WO2000071139A2 - Compositions de lactobacillus et procedes pour prevenir les infections de plaies et la formation d'un film biologique sur des dispositifs chirurgicaux implantables - Google Patents

Compositions de lactobacillus et procedes pour prevenir les infections de plaies et la formation d'un film biologique sur des dispositifs chirurgicaux implantables Download PDF

Info

Publication number
WO2000071139A2
WO2000071139A2 PCT/CA2000/000611 CA0000611W WO0071139A2 WO 2000071139 A2 WO2000071139 A2 WO 2000071139A2 CA 0000611 W CA0000611 W CA 0000611W WO 0071139 A2 WO0071139 A2 WO 0071139A2
Authority
WO
WIPO (PCT)
Prior art keywords
lactobacillus
effective amount
therapeutically effective
probiotic
fermentum
Prior art date
Application number
PCT/CA2000/000611
Other languages
English (en)
Other versions
WO2000071139A3 (fr
Inventor
Gregor Reid
Andrew W. Bruce
Original Assignee
Gregor Reid
Bruce Andrew W
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Application filed by Gregor Reid, Bruce Andrew W filed Critical Gregor Reid
Priority to CA002374938A priority Critical patent/CA2374938A1/fr
Priority to AU49054/00A priority patent/AU4905400A/en
Publication of WO2000071139A2 publication Critical patent/WO2000071139A2/fr
Publication of WO2000071139A3 publication Critical patent/WO2000071139A3/fr

Links

Classifications

    • 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
    • A61L31/00—Materials for other surgical articles, e.g. stents, stent-grafts, shunts, surgical drapes, guide wires, materials for adhesion prevention, occluding devices, surgical gloves, tissue fixation devices
    • A61L31/08—Materials for coatings
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00—Medicinal preparations containing organic active ingredients
    • A61K31/70—Carbohydrates; Sugars; Derivatives thereof
    • A61K31/715—Polysaccharides, i.e. having more than five saccharide radicals attached to each other by glycosidic linkages; Derivatives thereof, e.g. ethers, esters
    • A61K31/733—Fructosans, e.g. inulin
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K35/00—Medicinal preparations containing materials or reaction products thereof with undetermined constitution
    • A61K35/66—Microorganisms or materials therefrom
    • A61K35/74—Bacteria
    • A61K35/741—Probiotics
    • A61K35/744—Lactic acid bacteria, e.g. enterococci, pediococci, lactococci, streptococci or leuconostocs
    • A61K35/747—Lactobacilli, e.g. L. acidophilus or L. brevis
    • 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
    • A61L27/28—Materials for coating prostheses
    • 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
    • A61L27/36—Materials for grafts or prostheses or for coating grafts or prostheses containing ingredients of undetermined constitution or reaction products thereof, e.g. transplant tissue, natural bone, extracellular matrix
    • 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
    • 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
    • A61L31/00—Materials for other surgical articles, e.g. stents, stent-grafts, shunts, surgical drapes, guide wires, materials for adhesion prevention, occluding devices, surgical gloves, tissue fixation devices
    • A61L31/005—Ingredients of undetermined constitution or reaction products 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
    • A61L31/00—Materials for other surgical articles, e.g. stents, stent-grafts, shunts, surgical drapes, guide wires, materials for adhesion prevention, occluding devices, surgical gloves, tissue fixation devices
    • A61L31/14—Materials characterised by their function or physical properties, e.g. injectable or lubricating compositions, shape-memory materials, surface modified materials
    • A61L31/16—Biologically active materials, e.g. therapeutic substances
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P31/00—Antiinfectives, i.e. antibiotics, antiseptics, chemotherapeutics

Definitions

  • This invention relates to probiotic compositions and methods of employing said compositions for treating and preventing wound infections and biofilm formation on implantable surgical devices .
  • Wound infections represent a catchall category for a group of diverse anatomic problems ranging from superficial and cutaneous to infections involving tissue and muscle invasion and foreign implants. Wound infections are caused by accidental (e.g. burns) and intentional (e.g. surgical) trauma, nosocomial complications of surgery, hospitalization and insertion of implants. Wound infections are also caused by occupational and recreational activities.
  • the infection- causing organisms e.g. bacteria, viruses, yeasts
  • Infections associated with wounds at the skin-material interface are particularly difficult to prevent and treat.
  • the primary causes of wound infections are Bacteroides sp . , Enterocpccus faecalis (vancomycin- resistant - VRF are particularly problematic) , S. aurexxs (including methicillin-resistant - MRSA) , S. epi ⁇ ermidis (including methicillin-resistant) - M SE) , -Streptococcus pyogenes, Clostridum sp, E ⁇ cherichia coli , Pseudomonas aer g ⁇ nosa, Klebsiella sp, Proteus sp, and Peptostreptococcus sp. (Blatan, et al.
  • Microbial biofilms defined as an accumulation of microorganisms and connecting extracellular products on a surface or to each other at some distance away from a surface, (see Costerton GD, et al. (1989), "Microbial and foreign body factors in the pathogenesis of medical device infections In: Infections associated with indwelling medical devices.
  • Surgically implanted medical devices such as heart valves and artificial veins and joints, are especially vulnerable to microbial biofilm formation and disease.
  • Gristina (1987) , "Biomaterial-centered infection: microbial adhesion versus tissue integration", Science. 237:1588-1595.
  • the surfaces of such devices are not protected by host defenses and thus provide a focal point for infecting pathogens. Closed implants are frequently associated with life-threatening infections, with Staphylococcus epidermidis and S . aureus constituting the major pathogenic force, Costerton, et al. (1989) , supra.
  • Lactobacillus bacteria are found in the intestine and urogenital tract where they are part of the normal, healthy flora: they represent one example of probiotics.
  • the use of other organisms, such as avirulent skin flora, such as staphylococci, can also be applied to reduce the risk of wound infections.
  • the present invention provides methods and compositions for the use of Lactobacillus and associated by products including biosurfactants, avirulent staphylococci and other probiotic organisms, to prevent, treat, inhibit or reduce the risk of infections around wounds and at the site of implants.
  • the present invention demonstrates that lactobacilli with antagonistic properties against pathogens provide protection against infection when applied topically or after oral intake.
  • the Lactobacillus may be administered as viable whole cells.
  • the Lactobacillus species may be aerobically grown or microaerophilically grown and selected from Lactobacillus casei, L. acidophilus, L. plantarum, L. fermentum, L. brevis, L. jensenii, L. crispatus, L. rhamnosus, L. reuteri, L. paracasei, L. gasseri, L. cellobiosis, L. delbruckii, L. helveticus, L. salvarius, L. collinoides, L. buchneri, L. rogosae and L. bifidium.
  • the present invention provides a method for inhibiting the occurrence of wound infections in a mammal by administration of a probiotic organism.
  • the probiotic organism is a Lactobacillus .
  • the Lactobacillus species are preferred.
  • L rhamnosus GR-1 L . fermentum RC-14 and L . fermentum B-54.
  • the present invention provides a method for inhibiting the occurrence of wound infections in a mammal in need of such treatment by administration of Lactobacillus and a prebiotic compound.
  • the present invention describes a method for inhibiting or reducing the occurrence of biofilm formation in a mammal by coating at least part of a surgically implantable device with a therapeutically effective amount of a probiotic organism or a by product thereof including biosurfactant .
  • the probiotic organism is a Lactobacillus .
  • the Lactobacillus species are L . rhamnosus GR-1, L . fermentum RC-14 and L . fermentum B-54.
  • the present invention provides a pharmaceutical composition suitable for inhibiting the occurrence of wound infections in mammals which comprises a therapeutically effective amount of a probiotic organism and a pharmaceutically acceptable carrier.
  • a probiotic organism is a Lactobacillus .
  • the Lactobacillus species are L . rhamnosus GR-1, L. fermentum RC-14 and L . fermentum B-54.
  • Figure 1A is a bar graph showing the effect of L . fermentum RC-14 on surgical implant infection in Sprague Dawley rats. Surgical implants removed from RC-14 treated rats (see Table 2) were rinsed in PBS and then briefly sonicated (30 sec) to recover implant associated bacteria. PBS diluted bacterial suspensions (10 ⁇ 3 ) were then plated onto MRS agar plates and incubated overnight at 37°C in an anaerobic (5% C0 2 ) bacterial chamber. Colonies were then scored for each of the 7 groups of inoculated rats (9 rats/group) .
  • Figure IB is a bar graph showing the effect of L . rhamnosus GR-1 on surgical implant infection in
  • Sprague Dawley rats Surgical implants removed from GR-1 treated rats (see Table 2) were rinsed in PBS and then briefly sonicated (30 sec) to recover implant associated bacteria. PBS diluted bacterial suspensions (10 ⁇ 3 ) were then plated onto MRS agar plates and incubated overnight at 37°C in an anaerobic (5% C0 2 ) bacterial chamber. Colonies were then scored for each of the 7 groups of inoculated rats (9 rats/group) .
  • Figure 2 is a surface enhanced laser desorption/ionization (SELDI) mass profile of lactobacillus expression of collagen binding proteins in Lactobacillus acidophilus RC-14; L. rhamnosus GR-1 and L. rhamnosus 36W.
  • SELDI surface enhanced laser desorption/ionization
  • Figure 3 is a SELDI mass profile of lactobacillus expression of collagen binding proteins in Lactobacillus acidophilus RC-14 using protein chip PS-1/CN-III.
  • Figure 4 is a bar graph illustrating dose dependant S . aureus rates and associated S . aureus surgical implant colony forming units (CFUs) .
  • Silicone implants (1cm 2 ) placed within a surgically subcutaneous pocket located on the dorsum of male rats (Sprague Dawley, 300gm) were inoculated with the indicated number of colony forming units (CFUs) of S . aureus . Animals were sacrificed 3 days later. Acute infection rates were scored and the number of S . aureus [CFUs] per implant determined.
  • the 50% infectious dose (IDso) for S . aureus in this in vivo model was determined to be " 7x10 6 CFUs.
  • Figure 5A is a bar graph illustrating the effect of co-inoculation of S . aureus (IDso%) and L . fermentum RC-14 on the incidence of surgical implant infection in Sprague-Dawley rats.
  • Surgical implants were recovered from rats co-inoculated with S . aureus [ID50] and the indicated number of RC-14 CFUs 3 days after surgery (see Table 2) .
  • Implants were rinsed in PBS and then briefly sonicated (30 sec) to recover implant associated bacteria. The indicated dilutions of the implant associated bacterial suspensions were then plated on either MRS (RC-14) or BH agar (S . aureus ) plates and incubated overnight at 37°C. MRS plates were incubated in a specific anaerobic (5% C0 2 ) chamber. Colonies were then scored for each of the indicated co-inoculated groups of rats (9 rats/group) .
  • Figure 5B depicts the results of PCR-based identification of implant associated RC-14 CFUs from S . aureus and RC-14 co-inoculated rats.
  • Surgical implant associated L. fermentum RC-14 CFUs were positively identified using RC-14 specific primers that were designed to amplify a specific portion ( ⁇ 100bp) of the 16S-23S rDNA intergenic spacer region.
  • RC-14 specific primers that were designed to amplify a specific portion ( ⁇ 100bp) of the 16S-23S rDNA intergenic spacer region.
  • S . aureus positive PCR was performed on genomic DNA extracted from several single MRS colonies using both S . aureus and RC- 14 specific primers. PCR identification of the S .
  • aureus species was performed using Staphylococcus specific 16S- 23S rDNA intergenic spacer primers (Mendoza et al . (1998) International Journal of Systematic Bacteriology 48:1049- 1055, incorporated herein by reference).
  • ND* no DNA (negative control reaction)
  • M 1 Kb DNA ladder marker.
  • Figure 5C depicts the results of PCR-based identification of implant associated S . aureus CFUs from S . aureus and RC-14 co-inoculated rats.
  • Surgical implant associated S . aureus CFUs were positively identified using Staphylococcus specific 16S-23S rDNA intergenic spacer primers (see Mendoza et al . International Journal of Systematic Bacteriology 48:1049-1055, 1998, supra).
  • PCR was performed on genomic DNA extracted from single BH-agar colonies using both S . aureus and RC-14 specific primers.
  • M 1 Kb DNA ladder marker.
  • Figure 6 is a bar graph showing the effect of
  • Biosurfactants isolated from probiotic strain L . fermentum RC-14 on S . aureus [ID50] induced surgical implant infection in rats. Silicone implants (1cm 2 ) were pretreated (12 hrs, 4°C) with L . fermentum RC-14 BSF (1 mg/ml) prior to surgical placement in the animals as described above. The surgical site containing the silicone implant was then co-inoculated with S . aureus [ID50] and 100 g of L . fermentum RC-14 BSF, respectively. Animals were sacrificed 3 days after surgery. Acute surgical infection was scored and the number of surgical implant associated S . aureus CFUs determined, as described above. **P ⁇ 0.004
  • Figure 7 is a bar graph showing the effect of p29 collagen binding protein (p29CnB) on S . aureus [ID100] induced surgical implant infection in rats.
  • Silicone implants (1cm 2 ) were pretreated (12 hrs, 4°C) with recombinant p29CnB (His-tag) or BSA (1 mg/ml) prior to surgical placement in the animals as described above.
  • the surgical site containing the silicone implants were then co-inoculated with 10 8 CFUs of S . aureus [ID100] and either 100 g of p29CnB or BSA (negative control) , respectively.
  • Animals were sacrificed 3 days after surgery. Acute surgical infection was scored and wound tissue and implants collected for further analysis.** P ⁇ 0.05 (Infection rate and Implant S . aureus CFUs).
  • the present invention provides methods for the inhibition of wound infections of mammals including humans which comprises administering a therapeutically effective amount of a probiotic organism or a by product thereof to a wound infection site and/or a biocompatible medical device.
  • inhibittion is meant treating or reducing the occurrence of wound infections with the probiotic organisms of the present invention.
  • by product is meant biosurfactants, anti-adhesion molecules and immune modulators which exhibit the infection- inhibiting activity of probiotics.
  • the artificial implantation of non-pathogenic probiotics provides novel intervention strategies that reduce the risk of infections around wounds and other surgical sites.
  • probiotic an organism which has one or more of the following characteristics, an ability to facilitate or enhance wound healing comprising an ability to: adhere to epidermal or epithelial cells by electrostatic, hydrophobic or specific adhesins including a collagen binding protein; pass through the stomach and reach the small and large intestine; grow and persist in the gastrointestinal, urogenital tracts and at a wound surface interfaces; inhibit the adhesion of wound- associated pathogens including organisms which cause infection; coaggregate; produce acid and other substances such as hydrogen peroxide and/or bacteriocins and bacteriocin-like compounds which inhibit pathogen growth; produce biosurfactant or related by-products of growth which interfere with adhesion of pathogens to cells and materials; resist antimicrobial agents; and/or enhance the host's immune function to further inhibit pathogen growth.
  • a preferred probiotic bacteria is one or more species of lactobacillus or by-products thereof such as proteins or peptides or amino acids as identified using SELDI methodology. Separation and detection of biosurfactants produced by lactobacilli may be preferably accomplished by the SELDI technique (Surface Enhanced Laser desorption/ionization) .
  • SELDI Surface Enhanced Laser desorption/ionization
  • SELDI system is meant a method which uses protein chips which contain chemically or biologically treated surfaces that specifically interact with or bind the proteins of interest. The protein chips are inserted into a reader which provides an accurate mass profile of the proteins bound to each chip in just a few minutes.
  • a most preferred probiotic lactobacillus species is L. fermentum RC-14. Another preferred lactobacillus species is L. rhamnosus GR-1. Still another preferred lactobacillus species is L. fermentum B-54.
  • lactobacilli within the scope of this invention are anaerobic and microaerophilic isolates.
  • biosurfactant is meant a biological substance, for example a protein or peptide, produced by a lactobacillus having a molecular weight of about 5 kd to about 100 kd which inhibits the binding of pathogenic bacteria to surfaces. Components of the biosurfactant can stimulate immune defenses of a host to promote the reduction and prevention of wound infections.
  • the preferred lactobacillus by-product is a biosurfactant having a molecular weight of about 8 kd to about 90 kd.
  • anti-adhesin is meant a biological substance which inhibits, reduces or prevents adhesion of pathogenic bacteria to surfaces.
  • prebiotic is meant a nonmetabolized, nonabsorbed substrate that is useful for the host which selectively enhances the growth and/or the metabolic activity of a bacterium or a group of bacteria.
  • a prebiotic also includes a nutrient utilized by lactobacilli to stimulate and/or enhance growth of lactobacilli relative to pathogenic bacteria.
  • infected wound is meant an area of open and inflamed epidermal tissue with detectable levels of pathogens e.g., staphylococcus or Candida, present.
  • visual inspection of a wound can accurately determine indicia of infection, including but not limited to, inflammation, discharge, the patient's subjective pain assessments, bleeding, and presence of dead cells, for example.
  • a “biofilm” means an accumulation of microorganisms and connecting extracellular products on a surface or to each other at some distance away from a surface.
  • compositions suitable for inhibiting wound infections of mammals including humans which comprise one or more lactobacillus viable whole cells, non-viable whole cells or cell wall fragments and a pharmaceutically acceptable carrier.
  • the lactobacillus is aerobically, microaerophilically or anaerobically grown and may be selected from the group consisting of Lactobacillus casei, L. acidophilus, L. plantarum, L. fermentum, L. brevis, L. jensenii, L. crispatus, L. rhamnosus, L.reuteri, L. paracasei, L. gasseri, L. cellobiosis, L. delbruckii, L. helveticus, L. salvarius, L. collinoides, L. buchneri, L. rogosae and L. bifidium.
  • the lactobacillus may be microaerophilically or anaerobically grown and selected from the group consisting of Lactobacillus casei var rhamnosus (GR-1 (ATCC 55826) , L. casei var rhamnosus GR-2 (ATCC 55915) , L. casei var rhamnosus GR-3 (ATCC 55917) , L. casei var rhamnosus GR-4 (ATCC 55916), L. casei var rhamnosus RC-9, L. casei var rhamnosus RC-17 (ATCC 55825) , L. casei var alactosus RC-21, L. casei NRC 430, L. casei ATCC 7469, L.
  • casei var rhamnosus 81 L. casei var rhamnosus 76, L. casei var rhamnosus 36W, L. casei var rhamnosus 36g, L. casei RC-65, L. casei RC-15, L. casei 558, . casei, RC- 21, L. casei 55, L. casei 8, L. casei 43, L. plantaxrum RC-12 (ATCC 55895), L. acidophilus RC-25, L. plantarum RC-19, L. jensenii RC-11 (ATCC 55901), L. acidophilus ATCC 4357, L . acidophilus 2099B, L . acidophilus 2155C, L .
  • a pharmaceutical composition for inhibiting wound infections in humans and lower animals which comprises a therapeutically effective amount of one or more of the aforementioned lactobacilli, together with a pharmaceutically acceptable carrier.
  • Bacteria on surfaces such as mucosal tissues and skin, exist in biofilms and compete with other organisms for space and nutrients.
  • the lactobacillus compositions of the present invention at various doses significantly reduce surgical implant infection caused by S . aureus , staphylococci and Candida by competitively excluding such pathogens.
  • Non-pathogenic organisms such as viable or non-viable lactobacilli, or their by-products such as biosurfactants, anti-adhesins and immune modulators, are applied to the wound surface interface or the biocompatible device-host interface, to reduce the risk of infecting pathogens colonizing and infecting the host.
  • the lactobacilli form a barrier to biofilm formation and infection by organisms which infect the host, for example, opportunistic pathogens such as S . aureus .
  • Biosur actants produced by lactobacilli significantly inhibit the binding of pathogens to wound sites. These biosurfactants contain carbohydrate and proteinaceous compounds. Biochemical analysis using
  • a dosage of 10 3 to about 10 11 viable or non-viable lactobacilli per ml, and optionally about 0.1 to about lO ⁇ g/ml of biosurfactant or other anti-adhesion molecule is applied directly to the wound infection site on the skin or device interface.
  • This administration is embodied in a suitable carrier, such as a cream, paste, solution, hydrogel, liposome, for example.
  • the treatment using the organisms is preferably administered in a single dose.
  • curative treatment administration of the compositions contemplated by the present invention is from about 1 to about 3 times daily to about 1 to about 3 times weekly depending upon the severity of the infection.
  • the effect of the probiotic composition is further enhanced by stimulating lactobacilli growth over that of pathogens at a surface, using prebiotics, such as a natural sugar, inulin, extracted from chicory roots and found not to be metabolized by humans but to act as a substrate for growth of lactobacilli, for example.
  • prebiotics such as a natural sugar, inulin, extracted from chicory roots and found not to be metabolized by humans but to act as a substrate for growth of lactobacilli, for example.
  • compositions may be administered in the form of a cream, liquid, paste, or gel as desired.
  • a cream formulation comprising one or more lactobacillus viable whole cells, non-viable whole cells or in a base that is non-toxic nor irritating to the skin such as a hydrogel or liposome base.
  • a contemplated cream formulation includes cocoa butter.
  • Another preferred form of application involves the preparation of a freeze-dried capsule, taken orally comprising the composition of the present invention. It has been found that a capsule comprising about 10 3 to about l ⁇ n probiotic organisms is suitable.
  • a capsule may contain one single or two or more different species of probiotic organism (s) .
  • a probiotic organism e.g. , Lactobacillus
  • a therapeutically effective amount of lactobacillus will vary with the particular wound infection being treated, the age and physical condition of the patient being treated, the severity of the infection, the duration of treatment, the nature of concurrent therapy and the specific lactobacillus employed.
  • lactobacillus will thus be the minimum amount which will provide the desired attachment to epithelial and epidermal cells.
  • aceutically-acceptable carrier as used herein is meant one or more compatible solid or liquid filler diluents, or encapsulating substances.
  • compatible as used herein is meant that the components of the composition are capable of being comingled without interacting in a manner which would substantially decrease the pharmaceutical efficacy of the total composition under ordinary use situations.
  • the probiotic organism may be administered in a convenient manner such as by the topical, oral, intravenous (where non-viable) , or suppository (vaginal or rectal) routes.
  • the active ingredients which comprise probiotic organisms may be required to be coated in a material to protect said organisms from the action of enzymes, acids and other natural conditions which may inactivate said organisms.
  • they should be coated by, or administered with, a material to prevent inactivation.
  • probiotic organisms may be co-administered with enzyme inhibitors or in liposomes.
  • Enzyme inhibitors include pancreatic trypsin inhibitor, diisopropylfluorophosphate (DFP) and trasylol.
  • Liposomes include water-in-oil-in-water P40 emulsions as well as conventional and specifically designed liposomes which transport lactobacilli or their by-products to the urogenital surface.
  • the probiotic organisms may also be administered parenterally or intraperitoneally .
  • Dispersions can also be prepared, for example, in glycerol, liquid polyethylene glycols, and mixtures thereof, and in oils.
  • the pharmaceutical forms suitable for injectable use include sterile aqueous solutions (where water soluble) or dispersions and sterile powders for the extemporaneous preparation of sterile injectable solutions or dispersion.
  • the carrier can be a solvent or dispersion medium containing, for example, water, ethanol, polyol (for example, glycerol, propylene glycol, liquid polyethylene glycol, and the like) , suitable mixtures thereof and vegetable oils.
  • the proper fluidity can be maintained, for example, by the use of a coating such as lecithin, by the maintenance of the required particle size in the case of dispersion.
  • a coating such as lecithin
  • isotonic agents for example, sugars or sodium chloride.
  • Prolonged absorption of the injectable compositions can be brought about by the use in the compositions of agents delaying absorption, for example, aluminum monostearate and gelatin.
  • Sterile injectable solutions are prepared by incorporating the probiotic organisms in the required amount in the appropriate solvent with various of the other ingredients enumerated above, as required, followed by filtered sterilization.
  • dispersions are prepared by incorporating the various sterilized probiotic organisms into a sterile vehicle which contains the basic dispersion medium and the required other ingredients from those enumerated above.
  • the preferred methods of preparation are vacuum-drying and the freeze-drying technique which yield a powder of the active ingredient plus any additional desired ingredient from previously sterile-filtered solution thereof. Additional preferred methods of preparation include but are not limited to lyophilization and heat-drying.
  • the active compound may be orally administered, for example, with an inert diluent or with an assimilable edible carrier, or it may be enclosed in hard or soft shell gelatin capsule, or it may be compressed into tablets designed to pass through the stomach (i.e., enteric coated), or it may be incorporated directly with the food of the diet.
  • the probiotic organisms may be incorporated with excipients and used in the form of ingestible tablets, buccal tablets, troches, capsules, elixirs, suspensions, syrups, wafers, and the like.
  • Compositions or preparations according to the present invention are prepared so that an oral dosage unit form contains about 10 3 to about 10 11 viable or non-viable e.g., lactobacilli per ml.
  • the tablets, troches, pills, capsules or lactobacilli in suspension as described above, may also contain the following: a binder such as gum tragacanth, acacia, corn starch or gelatin; excipients such as dicalcium phosphate; a disintegrating agent such as corn starch, potato starch, alginic acid, and the like; a lubricant such as magnesium stearate; and a sweetening agent such as sucrose, lactose or saccharin may be added or a flavoring agent such as peppermint, oil or wintergreen or cherry flavoring.
  • a binder such as gum tragacanth, acacia, corn starch or gelatin
  • excipients such as dicalcium phosphate
  • a disintegrating agent such as corn starch, potato starch, alginic acid, and the like
  • a lubricant such as magnesium stearate
  • a sweetening agent such as sucrose, lactose or saccharin may be added
  • any material used in preparing any dosage unit form should be pharmaceutically pure and substantially non-toxic in the amounts employed.
  • the probiotic organism may be incorporated into sustained-release preparations and formulations .
  • Dosage unit form refers to physically discrete units suited as unitary dosages for the mammalian subjects to be treated; each unit containing a predetermined quantity of the probiotic organisms calculated to produce the desired therapeutic effect in association with the required pharmaceutical carrier.
  • the specification for the novel dosage unit forms of the invention are dictated by and directly depending on (a) the unique characteristics of the probiotic organism and the particular therapeutic effect to be achieved, and (b) the limitations inherent in the art of compounding such probiotic for the establishment and maintenance of a healthy urogenital flora.
  • the probiotic organism is compounded for convenient and effective administration in effective amounts with a suitable pharmaceutically or food acceptable carrier in dosage unit form as hereinbefore disclosed.
  • a unit dosage form can, for example, contain the principal active compound in an amount approximating 10 3 to about 10 11 viable or non-viable lactobacilli, per ml.
  • the dosages are determined by reference to the usual dose and manner of administration of the said ingredients.
  • the pharmaceutically acceptable carrier may be in the form of milk or portions thereof including yogurt. Skim milk, skim milk powder, non-milk or non-lactose containing products may also be employed.
  • the skim milk powder is conventionally suspended in phosphate buffered saline (PBS) , autoclaved or filtered to eradicate proteinaceous and living contaminants, then freeze dried heat dried, vacuum dried, or lyophilized.
  • PBS phosphate buffered saline
  • substances which can serve as pharmaceutical carriers are sugars, such as lactose, glucose and sucrose; starches such as corn starch and potato starch; cellulose and its derivatives such as sodium carboxymethycellulose, ethylcellulose and cellulose acetates; powdered tragancanth; malt; gelatin; talc; stearic acids; magnesium stearate; calcium sulfate; calcium carbonate; vegetable oils, such as peanut oils, cotton seed oil, sesame oil, olive oil, corn oil and oil of theobroma; polyols such as propylene glycol, glycerine, sorbitol, manitol, and polyethylene glycol; agar; alginic acids; pyrogen-free water; isotonic saline; cranberry extracts and phosphate buffer solution; skim milk powder; as well as other non-toxic compatible substances used in pharmaceutical formulations such as Vitamin C, estrogen and echinacea, for example.
  • the patient is administered a therapeutically effective amount of a lactobacillus composition in accordance with the present invention, for example, topically.
  • a most preferred composition comprises one or more lactobacillus viable whole cells and a pharmaceutically acceptable carrier.
  • This composition may be administered topically, orally or intravenously in the form of a cream, capsule, gel, liquid or paste.
  • the lactobacillus is selected from the group comprising L . casei ss rhamnosus , L . casei ss alactosus, L . fermentum and L . brevis .
  • the lactobacillus is either L . rhamnosus GR-1, L . fermentum B-54 or L . fermentum RC-14.
  • a method of treating or preventing biofilm formation involves coating a surgically implantable device with a therapeutically effective amount of one or more of lactobacillus viable whole cells, non-viable whole cells, or a cell wall fragment.
  • lactobacilli probiotics e.g., biosurfactants, acids, hydrogen peroxide, bacteriocins
  • host responses are stimulated which inhibit pathogens and/or create a microenvironment less conducive to pathogen spread.
  • a medical device is contacted or coated with lactobacillus at a concentration of about 10 3 to about 10 organisms/ml prior to introduction into a patient in need of such device.
  • the surgically implantable device may be composed of polymers such as fluorinated ethylene propylene, sulfonated polystyrene, polystyrene, polyvinyl chloride (PVC) , polyurethane or polyethylene terephthalate silicon rubber or other biomaterials and in addition, glass or hydrophilic substrata.
  • the devices contemplated by the present invention include, but are not limited to, artificial heart valves, artificial veins, artificial arteries, shunts, drainage tubes, joints or catheters, intrauterine devices, catheters, stents, intravenous lines, diaphragms, implants, screws, sutures, pads and tampons, for example.
  • a combination of coaggregation of lactobacillus and the production by lactobacillus of one or more inhibitory substances is responsible for excluding pathogens and/or reducing their numbers at the wound site.
  • a collagen binding protein is contemplated to transduce a signal which interferes with pathogen virulence.
  • the signal transduced by the collagen binding protein causes the pathogen to rapidly become avirulent or aseptic.
  • the present invention contemplates methods for inhibiting pathogenic colonization in a host at a wound-surface interface, for example, by the administration of a probiotic cultured to induce the production of a collagen binding protein.
  • lactobacillus acts as a block to pathogens by preventing access to receptor sites.
  • complete exclusion of pathogens theoretically can occur, the most common finding of the results of the present invention is that there is a reduction in pathogen numbers compared to lactobacilli.
  • some lactobacilli may not completely exclude pathogens, they are still capable of interfering with pathogen colonization in vivo,
  • Staphylococcus aureus (Oxford strain) was cultured in Brain Heart (BH) growth media overnight at 37°C and plated on BH agar culture plates to determine the colony-forming unit (CFU) activity of the bacteria.
  • BH Brain Heart
  • SA Staphylococcus aureus
  • PBS phosphate buffered saline
  • Lactobacillus fermentum RC-14 and Lactobacillus rhamnosus GR-1 were prepared using MRS Broth and MRS agar culture plates. Appropriate CFU measurements and inoculation dilutions were prepared as described above for S . aureus (SA) .
  • the bacteria and surfactant were separated by centrifugation (10,000 x g, 10 min, 10°C) , and the supernatant was removed, filtered (0.22mm) and dialyzed against double demineralized water at 4°C using spectrapor dialysis tubing (6000-8000 Da M.W. cutoff) .
  • the dialyzed BSF was either used directly or freeze-dried (-10°C , -5uM Hg, 1-2 days) followed by overnight drying (Savant speedvac, RCT4104) .
  • the biosurfactant powder was stored at -20°C. Protein concentrations were determined using a protein assay kit (Pierce, ON) .
  • the BSF was suspended in sterile PBS (2mg/ml) and incubated with the surgical implants for 12 hours at 4°C prior to placing them in the animals.
  • Sprague-Dawley rats were anaesthetized via peritoneal injection of a mixture of hydrochloride- keta ine (100 mg/ml) and xylazine (lOmg/ml) at the rate of O.lml per lOOg of body weight.
  • Each anaesthetized rat was clipped of dorsal hair at the surgical site and liberally swabbed with a povi-iodine antiseptic solution prior to surgery. A single 2cm incision was made along the dorsal skin.
  • a single (1cm x 1cm x 0.5 mm) sterile silicone implant (Dagnone Inc, Quebec) was then inserted into the subcutaneous pocket adjacent to the skin incision and inoculated with the indicated number of bacterial CFUs and/or BSF.
  • the incision was closed with 3.0 coated polygalactin 910 (vicryl) interrupted sutures and a post- operative analgesic (buprenorphine hydrochloride, 0.01 mg/kg subcutaneously) was administered to each animal.
  • mice were sacrificed by C0 2 asphyxiation.
  • Acute surgical infection was scored (large fluid-filled capsule containing the surgical implant) and the surgical implants, wound tissue and fluid harvested for further analysis. Implants from uninfected rats were free of any signs of inflammation.
  • the implant-associated bacteria [CFUs] numbers were quantified using standard microbiological techniques. Briefly, implants were rinsed in PBS and then subjected to a 30-second sonication treatment. Diluted bacterial suspensions were then plated on either MRS- or BH-agar plates.
  • CMS protocol a Gram staining kit
  • the staining procedure included: Crystal violet (1 min) , iodine solution (1 min) , decolorizing agent (30 sec) , sephranin (1 min) . Implants were then imaged using a light microscope (Axiophot, Zeiss) .
  • the PCR technique was used to unambiguously identify the types of bacterial CFUs obtained from each surgical implant. Briefly, bacteria colonies were picked, washed in Tris-EDTA buffer (pH 7.4), and lysed in 10% SDS for 30 minutes (Bollet et al . Nucleic Acid Research 19(8): 1955 ). The lysate was isolated by brief centrifugation (4000 rpm, 5min) and heated in a 300 Watt- microwave (Hi power) for 5 minutes. The pellets were subjected to one round of phenol/chloroform/ isoamyl alcohol (24:23:1) extraction (pH 8.0) and the DNA precipitated with two volumes of absolute ethanol.
  • aureus [denaturation (1 min at 94°C) , annealing (7 min at 52°C) and elongation (2 min at 68°C) ] or 40 cycles for RC-14 and GR-1 [denaturation (1 min / 94°C) , annealing (2 min / 52°C or
  • CFUs colony forming units
  • the percentage of acute surgical infection (Day 3) .
  • CFUs Lactobacilli colony forming units
  • lactobacillus compositions of the present invention at various doses (10 3 -10 n ) significantly reduced surgical implant infection caused by of S. aureus (See Figures 1A-1B and 6) .
  • EXAMPLE 2 Identification of Lactobacillus by-products-collagen binding protein
  • Proteins usually do not act alone and are often complexed with other important proteins.
  • the transcription and replication machinery, ribosomes and the cytoskeleton are all multiprotein complexes that control fundamental cellular processes. Identifying and characterizing the proteins present within these complexes is paramount to understanding how they function normally.
  • Recent advances in biological mass spectroscopy were used to analyze trace concentrations of proteins in their native environments.
  • the SELDI Surface .Enhanced Laser Desorption/Jonization ProteinChip technology of Ciphergen Biosystems combines the analytical sensitivity of mass spectroscopy (10 ⁇ 15 to 10 "12 range) with novel surface chemistry capable of either general or selective capture of proteins from small crude biological samples.
  • lactobacilli components were identified that promoted bacterial homeostasis in a wound. This identification was possible because lactobacilli and other probiotic organisms out-competed the pathogens, such as Staphylococcus aureus and S . epidermidis , and because the pathogens triggered Lactobacillus to produce a bioactive compound, i.e. a collagen binding protein that either directly or indirectly (immune stimulation) inhibited the growth of the pathogens. The effect was also aided by the probiotic organisms acting as chemotactic agents for immune defenses, such as neutrophils, macrophages, lymphocytes, antibodies and complement.
  • pathogens such as Staphylococcus aureus and S . epidermidis
  • a bioactive compound i.e. a collagen binding protein that either directly or indirectly (immune stimulation) inhibited the growth of the pathogens.
  • the effect was also aided by the probiotic organisms acting as chemotactic agents for immune
  • SELDI surface enhanced laser desorption/ionization
  • Lactobacilli were rapidly detected from surgical implant associated specimens via intergenic 16S-23S Ribosomal spacer PCR analysis using specific primers of L . fermentum RC14. The following method was employed:
  • Lactobacilli isolates were cultured at 37°C for 48 hours on an LBS plate in anaerobic chamber.
  • One loop of bacteria colonies was picked from the LBS plate and suspended in 1 ml of d 2 H 2 0, then centrifuged for 1 min at 12,000 rpm.
  • 200 ⁇ l of InstaGene matrix (Bio-Rad) was added to the pellet and incubated at 56°C in a water bath for 30 min.
  • the pellet was vortexed at high speed for 10 seconds keeping the sample in the boiling waterbath for 8 min.
  • the sample was vortexed at high speed again and spun at 12,000 rpm for 3 min.
  • the chromosomal DNA was stored at -20°C until used.
  • Optimal PCR conditions for different strains of Lactobacillus were established by using two universal primers from E . coli .
  • the DNA fragment containing the spacer regions between 16S rRNA and 23S rRNA genes of RC- 14 strains was amplified by using PCR with two universal primers Al and Bl from E . coli .
  • the 5' primer, 5 ⁇ GTCGTAACAAGGTAAGCCG3 • corresponds to a conserved sequence motif from the 3 ' end of 16S rRNAs [Primer Al, position 1493 - 1513 (Escherichia coli 16S rRNA numbering)] and the 3 1 primer, 5'C T/C A/G T/C TGCCAAGCATCCACT3 ' (SEQ ID NO: 8) was deduced from an alignment of the 13 23S 5' sequences [primer Bl, position 23 - 43 (Escherichia coli 23S rRNA numbering) ] , respectively.
  • DNA templates (1.6 ug, 40 ⁇ l) were amplified in a 100 ⁇ l reaction volume that contained 2.5 u Tag polymerase (Boehringer Mannheim) , 100 ng of each of the primers, 4 mM MgCl 2 ,0.2 mM of each of the four dNTPs (Pharmacia Biotech), 10 mM Tris-Cl (PH 8.0), 50 mM KCl and 1% (v/v) Triton X-100. Reaction mixtures were overlaid with 100 ⁇ l mini oil (liquired paraffin, VWR) and preheated at 95° for 5 min. Amplification was carried out in a AMPLITRON II Thermolyne for 40 cycles.
  • 2.5 u Tag polymerase Boehringer Mannheim
  • Each amplification cycle was as follows: 30 seconds at 95°C (denaturation) , 1 min. at 40°C, 45°C or 50°C.
  • the optimal annealing temperature was 40°C for RC-14, and 1 min at 72°C (extension) .
  • a QIAquick Gel Extraction Kit (Qiagen, Mississauga, Ontario) for extraction of DNA fragments 70bp-10kb from standard agrose gel in TAE or TBE buffer was used to purify PCR bands. Each of the two PCR DNA fragment bands were excised from the agarose gel with a scalpel and the gel slice was weighed. The protocol of QIAquick Gel Extraction Kit was then followed. The Kit system combined the spin-column with the silica-gel membrane. The DNA band was dissolved completely with solubilization buffer in 50°C for 10 min. DNA adsorbed to the silica membrane in the high salt conditions. Pure DNA was eluted with Tris buffer (PH 8.0). This pure PCR product was stored at -20°C for later use.
  • Tris buffer PH 8.0
  • Each PCR band product was ligated into pGEM-T vector (Promega) .
  • Each pGEM-T vector was transformed into E. coli JM 109 high efficiency competent cells by using Transformation Aid (MBI Fermentas Inc.) on the LB plate with 50 ug/ml ampicillin. Several white colonies or light blue colonies were selected as positive colonies which contained the PCR insert. Colonies were cultured on the LB-ampicillin plate. Each plate contained 32 different colonies. Colonies were cultured with LB-ampicillin broth. One part of culture was frozen quickly by using liquid nitrogen and was kept at -80°C. Another part of culture was used for further miniprep of plasmid DNA. The remainder of culture was kept at 4°C.
  • the QIAprep Spin Miniprep Kit (Qiagen, Mississauga, Ontario) was used to prepare plasmid DNA. Each of two PCR products was automatically sequenced by using T7 & SP6 promoter primers with two directions. Analysis of sequence was performed using the sequence analysis software package - DNA Star program.
  • DNA templates (1.6 ug, 40 ⁇ l) were amplified in a 100 ⁇ l reaction volume that contained 2.5 u Tag polymerase (Boehringer Mannheim) , 100 ng of each of the primer, 4 mM MgCl 2 , 0.2 mM of each of the four dNTPs

Landscapes

  • Health & Medical Sciences (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Chemical & Material Sciences (AREA)
  • Veterinary Medicine (AREA)
  • Public Health (AREA)
  • General Health & Medical Sciences (AREA)
  • Animal Behavior & Ethology (AREA)
  • Epidemiology (AREA)
  • Medicinal Chemistry (AREA)
  • Molecular Biology (AREA)
  • Heart & Thoracic Surgery (AREA)
  • Transplantation (AREA)
  • Surgery (AREA)
  • Biomedical Technology (AREA)
  • Engineering & Computer Science (AREA)
  • Pharmacology & Pharmacy (AREA)
  • Chemical Kinetics & Catalysis (AREA)
  • Dermatology (AREA)
  • Oral & Maxillofacial Surgery (AREA)
  • Vascular Medicine (AREA)
  • Microbiology (AREA)
  • Mycology (AREA)
  • Communicable Diseases (AREA)
  • Oncology (AREA)
  • General Chemical & Material Sciences (AREA)
  • Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
  • Organic Chemistry (AREA)
  • Botany (AREA)
  • Medicines Containing Material From Animals Or Micro-Organisms (AREA)
  • Medicines That Contain Protein Lipid Enzymes And Other Medicines (AREA)

Abstract

L'invention porte sur des compositions probiotiques et sur leurs sous-produits, ainsi que sur des procédés d'utilisation de ces compositions pour inhiber, traiter ou prévenir les infections de plaies et la formation d'un film biologique sur des dispositifs chirurgicaux implantables.
PCT/CA2000/000611 1999-05-25 2000-05-25 Compositions de lactobacillus et procedes pour prevenir les infections de plaies et la formation d'un film biologique sur des dispositifs chirurgicaux implantables WO2000071139A2 (fr)

Priority Applications (2)

Application Number Priority Date Filing Date Title
CA002374938A CA2374938A1 (fr) 1999-05-25 2000-05-25 Compositions de lactobacillus et procedes pour prevenir les infections de plaies et la formation d'un film biologique sur des dispositifs chirurgicaux implantables
AU49054/00A AU4905400A (en) 1999-05-25 2000-05-25 Lactobacillus compositions and methods for preventing wound infections and biofilm formation on implantable surgical devices

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
US13595499P 1999-05-25 1999-05-25
US60/135,954 1999-05-25

Publications (2)

Publication Number Publication Date
WO2000071139A2 true WO2000071139A2 (fr) 2000-11-30
WO2000071139A3 WO2000071139A3 (fr) 2001-03-08

Family

ID=22470552

Family Applications (1)

Application Number Title Priority Date Filing Date
PCT/CA2000/000611 WO2000071139A2 (fr) 1999-05-25 2000-05-25 Compositions de lactobacillus et procedes pour prevenir les infections de plaies et la formation d'un film biologique sur des dispositifs chirurgicaux implantables

Country Status (3)

Country Link
AU (1) AU4905400A (fr)
CA (1) CA2374938A1 (fr)
WO (1) WO2000071139A2 (fr)

Cited By (28)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2003009879A1 (fr) * 2001-07-23 2003-02-06 Scimed Life Systems, Inc. Dispositifs medicaux a agents antimicrobiens et surface tensioactive
EP1283010A1 (fr) * 2001-08-10 2003-02-12 Cobiotex Compositions à base de complexes bactériens et leurs applications pour la prévention des infections nosocomiales.
WO2003068250A1 (fr) * 2002-02-15 2003-08-21 Dermavri Pty Ltd. Compositions et methodes de traitement de troubles cutanes
EP1492549A1 (fr) * 2002-03-13 2005-01-05 Kibow Biotech, Inc. Compositions et procedes pour augmenter les fonctions renales
WO2005021575A2 (fr) * 2003-08-29 2005-03-10 Gregor Reid Molecules de signalisation bacterienne regulant en les diminuant les proprietes de virulence pathogene bacterienne
DE102004023612A1 (de) * 2004-05-10 2005-12-01 Helmut Meusel Verwendung von microbiellen Mischbiozönosen zur Behandlung von Entzündungen der Haut
WO2006136420A3 (fr) * 2005-06-22 2007-04-19 Organobalance Gmbh Procedes et moyens de protection de la peau contre des micro-organismes pathogenes
AU2003245473B2 (en) * 2002-02-15 2007-09-06 Bioxyne Ltd Compositions and methods for treatment of skin disorders
US7332179B2 (en) 2003-12-12 2008-02-19 Kimberly-Clark Worldwide, Inc. Tissue products comprising a cleansing composition
WO2011000123A1 (fr) 2009-07-01 2011-01-06 Universidad De Concepcion Patchs hydrocolloïdes renfermant des souches viables probiotiques de lactobacillus spp. et utilisés pour le traitement de blessures de diverses natures, dans des tissus infectés, des plaies sèches et des plaies profondes
BE1018502A3 (nl) * 2009-02-17 2011-02-01 Metatecta Nv Probiotische oplossing voor het beschermen van een oppervlak en werkwijze die gebruik maakt van zulke oplossing.
US7998470B2 (en) 1999-04-30 2011-08-16 Kibow Biotech, Inc. Compositions and methods improving renal function
US8257693B2 (en) 2002-03-13 2012-09-04 Kibow Biotech, Inc. Composition for maintaining healthy kidney function
US8343536B2 (en) 2007-01-25 2013-01-01 Cook Biotech Incorporated Biofilm-inhibiting medical products
EP2596776A1 (fr) * 2011-11-25 2013-05-29 Micuri ApS Composition probiotique emballée et utilisations associées
WO2013180585A1 (fr) * 2012-05-29 2013-12-05 Danuta Kruszewska Nano-produit comportant lactobacillus reuteri dan080, utile en prophylaxie et en médecine humaine et vétérinaire, et son utilisation médicale
EP2674162A1 (fr) * 2012-05-29 2013-12-18 Danuta Kruszewska Nano-produit, comprenant des lactobacillus reuteri dan080, utile dans la prophylaxie, diagnostic et traitement médical et vétérinaire
JP2014516957A (ja) * 2011-05-16 2014-07-17 オルガノバランス メディカル アーゲー 細菌性感冒に対する新規な乳酸菌及びそれを含有する組成物
JP2016522810A (ja) * 2013-04-23 2016-08-04 テラジェン ホールディングス リミテッドＴｅｒｒａｇｅｎ Ｈｏｌｄｉｎｇｓ Ｌｉｍｉｔｅｄ 抗菌活性を有する細菌株およびそれを含む生物的防除組成物
WO2017015208A1 (fr) * 2015-07-17 2017-01-26 Hollister Incorporated Pansement et méthode de traitement de plaies aiguës et chroniques
US9655932B2 (en) 2002-03-13 2017-05-23 Kibow Biotech, Inc. Composition and method for preventing or treating gout or hyperuricemia
ITUA20164065A1 (it) * 2016-06-01 2017-12-01 Probiotical Spa Composizioni in gel per applicazioni topiche a base di batteri, preparazione delle stesse e loro usi.
US9980988B2 (en) 2002-03-13 2018-05-29 Kibow Biotech, Inc. Compositions and methods for augmenting kidney function
IT201800004036A1 (it) * 2018-03-28 2019-09-28 Teres Srl Gel transdermico
US10953051B2 (en) 2005-06-22 2021-03-23 Novozymes A/S Methods and means for protecting the skin against pathogenic microorganisms
US11103542B2 (en) 2002-03-13 2021-08-31 Kibow Biotech, Inc. Composition and method for maintaining healthy kidney function
US11179426B2 (en) 2016-12-29 2021-11-23 Kibow Biotech, Inc. Composition and method for maintaining healthy kidney function
CN114767936A (zh) * 2022-05-10 2022-07-22 武汉理工大学 复合干酪乳杆菌的皮肤损伤修复支架材料制备方法

Families Citing this family (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CZ201523A3 (cs) 2015-01-16 2016-07-27 Bio Agens Research And Development - Bard, S.R.O. Duální mikrobiální preparát pro dlouhodobé potlačení nebo prevenci symptomů oportunních mikrobiálních infekcí

Citations (5)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
DE2738652A1 (de) * 1977-08-26 1979-03-15 Seiken Kai Foundational Juridi Lactobacillus-praeparat und seine verwendung zur behandlung bakterieller infektionen
WO1997014802A1 (fr) * 1995-10-20 1997-04-24 Nederlandse Organisatie Voor Toegepast-Natuurwetenschappelijk Onderzoek Tno Nouveaux facteurs d'adherence de micro-organismes non pathogenes et leurs applications pour le criblage de micro-organismes a la recherche de proprietes probiotiques specifiques; nouvelles compositions pharmaceutiques et additifs alimentaires comportant ces micro-organismes et comprenant des facteurs d'adherence
US5645830A (en) * 1991-11-15 1997-07-08 Research Corporation Technologies, Inc. Lactobacillus and skim milk compositions and methods for preventing microbial urogenital infections
WO1998023727A1 (fr) * 1996-11-29 1998-06-04 Bio K + International Inc. Ferment lactique contenant une souche de lactobacillus acidophilus et utilisation de celui-ci
US5804179A (en) * 1985-12-31 1998-09-08 Research Corporation Technologies, Inc. Lactobacillus compositions and methods for treating urinary tract infections

Patent Citations (5)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
DE2738652A1 (de) * 1977-08-26 1979-03-15 Seiken Kai Foundational Juridi Lactobacillus-praeparat und seine verwendung zur behandlung bakterieller infektionen
US5804179A (en) * 1985-12-31 1998-09-08 Research Corporation Technologies, Inc. Lactobacillus compositions and methods for treating urinary tract infections
US5645830A (en) * 1991-11-15 1997-07-08 Research Corporation Technologies, Inc. Lactobacillus and skim milk compositions and methods for preventing microbial urogenital infections
WO1997014802A1 (fr) * 1995-10-20 1997-04-24 Nederlandse Organisatie Voor Toegepast-Natuurwetenschappelijk Onderzoek Tno Nouveaux facteurs d'adherence de micro-organismes non pathogenes et leurs applications pour le criblage de micro-organismes a la recherche de proprietes probiotiques specifiques; nouvelles compositions pharmaceutiques et additifs alimentaires comportant ces micro-organismes et comprenant des facteurs d'adherence
WO1998023727A1 (fr) * 1996-11-29 1998-06-04 Bio K + International Inc. Ferment lactique contenant une souche de lactobacillus acidophilus et utilisation de celui-ci

Non-Patent Citations (5)

* Cited by examiner, † Cited by third party
Title
GRISTINA A G: "BIOMATERIAL-CENTERED INFECTION MICROBIAL ADHESION VERSUS TISSUE INTEGRATION" SCIENCE (WASHINGTON D C), vol. 237, no. 4822, 1987, pages 1588-1595, XP000971033 ISSN: 0036-8075 cited in the application *
HAWTHORN L A ET AL: "EXCLUSION OF UROPATHOGEN ADHESION TO POLYMER SURFACES BY LACTOBACILLUS-ACIDOPHILUS" JOURNAL OF BIOMEDICAL MATERIALS RESEARCH, vol. 24, no. 1, 1990, pages 39-46, XP000961246 ISSN: 0021-9304 *
HEINEMANN CHRISTINE ET AL: "Purification and characterization of a surface-binding protein from Lactobacillus fermentum RC-14 that inhibits adhesion of Enterococcus faecalis 1131." FEMS MICROBIOLOGY LETTERS, vol. 190, no. 1, 2000, pages 177-180, XP000971010 ISSN: 0378-1097 *
HORIKAWA Y: "EFFECTS OF LACTOBACILLUS-CASEI CONTAINING OINTMENT ON THE HEALING AND PROTECTION AGAINST OPPORTUNISTIC INFECTION OF THERMAL INJURY WOUNDS IN MICE" HIROSHIMA JOURNAL OF MEDICAL SCIENCES, vol. 35, no. 1, 1986, pages 1-14, XP000961244 ISSN: 0018-2052 *
RASIC J LJ & KURMANN J A: "Yoghurt" SCIENTIFIC GROUNDS, TECHNOLOGY, MANUFACTURE AND PREPARATIONS,COPENHAGEN, TECHNICAL DAIRY PUBLISHING,DK, 1978, pages 120-137, XP002142806 *

Cited By (44)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US7998470B2 (en) 1999-04-30 2011-08-16 Kibow Biotech, Inc. Compositions and methods improving renal function
WO2003009879A1 (fr) * 2001-07-23 2003-02-06 Scimed Life Systems, Inc. Dispositifs medicaux a agents antimicrobiens et surface tensioactive
US7749203B2 (en) 2001-07-23 2010-07-06 Boston Scientific Scimed, Inc. Long-term indwelling medical devices containing slow-releasing antimicrobial agents and having a surfactant surface
US6921390B2 (en) 2001-07-23 2005-07-26 Boston Scientific Scimed, Inc. Long-term indwelling medical devices containing slow-releasing antimicrobial agents and having a surfactant surface
EP1283010A1 (fr) * 2001-08-10 2003-02-12 Cobiotex Compositions à base de complexes bactériens et leurs applications pour la prévention des infections nosocomiales.
FR2828407A1 (fr) * 2001-08-10 2003-02-14 Cobiotex Compositions a base de complexes bacteriens et leurs applications pour la prevention des infections nosocomiales
AU2003245473B2 (en) * 2002-02-15 2007-09-06 Bioxyne Ltd Compositions and methods for treatment of skin disorders
WO2003068250A1 (fr) * 2002-02-15 2003-08-21 Dermavri Pty Ltd. Compositions et methodes de traitement de troubles cutanes
US8481025B2 (en) 2002-03-13 2013-07-09 Kibow Biotech, Inc. Composition for maintaining healthy kidney function
EP1492549A1 (fr) * 2002-03-13 2005-01-05 Kibow Biotech, Inc. Compositions et procedes pour augmenter les fonctions renales
US11103542B2 (en) 2002-03-13 2021-08-31 Kibow Biotech, Inc. Composition and method for maintaining healthy kidney function
EP1492549A4 (fr) * 2002-03-13 2005-10-26 Kibow Biotech Inc Compositions et procedes pour augmenter les fonctions renales
US9980988B2 (en) 2002-03-13 2018-05-29 Kibow Biotech, Inc. Compositions and methods for augmenting kidney function
US9655932B2 (en) 2002-03-13 2017-05-23 Kibow Biotech, Inc. Composition and method for preventing or treating gout or hyperuricemia
US8257693B2 (en) 2002-03-13 2012-09-04 Kibow Biotech, Inc. Composition for maintaining healthy kidney function
WO2005021575A3 (fr) * 2003-08-29 2005-05-19 Gregor Reid Molecules de signalisation bacterienne regulant en les diminuant les proprietes de virulence pathogene bacterienne
WO2005021575A2 (fr) * 2003-08-29 2005-03-10 Gregor Reid Molecules de signalisation bacterienne regulant en les diminuant les proprietes de virulence pathogene bacterienne
US7332179B2 (en) 2003-12-12 2008-02-19 Kimberly-Clark Worldwide, Inc. Tissue products comprising a cleansing composition
DE102004023612A1 (de) * 2004-05-10 2005-12-01 Helmut Meusel Verwendung von microbiellen Mischbiozönosen zur Behandlung von Entzündungen der Haut
KR101398347B1 (ko) 2005-06-22 2014-05-22 오르가노발란스 게엠베하 병원성 미생물에 대해 피부를 보호하기 위한 방법 및 수단
CN101203600B (zh) * 2005-06-22 2015-05-13 有机金属平衡有限公司 保护皮肤对抗病原微生物的方法和手段
WO2006136420A3 (fr) * 2005-06-22 2007-04-19 Organobalance Gmbh Procedes et moyens de protection de la peau contre des micro-organismes pathogenes
US10953051B2 (en) 2005-06-22 2021-03-23 Novozymes A/S Methods and means for protecting the skin against pathogenic microorganisms
EP1995307A1 (fr) * 2005-06-22 2008-11-26 OrganoBalance GmbH Procédés et supports de protection de la peau contre des micro-organismes pathogènes
US8343536B2 (en) 2007-01-25 2013-01-01 Cook Biotech Incorporated Biofilm-inhibiting medical products
BE1018502A3 (nl) * 2009-02-17 2011-02-01 Metatecta Nv Probiotische oplossing voor het beschermen van een oppervlak en werkwijze die gebruik maakt van zulke oplossing.
WO2011000123A1 (fr) 2009-07-01 2011-01-06 Universidad De Concepcion Patchs hydrocolloïdes renfermant des souches viables probiotiques de lactobacillus spp. et utilisés pour le traitement de blessures de diverses natures, dans des tissus infectés, des plaies sèches et des plaies profondes
JP2014516957A (ja) * 2011-05-16 2014-07-17 オルガノバランス メディカル アーゲー 細菌性感冒に対する新規な乳酸菌及びそれを含有する組成物
RU2651466C2 (ru) * 2011-05-16 2018-04-19 ОРГАНОБАЛАНС Медикал АГ Новые молочнокислые бактерии и содержащие их составы против бактериальных простудных заболеваний
CN104159548A (zh) * 2011-11-25 2014-11-19 米库瑞公司 包装的益生菌组合物及其用途
CN104159548B (zh) * 2011-11-25 2017-02-22 米库瑞公司 包装的益生菌组合物及其用途
EP2596776A1 (fr) * 2011-11-25 2013-05-29 Micuri ApS Composition probiotique emballée et utilisations associées
WO2013076272A1 (fr) * 2011-11-25 2013-05-30 Micuri Aps Composition probiotique emballée et ses utilisations
EP2674162A1 (fr) * 2012-05-29 2013-12-18 Danuta Kruszewska Nano-produit, comprenant des lactobacillus reuteri dan080, utile dans la prophylaxie, diagnostic et traitement médical et vétérinaire
CN104684565A (zh) * 2012-05-29 2015-06-03 达努塔·克鲁谢夫斯卡 在人类和兽医的预防和医疗中有用的、包含罗伊氏乳杆菌dan080的纳米产品及其医疗用途
WO2013180585A1 (fr) * 2012-05-29 2013-12-05 Danuta Kruszewska Nano-produit comportant lactobacillus reuteri dan080, utile en prophylaxie et en médecine humaine et vétérinaire, et son utilisation médicale
JP2016522810A (ja) * 2013-04-23 2016-08-04 テラジェン ホールディングス リミテッドＴｅｒｒａｇｅｎ Ｈｏｌｄｉｎｇｓ Ｌｉｍｉｔｅｄ 抗菌活性を有する細菌株およびそれを含む生物的防除組成物
EP2988763A4 (fr) * 2013-04-23 2016-11-16 Terragen Holdings Ltd Souches bactériennes possédant une activité antimicrobienne et compositions de lutte biologique les comprenant
WO2017015208A1 (fr) * 2015-07-17 2017-01-26 Hollister Incorporated Pansement et méthode de traitement de plaies aiguës et chroniques
WO2017208172A1 (fr) * 2016-06-01 2017-12-07 Probiotical S.P.A. Compositions de gel à base de bactéries pour application topique et utilisations associées
ITUA20164065A1 (it) * 2016-06-01 2017-12-01 Probiotical Spa Composizioni in gel per applicazioni topiche a base di batteri, preparazione delle stesse e loro usi.
US11179426B2 (en) 2016-12-29 2021-11-23 Kibow Biotech, Inc. Composition and method for maintaining healthy kidney function
IT201800004036A1 (it) * 2018-03-28 2019-09-28 Teres Srl Gel transdermico
CN114767936A (zh) * 2022-05-10 2022-07-22 武汉理工大学 复合干酪乳杆菌的皮肤损伤修复支架材料制备方法

Also Published As

Publication number Publication date
WO2000071139A3 (fr) 2001-03-08
AU4905400A (en) 2000-12-12
CA2374938A1 (fr) 2000-11-30

Similar Documents

Publication Publication Date Title
WO2000071139A2 (fr) Compositions de lactobacillus et procedes pour prevenir les infections de plaies et la formation d'un film biologique sur des dispositifs chirurgicaux implantables
US20230285475A1 (en) Probiotic formulations and methods for use
US6479051B1 (en) Oral administration of lactobacillus for the treatment and prevention of urogenital infection
US6849256B1 (en) Inhibition of pathogens by probiotic bacteria
AU785159B2 (en) Inhibition of pathogens by bacillus coagulans strains
US20030118571A1 (en) Oral administration of lactobacillus for the maintenance of health in women during pregnancy and at other life stages, to reduce the risk of urogenital diseases
EP2101796B1 (fr) Souche de bacillus smithii tbmi12 mscl p737 et utilisation d'endospores de celle-ci en tant que probiotique ou supplément alimentaire
ES2545209T3 (es) Método de obtención de una nueva cepa de Bifidobacterium bifidum con actividad frente a la infección por Helicobacter pylori
PT986314E (pt) Bactéria láctica probiótica para tratar infecções bacterianas associadas à sids
US20230141737A1 (en) Compositions and methods for preventing and treating antibiotic induced pathologies using probiotics in the biofilm state
JP2005507670A (ja) プロバイオティック組成物
EP0363491B1 (fr) Preparation bacterienne pour prophylaxie et traitement de processus inflammatoires et d'affections allergiques
CN116103197B (zh) 一种对幽门螺旋杆菌有抑制作用的嗜热链球菌及其应用
RU2302877C2 (ru) Биологическая активная добавка к пище для лечения и профилактики кишечных инфекций, осложненных дисбактериозом, и повышения неспецифической резистентности организма
Al-Aabideen Effect of Probiotic on Motility Factors and Swarming Phenomenon of Proteus mirabilis
Al-Amean Investigation on Colanic acid and Molecular Detection of Cytotoxic Necrotizing Factor in Bacteria Associated with Post-Operative Wound Infections
EA043487B1 (ru) Штамм bifidobacterium bifidum 8, используемый в качестве пробиотика

Legal Events

Date Code Title Description
AK Designated states

Kind code of ref document: A2

Designated state(s): AE AL AM AT AU AZ BA BB BG BR BY CA CH CN CR CU CZ DE DK DM EE ES FI GB GD GE GH GM HR HU ID IL IN IS JP KE KG KP KR KZ LC LK LR LS LT LU LV MA MD MG MK MN MW MX NO NZ PL PT RO RU SD SE SG SI SK SL TJ TM TR TT TZ UA UG UZ VN YU ZA ZW

AL Designated countries for regional patents

Kind code of ref document: A2

Designated state(s): GH GM KE LS MW MZ SD SL SZ TZ UG ZW AM AZ BY KG KZ MD RU TJ TM AT BE CH CY DE DK ES FI FR GB GR IE IT LU MC NL PT SE BF BJ CF CG CI CM GA GN GW ML MR NE SN TD TG

121 Ep: the epo has been informed by wipo that ep was designated in this application
AK Designated states

Kind code of ref document: A3

Designated state(s): AE AL AM AT AU AZ BA BB BG BR BY CA CH CN CR CU CZ DE DK DM EE ES FI GB GD GE GH GM HR HU ID IL IN IS JP KE KG KP KR KZ LC LK LR LS LT LU LV MA MD MG MK MN MW MX NO NZ PL PT RO RU SD SE SG SI SK SL TJ TM TR TT TZ UA UG UZ VN YU ZA ZW

AL Designated countries for regional patents

Kind code of ref document: A3

Designated state(s): GH GM KE LS MW MZ SD SL SZ TZ UG ZW AM AZ BY KG KZ MD RU TJ TM AT BE CH CY DE DK ES FI FR GB GR IE IT LU MC NL PT SE BF BJ CF CG CI CM GA GN GW ML MR NE SN TD TG

DFPE Request for preliminary examination filed prior to expiration of 19th month from priority date (pct application filed before 20040101)
ENP Entry into the national phase

Ref document number: 2374938

Country of ref document: CA

Kind code of ref document: A

Ref document number: 2374938

REG Reference to national code

Ref country code: DE

Ref legal event code: 8642

122 Ep: pct application non-entry in european phase
NENP Non-entry into the national phase

Ref country code: JP