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1.
Proc Natl Acad Sci U S A ; 119(3)2022 01 18.
Artigo em Inglês | MEDLINE | ID: mdl-35012978

RESUMO

Preventing pathogenic viral and bacterial transmission in the human environment is critical, especially in potential outbreaks that may be caused by the release of ancient bacteria currently trapped in the permafrost. Existing commercial disinfectants present issues such as a high carbon footprint. This study proposes a sustainable alternative, a bioliquid derived from biomass prepared by hydrothermal liquefaction. Results indicate a high inactivation rate of pathogenic virus and bacteria by the as-prepared bioliquid, such as up to 99.99% for H1N1, H5N1, H7N9 influenza A virus, and Bacillus subtilis var. niger spores and 99.49% for Bacillus anthracis Inactivation of Escherichia coli and Staphylococcus epidermidis confirmed that low-molecular-weight and low-polarity compounds in bioliquid are potential antibacterial components. High temperatures promoted the production of antibacterial substances via depolymerization and dehydration reactions. Moreover, bioliquid was innoxious as confirmed by the rabbit skin test, and the cost per kilogram of the bioliquid was $0.04427, which is notably lower than that of commercial disinfectants. This study demonstrates the potential of biomass to support our biosafety with greater environmental sustainability.


Assuntos
Biomassa , Contenção de Riscos Biológicos , Meio Ambiente , Energia Renovável , Antibacterianos/farmacologia , Escherichia coli/efeitos dos fármacos , Escherichia coli/ultraestrutura , Humanos , Testes de Sensibilidade Microbiana , Peso Molecular , Pandemias , Staphylococcus epidermidis/efeitos dos fármacos , Staphylococcus epidermidis/ultraestrutura
2.
Med Microbiol Immunol ; 209(6): 669-680, 2020 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-32880037

RESUMO

The ability of bacteria to aggregate and form biofilms impairs phagocytosis by polymorphonuclear leukocytes (PMNs). The aim of this study was to examine if the size of aggregates is critical for successful phagocytosis and how bacterial biofilms evade phagocytosis. We investigated the live interaction between PMNs and Pseudomonas aeruginosa, Staphylococcus aureus, Escherichia coli and Staphylococcus epidermidis using confocal scanning laser microscopy. Aggregate size significantly affected phagocytosis outcome and larger aggregates were less likely to be phagocytized. Aggregates of S. epidermidis were also less likely to be phagocytized than equally-sized aggregates of the other three species. We found that only aggregates of approx. 5 µm diameter or smaller were consistently phagocytosed. We demonstrate that planktonic and aggregated cells of all four species significantly reduced the viability of PMNs after 4 h of incubation. Our results indicate that larger bacterial aggregates are less likely to be phagocytosed by PMNs and we propose that, if the aggregates become too large, circulating PMNs may not be able to phagocytose them quickly enough, which may lead to chronic infection.


Assuntos
Biofilmes , Escherichia coli/fisiologia , Neutrófilos/fisiologia , Fagocitose , Pseudomonas aeruginosa/fisiologia , Staphylococcus aureus/fisiologia , Staphylococcus epidermidis/fisiologia , Escherichia coli/ultraestrutura , Humanos , Pseudomonas aeruginosa/ultraestrutura , Pele/microbiologia , Staphylococcus aureus/ultraestrutura , Staphylococcus epidermidis/ultraestrutura
3.
Arch Microbiol ; 202(6): 1327-1340, 2020 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-32146497

RESUMO

Staphylococcus epidermidis is well recognized nosocomial pathogen in clinical settings for their implants associated infections. Biofilm and virulence production executes a S. epidermidis pathogenesis against host. Hence, interfering of biofilm formation has become an auspicious to control the pathogenesis of S. epidermidis. The present study evaluates antibiofilm potential of Rhizophora mucronata against S. epidermidis biofilms. Rhizophora mucronata leaves extract significantly inhibited the biofilm formation and quebrachitol was identified as an active compound responsible for the biofilm inhibition. Quebrachitol significantly inhibited biofilm formation at concentration dependent manner without exhibit non-bactericidal property. And, quebrachitol reduced the biofilm building components such as exopolysaccharides, lipase and proteins production. Confocal laser scanning microscopic studies obtained quebrachitol surface independent biofilm efficacy against S. epidermidis. Notably, quebrachitol significantly reduced S. epidermidis adherence on biotic (coated with type I collagen and fibrinogen) and abiotic (hydrophobic and hydrophilic) surfaces. Addition of quebrachitol inhibits autolysis mediated initial attachment and accumulation associated aggregation process. Moreover, quebrachitol significantly reduced the hydrolases virulence production which supports S. epidermidis invasion into the host. Furthermore, gene expression analysis revealed the ability of quebrachitol to downregulate the virulence genes expression which are mainly involved in biofilm formation and virulence production. The results obtained from the present study suggest that quebrachitol as an ideal candidate for the therapeutic action against S. epidermidis pathogenesis.


Assuntos
Aderência Bacteriana/efeitos dos fármacos , Biofilmes/efeitos dos fármacos , Inositol/análogos & derivados , Extratos Vegetais/farmacologia , Staphylococcus epidermidis/efeitos dos fármacos , Inositol/farmacologia , Microscopia Eletrônica de Varredura , Rhizophoraceae/química , Staphylococcus epidermidis/metabolismo , Staphylococcus epidermidis/fisiologia , Staphylococcus epidermidis/ultraestrutura , Virulência/efeitos dos fármacos
4.
Spine Deform ; 8(3): 351-359, 2020 06.
Artigo em Inglês | MEDLINE | ID: mdl-32096135

RESUMO

STUDY DESIGN: A microscopy-based investigation of the permissive factors leading towards bacterial adherence on commonly utilized spinal implants. OBJECTIVE: The adherence and subsequent colonization and biofilm formation of bacteria on orthopaedic implants represents one of the most serious problems facing orthopaedic surgeons. Once a biofilm is formed, surgeons may have to resort to implant removal, a strategy that may cause substantial patient morbidity and lead to additional cost to the healthcare system. This problem has been further compounded by the rise of antibiotic-resistant strains of bacterial pathogens. In this study, two commonly encountered bacterial pathogens in surgical site infections (SSI) were characterized for adherence pattern, density, and propagation on five commonly used spinal implant materials via scanning electron microscopy (SEM) and confocal laser scanning microscopy (CLSM). The results show that bacterial adherence is largely dependent on the microtopographical features observed on the surface of the materials tested. METHODS: Five commonly utilized spinal implant materials were inoculated with two of the most common nosocomial pathogens and visualized via scanning electron microscopy and confocal laser scanning microscopy. RESULTS: Analysis of 90 spinal implant pieces showed that even though no material showed the ability to prevent adherence of both pathogens tested, the presence of surface imperfections and rougher microtopography was found to harbor the most bacterial presence. CONCLUSION: Our data suggests that implants materials with uniform surface and minimal imperfections may reduce the ability of bacterial to adhere to implants. LEVEL OF EVIDENCE: Level I evidence: "Investigation of a diagnostic test".


Assuntos
Biofilmes/crescimento & desenvolvimento , Procedimentos Ortopédicos , Próteses e Implantes/microbiologia , Staphylococcus aureus/crescimento & desenvolvimento , Staphylococcus epidermidis/crescimento & desenvolvimento , Infecção da Ferida Cirúrgica/microbiologia , Aderência Bacteriana , Microscopia Confocal , Microscopia Eletrônica de Varredura , Próteses e Implantes/ultraestrutura , Coluna Vertebral/cirurgia , Staphylococcus aureus/fisiologia , Staphylococcus aureus/ultraestrutura , Staphylococcus epidermidis/fisiologia , Staphylococcus epidermidis/ultraestrutura
5.
PLoS One ; 14(4): e0214744, 2019.
Artigo em Inglês | MEDLINE | ID: mdl-30946772

RESUMO

For combating multidrug-resistant microorganisms, exploration of natural compounds from plant endophytes increases the chance of finding novel compounds. An efficient bioactive metabolites producing endophytic fungal strain AE1 was isolated from leaves of Azadirachta indica A. Juss. The metabolites were found to be thermostable, non-proteinacious and produced prominent zones of inhibition against numbers of Gram positive and Gram negative bacteria. Based on 28S rDNA (D1/D2) sequence homology the isolate AE1 was identified as Alternaria alternata. Malt extract broth was found effective for the maximum production of bioactive metabolites by the isolate and was subjected for solvent extraction. The Ethyl acetate (EA) fraction of AE1 showed MIC values of 300-400 µg/ml against Gram positive and Gram negative bacteria tested. The cidal mode of action of EA fraction was detected by treating bacterial cultures at mid log phase. Scanning electron microscopic study supported morphological disintegration of bacterial cells. Release of nucleic acid, protein and potassium ions (K+) also suggested lysis of bacterial cells or leakage of cell membrane upon treatment. In addition, reduction of the activity of EMP pathway, TCA cycle and gluconeogenic enzymes in all bacteria suggested the interference of antibacterial principles with central carbohydrate metabolic pathways. Thin layer chromatographic separation followed by GC-MS analysis of EA fraction suggested numbers of antimicrobial compound production by AE1. In addition, DPPH free radical as well as superoxide radical scavenging assay also suggested strong antioxidant potential of AE1 with an IC50 value of 38.0±1.7 µg/ml and 11.38±1.2 µg/ml respectively. On the basis of above facts it can be concluded that the strain AE1 will be a good source of bioactive compounds having medicinal importance.


Assuntos
Alternaria/metabolismo , Antibacterianos/biossíntese , Antioxidantes/metabolismo , Azadirachta/microbiologia , Antibacterianos/isolamento & purificação , Antibacterianos/farmacologia , Antioxidantes/isolamento & purificação , Antioxidantes/farmacologia , Bacillus subtilis/efeitos dos fármacos , Bacillus subtilis/ultraestrutura , Endófitos/metabolismo , Escherichia coli/efeitos dos fármacos , Escherichia coli/ultraestrutura , Sequestradores de Radicais Livres/isolamento & purificação , Sequestradores de Radicais Livres/metabolismo , Sequestradores de Radicais Livres/farmacologia , Cromatografia Gasosa-Espectrometria de Massas , Listeria monocytogenes/efeitos dos fármacos , Listeria monocytogenes/ultraestrutura , Testes de Sensibilidade Microbiana , Folhas de Planta/microbiologia , Pseudomonas/efeitos dos fármacos , Pseudomonas/ultraestrutura , Salmonella typhimurium/efeitos dos fármacos , Salmonella typhimurium/ultraestrutura , Staphylococcus aureus/efeitos dos fármacos , Staphylococcus aureus/ultraestrutura , Staphylococcus epidermidis/efeitos dos fármacos , Staphylococcus epidermidis/ultraestrutura
6.
J Hazard Mater ; 368: 52-62, 2019 04 15.
Artigo em Inglês | MEDLINE | ID: mdl-30665108

RESUMO

Effect of Staphylococcus epidermidis (S. epidermidis) on U(VI) sequestration by Al-goethite were conducted under different geologic conditions. The batch experiments showed that S. epidermidis significantly enhanced the adsorption rates of U(VI) at pH < 9.0 due to the additional metal binding sites. The maximum adsorption capacities of U(VI) on Al-goethite and Al-goethite +S. epidermidis at pH 4.0 and 310 K were calculated from Langmuir equation to be 13.16 and 47.86 mg/g, respectively. The decreased adsorption of U(VI) on Al-goethite+ S. epidermidis at high carbonate and pH conditions were primarily driven by the electrostatic repulsion between negatively charged U(VI)-carbonate complexes and the negatively charged adsorbents. According to XPS analysis, the adsorbed U(VI) can be reduced to U(IV) by S. epidermidis, whereas inhibited reduction of U(VI) on Al-goethite + S. epidermidis at high pH could be attributed to the complexation of structural Fe(III) with the oxygen-containing functional groups of S. epidermidis. FT-IR analysis further demonstrated that the bonding of structural Fe(III) with functional groups (e.g., carboxyl and phosphate groups) of S. epidermidis. These results herein are important to understand the fate and transport of U(VI) on the mineral-bacteria ternary systems in the near-surface environment.


Assuntos
Alumínio/química , Compostos de Ferro/química , Minerais/química , Staphylococcus epidermidis , Urânio/química , Poluentes Radioativos da Água/química , Adsorção , Concentração de Íons de Hidrogênio , Ferro/química , Oxirredução , Staphylococcus epidermidis/ultraestrutura
7.
Sensors (Basel) ; 18(12)2018 Nov 22.
Artigo em Inglês | MEDLINE | ID: mdl-30469521

RESUMO

The biofilm-forming microbial species Candida parapsilosis and Staphylococcus epidermidis have been recently linked to serious infections associated with implanted medical devices. We studied microbial biofilms by high resolution scanning electron microscopy (SEM), which allowed us to visualize the biofilm structure, including the distribution of cells inside the extracellular matrix and the areas of surface adhesion. We compared classical SEM (chemically fixed samples) with cryogenic SEM, which employs physical sample preparation based on plunging the sample into various liquid cryogens, as well as high-pressure freezing (HPF). For imaging the biofilm interior, we applied the freeze-fracture technique. In this study, we show that the different means of sample preparation have a fundamental influence on the observed biofilm structure. We complemented the SEM observations with Raman spectroscopic analysis, which allowed us to assess the time-dependent chemical composition changes of the biofilm in vivo. We identified the individual spectral peaks of the biomolecules present in the biofilm and we employed principal component analysis (PCA) to follow the temporal development of the chemical composition.


Assuntos
Infecções Bacterianas/diagnóstico , Biofilmes/crescimento & desenvolvimento , Candida parapsilosis/isolamento & purificação , Staphylococcus epidermidis/isolamento & purificação , Infecções Bacterianas/microbiologia , Candida parapsilosis/patogenicidade , Candida parapsilosis/ultraestrutura , Humanos , Microscopia Eletrônica de Varredura , Análise Espectral Raman , Staphylococcus epidermidis/patogenicidade , Staphylococcus epidermidis/ultraestrutura
8.
Int J Nanomedicine ; 13: 6575-6583, 2018.
Artigo em Inglês | MEDLINE | ID: mdl-30410338

RESUMO

BACKGROUND: Many cancer patients suffer from cancer-related life-threatening infections due to immune system damage. Therefore, researchers are continuously looking for new options to treat cancer-related infections. As nanotechnology has gained tremendous interest over the past several decades, silver nanoparticles have been investigated as an effective antimicrobial agent. Here, silver-coated gold nanorods were synthesized to share similar optical properties as gold nanopar-ticles for cancer diagnosis and treatment, with an added advantage of antibacterial properties. RESULTS: Their dose-dependent antimicrobial properties were demonstrated on both Gram-negative and Gram-positive bacteria species. These nanorods were found to be highly efficient in killing bacteria and suppressing biofilm formation. CONCLUSION: Collectively, such results suggest that silver-coated gold nanorods should be further investigated as a novel material, which can both decrease cancer cell functions and reduce the risk of infection for cancer patients.


Assuntos
Anti-Infecciosos/uso terapêutico , Doenças Transmissíveis/tratamento farmacológico , Ouro/química , Nanopartículas Metálicas/química , Nanotubos/química , Neoplasias/complicações , Prata/química , Anti-Infecciosos/farmacologia , Doenças Transmissíveis/microbiologia , Humanos , Nanopartículas Metálicas/ultraestrutura , Nanotubos/ultraestrutura , Staphylococcus aureus/efeitos dos fármacos , Staphylococcus aureus/crescimento & desenvolvimento , Staphylococcus aureus/ultraestrutura , Staphylococcus epidermidis/efeitos dos fármacos , Staphylococcus epidermidis/crescimento & desenvolvimento , Staphylococcus epidermidis/ultraestrutura
9.
ACS Appl Mater Interfaces ; 10(42): 35784-35793, 2018 Oct 24.
Artigo em Inglês | MEDLINE | ID: mdl-30273480

RESUMO

Biomaterials which promote tissue integration and resist microbial colonisation are required in bone tissue engineering to prevent biomaterial-associated infections. Surface modification of established materials for bone tissue engineering, such as TiO2, have emerged as promising anti-infective strategies. Interestingly, the antibacterial activity of TiO2 in the form of particles can be enhanced by combining it with H2O2, even in the absence of irradiation. However, it remains unknown whether TiO2 surfaces elicit a similar effect. In this study, the antibacterial effect of porous TiO2 scaffolds generated by the catalytic decomposition of H2O2 in the absence of light (dark catalysis) was investigated. Porous ceramic foams were fabricated and sol-gel coated for high catalytic activity. Degradation of methylene blue in the presence of 3% H2O2 increased by 80% for the sol-gel-coated surfaces. The degradation kinetics indicate that intermediate free radicals that form at the liquid-TiO2 interface are responsible for the oxidative behavior of the surface. TiO2 surfaces were further pretreated with 30% H2O2 for prolonged oxidative behavior. The biological response toward such surfaces was assessed in vitro. S. epidermidis biofilms formed on modified surfaces showed reduced viability compared to nonmodified surfaces. Further, the same surface modification showed no cytotoxic effects on MC3T3 preosteoblasts. However, the results from the conducted genotoxicity assay were inconclusive, and further studies are needed to exclude ROS-mediated DNA damage. To conclude, this study provides evidence that a simple surface modification based on the dark catalytic effect of TiO2 can be used to create antibacterial surface properties for ceramic bone scaffolds.


Assuntos
Antibacterianos/farmacologia , Osso e Ossos/fisiologia , Materiais Revestidos Biocompatíveis/farmacologia , Escuridão , Alicerces Teciduais/química , Titânio/farmacologia , Animais , Biofilmes/efeitos dos fármacos , Catálise , Adesão Celular/efeitos dos fármacos , Morte Celular/efeitos dos fármacos , Linhagem Celular , Forma Celular/efeitos dos fármacos , Luminescência , Camundongos , Testes de Sensibilidade Microbiana , Mutagênicos/toxicidade , Staphylococcus epidermidis/efeitos dos fármacos , Staphylococcus epidermidis/fisiologia , Staphylococcus epidermidis/ultraestrutura , Propriedades de Superfície
10.
Nanomedicine (Lond) ; 13(13): 1583-1594, 2018 07.
Artigo em Inglês | MEDLINE | ID: mdl-30028247

RESUMO

AIM: A novel electrospun biocompatible nanofibrous material loaded with commensal bacteria for potential preventive treatment of the diabetic foot was developed. MATERIALS & METHODS: Two biocompatible polymers (carboxymethylcellulose and polyethylene oxide) were combined with a bacterium isolate from the skin located between the toes of a healthy adult (identified using a matrix-assisted laser desorption/ionization mass spectrometry-based method as a strain of Staphylococcus epidermidis). Higher bacteria loads in the material were assured through their encapsulation in polyethylenimine. The nanofibrous material was characterized using scanning electron microscopy, zeta-potential measurements and through evaluation of cell growth and viability. RESULTS & DISCUSSION: nanometer formation was confirmed using scanning electron microscopy, while the zeta-potential measurements revealed successful bacteria encapsulation. Viable and sufficiently growing cells were confirmed prior and after their incorporation. CONCLUSION: The prepared materials were proven suitable to deliver viable commensal bacteria in a comparable share to the Staphylococcaceae in the foot microbiome making this approach promising for preventive diabetic foot treatment.


Assuntos
Pé Diabético/tratamento farmacológico , Nanofibras/administração & dosagem , Staphylococcus epidermidis/crescimento & desenvolvimento , Simbiose , Materiais Biocompatíveis/administração & dosagem , Materiais Biocompatíveis/química , Carboximetilcelulose Sódica/administração & dosagem , Carboximetilcelulose Sódica/química , Pé Diabético/microbiologia , Pé Diabético/patologia , Humanos , Microbiota/efeitos dos fármacos , Microscopia Eletrônica de Varredura , Nanofibras/química , Nanofibras/microbiologia , Polietilenoglicóis/química , Polietilenoglicóis/farmacologia , Espectrometria de Massas por Ionização e Dessorção a Laser Assistida por Matriz , Staphylococcus epidermidis/química , Staphylococcus epidermidis/ultraestrutura
11.
Micron ; 110: 28-35, 2018 07.
Artigo em Inglês | MEDLINE | ID: mdl-29715620

RESUMO

In this study we present an innovative method for the preparation of fully hydrated samples of microbial biofilms of cultures Staphylococcus epidermidis, Candida parapsilosis and Candida albicans. Cryo-scanning electron microscopy (cryo-SEM) and high-pressure freezing (HPF) rank among cutting edge techniques in the electron microscopy of hydrated samples such as biofilms. However, the combination of these techniques is not always easily applicable. Therefore, we present a method of combining high-pressure freezing using EM PACT2 (Leica Microsystems), which fixes hydrated samples on small sapphire discs, with a high resolution SEM equipped with the widely used cryo-preparation system ALTO 2500 (Gatan). Using a holder developed in house, a freeze-fracturing technique was applied to image and investigate microbial cultures cultivated on the sapphire discs. In our experiments, we focused on the ultrastructure of the extracellular matrix produced during cultivation and the relationships among microbial cells in the biofilm. The main goal of our investigations was the detailed visualization of areas of the biofilm where the microbial cells adhere to the substrate/surface. We show the feasibility of this technique, which is clearly demonstrated in experiments with various freeze-etching times.


Assuntos
Candida albicans/ultraestrutura , Candida parapsilosis/ultraestrutura , Microscopia Crioeletrônica/métodos , Matriz Extracelular/ultraestrutura , Técnica de Fratura por Congelamento/métodos , Microscopia Eletrônica de Varredura/métodos , Staphylococcus epidermidis/ultraestrutura , Biofilmes
12.
Sci Rep ; 8(1): 192, 2018 01 09.
Artigo em Inglês | MEDLINE | ID: mdl-29317687

RESUMO

In this study, the synergistic effect of vancomycin, a cell wall synthesis inhibitor, and ultrasound-targeted microbubble destruction (UTMD), on cell viability of Staphylococcus epidermidis, embedded in biofilm, was investigated. Biofilms are the leading causes of antibiotic-resistant bacterial infections of medical implants and prosthetics worldwide. The antibiotic-resistant nature of biofilm-embedded pathogens poses a critical challenge to the medical community. Previously, studies have demonstrated the efficacy of using ultrasound waves and UTMD in circumventing this problem. However, the mechanism(s) underlying this phenomenon was not clear. Here, the present study showed that both ultrasound and UTMD damaged the cell wall structure of S. epidermidis, and floccules and fragments from damaged cells were observed on transmission electron microscope micrograph. However, the cell membrane integrity was not seriously affected by treatments, and the treatment increased the metabolic activity levels of the dormant biofilm-embedded bacteria, detected by confocal laser scanning microscope and flow cytometry, which could make them susceptible to the effect of the antibiotic. Thus, the biological mechanism underlying the efficacy of the combined treatment involving UTMD and vancomycin in the case of S. epidermidis biofilm was dissected, which may be utilized for further investigations on other biofilm pathogens before clinical use.


Assuntos
Antibacterianos/farmacologia , Biofilmes/efeitos dos fármacos , Desinfecção/métodos , Staphylococcus epidermidis/efeitos dos fármacos , Ondas Ultrassônicas , Vancomicina/farmacologia , Biofilmes/efeitos da radiação , Membrana Celular/efeitos dos fármacos , Membrana Celular/efeitos da radiação , Sonicação/métodos , Staphylococcus epidermidis/efeitos da radiação , Staphylococcus epidermidis/ultraestrutura
13.
Int J Biol Macromol ; 110: 150-156, 2018 Apr 15.
Artigo em Inglês | MEDLINE | ID: mdl-28834707

RESUMO

Polymicrobial biofilms with fungi and bacteria are the leading cause for the failure of medical devices and related infections. In this study, antibiofilm activities of carboxymethyl chitosan (CM-chitosan) on monomicrobial and polymicrobial biofilms of Staphylococcus epidermidis and Candida tropicalis in vitro were evaluated. CM-chitosan was effective as a sole agent, inhibiting both monomicrobial and polymicrobial biofilms in microplates and also on the silicone surface in short- and long-term periods. Biofilm architecture was investigated by scanning electron microscopy and confocal laser scanning microscopy was used to examine living/dead organisms within biofilm. CM-chitosan inhibited planktonic growth as well as adhesion. Further biofilm formation was inhibited by CM-chitosan added at 90min or 12h after biofilm initiation. CM-chitosan may serve as a possible antibiofilm agent to limit monomicrobial and polymicrobial biofilm.


Assuntos
Biofilmes/efeitos dos fármacos , Candida tropicalis/fisiologia , Quitosana/análogos & derivados , Quitosana/farmacologia , Staphylococcus epidermidis/fisiologia , Biofilmes/crescimento & desenvolvimento , Candida tropicalis/ultraestrutura , Quitosana/química , Staphylococcus epidermidis/ultraestrutura
14.
Carbohydr Polym ; 181: 345-350, 2018 Feb 01.
Artigo em Inglês | MEDLINE | ID: mdl-29253982

RESUMO

Development of advanced dressings with antimicrobial properties for the treatment of infected wounds is an important approach in the fight against evolution of antibiotic resistant bacterial strains. Herein, the effects of ion-crosslinked nanocellulose hydrogels on bacteria commonly found in infected wounds were investigated in vitro. By using divalent calcium or copper ions as crosslinking agents, different antibacterial properties against the bacterial strains Staphylococcus epidermidis and Pseudomonas aeruginosa were obtained. Calcium crosslinked hydrogels were found to retard S. epidermidis growth (up to 266% increase in lag time, 36% increase in doubling time) and inhibited P. aeruginosa biofilm formation, while copper crosslinked hydrogels prevented S. epidermidis growth and were bacteriostatic towards P. aeruginosa (49% increase in lag time, 78% increase in doubling time). The wound dressing candidates furthermore displayed barrier properties towards both S. epidermidis and P. aeruginosa, hence making them interesting for further development of advanced wound dressings with tunable antibacterial properties.


Assuntos
Antibacterianos/farmacologia , Celulose/farmacologia , Reagentes de Ligações Cruzadas/química , Hidrogéis/farmacologia , Nanofibras/química , Madeira/química , Cicatrização/efeitos dos fármacos , Íons , Pseudomonas aeruginosa/efeitos dos fármacos , Pseudomonas aeruginosa/crescimento & desenvolvimento , Pseudomonas aeruginosa/ultraestrutura , Staphylococcus epidermidis/efeitos dos fármacos , Staphylococcus epidermidis/crescimento & desenvolvimento , Staphylococcus epidermidis/ultraestrutura
15.
Sci Rep ; 7(1): 15298, 2017 11 10.
Artigo em Inglês | MEDLINE | ID: mdl-29127333

RESUMO

Evidence has shown that environmental surfaces play an important role in the transmission of nosocomial pathogens. Deploying antimicrobial surfaces in hospital wards could reduce the role environmental surfaces play as reservoirs for pathogens. Herein we show a significant reduction in viable counts of Staphylococcus epidermidis, Saccharomyces cerevisiae, and MS2 Bacteriophage after light treatment of a medical grade silicone incorporating crystal violet, methylene blue and 2 nm gold nanoparticles. Furthermore, a migration assay demonstrated that in the presence of light, growth of the fungus-like organism Pythium ultimum and the filamentous fungus Botrytis cinerea was inhibited. Atomic Force Microscopy showed significant alterations to the surface of S. epidermidis, and electron microscopy showed cellular aggregates connected by discrete surface linkages. We have therefore demonstrated that the embedded surface has a broad antimicrobial activity under white light and that the surface treatment causes bacterial envelope damage and cell aggregation.


Assuntos
Anti-Infecciosos/química , Fungos/crescimento & desenvolvimento , Levivirus/crescimento & desenvolvimento , Luz , Staphylococcus epidermidis/crescimento & desenvolvimento , Fungos/ultraestrutura , Violeta Genciana/química , Ouro/química , Levivirus/ultraestrutura , Nanopartículas Metálicas/química , Azul de Metileno/química , Staphylococcus epidermidis/ultraestrutura , Propriedades de Superfície
16.
Eur Cell Mater ; 34: 321-340, 2017 11 21.
Artigo em Inglês | MEDLINE | ID: mdl-29160896

RESUMO

Fracture-related infection (FRI) is a major complication in surgically fixed fractures. Instability of the fracture after fixation is considered a risk factor for infection; however, few experimental data are available confirming this belief. To study whether stable fractures led to higher infection clearance, mouse femoral osteotomies were fixed with either stable or unstable fixation and the surgical site was contaminated with either Staphylococcus epidermidis (S. epidermidis)or Staphylococcus aureus (S. aureus)clinical isolates. Infection progression was assessed at different time points by quantitative bacteriology, total cell counts in spleen and lymph node and histological analysis. Operated, non-inoculated mice were used as controls. Two inbred mouse strains (C57BL/6 and BALB/c) were included in the study to determine the influence of different host background in the outcome. Stable fixation allowed a higher proportion of C57BL/6 mice to clear S. epidermidis inoculation in comparison to unstable fixation. No difference associated with fixation type was observed for BALB/c mice. Inoculation with S. aureus resulted in a more severe infection for both stable and unstable fractures in both mouse strains; however, significant osteolysis around the screws rendered the stable group functionally unstable. Our results suggested that fracture stability could have an influence on S. epidermidis infection, although host factors also played a role. No differences were observed when using S. aureus, due to a more severe infection, leading to osteolysis and loss of stability in both groups. Further studies are required in order to address the biological features underlying the differences observed.


Assuntos
Fraturas do Fêmur/cirurgia , Fixação de Fratura/métodos , Staphylococcus aureus/crescimento & desenvolvimento , Staphylococcus epidermidis/crescimento & desenvolvimento , Animais , Carga Bacteriana , Biofilmes/crescimento & desenvolvimento , Feminino , Fraturas do Fêmur/microbiologia , Fixação de Fratura/efeitos adversos , Fixação de Fratura/instrumentação , Interações Hospedeiro-Patógeno , Camundongos , Camundongos Endogâmicos BALB C , Camundongos Endogâmicos C57BL , Microscopia Eletrônica de Varredura , Osteólise/microbiologia , Especificidade da Espécie , Infecções Estafilocócicas/microbiologia , Staphylococcus aureus/fisiologia , Staphylococcus aureus/ultraestrutura , Staphylococcus epidermidis/fisiologia , Staphylococcus epidermidis/ultraestrutura , Infecção da Ferida Cirúrgica/etiologia , Infecção da Ferida Cirúrgica/microbiologia
17.
Mol Med Rep ; 15(4): 1624-1630, 2017 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-28260022

RESUMO

Implant-associated infections commonly result from biofilm­forming bacteria and present severe complications in total joint arthroplasty. Therefore, there is a requirement for the development of biocompatible implant surfaces that prevent bacterial biofilm formation. The present study coated titanium samples with a thin, rapidly corroding layer of magnesium, which were subsequently investigated with respect to their antibacterial and cytotoxic surface properties using a Staphylococcus epidermidis (S. epidermidis) and human osteoblast (hOB) co­culture model. Primary hOBs and S. epidermidis were co­cultured on cylindrical titanium samples (Ti6Al4V) coated with pure magnesium via magnetron sputtering (5 µm thickness) for 7 days. Uncoated titanium test samples served as controls. Vital hOBs were identified by trypan blue staining at days 2 and 7. Planktonic S. epidermidis were quantified by counting the number of colony forming units (CFU). The quantification of biofilm­bound S. epidermidis on the surfaces of test samples was performed by ultrasonic treatment and CFU counting at days 2 and 7. The number of planktonic and biofilm­bound S. epidermidis on the magnesium­coated samples decreased by four orders of magnitude when compared with the titanium control following 7 days of co­culture. The number of vital hOBs on the magnesium­coated samples was observed to increase (40,000 cells/ml) when compared with the controls (20,000 cells/ml). The results of the present study indicate that rapidly corroding magnesium­coated titanium may be a viable coating material that possesses antibacterial and biocompatible properties. A co­culture test is more rigorous than a monoculture study, as it accounts for confounding effects and assesses additional interactions that are more representative of in vivo situations. These results provide a foundation for the future testing of this type of surface in animals.


Assuntos
Antibacterianos/farmacologia , Materiais Revestidos Biocompatíveis/farmacologia , Técnicas de Cocultura/métodos , Magnésio/farmacologia , Modelos Biológicos , Próteses e Implantes , Idoso , Ligas , Sobrevivência Celular/efeitos dos fármacos , Meios de Cultura/química , Feminino , Humanos , Concentração de Íons de Hidrogênio , Íons , Masculino , Osteoblastos/citologia , Osteoblastos/efeitos dos fármacos , Osteoblastos/ultraestrutura , Plâncton/efeitos dos fármacos , Staphylococcus epidermidis/citologia , Staphylococcus epidermidis/efeitos dos fármacos , Staphylococcus epidermidis/ultraestrutura , Titânio/farmacologia
18.
Appl Microbiol Biotechnol ; 101(8): 3349-3359, 2017 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-28343241

RESUMO

Alpha-mangostin (α-MG) has been reported to be an effective antibacterial agent against planktonic cells of many Gram-positive bacteria. However, the antibiofilm potency of α-MG remains unexplored till date. In this study, the antibiofilm and mature biofilm eradication ability of α-MG against Staphylococcus epidermidis RP62A (ATCC 35984) biofilms were evaluated. The minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC) of α-MG against S. epidermidis RP62A were found to be 1.25 and 5 µg/mL, respectively. α-MG exhibited a phenomenal concentration dependent rapid bactericidal activity (>4-log reduction within 5 min). In a multi-passage resistance analysis using S. epidermidis, no development of resistance to α-MG as well as antibiotics was observed in its habituation. α-MG at its 1/2 MIC effectively inhibited the initial biofilm formation of S. epidermidis, which was further confirmed through scanning electron microscopic (SEM) analysis that portrayed a lucid reduction in the aggregation and the spread of biofilm. The crystal violet staining and viable cell quantification results confirmed the eradication of preformed immature and mature biofilms of S. epidermidis by α-MG in a concentration dependent manner. Besides, the biofilm eradication ability was also confirmed through SEM and live/dead BacLight staining using confocal laser scanning microscopy (CLSM). Thus, the present study exemplifies that α-MG could plausibly assist to eliminate biofilm infections associated with multidrug-resistance staphylococci.


Assuntos
Biofilmes/efeitos dos fármacos , Staphylococcus epidermidis/efeitos dos fármacos , Xantonas/farmacologia , Antibacterianos/farmacologia , Biofilmes/crescimento & desenvolvimento , Testes de Sensibilidade Microbiana , Viabilidade Microbiana/efeitos dos fármacos , Microscopia Confocal , Microscopia Eletrônica de Varredura , Plâncton/efeitos dos fármacos , Staphylococcus epidermidis/ultraestrutura , Vancomicina/farmacologia
19.
J Biomed Mater Res B Appl Biomater ; 105(6): 1432-1437, 2017 08.
Artigo em Inglês | MEDLINE | ID: mdl-27086715

RESUMO

In this study, we developed a scanning transmission electron microscopy (STEM) method for imaging the antibacterial activity of organic polymeric nanoparticles (NPs) toward biofilms formed by Staphylococcus epidermidis bacterial cells, for optimizing NPs to treat biofilm infections. The combination of sample preparation method using a hydrophilic ionic liquid (IL) and STEM observation using the cooling holder eliminates the need for specialized equipment and techniques for biological sample preparation. The annular dark-field STEM results indicated that the two types of biodegradable poly-(DL-lactide-co-glycolide) (PLGA) NPs: PLGA modified with chitosan (CS), and clarithromycin (CAM)-loaded + CS-modified PLGA, prepared by emulsion solvent diffusion exhibited different antibacterial activities in nanoscale. To confirm damage to the sample during STEM observation, we observed the PLGA NPs and the biofilm treated with PLGA NPs by both the conventional method and the newly developed method. The optimized method allows microstructure of the biofilm treated with PLGA NPs to be maintained for 25 min at a current flow of 40 pA. The developed simple sample preparation method would be helpful to understand the interaction of drugs with target materials. In addition, this technique could contribute to the visualization of other deformable composite materials at the nanoscale level. © 2016 Wiley Periodicals, Inc. J Biomed Mater Res Part B: Appl Biomater, 105B: 1432-1437, 2017.


Assuntos
Antibacterianos , Biofilmes/efeitos dos fármacos , Líquidos Iônicos , Microscopia Eletrônica de Transmissão e Varredura , Nanopartículas/química , Staphylococcus epidermidis , Antibacterianos/química , Antibacterianos/farmacologia , Quitosana/química , Quitosana/farmacologia , Líquidos Iônicos/química , Líquidos Iônicos/farmacologia , Ácido Láctico/química , Ácido Láctico/farmacologia , Nanopartículas/ultraestrutura , Ácido Poliglicólico/química , Ácido Poliglicólico/farmacologia , Copolímero de Ácido Poliláctico e Ácido Poliglicólico , Staphylococcus epidermidis/fisiologia , Staphylococcus epidermidis/ultraestrutura
20.
Langmuir ; 32(31): 7866-74, 2016 08 09.
Artigo em Inglês | MEDLINE | ID: mdl-27397718

RESUMO

The surfaces or coatings that can effectively suppress bacterial adhesion in the long term are of critical importance for biomedical applications. Herein, a group of amino acid-based zwitterionic polymers (pAAZ) were investigated for their long-term resistance to bacterial adhesion. The polymers were derived from natural amino acids including serine, ornithine, lysine, aspartic acid, and glutamic acid. The pAAZ brushes were grafted on gold via the surface-initiated photoiniferter-mediated polymerization (SI-PIMP). Results show that the pAAZ coatings highly suppressed adsorption from the undiluted human serum and plasma. Long-term bacterial adhesion on these surfaces was investigated, using two kinds of representative bacteria [Gram-positive Staphylococcus epidermidis and Gram-negative Pseudomonas aeruginosa] as the model species. Results demonstrate that the pAAZ surfaces were highly resistant to bacterial adhesion after culturing for 1, 5, 9, or even 14 days, representing at least 95% reduction at all time points compared to the control unmodified surfaces. The bacterial accumulation on the pAAZ surfaces after 9 or 14 days was even lower than on the surfaces grafted with poly[poly(ethyl glycol) methyl ether methacrylate] (pPEGMA), one of the most common antifouling materials known to date. The pAAZ brushes also exhibited excellent structural stability in phosphate-buffered saline after incubation for 4 weeks. The bacterial resistance and stability of pAAZ polymers suggest they have good potential to be used for those applications where long-term suppression to bacterial attachment is desired.


Assuntos
Aminoácidos/química , Aderência Bacteriana , Materiais Revestidos Biocompatíveis/química , Metacrilatos/química , Polietilenoglicóis/química , Pseudomonas aeruginosa/crescimento & desenvolvimento , Staphylococcus epidermidis/crescimento & desenvolvimento , Humanos , Pseudomonas aeruginosa/ultraestrutura , Soro/química , Staphylococcus epidermidis/ultraestrutura
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