care: V.A.C.® Granufoam Silver™ and V.A.C.® Granufoam™. We observed statistically significant reduction of methicillin-resistant Staphylococcus aureus (MRSA) and Acinetobacter baumannii biofilms with the CZ-01179 antibiofilm foam in comparison to current standard of care foams. These findings motivate further development of an antibiofilm PU foam loaded with CZ-01179.

Chitosan-based emulgel and xerogel film containing Thymus pubescens essential oil as a potential wound dressing

Controlling the wound exudates accompanied by microbial wound infections has still remained as one the most challenging clinical issues. Herein, a chitosan/gelatin/polyvinyl alcohol xerogel film containing Thymus pubescens essential oil is fabricated for antimicrobial wound dressing application. The chemical and physical characteristics of the devised formulation is characterized by Fourier transform infrared spectroscopy, scanning electron microscopy, atomic force microscope, and tensile tests. Moreover, swelling capability, water vapour transmission rate, water contact angle, solubility, moisture content, and release properties are also studied. The antimicrobial and antibiofilm tests are performed using the broth microdilution and XTT assay, respectively. The produced formulation shows excellent antimicrobial efficacy against Escherichia coli, Staphylococcus aureus, Pseudomonas aeruginosa and Candida species. It is also demonstrated that the obtained film can reduce (∼80 %) Candida albicans biofilm formation, and its biocompatibility is confirmed with MTT (∼100 %) and hemolysis tests. The antimicrobial activity can be correlated to the microbial membrane attraction for Candida albicans cells, illustrated by flow cytometry. This proposed film with appropriate mechanical strength, high swelling capacity in different pH values (∼200–700 %), controlled release property, and antimicrobial and antioxidant activities as well as biocompatibility can be used as a promising candidate for antimicrobial wound dressing applications.

Antibacterial Activity of Dissolved Silver Fractions Released from Silver-Coated Titanium Dental Implant Abutments: A Study on Streptococcus mutans Biofilm Formation.

(1) Background: The aim of this research was to investigate the antibacterial activity of dissolved silver from silver-coated titanium implants against Streptococcus mutans. (2) Methodology: Silver-coated titanium implant discs were immersed in 1.8 mL of brain heart infusion broth (BHIB) and incubated for 24 h in order to release the silver ions into the broth. The coating quality was confirmed via EDS, and the dissolved silver was measured via inductively coupled plasma mass spectrometry (ICP-MS). The experimental design used unconditioned broth (control) and broth conditioned with silver released from silver-coated titanium implants (n = 6). Regarding the antibacterial activity, isolated Streptococcus mutans was used. A turbidity test and lactate production test were performed to determine the effect of dissolved silver on bacterial growth in a suspension and biofilm formation. (3) Result: The results showed that the coating was successfully applied on the substrate. There was around 0.3 mg/L of silver released into the BHIB, and the turbidity of the control group was significantly higher than the treatment, with measured absorbance values of 1.4 and 0.8, respectively, indicating that the dissolved silver ions from the silver-coated titanium discs exhibited some degree of antibacterial activity by preventing the growth of Streptococcus mutans. However, the results of the antibiofilm activity test did not show any significant difference between the groups. (4) Conclusion: The dissolved silver from silver-coated titanium implants has an antibacterial activity but not a significant antimicrobial activity, indicating that the dissolved silver from silver-coated titanium abutments can significantly reduce the incidence of peri-implant mucositis.