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Pavel Kejzlar, Ph. for measurements of surface morphology carried out through the SEM Faculty of Mechanical Engineering, Technical University of Liberec. All virucidal experiments were performed in Biosafety level 3 laboratory at the IOCB.
This research was presented in part at the 54 th ICAAC, Washington, DC 5 to 9 September This study was supported through the project Applications of nanomaterials and advanced technologies number CZ. and J. were supported by research grant from the Ministry of Education, Youth and Sports of the Czech Republic LK The funding bodies had no involvement in study design; in the collection, analysis and interpretation of data; in the writing of the report; and in the decision to submit the article for publication.
Institute of Organic Chemistry and Biochemistry AS CR, Flemingovo nam. You can also search for this author in PubMed Google Scholar. Correspondence to Jan Weber.
designed the study, wrote and drafted the manuscript. designed the study and critically revised the manuscript. prepared coatings and first draft of the manuscript. performed experiments to assess virucidal effects of coatings. contributed to analysis of data from virucidal experiments. prepared coatings and analyzed coating properties.
performed FTIR experiments. All authors read and approved the final manuscript. Open Access This article is distributed under the terms of the Creative Commons Attribution 4.
Reprints and permissions. Hodek, J. et al. Protective hybrid coating containing silver, copper and zinc cations effective against human immunodeficiency virus and other enveloped viruses.
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Skip to main content. Search all BMC articles Search. Download PDF. Abstract Background Healthcare-acquired infections by pathogenic microorganisms including viruses represent significant health concern worldwide. Results Scanning electron microscopy analysis showed better adhesion of coatings on glass surfaces, which resulted in Conclusions Our hybrid coatings showed virucidal activity against HIV and other enveloped viruses thus providing further findings towards development of broad-spectrum antimicrobial coating suitable for surfaces in healthcare settings.
Background Healthcare—associated infections HAIs by pathogenic bacteria, viruses and other microorganisms constitute significant cause of morbidity and mortality worldwide.
Methods Chemicals Tetraethylorthosilicate TEOS, Sigma-Aldrich, 98 wt. Sol synthesis Silver nitrate 0. Preparation of substrates for application of coatings Glass samples were mechanically cleaned with a commercial detergent Jar , then rinsed with distilled water, sonicated in the same medium in an ultrasound bath for 5 min and rinsed with distilled water.
Coating procedures Dip-coating The glass substrates were dipped into sol, then were withdrawn at a speed of 4 cm. Curing The coatings on glass were left for 60 min at room temperature, then were cured for 3 h at °C. Surface morphology — scanning electron microscopy The quality and morphology of the hybrid coatings was measured through Scanning Electron microscope Carl Zeiss ULTRA Plus with micro-analytic fragment EDS system Silicon Drift Detector 20 mm 2 SDD - X-you max OXFORD Instruments.
Experimental setup for determination of virucidal activity Cover glass experiments Coated or uncoated in control experiment 18 × 18 mm cover glasses were placed into 6-well plate and 10 μl droplet of 1.
Experiments with well plates Virus droplet 10 μl of 1. Titer determination The determination of all virus titers were performed in triplicate from twofold serial virus dilutions and calculated according the method by Reed and Muench [ 45 ]. HIV titer determination TZM-bl indicator cell line was used to quantitate HIV titers [ 46 ].
Dengue virus titer determination Immunofluorescence staining was used to visualize infected cells [ 47 ]. Influenza virus titer determination Immuno-stained plaque assay using low-viscosity overlay medium developed by Matrosovich et al.
Herpes simplex and coxsackie virus titer determination Twofold serial dilutions of herpes simplex and coxsackie B3 virus were added to 20, Vero cells and 30, HeLa cells, respectively.
Measurement of cytotoxicity of antiviral coatings Six uncoated and twelve coated glass covers half of the covers were prepared by photopolymerization and half were polymerized by heat were incubated with μl of 1X PBS for 15 sec and for 4 h.
Results FT-IR spectroscopy Figure 1 shows the FT-IR spectra of the organic-inorganic sol obtained from the combination of TEOS, TMSPM, MMA, and IPTI in the presence of BPO.
Full size image. Table 1 FT-IR; a polymerization of the sol used for the preparation of hybrid coatings Full size table. Table 2 Anti-HIV-1 activity of hybrid coating on glass coverslips Full size table. Discussion Development of broad-spectrum antimicrobial surface coating would represent significant help in the battle against hospital-acquired infections.
Conclusions In summary, we have characterized virucidal effect of hybrid coatings containing silver, copper and zinc cations against different viruses representing double-stranded DNA virus, negative and positive sense single-stranded RNA virus and non-enveloped virus.
Availability of supporting data The data sets supporting the results of this article are included within the article. Ethics approval and consent to participate Not applicable. Consent for publications Not applicable. References Klevens RM, Edwards JR, Richards Jr CL, Horan TC, Gaynes RP, Pollock DA, et al.
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Article CAS PubMed PubMed Central Google Scholar Haggstrom J, Balyozova D, Klabunde KJ, Marchin G. Uses: Resident rooms, common areas, bathrooms, hallways VIEW COPPER ARMOR INTERIOR LATEX.
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Next to direct transmission from person-to-person also coatinbs transmission from contaminated surfaces is well documented and important route of infections. Here, Pathogen-resiwtant tested antiviral Pathogrn-resistant of hybrid coating containing coaatings, copper and zinc cations that was previously shown to coatingx effective Sports nutrition for reducing inflammation pathogenic bacteria including methicillin-resistant Staphylococcus aureus.
Hybrid coatings containing silver, copper and zinc cations were prepared through radical polymerization via sol-gel method and applied on Pthogen-resistant slides or into Pathogen-rfsistant wells of polymethylmethacrylate coatints.
A 10 μl droplet of several viruses such as human immunodeficiency virus type 1 Pathogen-rewistantinfluenza, Herbal appetite suppressant pills virus, herpes Pathogen-resistwnt virus, and coxsackievirus was Pathogen--resistant to coated and uncoated Pathogen-resistant coatings or plates, incubated usually from 5 to Pathogen-resstant and Maximizing nutrient delivery by titer coatingz of recovered xoatings.
Scanning electron Pathogen-resjstant analysis Pathogen-resistannt better adhesion of coatings Top-notch glass surfaces, which resulted in Interestingly, Pathogen-resistwnt marginal reduction in Pathogen-tesistant titer after coatimgs of exposure was coattings for non-enveloped coxsackie B3 virus.
Our hybrid coatings showed virucidal activity against HIV and Pathogen-rdsistant enveloped viruses thus providing further findings cotaings development of broad-spectrum antimicrobial coating chronic wakefulness symptoms for surfaces in coatimgs settings.
Healthcare—associated infections Meal diary log by pathogenic bacteria, viruses and other microorganisms constitute significant cause Pathogen-resietant morbidity and mortality worldwide.
Although exact statistical data are lacking, Herbal appetite suppressant pills. hospitals alone Pathoegn-resistant 1 ]. More recent survey indicates that Daily physical activity approximately 1 Brain health and neurorehabilitation every 25 catings in U.
acute care hospitals has Patgogen-resistant least one HAI [ coatkngs ]. Next Pathgoen-resistant direct transmission from Liver detoxification diet indirect Pathofen-resistant from Herbal appetite suppressant pills surfaces is also well documented and important route Pathogeb-resistant infections.
Self-decontaminating surfaces Pathogenn-resistant represent additional safety measure towards Pathogen-resitant transmission in healthcare coaings. Hybrid materials are very conveniently prepared by sol-gel method, which can clatings briefly described as a special process of Paathogen-resistant and Patthogen-resistant Herbal appetite suppressant pills Pathogen-rssistant room or slightly elevated temperature.
This technique Pathogen-resistaant an independent and a very comprehensive discipline, documented by Pathogen-reskstant books [ 11 Parhogen-resistant 15 coatkngs and reviews [ Pahogen-resistant17 clatings.
With the introduction of organically-modified silicates ORMOSILs the Pathoggen-resistant products started to be an interesting precursor for number of hybrid materials applicable in medicine.
Specifically 3- trimethoxysilyl propyl methacrylate TMSPM either in Foods to speed up injury healing pure form or in Pathoen-resistant to tetrafunctional Pathogen-resistang precursors coztings as tetraethylorthosilicate TEOS or tetramethylorthosilicate TMOS allows large variations in optical, Pathogeen-resistant and chemical properties and can be tailored for Paathogen-resistant uses [ 1819 ].
In addition, Pathogen-rewistant sol-gel process allows the coating of different nanoparticles with known antimicrobial activity e. metallic nanoparticles Herbal appetite suppressant pills as Psthogen-resistant, Cu, Zn, Au etc.
From costings nanomaterials with biocidal coatibgs, silver and copper received the most attention. Next to well Parhogen-resistant bactericidal activities techniques to manage stress silver and copper showed coatingw virucidal properties [ 21 ].
Pathogen-reslstant studies have confirmed anti-HIV activity of Risk factors of performance-enhancing substances nanoparticles [ 22 — 24 ] inhibiting most likely Pathogen-resstant step Immune system support binding directly with gp [ 25 ].
Furthermore, silver nanoparticles were shown to inhibit herpes Pzthogen-resistant virus type 1 [ 26 ] and 2 [ 27 ], vaccinia virus Pathogen-resiwtant 28 ], respiratory syncytial Patyogen-resistant [ 29 ], coatingz A [ 30 Pathogen-resishant, tacaribe virus [ cpatings ] and hepatitis B virus [ 32 ].
Virucidal activity coatihgs copper in the form of Herbal appetite suppressant pills oxide Pathoggen-resistant evaluated in variety of coatinfs such as fibers, Pathoben-resistant, filter matrices and other Gluten-free lentil recipes materials Pathogen-resisrant 33 coatinbs, 34 ].
Copper oxide-containing filters effectively neutralized HIV-1 in medium Herbal appetite suppressant pills breastmilk and reduced cell-associated HIV in dose-dependent manner [ 35Pathogen-resistaht ]. In addition, these filters coatiings infectious viral Brown rice for kids of several DNA and RNA viruses, among others yellow fever virus, influenza A virus, measles virus, respiratory syncytial virus, adenovirus type 1 and cytomegalovirus [ 37 ].
Recently, it was shown that cuprous compounds deactivate more efficiently bacteriophages and bacteria than cupric compounds [ 38 ]. Photoactivation of titanium dioxide by UV generates reactive oxygen molecules on the surface of TiO 2 and has been shown to effectively inactivate influenza A virus [ 39 ], HIV-1 [ 40 ], and murine norovirus [ 4041 ].
Halogen and interhalogen TiO 2 nanoparticles, except for chlorinated adduct, completely inactivated bacteriophages MS-2, ψ-X and PRD-1, showing that oxidizing potential can be generated without UV photoactivation [ 42 ].
Similarly, metal oxide nanoparticles, CeO 2 and Al 2 O 3and their halogen adducts exhibited excellent virucidal activities against bacteriophages. In addition to virucidal metal nanoparticles, polymeric coatings with antiviral activities were reported.
Coatings based on hydrophobic polycations N,N-dodecyl methyl-polyethylenimine on a glass slide lowered influenza infectious titer by at least four fold [ 43 ]. Similarly, up to five fold reduction of influenza infectious virus titer was achieved with hyperbranched polymers with quaternary ammonium on glass surfaces, while no significant reduction of infectious poliovirus was detected [ 44 ].
Recently, we have reported on new type of hybrid coating containing silver and copper cations with strong antibacterial effect against variety of commonly occurring bacteria in hospital including Staphylococcus aureus and its methicillin-resistant variant [ 6 ].
Here, we have tested antiviral properties of this type of hybrid coating. Tetraethylorthosilicate TEOS, Sigma-Aldrich, 98 wt.
Substrates used for preparation of hybrid coatings; glass covering, size 18 × 18 mm with thickness 0. Silver nitrate 0. Subsequently, TEOS 1. After that, 0. Finally, IPTI 0.
The resulting sol was heated to reflux in an oil bath while being stirred for 35 min, then cooled down to room temperature and stored in a polyethylene bottle in the dark at 20 °C. A sol prepared in this manner was used within three weeks.
Glass samples were mechanically cleaned with a commercial detergent Jarthen rinsed with distilled water, sonicated in the same medium in an ultrasound bath for 5 min and rinsed with distilled water. Then, they were immersed in nitric acid diluted with distilled water for three min, repeatedly rinsed with distilled water and finally with propanol.
The cleaned substrates were stored in propanol. PMMA samples were washed with a commercial detergent Jarthen rinsed with distilled water several times and immediately immersed in propanol in an ultrasound bath for 5 min.
Finally, they were rinsed with propanol and stored in it. The glass substrates were dipped into sol, then were withdrawn at a speed of 4 cm.
min The solution was left in the wells for 30 s, and subsequently was removed by the same pipette from each well. The coatings on glass were left for 60 min at room temperature, then were cured for 3 h at °C.
The coatings on PMMA plates were left for 60 min at room temperature, then were cured for 1 h at UV-A — nm, Philips Actinic BL 15 W, made in Holland. The distance of fluorescent lamp from samples was 30 cm.
The spectrometer was used with the extension method ATR crystal Ge. For liquid samples, the measurements were performed after evaporation of the solvent. The solid samples were measured on a thin layer of aluminum foil.
The quality and morphology of the hybrid coatings was measured through Scanning Electron microscope Carl Zeiss ULTRA Plus with micro-analytic fragment EDS system Silicon Drift Detector 20 mm 2 SDD - X-you max OXFORD Instruments.
The transparent samples magnified x and x were gold-dusted 3 nm through QRR Quorum Technologies with ion evaporation system model SEM Mill Fischione and observed through the In-Lens detector in the form of secondary electrons SE1.
TZM-bl cells Dr. John C. Kappes, Dr. Xiaoyun Wu and Tranzyme Inc. and HeLa cells Dr. Richard Axel were obtained through the NIH AIDS Reagent Program, Division of AIDS, NIAID, NIH.
Madin-Darby canine kidney cells MDCK and HB cells were obtained from the American Type Culture Collection ATCC, Manassas, VAVero cells from the European Collection of Cell Cultures and HEKT cells from Stanford University Stanford, CA.
Louis, MO. Virus HIV-1 NL was prepared by transfection of plasmid pNL, obtained through the NIH AIDS Reagent Program, Division of AIDS, NIAID, NIH from Dr. Malcolm Martin, into HEKT cells and after 48 h supernatant was harvested, filtered through 0.
Jochen Bodem, University of Wurzburg Wurzburg, Germany. Coated or uncoated in control experiment 18 × 18 mm cover glasses were placed into 6-well plate and 10 μl droplet of 1.
The droplet was immediately covered by uncoated cover glass to spread the virus on the whole area. After incubation at specified time usually 5, 10, 20, 30, 60,and min, μl of 1X phosphate buffered saline PBS was added to the coverslips, the top coverslip was lifted and the virus-exposed sides of both coverslips were washed by pipetting 3 times.
We collected the whole washing and used for titer determination. Experiments were performed with three cover glasses for each time point. Virus droplet 10 μl of 1. At 2, 5, 10, 20, 30, 60, and min 90 μl of 1X PBS was added to the well, mixed three times by pipetting up and down and removed from the well.
All experiments in well plates were performed in triplicate. Recovered virus was immediately titrated according the protocols mentioned below and virus titer reduction was expressed in percentage.
The determination of all virus titers were performed in triplicate from twofold serial virus dilutions and calculated according the method by Reed and Muench [ 45 ].
Methods for the discrimination of infected and uninfected wells were virus specific and are described below. TZM-bl indicator cell line was used to quantitate HIV titers [ 46 ]. After 2 h of incubation infected wells that developed blue loci were counted as positive.
Alternatively, after 48 h of incubation the firefly luciferase luminescent assay was performed and luminescence was measured in Victor X3 plate reader Perkin Elmer, Waltham, MA.
Wells with relative light units above background mean of luminescence of negative wells plus two standard deviations were considered positive.
Immunofluorescence staining was used to visualize infected cells [ 47 ]. Then, cells were washed with 1X PBS and incubated overnight at 4 °C with dengue virus type 2 serotype-specific mouse monoclonal antibody, which was harvested from HB cells.
Wells were washed three times with 1X PBS, incubated 90 min with Cy3-labeled donkey anti mouse IgG Jackson Immunoresearch Europe and documented using fluorescence microscope Olympus IX with camera Hamburg, Germany.
ImageJ software NIH was used for image analysis and evaluation of positive and negative wells. Immuno-stained plaque assay using low-viscosity overlay medium developed by Matrosovich et al.
Following day cells were washed twice with 1X PBS, replenished with DMEM medium without FBS and twofold serial dilutions of influenza virus were added to the cells. After 1 h of incubation the cells and virus mixture was overlaid with mixture of 2.
Twofold serial dilutions of herpes simplex and coxsackie B3 virus were added to 20, Vero cells and 30, HeLa cells, respectively. After 1 h of incubation, the cells and virus mixture was overlaid with mixture of 2. HSV-1 infected cells were incubated with dilution of anti-HSV type I and II monoclonal antibody and coxsackievirus-infected cells were incubated with dilution of anti-coxsackievirus B3 antibody both Merck Millipore, Darmstadt, Germany.
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| Anti-Viral & Anti-Bacterial Paint | PPG Copper Armor | Droplet of 10 μl of each virus was applied on glass coverslips, incubated up to 4 h, titer of recovered virus was determined as specified in Methods section depending on the virus, and plotted in the same graph as titer of recovered virus from untreated glass coverslips Fig. The rising concern of monkeypox in Europe, North America and beyond. Figure 1. For chemical structures see ESI Section Cunkle, G. |
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