WO2015011327A1 - Method for inhibiting microbial growth - Google Patents

Method for inhibiting microbial growth Download PDF

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Publication number
WO2015011327A1
WO2015011327A1 PCT/ES2014/070595 ES2014070595W WO2015011327A1 WO 2015011327 A1 WO2015011327 A1 WO 2015011327A1 ES 2014070595 W ES2014070595 W ES 2014070595W WO 2015011327 A1 WO2015011327 A1 WO 2015011327A1
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WO
WIPO (PCT)
Prior art keywords
glass
microbial growth
inhibiting microbial
weight
biocidal
Prior art date
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PCT/ES2014/070595
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Spanish (es)
French (fr)
Inventor
José Serafín MOYA CORRAL
Ramón TORRECILLAS SAN MILLÁN
María Belén CABAL ÁLVAREZ
Eduardo ÁLVAREZ DÍAZ
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Consejo Superior De Investigaciones Científicas (Csic)
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Publication of WO2015011327A1 publication Critical patent/WO2015011327A1/en

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Classifications

    • A—HUMAN NECESSITIES
    • A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
    • A01N—PRESERVATION OF BODIES OF HUMANS OR ANIMALS OR PLANTS OR PARTS THEREOF; BIOCIDES, e.g. AS DISINFECTANTS, AS PESTICIDES OR AS HERBICIDES; PEST REPELLANTS OR ATTRACTANTS; PLANT GROWTH REGULATORS
    • A01N59/00—Biocides, pest repellants or attractants, or plant growth regulators containing elements or inorganic compounds
    • A—HUMAN NECESSITIES
    • A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
    • A01N—PRESERVATION OF BODIES OF HUMANS OR ANIMALS OR PLANTS OR PARTS THEREOF; BIOCIDES, e.g. AS DISINFECTANTS, AS PESTICIDES OR AS HERBICIDES; PEST REPELLANTS OR ATTRACTANTS; PLANT GROWTH REGULATORS
    • A01N59/00—Biocides, pest repellants or attractants, or plant growth regulators containing elements or inorganic compounds
    • A01N59/14—Boron; Compounds thereof
    • A—HUMAN NECESSITIES
    • A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
    • A01N—PRESERVATION OF BODIES OF HUMANS OR ANIMALS OR PLANTS OR PARTS THEREOF; BIOCIDES, e.g. AS DISINFECTANTS, AS PESTICIDES OR AS HERBICIDES; PEST REPELLANTS OR ATTRACTANTS; PLANT GROWTH REGULATORS
    • A01N59/00—Biocides, pest repellants or attractants, or plant growth regulators containing elements or inorganic compounds
    • A01N59/16—Heavy metals; Compounds thereof
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61L—METHODS OR APPARATUS FOR STERILISING MATERIALS OR OBJECTS IN GENERAL; DISINFECTION, STERILISATION OR DEODORISATION OF AIR; CHEMICAL ASPECTS OF BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES; MATERIALS FOR BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES
    • A61L27/00—Materials for grafts or prostheses or for coating grafts or prostheses
    • A61L27/02—Inorganic materials
    • A61L27/10—Ceramics or glasses
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61L—METHODS OR APPARATUS FOR STERILISING MATERIALS OR OBJECTS IN GENERAL; DISINFECTION, STERILISATION OR DEODORISATION OF AIR; CHEMICAL ASPECTS OF BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES; MATERIALS FOR BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES
    • A61L27/00—Materials for grafts or prostheses or for coating grafts or prostheses
    • A61L27/50—Materials characterised by their function or physical properties, e.g. injectable or lubricating compositions, shape-memory materials, surface modified materials
    • A61L27/54—Biologically active materials, e.g. therapeutic substances
    • C—CHEMISTRY; METALLURGY
    • C03—GLASS; MINERAL OR SLAG WOOL
    • C03C—CHEMICAL COMPOSITION OF GLASSES, GLAZES OR VITREOUS ENAMELS; SURFACE TREATMENT OF GLASS; SURFACE TREATMENT OF FIBRES OR FILAMENTS MADE FROM GLASS, MINERALS OR SLAGS; JOINING GLASS TO GLASS OR OTHER MATERIALS
    • C03C3/00—Glass compositions
    • C03C3/04—Glass compositions containing silica
    • C03C3/062—Glass compositions containing silica with less than 40% silica by weight
    • C03C3/064—Glass compositions containing silica with less than 40% silica by weight containing boron
    • C03C3/068—Glass compositions containing silica with less than 40% silica by weight containing boron containing rare earths
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61L—METHODS OR APPARATUS FOR STERILISING MATERIALS OR OBJECTS IN GENERAL; DISINFECTION, STERILISATION OR DEODORISATION OF AIR; CHEMICAL ASPECTS OF BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES; MATERIALS FOR BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES
    • A61L2300/00—Biologically active materials used in bandages, wound dressings, absorbent pads or medical devices
    • A61L2300/10—Biologically active materials used in bandages, wound dressings, absorbent pads or medical devices containing or releasing inorganic materials
    • A61L2300/102—Metals or metal compounds, e.g. salts such as bicarbonates, carbonates, oxides, zeolites, silicates
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61L—METHODS OR APPARATUS FOR STERILISING MATERIALS OR OBJECTS IN GENERAL; DISINFECTION, STERILISATION OR DEODORISATION OF AIR; CHEMICAL ASPECTS OF BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES; MATERIALS FOR BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES
    • A61L2300/00—Biologically active materials used in bandages, wound dressings, absorbent pads or medical devices
    • A61L2300/40—Biologically active materials used in bandages, wound dressings, absorbent pads or medical devices characterised by a specific therapeutic activity or mode of action
    • A61L2300/404—Biocides, antimicrobial agents, antiseptic agents

Definitions

  • the present invention is related to a process for preventing the growth of microorganisms through the use of inorganic materials, and more specifically of compounds that include boron.
  • the invention is related to the use of said method in the manufacture of packages, drugs, medical devices, surgical implants, tissues, means of transport, air installations or water installations.
  • the antimicrobial capacity of these glasses is mainly due to the increase in pH and osmotic pressure of the medium caused by the leaching of sodium, calcium, phosphate and silicate ions, although the disturbing effect caused by the high concentration of ions is also known. alkalis leached in the membrane potential and therefore may have consequences on the antimicrobial capacity of biovidrios.
  • sodoccalcic glasses have been described in which the biocidal activity is associated with the presence in calcium oxide (CaO) glass [ES2361695 of 04/27/2012].
  • CaO calcium oxide
  • the inventors establish a minimum content of 10% by weight of CaO as a requirement for the glass to exhibit biocidal activity.
  • biocidal activity of this sodoccalcic glass is attributed to the direct interaction between the glass particle and the cell membrane of the bacteria or fungus that would produce a partial or total interference with the metabolism of said bacterium or fungus.
  • glasses with antimicrobial properties characterized by containing metals such as copper or silver in the form of salts [patent ES2190732 of 06/16/2004], [patent US6921546 of 07/26/2005], [ patent application WO2005030665 of 07/04/2005] or as nanoparticles [Esteban-Tejeda L, Malpartida F, Esteban-Cubillo A, Pecharromán C, Moya JS Nanotechnology 25; 20 (8): 085103 (2009)], [Esteban-Tejeda L, Malpartida F, Esteban-Cubillo A, Pecharromán C, Moya JS, Nanotechnology 16; 20 (50): 505701 (2009)].
  • the activity in this type of glass is attributed to the presence of these metallic elements. It is known to use these ions or metal nanoparticles as bactericides, fungicides and algaecides.
  • a method has been developed that allows the inhibition of the growth of microorganisms in a lasting way by using a glass with a particular composition that includes a zinc oxide content greater than 15%.
  • the glass used in the invention does not require the presence of metals such as silver or copper and does not belong to the family of phosphate or sodoccalc glass.
  • glass is understood as an inorganic amorphous material and therefore has no crystalline phases.
  • Biocide means any synthetic or natural substance intended to destroy, counteract, neutralize, prevent the action or exert control of any other type on any microbial organism considered harmful to man, including bacteria, yeasts, fungi and viruses.
  • Bactericidal means any substance of natural or synthesized origin that is capable of destroying bacteria.
  • material is meant a substance with some useful property, whether mechanical, electrical, optical, thermal or magnetic, such as, for example, the material can be or be part of a metallic, ceramic, agglomerated, textile, plastic or plastic fibers. polymeric materials or composite materials, among others.
  • the object of the invention consists in this way in a method for the inhibition of microbial growth characterized by the use of a glass added to a material, hereinafter glass of the invention, which does not contain any of the following elements: phosphorus, silver, copper and which includes in its composition at least silicon oxide (Si0 2 ), boron oxide (B 2 0 3 ), zinc oxide (ZnO) with a zinc oxide content exceeding 15% by weight and calcium oxide with a content less than 10% by weight.
  • Si0 2 silicon oxide
  • B 2 0 3 boron oxide
  • ZnO zinc oxide
  • a preferred aspect of the invention is the presence in the glass used for the inhibition of microbial growth of the invention of a percentage of zinc oxide less than 55% by weight.
  • the glass of the invention has a silicon oxide content of between 16% and 30% by weight. In an even more preferred embodiment, the glass of the invention does not contain calcium oxide or contains traces (less than 0.1% by weight).
  • a preferred aspect of the invention consists in the use of the glass of the invention in particulate, fiber or sphere form.
  • a particular embodiment consists in incorporating the glass of the invention as a filler material in polymeric materials.
  • the biocidal effect of coatings made with the glass of the invention has been verified. Therefore, another particular embodiment is the use for biocidal purposes of coatings formed with the glass of the invention.
  • the method object of the invention can be used in various applications.
  • this method for the inhibition of microbial growth can be applied to packages, drugs, medical devices, surgical implants, tissues, means of transport (for example for devices or structural elements), air installations (for example for devices or structural elements , such as air purification filters) or water installations (for example for devices or structural elements, such as water purification filters).
  • Example 1 Test of biocidal activity of the glass of the invention in particulate form
  • a glass of chemical composition (% weight): 4.97% AI2O3 was prepared; 5.38% Na 2 0; 18.7% of Si02; 0, 10% of K 2 0; 0.17% Ti0 2 ; 34.7% ZnO; 0.0024% SrO; 33.2% of B 2 0 3 ; 0.45% of Ce0 2 ; 2.25% of Zr0 2 and 0.056% of others.
  • the bacteria were seeded in solid medium, Petri dishes, from Luria Bertani (LB) whose composition is (% weight): 1% tryptone, 0.5% yeast extract, 1% NaCl and 1.5% agar. The plates were incubated 18 hours at 37 ° C.
  • Yeasts in solid medium, Petri dishes of (YEPD) whose composition is (% weight): Bacto-pectone 1%, yeast extract 1%, glucose 2% and agar 2%. The plates were incubated 18 hours at 37 ° C.
  • colonies isolated from the above plates were inoculated in 1 mL of LB or liquid YEPD and cultured at 37 ° C for 6 hours to obtain precultures at a density of between 10 7 to 10 8 viable colonies per milliliter of culture.
  • a 200 mg / mL suspension in glass powder water will be prepared and sterilized for 30 minutes at 125 ° C.
  • 10 ⁇ _ of the bacterial preculture was inoculated in 1 ml_ of LB
  • 10 ⁇ _ of the yeast preculture was inoculated in 1 ml_ of YEPD.
  • 75 ⁇ _ of the glass suspension to be tested was added to this culture.
  • the same medium in which the amount of glass was replaced by an equivalent volume of water was prepared as a control.
  • the different cultures were incubated in horizontal agitation, making extractions every 24 hours in order to proceed to title the surviving microorganisms by plating in solid LB medium by serial dilutions from 10 ⁇ 1 to 10 ⁇ 8 .
  • Example 2 Biocidal activity test of the glass of the invention applied as a coating on titanium alloys
  • the coatings were carried out with the particulate biocide glass described in example 1, on plates of the titanium alloy Ti- 6AI-4V, of dimensions 12.5 x 8.3 x 1 mm and with 99.0 % purity
  • antimicrobial efficacy was determined following the procedure described in ISO 22196. This standard has been adopted internationally as a reference standard to evaluate the antimicrobial efficacy of non-porous, ceramic, plastic surfaces, etc. According to this standard, antimicrobial activity is determined by comparing the results obtained between the treated surface and a control surface, without biocidal agent (blank), after an incubation period of 48 hours at 37 ° C, optimum temperature for the growth of the selected microorganism.
  • ISO 22196 defines the antimicrobial activity of a surface as the difference between the number of bacteria in the control sample (blank, A) and the number of bacteria in the treated sample (B), according to the equation:
  • Example 3 Biocidal activity test of a polymer which contains the glass of the invention in particulate form as filler material
  • thermoplastic polymer polyurethane was selected as the polymer.
  • filler material a particulate glass similar to that described in example 1 was selected.
  • a weight load of 50% biocidal glass was added. Homogeneous mixing of all the members of the formulation was performed hot. Using an extruder, the formulation was obtained in the form of threads, which were subsequently processed in the form of sheets by using a hot plate hydraulic press. Similarly, controls were obtained containing only the polymer without biocide additive.
  • bactericidal tests were performed against the Gram-negative Escherichia coli DH10B bacteria.
  • the value of antimicrobial efficacy was determined following the procedure described in ISO 22196. The standard indicates that the bactericidal efficacy index must be greater than 2 to be considered as such. In all In the cases studied, the value obtained was greater than 5. This means a decrease in the number of bacteria greater than 99.999% compared to the initial population.

Abstract

The invention relates to a method for inhibiting microbial growth using glass with a composition containing various oxides of elements such as calcium, zinc, silicon and boron. The invention also relates to the use of the method for producing containers, drugs, medical devices, surgical implants, fabrics, transportation means, air systems and water systems.

Description

PROCEDIMIENTO PARA LA INHIBICIÓN DEL CRECIMIENTO  PROCEDURE FOR INHIBITION OF GROWTH
MICROBIANO  MICROBIAL
SECTOR DE LA TÉCNICA SECTOR OF THE TECHNIQUE
La presente invención está relacionada con un procedimiento para impedir el crecimiento de microorganismos mediante el empleo de materiales inorgánicos, y más concretamente de compuestos que incluyen boro. The present invention is related to a process for preventing the growth of microorganisms through the use of inorganic materials, and more specifically of compounds that include boron.
Asimismo, la invención está relacionada con el uso de dicho procedimiento en la fabricación de envases, fármacos, dispositivos médicos, implantes quirúrgicos, tejidos, medios de transporte, instalaciones de aire o instalaciones de agua. Likewise, the invention is related to the use of said method in the manufacture of packages, drugs, medical devices, surgical implants, tissues, means of transport, air installations or water installations.
ESTADO DE LA TÉCNICA. STATE OF THE TECHNIQUE.
El empleo de vidrios con capacidad antimicrobiana es conocido. Así, existen numerosas referencias en el estado del arte que reivindican la capacidad antimicrobiana de estos vidrios de fosfato, conocidos como biovidrios [Munukka E, Leppáranta O, Korkeamáki M, Vaahtio M, Peltola T, Zhang D, Hupa L, Ylánen H, Salonen Jl, Viljanen MK, Eerola E.J. Mater. Sci: Mater. Med. 19: 27-32 (2008)], [Patente EP1601623 de 1 1/04/2007]. La capacidad antimicrobiana estos vidrios se debe fundamentalmente al aumento del pH y de la presión osmótica del medio provocado por la lixiviación de iones de sodio, calcio, fosfato y silicato, si bien también es conocido el efecto perturbador que provoca la alta concentración de los iones alcalinos lixiviados en el potencial de membrana y que por tanto pueden tener consecuencias en la capacidad antimicrobiana de los biovidrios. The use of glasses with antimicrobial capacity is known. Thus, there are numerous references in the state of the art that claim the antimicrobial capacity of these phosphate glasses, known as bio-glass [Munukka E, Leppáranta O, Korkeamáki M, Vaahtio M, Peltola T, Zhang D, Hupa L, Ylánen H, Salonen Jl, Viljanen MK, Eerola EJ Mater. Sci: Mater. Med. 19: 27-32 (2008)], [Patent EP1601623 of 1 04/01/2007]. The antimicrobial capacity of these glasses is mainly due to the increase in pH and osmotic pressure of the medium caused by the leaching of sodium, calcium, phosphate and silicate ions, although the disturbing effect caused by the high concentration of ions is also known. alkalis leached in the membrane potential and therefore may have consequences on the antimicrobial capacity of biovidrios.
Por otro lado se ha descrito el uso biocida de vidrios sodocálcicos en los que la actividad biocida se asocia a la presencia en el vidrio de óxido de calcio (CaO)[patente ES2361695 de 27/04/2012]. En esta patente los inventores establecen un contenido mínimo del 10% en peso de CaO como requisito para que el vidrio presente actividad biocida. Asimismo la actividad biocida de este vidrio sodocálcico es atribuida a la interacción directa entre la partícula de vidrio y la membrana celular de la bacteria u hongo que produciría una interferencia, parcial o total, con el metabolismo de dicha bacteria u hongo. On the other hand, the biocidal use of sodoccalcic glasses has been described in which the biocidal activity is associated with the presence in calcium oxide (CaO) glass [ES2361695 of 04/27/2012]. In this patent the inventors establish a minimum content of 10% by weight of CaO as a requirement for the glass to exhibit biocidal activity. Also the biocidal activity of this sodoccalcic glass is attributed to the direct interaction between the glass particle and the cell membrane of the bacteria or fungus that would produce a partial or total interference with the metabolism of said bacterium or fungus.
Por otro lado, es conocido el uso de vidrios con propiedades antimicrobianas caracterizados por contener metales como cobre o plata ya sea en forma de sales [patente ES2190732 de 16/06/2004], [patente US6921546 de 26/07/2005], [solicitud de patente WO2005030665 de 07/04/2005] o como nanopartículas [Esteban-Tejeda L, Malpartida F, Esteban-Cubillo A, Pecharromán C, Moya JS Nanotechnology 25; 20(8):085103 (2009)], [Esteban-Tejeda L, Malpartida F, Esteban-Cubillo A, Pecharromán C, Moya JS, Nanotechnology 16;20(50):505701 (2009)]. La actividad en este tipo de vidrios viene atribuida a la presencia de estos elementos metálicos. Es sabido el uso de estos iones o nanopartículas metálicas como bactericidas, fungicidas y alguicidas. On the other hand, it is known to use glasses with antimicrobial properties characterized by containing metals such as copper or silver in the form of salts [patent ES2190732 of 06/16/2004], [patent US6921546 of 07/26/2005], [ patent application WO2005030665 of 07/04/2005] or as nanoparticles [Esteban-Tejeda L, Malpartida F, Esteban-Cubillo A, Pecharromán C, Moya JS Nanotechnology 25; 20 (8): 085103 (2009)], [Esteban-Tejeda L, Malpartida F, Esteban-Cubillo A, Pecharromán C, Moya JS, Nanotechnology 16; 20 (50): 505701 (2009)]. The activity in this type of glass is attributed to the presence of these metallic elements. It is known to use these ions or metal nanoparticles as bactericides, fungicides and algaecides.
Sin embargo no son conocidos vidrios con uso antimicrobiano cuya eficacia se pueda atribuir exclusivamente al óxido de cinc (ZnO). However, glasses with antimicrobial use whose efficacy can be attributed exclusively to zinc oxide (ZnO) are not known.
DESCRIPCIÓN DE LA INVENCIÓN DESCRIPTION OF THE INVENTION
Se ha desarrollado un método que permite la inhibición del crecimiento de microorganismos de una forma duradera mediante el empleo de un vidrio con una particular composición que incluye un contenido en óxido de cinc superior al 15%. A diferencia de los vidrios conocidos el vidrio empleado en la invención no requiere la presencia de metales como plata o cobre y no pertenece a la familia de vidrios de fosfato ni sodocálcicos.  A method has been developed that allows the inhibition of the growth of microorganisms in a lasting way by using a glass with a particular composition that includes a zinc oxide content greater than 15%. Unlike the known glasses, the glass used in the invention does not require the presence of metals such as silver or copper and does not belong to the family of phosphate or sodoccalc glass.
En la presente invención se entiende por "vidrio" un material inorgánico amorfo y que por tanto no presenta fases cristalinas. Por "biocida" se entiende cualquier sustancia sintética o de origen natural destinada a destruir, contrarrestar, neutralizar, impedir la acción o ejercer un control de otro tipo sobre cualquier organismo microbiano considerado nocivo para el hombre incluyendo bacterias, levaduras, hongos y virus. In the present invention, "glass" is understood as an inorganic amorphous material and therefore has no crystalline phases. "Biocide" means any synthetic or natural substance intended to destroy, counteract, neutralize, prevent the action or exert control of any other type on any microbial organism considered harmful to man, including bacteria, yeasts, fungi and viruses.
Por "bactericida" se entiende cualquier sustancia de origen natural o sintetizada que es capaz de destruir bacterias. "Bactericidal" means any substance of natural or synthesized origin that is capable of destroying bacteria.
Por "material" se entiende a una sustancia con alguna propiedad útil ya sea esta mecánica, eléctrica, óptica, térmica o magnética, como por ejemplo el material puede ser o formar parte de un material metálico, cerámico, aglomerados, fibras textiles, plásticos o materiales poliméricos o materiales compuestos, entre otros. El objeto de la invención consiste de esta forma en un método para la inhibición del crecimiento microbiano caracterizado por el uso de un vidrio adicionado a un material, en adelante vidrio de la invención, que no contiene ninguno de los siguientes elementos: fósforo, plata, cobre y que incluye en su composición al menos óxido de silicio (Si02), óxido de boro (B203), óxido de cinc (ZnO) con un contenido en óxido de zinc superior a 15% en peso y óxido de calcio con un contenido inferior al 10% en peso. By "material" is meant a substance with some useful property, whether mechanical, electrical, optical, thermal or magnetic, such as, for example, the material can be or be part of a metallic, ceramic, agglomerated, textile, plastic or plastic fibers. polymeric materials or composite materials, among others. The object of the invention consists in this way in a method for the inhibition of microbial growth characterized by the use of a glass added to a material, hereinafter glass of the invention, which does not contain any of the following elements: phosphorus, silver, copper and which includes in its composition at least silicon oxide (Si0 2 ), boron oxide (B 2 0 3 ), zinc oxide (ZnO) with a zinc oxide content exceeding 15% by weight and calcium oxide with a content less than 10% by weight.
La presencia de ZnO inferior al 15% no garantiza la capacidad antimicrobiana de los vidrios de la invención y podrían afectar a la estabilidad del vidrio y a su uso. The presence of ZnO less than 15% does not guarantee the antimicrobial capacity of the glasses of the invention and could affect the stability of the glass and its use.
Un aspecto preferente de la invención es la presencia en el vidrio empleado para la inhibición del crecimiento microbiano de la invención de un porcentaje de óxido de zinc inferior al 55% en peso. A preferred aspect of the invention is the presence in the glass used for the inhibition of microbial growth of the invention of a percentage of zinc oxide less than 55% by weight.
Otro aspecto preferente es la presencia en el vidrio de la invención de un contenido en óxido de boro comprendido entre el 30 y 40% en peso. En una realización preferida, el vidrio de la invención tiene un contenido de óxido de silicio de entre 16% y 30% en peso. En una realización aún más preferida el vidrio de la invención no contiene óxido de calcio o contiene trazas (menos del 0,1 % en peso). Another preferred aspect is the presence in the glass of the invention of a boron oxide content between 30 and 40% by weight. In a preferred embodiment, the glass of the invention has a silicon oxide content of between 16% and 30% by weight. In an even more preferred embodiment, the glass of the invention does not contain calcium oxide or contains traces (less than 0.1% by weight).
El método de inhibición del crecimiento microbiano reivindicado en la invención puede ser aplicado siguiendo diferentes procedimientos. Así un aspecto preferente de la invención consiste en el empleo del vidrio de la invención en forma particulada, de fibra o de esfera. Un modo particular de realización consiste en la incorporación del vidrio de la invención como material de relleno en materiales poliméricos. Asimismo, se ha comprobado el efecto biocida de recubrimientos realizados con el vidrio de la invención. Por ello, otro modo de realización particular lo constituye el empleo con fines biocidas de recubrimientos formados con el vidrio de la invención. El método objeto de la invención puede ser empleado en diversas aplicaciones. De forma preferente este método para la inhibición del crecimiento microbiano puede aplicarse a envases, fármacos, dispositivos médicos, implantes quirúrgicos, tejidos, medios de transporte (por ejemplo para dispositivos o elementos estructurales), instalaciones de aire (por ejemplo para dispositivos o elementos estructurales, como pueden ser los filtro de depuración del aire) o instalaciones de agua (por ejemplo para dispositivos o elementos estructurales, como pueden ser los filtros de depuración del agua). The microbial growth inhibition method claimed in the invention can be applied following different procedures. Thus, a preferred aspect of the invention consists in the use of the glass of the invention in particulate, fiber or sphere form. A particular embodiment consists in incorporating the glass of the invention as a filler material in polymeric materials. Likewise, the biocidal effect of coatings made with the glass of the invention has been verified. Therefore, another particular embodiment is the use for biocidal purposes of coatings formed with the glass of the invention. The method object of the invention can be used in various applications. Preferably, this method for the inhibition of microbial growth can be applied to packages, drugs, medical devices, surgical implants, tissues, means of transport (for example for devices or structural elements), air installations (for example for devices or structural elements , such as air purification filters) or water installations (for example for devices or structural elements, such as water purification filters).
EJEMPLOS DE REALIZACIÓN EXAMPLES OF REALIZATION
Los siguientes ejemplos se presentan como guía adicional para el experto medio en la materia y en ningún caso deben considerarse como una limitación de la invención. En estos ejemplos se muestran resultados de ensayos realizados que ponen de manifiesto la especificidad y efectividad del vidrio biocida, así como de resinas en las que se introduce el vidrio como material de relleno y de recubrimientos realizados con el vidrio biocida. The following examples are presented as additional guidance for the average expert in the field and in no case should they be considered as a limitation of the invention These examples show results of tests carried out that show the specificity and effectiveness of biocide glass, as well as of resins in which glass is introduced as a filler material and of coatings made with biocide glass.
Ejemplo 1.- Ensayo de actividad biocida del vidrio de la invención en forma particulada Example 1.- Test of biocidal activity of the glass of the invention in particulate form
Se preparó un vidrio de composición química (% peso): 4,97% de AI2O3; 5,38% de Na20; 18,7% de Si02; 0, 10% de K20; 0, 17% de Ti02; 34,7% de ZnO; 0,0024% de SrO; 33,2% de B203; 0,45% de Ce02; 2,25% de Zr02 y 0,056% de otros. A glass of chemical composition (% weight): 4.97% AI2O3 was prepared; 5.38% Na 2 0; 18.7% of Si02; 0, 10% of K 2 0; 0.17% Ti0 2 ; 34.7% ZnO; 0.0024% SrO; 33.2% of B 2 0 3 ; 0.45% of Ce0 2 ; 2.25% of Zr0 2 and 0.056% of others.
Para evaluar el efecto microbiano del polvo de vidrio se realizaron ensayos frente la bacteria Gram-negativa Escherichia coli DH10B y la levadura Candida Krusei. El valor de la eficacia antimicrobiana se determinó siguiendo el procedimiento descrito en la norma ASTM E2149. Este método está diseñado para determinar la actividad biocida en condiciones dinámicas. Asegura un buen contacto entre los microorganismos y el material biocida mediante agitación constante de la muestra en una suspensión del microorganismo seleccionado durante un periodo de tiempo determinado. La actividad antimicrobiana se determina comparando el número de microorganismos viables en la suspensión, antes y después del tiempo de contacto estipulado. Las bacterias se sembraron en medio sólido, placas Petri, de Luria Bertani (LB) cuya composición es (%peso): Triptona 1 %, extracto de levadura 0.5%, NaCI 1 % y agar 1 .5%. Las placas se incubaron 18 horas a 37°C. Las levaduras en medio sólido, placas Petri, de (YEPD) cuya composición es (%peso): Bacto-pectona 1 %, extracto de levadura 1 %, glucosa 2% y agar 2%. Las placas se incubaron 18 horas a 37°C. A continuación, colonias aisladas de las placas anteriores se inocularon en 1 mL de LB ó YEPD líquido y se cultivaron a 37°C durante 6 horas para obtener los precultivos a una densidad de entre 107 a 108 colonias viables por mililitro de cultivo. Paralelamente se preparará una suspensión de 200 mg/mL en agua del polvo de vidrio y se esterelizó durante 30 minutos a 125°C. Finalmente 10 μΙ_ del precultivo de bacterias se inoculó en 1 ml_ de LB, y 10 μΙ_ del precultivo de levaduras se inoculó en 1 ml_ de YEPD. Por último, a este cultivo se añadió 75 μΙ_ de la suspensión del vidrio a ensayar. Asimismo, se preparó como control el mismo medio en el que la cantidad de vidrio fue sustituida por un volumen equivalente de agua. Los distintos cultivos se incubaron en agitación horizontal, realizando extracciones cada 24 horas con el fin de proceder a titular los microorganismos sobrevivientes mediante plaqueo en medio LB sólido mediante diluciones seriadas desde 10~1 hasta 10~8. To evaluate the microbial effect of glass dust, tests were carried out against the Gram-negative bacteria Escherichia coli DH10B and Candida Krusei yeast. The value of antimicrobial efficacy was determined following the procedure described in ASTM E2149. This method is designed to determine the biocidal activity under dynamic conditions. It ensures a good contact between the microorganisms and the biocidal material by constant agitation of the sample in a suspension of the selected microorganism for a certain period of time. The antimicrobial activity is determined by comparing the number of viable microorganisms in the suspension, before and after the stipulated contact time. The bacteria were seeded in solid medium, Petri dishes, from Luria Bertani (LB) whose composition is (% weight): 1% tryptone, 0.5% yeast extract, 1% NaCl and 1.5% agar. The plates were incubated 18 hours at 37 ° C. Yeasts in solid medium, Petri dishes of (YEPD) whose composition is (% weight): Bacto-pectone 1%, yeast extract 1%, glucose 2% and agar 2%. The plates were incubated 18 hours at 37 ° C. Then, colonies isolated from the above plates were inoculated in 1 mL of LB or liquid YEPD and cultured at 37 ° C for 6 hours to obtain precultures at a density of between 10 7 to 10 8 viable colonies per milliliter of culture. In parallel, a 200 mg / mL suspension in glass powder water will be prepared and sterilized for 30 minutes at 125 ° C. Finally 10 μΙ_ of the bacterial preculture was inoculated in 1 ml_ of LB, and 10 μΙ_ of the yeast preculture was inoculated in 1 ml_ of YEPD. Finally, 75 μΙ_ of the glass suspension to be tested was added to this culture. Likewise, the same medium in which the amount of glass was replaced by an equivalent volume of water was prepared as a control. The different cultures were incubated in horizontal agitation, making extractions every 24 hours in order to proceed to title the surviving microorganisms by plating in solid LB medium by serial dilutions from 10 ~ 1 to 10 ~ 8 .
Todas las muestras ensayadas mostraron una desinfección total a las 24 horas, presentado un logaritmo de reducción superior a 5. Esto significa una disminución del número de bacterias o levaduras superior al 99,999% frente a la población inicial. All samples tested showed a total disinfection at 24 hours, presenting a log reduction greater than 5. This means a decrease in the number of bacteria or yeasts greater than 99.999% compared to the initial population.
Ejemplo 2. -Ensayo de actividad biocida del vidrio de la invención aplicado como recubrimiento sobre aleaciones de titanio Example 2. - Biocidal activity test of the glass of the invention applied as a coating on titanium alloys
Los recubrimientos se llevaron a cabo con el vidrio biocida en forma particulada descrito en el ejemplo 1 , sobre placas de la aleación de titanio Ti- 6AI-4V, de dimensiones 12,5 x 8,3 x 1 mm y con un 99,0% de pureza. The coatings were carried out with the particulate biocide glass described in example 1, on plates of the titanium alloy Ti- 6AI-4V, of dimensions 12.5 x 8.3 x 1 mm and with 99.0 % purity
Para evaluar el efecto bactericida del polvo de vidrio se realizaron ensayos bactericidas frente a la bacteria Gram-negativa Escherichia coli DH10B. El valor de la eficacia antimicrobiana se determinó siguiendo el procedimiento descrito en la norma ISO 22196. Esta norma ha sido adoptada a nivel internacional como norma de referencia para evaluar la eficacia antimicrobiana de superficies no porosas, cerámicas, plásticos, etc. Según esta norma, la actividad antimicrobiana se determina comparando los resultados obtenidos entre la superficie tratada y una superficie control, sin agente biocida (blanco), después de un periodo de incubación de 48 horas a 37°C, temperatura óptima para el crecimiento del microorganismo seleccionado. To evaluate the bactericidal effect of glass dust, bactericidal tests were carried out against the Gram-negative bacteria Escherichia coli DH10B. The value of antimicrobial efficacy was determined following the procedure described in ISO 22196. This standard has been adopted internationally as a reference standard to evaluate the antimicrobial efficacy of non-porous, ceramic, plastic surfaces, etc. According to this standard, antimicrobial activity is determined by comparing the results obtained between the treated surface and a control surface, without biocidal agent (blank), after an incubation period of 48 hours at 37 ° C, optimum temperature for the growth of the selected microorganism.
La norma ISO 22196 define la actividad antimicrobiana de una superficie como la diferencia entre el número de bacterias en la muestra control (blanco, A) y el número de bacterias en la muestra tratada (B), según la ecuación: ISO 22196 defines the antimicrobial activity of a surface as the difference between the number of bacteria in the control sample (blank, A) and the number of bacteria in the treated sample (B), according to the equation:
log η = log A - log B Los resultados obtenidos ponen de manifiesto la capacidad biocida del recubrimiento. A las 48 horas se alcanzó en todos los muestras biocidas ensayadas un logaritmo de reducción superior a 5. Esto significa una disminución del número de bacterias superior al 99,999% frente a la población inicial.  log η = log A - log B The results obtained show the biocidal capacity of the coating. At 48 hours, a reduction log greater than 5 was achieved in all the biocidal samples tested. This means a decrease in the number of bacteria greater than 99.999% compared to the initial population.
Ejemplo 3.- Ensayo de actividad biocida de un polímero el cual contiene el vidrio de la invención en forma particulada como material de rellenoExample 3.- Biocidal activity test of a polymer which contains the glass of the invention in particulate form as filler material
Como polímero se seleccionó un polímero termoplástico: poliuretano. Como material de relleno se seleccionó un vidrio en forma particulada similar al que se describe en el ejemplo 1 . Se adicionó una carga en peso del vidrio biocida del 50%. El mezclado homogéneo de todos los integrantes de la formulación se realizó en caliente. Mediante una extrusora, la formulación se obtuvo en forma de hilos, los cuales fueron posteriormente procesados en forma de láminas mediante la utilización de una prensa hidráulica de platos calientes. De manera análoga se obtuvieron controles que sólo contienen el polímero sin aditivo biocida. A thermoplastic polymer: polyurethane was selected as the polymer. As filler material, a particulate glass similar to that described in example 1 was selected. A weight load of 50% biocidal glass was added. Homogeneous mixing of all the members of the formulation was performed hot. Using an extruder, the formulation was obtained in the form of threads, which were subsequently processed in the form of sheets by using a hot plate hydraulic press. Similarly, controls were obtained containing only the polymer without biocide additive.
Para evaluar el efecto biocida del polvo de vidrio se realizaron ensayos bactericidas frente a la bacteria Gram-negativa Escherichia coli DH10B. El valor de la eficacia antimicrobiana se determinó siguiendo el procedimiento descrito en la norma ISO 22196. La norma indica que el índice de eficacia bactericida debe ser superior a 2 para ser considerado como tal. En todos los casos estudiados el valor obtenido fue superior a 5. Esto significa una disminución del número de bacterias superior al 99.999% frente a la población inicial. To evaluate the biocidal effect of glass dust, bactericidal tests were performed against the Gram-negative Escherichia coli DH10B bacteria. The value of antimicrobial efficacy was determined following the procedure described in ISO 22196. The standard indicates that the bactericidal efficacy index must be greater than 2 to be considered as such. In all In the cases studied, the value obtained was greater than 5. This means a decrease in the number of bacteria greater than 99.999% compared to the initial population.

Claims

REIVINDICACIONES
1 . Método para la inhibición del crecimiento microbiano en un material, donde el método comprende la adición de un vidrio al material, caracterizado porque dicho vidrio no contiene ninguno de los siguientes elementos: fósforo, plata, cobre y que comprende, óxido de calcio en un contenido entre 0 y 10% en peso, óxido de zinc en un contenido superior al 15% e inferior al 55% en peso, óxido de silicio y óxido de boro en un contenido entre 30 y 40% en peso. one . Method for inhibiting microbial growth in a material, where the method comprises adding a glass to the material, characterized in that said glass does not contain any of the following elements: phosphorus, silver, copper and comprising, calcium oxide in a content between 0 and 10% by weight, zinc oxide in a content greater than 15% and less than 55% by weight, silicon oxide and boron oxide in a content between 30 and 40% by weight.
2. Método para la inhibición del crecimiento microbiano según la reivindicación 1 , caracterizado porque el vidrio se emplea en forma particulada, de fibra o de esfera. 2. Method for inhibiting microbial growth according to claim 1, characterized in that the glass is used in particulate, fiber or sphere form.
3. Método para la inhibición del crecimiento microbiano según una cualquiera de las reivindicaciones 1 a 2, caracterizado porque el material con el vidrio incorporado es un material de relleno en materiales poliméricos. 3. Method for inhibiting microbial growth according to any one of claims 1 to 2, characterized in that the material with the incorporated glass is a filler material in polymeric materials.
4. Método para la inhibición del crecimiento microbiano según una cualquiera de las reivindicaciones 1 a 2, caracterizado porque el material que comprende el vidrio es un recubrimiento. 4. Method for inhibiting microbial growth according to any one of claims 1 to 2, characterized in that the material comprising the glass is a coating.
5. Uso del material que comprende el vidrio descrito en cualquiera de las reivindicaciones 1 a 4, como biocida en envases, dispositivos médicos, implantes quirúrgicos, tejidos, medios de transporte, instalaciones de aire o instalaciones de agua. 5. Use of the material comprising the glass described in any of claims 1 to 4, as a biocide in packages, medical devices, surgical implants, tissues, means of transport, air installations or water installations.
Uso del material que comprende el vidrio descrito según cualquiera de las reivindicaciones 1 a 4, para la fabricación de un fármaco. Use of the material comprising the glass described according to any of claims 1 to 4, for the manufacture of a drug.
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Citations (4)

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US4098610A (en) * 1977-05-23 1978-07-04 Corning Glass Works Biocidal glass additive for marine paints
JPH1129343A (en) * 1997-07-07 1999-02-02 Nippon Electric Glass Co Ltd Antimicrobial glass powder and resin molded form
US6410633B1 (en) * 1997-08-20 2002-06-25 Nippon Electric Glass Co., Ltd. Antibacterial glass and resin composite comprising the same
US20060142413A1 (en) * 2003-02-25 2006-06-29 Jose Zimmer Antimicrobial active borosilicate glass

Patent Citations (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US4098610A (en) * 1977-05-23 1978-07-04 Corning Glass Works Biocidal glass additive for marine paints
JPH1129343A (en) * 1997-07-07 1999-02-02 Nippon Electric Glass Co Ltd Antimicrobial glass powder and resin molded form
US6410633B1 (en) * 1997-08-20 2002-06-25 Nippon Electric Glass Co., Ltd. Antibacterial glass and resin composite comprising the same
US20060142413A1 (en) * 2003-02-25 2006-06-29 Jose Zimmer Antimicrobial active borosilicate glass

Non-Patent Citations (1)

* Cited by examiner, † Cited by third party
Title
DATABASE WPI Week 199916, 6 October 2014 Derwent World Patents Index; Class D22, AN 1999-183564 *

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