COMPOSITIONS AND METHODS FOR INHIBITION AND INTERRUPTION OF BIOFILM FORMATION

Abstract

Compositions and methods for inhibiting and interrupting biofilm formation, and for destabilizing established biofilms are provided, the novel compositions including polymeric resins and monomeric non-polymerizable and polymerizable resins. More particularly, the compositions and methods enable the protection and removal of biofilms from surfaces in the context of medical, consumer, domestic, food service, environmental and industrial applications, where the effects constitute beneficial and desirable biofilm attenuating activity.

Claims

1. A polymerizable and/or polymerized composition for attenuating biofilms, comprising: one or more of non-polymerizable and polymerizable mixtures of quaternary ammonium and phosphonium compounds selected from (1) non-polymerizable antimicrobial mixtures containing a combination of at least one antimicrobially active quaternary ammonium compound, or at least one antimicrobially active quaternary phosphonium compound, or wherein, the combination of components a) and b) are present in a ratio by weight from 1:9 to 9:1. wherein the antimicrobially active quaternary ammonium compounds, including imidazolium, ammonium, pyrrolidinium, etc., (component a)) are represented by the formula
[R—N.sup.+R.sub.1R.sub.2R.sub.3]X.sup.−  (I) wherein R, R.sub.1, R.sub.2, and R.sub.3 are a preferably straight-chain or branched or cyclic of C2-C20 alkyl radical as same or different length independently; also be as fused cyclic or aromatic ring such as aziridine, azirine, oxaziridine, diazirine, azetidine, azete, diazetidine, pyrrolidine, pyrrole, imidazolidine, imidazole, pyrazolidine, pyrazole, thiazolidine, thiazole, isothioazolidine, isothiazole, piperdine, pyridine, piperzine, diazine, morpholinem oxazine, thiomopholine, thiazine, triazine, triazoles, furanzan, oxadiazole, thiadizole, dithozole, tetrazole, azepane, azepine, diazepine, thiazepine, azocane, azocine, azonane, azonine, etc. wherein X.sup.− is a counter anion, which can be inorganic, anions (Cl.sup.−, AlCl.sub.4.sup.−, PF.sub.6.sup.−, BF.sub.4.sup.−, NTf.sub.2.sup.−, DCA.sup.−, etc.) or organic anions (CH.sub.3COO.sup.−, CH.sub.3SO.sub.3.sup.−, etc.). These quaternary ammonium compounds can be present in the mixtures according to the invention either individually or in admixture with one another. wherein antimicrobially active quaternary phosphonium compounds (component b)) are, in particular, compounds corresponding to the following formulae:
[RP.sup.+R.sub.1R.sub.2R.sub.3]Y.sup.−  (II) in which R, R.sub.1, R.sub.2, and R.sub.3 are a preferably straight-chain, branched or cyclic of C2-C20 alkyl radical as same or different length independently; Y— is a counter anion, which can be inorganic, anions (Cl.sup.−, AlCl.sub.4.sup.−, PF.sub.6.sup.−, BF.sub.4.sup.−, NTf.sub.2.sup.−, DCA.sup.−, etc.) or organic anions (CH.sub.3COO.sup.−, CH.sub.3SO.sub.3.sup.−, etc.). and
[(R′).sub.3P.sup.+R″]Y.sup.−  (III) in which R′ is a C1-C5 alkyl radical, a C1-C6 hydroxyalkyl radical or a phenyl radical, R″ is a C3-C18 alkyl radical and Y.sup.− is a halide anion, more especially a chloride anion or a bromide anion. The radicals R″ and R′″ in formula II are preferably straight-chain or branched or cyclic radicals. The quaternary phosphonium compounds can be present in the mixtures of the invention either individually or in admixture with one another. Examples of quaternary phosphonium compounds of the above type are trimethyl-n-dodecyl phosphonium chloride, triethyl-n-decyl phosphonium bromide, tri-n-propyl-n-tetradecyl phosphonium chloride, trimethylol-n-hexadecyl phosphonium chloride, tri-n-butyl-n-decyl phosphonium chloride, tri-n-butyl-n-dodecyl phosphonium bromide, tri-n-butyl-n-tetradecyl phosphonium chloride, tri-n-butyl-n-hexadecyl phosphonium bromide, tri-n-hexyl-n-decylphosphonium chloride, triphenyl-n-dodecyl phosphonium chloride, triphenyl-n-tetradecyl phosphonium bromide and triphenyl-n-octadecyl phosphonium chloride. and (2) polymerizable antimicrobial mixtures containing at least one type of moiety selected from
[R—N.sup.+R.sub.1R.sub.2R.sub.3]X.sup.−  (I) in which R, R.sub.1, R.sub.2, and R.sub.3 are a preferably straight-chain or branched or cyclic of C2-C20 alkyl radical as same or different length independently; also be as fused cyclic or aromatic ring such as aziridine, azirine, oxaziridine, diazirine, azetidine, azete, diazetidine, pyrrolidine, pyrrole, imidazolidine, imidazole, pyrazolidine, pyrazole, thiazolidine, thiazole, isothioazolidine, isothiazole, piperdine, pyridine, piperzine, diazine, morpholinem oxazine, thiomopholine, thiazine, triazine, triazoles, furanzan, oxadiazole, thiadizole, dithozole, tetrazole, azepane, azepine, diazepine, thiazepine, azocane, azocine, azonane, azonine, etc. where X.sup.− is a halide anion, such as chloride, bromide or iodine anion. and
[RP.sup.+R.sub.1R.sub.2R.sub.3]Y.sup.−  (II) in which R, R.sub.1, R.sub.2, and R.sub.3 are a preferably straight-chain, branched or cyclic of C6-C20 alkyl radical as same or different length independently; Y— is a counter anion, which can be inorganic, anions (Cl.sup.−, AlCl.sub.4.sup.−, PF.sub.6.sup.−, BF.sub.4.sup.−, NTf.sub.2.sup.−/trifluoromethanesulfonyl, DCA.sup.−/dicyanamide, etc.) or organic anions (CH.sub.3COO.sup.−, CH.sub.3SO.sub.3.sup.−, etc.). And
(R′).sub.3P.sup.+R″]Y.sup.−  (III) in which R′ is a C1-C5 alkyl radical, a C1-C6 hydroxyalkyl radical or a phenyl radical, R″ is a C3-C18 alkyl radical and Y.sup.− is a halide anion, more especially a chloride anion or a bromide anion. The radicals R″ and R′″ in formula II are preferably straight-chain or branched or cyclic radicals. The quaternary phosphonium compounds can be present in the mixtures of the invention either individually or in admixture with one another. Examples of quaternary phosphonium compounds of the above type are trimethyl-n-dodecyl phosphonium chloride, triethyl-n-decyl phosphonium bromide, tri-n-propyl-n-tetradecyl phosphonium chloride, trimethylol-n-hexadecyl phosphonium chloride, tri-n-butyl-n-decyl phosphonium chloride, tri-n-butyl-n-dodecyl phosphonium bromide, tri-n-butyl-n-tetradecyl phosphonium chloride, tri-n-butyl-n-hexadecyl phosphonium bromide, tri-n-hexyl-n-decylphosphonium chloride, triphenyl-n-dodecyl phosphonium chloride, triphenyl-n-tetradecyl phosphonium bromide and triphenyl-n-octadecyl phosphonium chloride further comprising at least one polymerizable group such as, but not limited to, acrylate, methacrylate, acrylamide, vinyl, vinyl-ether, cyclic ether(epoxy) or cyclic amines and cyclic imine, of which presented as modified R, R.sub.1, R.sub.2, R.sub.3, R′, and R″.

2. A composition according to claim 1, wherein the moiety III comprises tri-n-butyl-n-tetradecyl phosphonium chloride is preferred.

3. A composition according to claim 1, wherein the quaternary ammonium compounds can be present in the mixtures individually or in admixture with one another.

4. A composition according to claims 1 and 3, wherein the moiety I comprises imidazolium or substituted imidazolium moiety is preferred.

5. A composition according to claim 1, wherein any one or more of the non-polymerizable and polymerizable mixtures of quaternary ammonium and phosphonium compounds are non-cleavable for long-last effectiveness.

6. A composition according to claim 1, wherein any one or more of the non-polymerizable and polymerizable mixtures of quaternary ammonium and phosphonium compounds is loaded in final composition in 0.1-10% wt/wt or more and up to 50% wt percent for balanced antibacterial activity, cytotoxicity and mechanical property.

7. A composition according to claim 1, wherein any one or more of the non-polymerizable and polymerizable mixtures of quaternary ammonium and phosphonium compounds is present in compositions, articles and coatings in amounts of from about 0.1 weight percent to about 10 weight percent, the amount selected to achieve balanced biofilm attenuating activity, antibacterial activity/microbial cytotoxicity and mechanical properties of the compositions, articles and coatings.

8. A composition according to claim 1, wherein any one or more of the non-polymerizable and polymerizable mixtures of quaternary ammonium and phosphonium compounds is present in amounts from about 0.1 weight percent to about 10 weight percent, and in some embodiments up to 50 weight percent or more, including 0.1, 0.2, 0.3, 0.4, 0.5, 0.6, 0.7, 0.8, 0.9, 1.0, 2.0, 3.0, 4.0, 5.0, 6.0, 7.0, 8.0, 9.0, 10.0, 20.0, 30.0, 40.0 and 50.0 and fractional increments there between.

9. A composition according to claim 1, wherein any one or more of the non-polymerizable and polymerizable mixtures of quaternary ammonium and phosphonium compounds is formed into solid articles, applied as solid or film coatings on the surfaces of solid articles, or dispersed on, in or throughout other resins and composites, or coated on or dispersed in small particles that are used in fluid suspensions or in filtration, and they may be dispersed free in fluid suspensions.

10. A composition according to claim 1, wherein any one or more of the non-polymerizable and polymerizable mixtures of quaternary ammonium and phosphonium compounds is included in one or more of articles of manufacture, components, reagents, and kits.

11. A composition according to claim 1, wherein any one or more of the non-polymerizable and polymerizable mixtures of quaternary ammonium and phosphonium compounds is formed in an article and may be reactivated chemically or by abrasion/heating other treatment after a period of wear or exposure to fluids or other materials that may comprise microbes.

12. A composition according to claim 1, wherein any one or more of the non-polymerizable and polymerizable mixtures of quaternary ammonium and phosphonium compounds are formulated for providing one more of coating on to, infusion into, dispersion within, or formation of articles of manufacture for Dental Composite, Dental Adhesive, Dental Cement, Dental Sealant, Dental Liner, Dental Varnish, Denture, Root Canal Sealer, Implant Cement, Orthodontic Cement, Self-disinfected Dental Impression Material, Wearable or removable dental plaque treatment device (Antibacterial Night Guard). According to such embodiments, the compositions can be used in Resin Composite-based CAD/CAM Blocks; for Temporary Crown-bridge Composite; for Pediatric Crown; for Esthetic Orthodontic Aligner; for Esthetic Polymer based Orthodontic Bracket (and maybe coating for metal/ceramic bracket); and in some particular embodiments, the compositions can be used in Coating for Dental Implant Abutment. And according to other such embodiments, the compositions may be provided in suspension or coated on micro or nanoparticles for use in mouthwashes, dental strips, dental films and gels, toothpaste and other dental care items.

13. A composition according to claim 1, wherein any one or more of the non-polymerizable and polymerizable mixtures of quaternary ammonium and phosphonium compounds is formulated for providing one or more of coating on to, infusion into, dispersion within, or formation of articles of manufacture for medical and personal care applications, including continuous positive airway pressure (CPAP) device, Ventilation equipment, Central lines, Kwires and screws for fracture fixation, and orthopedic reduction or distraction and other medical implants, catheters, intravascular catheters, dialysis shunts, wound drainage tubes, skin sutures, vascular grafts, implantable meshes, intraocular devices, heart valves, graft materials, needles, transdermal and transmucosal patches, sponges, and personal care and hygiene products selected from but not limited to tampons, sponges, intrauterine devices, diaphragms, condoms, gloves, drapes and films, wound dressings, tapes and dressings, and the like.

14. A composition according to claim 1, wherein any one or more of the non-polymerizable and polymerizable mixtures of quaternary ammonium and phosphonium compounds is formulated for providing one or more of coating on to, infusion into, dispersion within, or formation of articles of manufacture for the inner surface of oil pipelines for reduced biofilm build-up.

15. A composition according to claim 1, wherein any one or more of the non-polymerizable and polymerizable mixtures of quaternary ammonium and phosphonium compounds is formulated for providing one or more of coating on to, infusion into, dispersion within, or formation of articles of manufacture for food service, home goods, and other general use goods, including but not limited to drink dispenser tubing, disposable and reusable drink wear and straws, water, food, and beverage coolers, Denture holders, Mouthguards, sports and Diving/Scuba/swim gear, appliances, and the like.

16. An article of manufacture comprising a kit for attenuating biofilms, the kit comprising: one or more individually packaged treatment formulations, each comprising one or more of treatment implements, such as brushes or other applicators and suspensions comprising the compositions, and, one or more removal implements for mechanical removal of biofilms from the surface after application of the treatment formulation.

Description

BRIEF DESCRIPTION OF THE DRAWINGS

[0117] FIG. 1 shows graphical results for Site Specificity of Predominant Bacterial Species in the Oral Cavity.

[0118] FIG. 2 shows graphical results for spirochetes and P. gingivalis analyses.

[0119] FIG. 3 shows a representative sample of human host subject levels of microbes in dental plaque.

[0120] FIG. 4A provides a visual representation of biofilm treatment and removal.

[0121] FIG. 4B provides the chemical structures of some exemplary embodiments of compositions according to the disclosure.

[0122] FIG. 5 shows a graphic of a Biofilm growth protocol.

[0123] FIG. 6 shows show the 3D architecture of 67 h-old biofilms formed on each surface.

[0124] FIG. 7 shows the quantitative data of biomass from each surface.

[0125] FIG. 8 shows the results of pH analysis of supernatant surrounding test composite and control composite.

[0126] FIG. 9 shows images of the supernatant during biofilm growth.

[0127] FIG. 10 shows the remained biomass from each composite surface after applying shear stress (n>=12).

[0128] FIG. 11 shows the representative confocal image of 67 h biofilms after exposure to shear stress of 0.804 N/m.sup.2.

[0129] FIG. 12 shows EPS-matrix in 2D Cartesian coordinate system (XY, YZ, and XZ planes)

[0130] FIG. 13 shows projection image of skeletonized EPS-matrix.

[0131] Wherever possible, the same reference numbers will be used throughout the drawings to represent the same parts.

DETAILED DESCRIPTION OF THE INVENTION

[0132] This description provides exemplary embodiments in accordance with the general inventive concepts and is not intended to limit the scope of the invention in any way. Indeed, the invention as described herein is broader than and not intended to be limited by the exemplary embodiments, drawing set forth herein, and the terms as used herein have their full ordinary meaning and as described herein.

[0133] As described herein in the examples in the context of in vitro study on biofilm formation, development and detachment, the inventors unexpectedly discovered that S. mutans biofilm on the surface of an antibacterial composite according to the disclosure was significantly reduced in comparison to a control composite and hydroxyapatite (HA). It was further discovered that the mechanical stability of the S. mutans biofilm formed on such antibacterial surface was significantly disrupted as evidenced by complete removal of the biofilm with moderate shear force from the inventive composite. In contrast, the biofilm formed on the control composite and the HA proved to be not susceptible to removal.

[0134] Here it is disclosed an effective methodology to remove biofilms in general. Active surfaces could effectively inhibit not only the initial biofilm formation but also further biofilm development. Total biomass formed on such active surfaces would be significantly reduced at least by 50%. The mechanical stability of the biofilm formed on such active surfaces could be significantly weakened and much less effort could be needed for a complete removal with moderate shear force as applied by a tooth brush, water jet, or ultrasonic treatment.

[0135] In accordance with various embodiments, such active surfaces could be formed in bulk from compositions formulated with a variety of antibacterial/antimicrobial components, including but not limited to polymerizable resins or additives, non-polymerizable additives, or particles/fillers or a combination of both.

[0136] In accordance with some embodiments, such active surfaces could be formed into a coating with a range of thicknesses from compositions formulated with a variety of antibacterial/antimicrobial components, including but not limited to polymerizable resins or additives, non-polymerizable additives, or particles/fillers or a combination of both.

[0137] In accordance with some embodiments, the antibacterial/antimicrobial components could be non-cleavable for long-lasting effectiveness.

[0138] In accordance with the various embodiments, the antibacterial/antimicrobial components will be loaded in a final composition of 0.1-10% wt/wt or more and up to 50% wt percent for balanced antibacterial activity, cytotoxicity and mechanical property.

[0139] In accordance with some embodiments, articles of manufacture, composite articles and materials and coated surfaces comprising any one or more of the non-polymerizable and polymerizable mixtures of quaternary ammonium and phosphonium compounds can be reactivated chemically or by abrasion/heating or other treatment after a period of wear or exposure to fluids or other materials that may comprise microbes. These are non-leachable components and thus it is expected that such an active surface can be readily regenerated as needed.

[0140] In some embodiments, the compositions are formulated for providing one or more of coating onto, infusion into, dispersion within, or formation of articles of manufacture for Dental Composite, Dental Adhesive, Dental Cement, Dental Sealant, Dental Liner, Dental Varnish, Denture, Root Canal Sealer, Implant Cement, Orthodontic Cement, Self-disinfected Dental Impression Material, Wearable or removable dental plaque treatment device (Antibacterial Night Guard). According to such embodiments, the compositions can be used in Resin Composite-based CAD/CAM Blocks; for Temporary Crown-bridge Composite; for Pediatric Crown; for Esthetic Orthodontic Aligner; for Esthetic Polymer based Orthodontic Bracket (and coating for metal/ceramic bracket); and in some particular embodiments, the compositions can be used in Coating for Dental Implant Abutment. And according to other such embodiments, the compositions may be provided in suspension or coated on micro or nanoparticles for use in mouthwashes, dental strips, dental films and gels, toothpaste and other dental care items.

[0141] Such an active surface can be readily formed on top of any non-active bulk substrates, metal, polymer or ceramic, etc., in a form of coating to cover such a non-active material to generate an active surface accordingly.

[0142] In other embodiments, the compositions are formulated for providing one or more of coating onto, infusion into, dispersion within, or formation of articles of manufacture for medical and personal care applications, including continuous positive airway pressure (CPAP) device, Ventilation equipment, Central lines, Kwires and screws for fracture fixation, and orthopedic reduction or distraction and other medical implants, catheters, intravascular catheters, dialysis shunts, wound drainage tubes, skin sutures, vascular grafts, implantable meshes, intraocular devices, heart valves, graft materials, needles, transdermal and transmucosal patches, sponges, and personal care and hygiene products selected from but not limited to tampons, sponges, intrauterine devices, diaphragms, condoms, gloves, drapes and films, wound dressings, tapes and dressings, and the like.

[0143] In yet other embodiments, the compositions are formulated for providing one or more of coating onto, infusion into, dispersion within, or formation of articles of manufacture for the inner surface of oil pipelines for reduced biofilm build-up, and likewise for containment and shipping vessels for oil and petrochemical products generally. In other examples, the compositions are formulated for use in connection with storage and shipment of paints and other organic based materials for domestic and/or industrial use. In certain embodiments, the compositions may provide protective effects for reducing rust and general degradation of metal storage and transport materials, and likewise for containment and shipping vessels for oil and petrochemical products generally. In other examples, the compositions are formulated for use in connection with storage and shipment of paints and other organic based materials for domestic and/or industrial use. In certain embodiments, the compositions may provide protective effects for reducing rust and general degradation of metal storage and transport materials.

[0144] In yet other embodiments, the compositions are formulated for providing one or more of coating onto, infusion into, dispersion within, or formation of articles of manufacture for food service, home goods, and other general use goods, including but not limited to drink dispenser tubing, disposable and reusable drink wear and straws, water, food, and beverage coolers, Denture holders, Mouthguards, sports and Diving/Scuba/swim gear, appliances, and the like.

[0145] In accordance with some embodiments, reagents, self-care formulations and kits comprising the compositions may be provided according to the invention. According to some such embodiments, kits comprising one or more individually packaged treatment formulations may be provided, each comprising one or more of treatment implements, such as brushes or other applicators and suspensions comprising the compositions, the treatment formulations provided for application to a surface for applicant to prevent biofilm formation or to treat existing biofilms. And also provided are one or more removal implements, for mechanical removal of biofilms from the surface after application of the treatment formulation. In some examples, the kits are directed to dental care. In other embodiments, the kits are directed to the care of household or consumer products. Accordingly, the kits may further comprise other conventional treatment formulations suited to a particular application.

[0146] Compositions

[0147] The compositions include, in some embodiments, non-polymerizable antimicrobial mixtures containing a combination of

[0148] a) at least one antimicrobially active quaternary ammonium compound, and

[0149] b) at least one antimicrobially active quaternary phosphonium compound,

[0150] wherein, the combination of components a) and b) are present in a ratio by weight from 1:9 to 9:1.

[0151] And wherein the antimicrobially active quaternary ammonium compounds (component a)) are represented by the formula

[R—N.sup.+R.sub.1R.sub.2R.sub.3]X.sup.−  (I)

[0152] in which R, R.sub.1, R.sub.2, and R.sub.3 are a preferably straight-chain or branched or cyclic of C2-C20 alkyl radical as same or different length independently; also be as fused cyclic or aromatic ring such as aziridine, azirine, oxaziridine, diazirine, azetidine, azete, diazetidine, pyrrolidine, pyrrole, imidazolidine, imidazole, pyrazolidine, pyrazole, thiazolidine, thiazole, isothioazolidine, isothiazole, piperdine, pyridine, piperzine, diazine, morpholinem oxazine, thiomopholine, thiazine, triazine, triazoles, furanzan, oxadiazole, thiadizole, dithozole, tetrazole, azepane, azepine, diazepine, thiazepine, azocane, azocine, azonane, azonine, etc.

[0153] where X.sup.− is a -counter anion, which can be inorganic, anions (Cl.sup.−, AlCl.sub.4.sup.−, PF.sub.6.sup.−, BF.sub.4.sup.−, NTf.sub.2.sup.−, DCA.sup.−, etc.) or organic anions (CH.sub.3COO.sup.−, CH.sub.3SO.sub.3.sup.−, etc.). These quaternary ammonium compounds can be present in the mixtures according to the invention either individually or in admixture with one another.

[0154] And wherein antimicrobially active quaternary phosphonium compounds (component b)) are, in particular, compounds corresponding to the following formula

[RP.sup.+R.sub.1R.sub.2R.sub.3]Y.sup.−  (II)

[0155] in which R, R.sub.1, R.sub.2, and R.sub.3 are a preferably straight-chain, branched or cyclic of C2-C20 alkyl radical as same or different length independently;

[0156] Y.sup.− is a halide anion, such as chloride, bromide or iodine anion.

[0157] Or according to the formula

[(R′).sub.3P.sup.+R″]Y.sup.−  (III)

[0158] in which R′ is a C1-C5 alkyl radical, a C1-C6 hydroxyalkyl radical or a phenyl radical, R″ is a C3-C18 alkyl radical and Y— is a halide anion, more especially a chloride anion or a bromide anion. The radicals R″ and R′″ in formula II are preferably straight-chain or branched or cyclic radicals. The quaternary phosphonium compounds can be present in the mixtures of the invention either individually or in admixture with one another. Examples of quaternary phosphonium compounds of the above type are trimethyl-n-dodecyl phosphonium chloride, triethyl-n-decyl phosphonium bromide, tri-n-propyl-n-tetradecyl phosphonium chloride, trimethylol-n-hexadecyl phosphonium chloride, tri-n-butyl-n-decyl phosphonium chloride, tri-n-butyl-n-dodecyl phosphonium bromide, tri-nbutyl-n-tetradecyl phosphonium chloride, tri-n-butyl-n-hexadecyl phosphonium bromide, tri-n-hexyl-n-decylphosphonium chloride, triphenyl-n-dodecyl phosphonium chloride, triphenyl-n˜tetradecyl phosphonium bromide and triphenyl-n-octadecyl phosphonium chloride. Tri-n-butyl-n-tetradecyl phosphonium chloride is preferred.

[0159] The compositions also include, in other embodiments, polymerizable antimicrobial mixtures containing at least one type of moieties as defined in I, II, III, the moieties further comprising at least one polymerizable group such as, but not limited to, acrylate, methacrylate, acrylamide, vinyl, vinyl-ether, cyclic ether(epoxy) or cyclic amines and cyclic imine, of which presented as modified R, R.sub.1, R.sub.2, R.sub.3, R′, and R″.

[0160] These quaternary ammonium and phosphonium compounds can be present in the mixtures according to the invention either individually or in admixture with one another.

[0161] Some specific examples of monomers in accordance with the embodiments hereof are shown in FIG. 4B, wherein:

[0162] n, m: same or independently as 0, 1, 2, 3 . . . .

[0163] P: 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15 . . . .

[0164] R, R′: same or independently as H, CH.sub.3, C.sub.2H.sub.5CH.sub.2C.sub.6H.sub.5

[0165] X: halide, carboxylic acid, sulfonic acid, phosphoric acid, other Lewis acid

[0166] Y: direct link, O, S, COO, CONN, CONR, OOCO, OCONH, NHCONH

[0167] Monomeric and polymeric resins as disclosed herein may be composed of, in some embodiments, the functional non-polymerizable resins containing at least one of each of antimicrobially active quaternary ammonium and phosphonium compounds, and in other embodiments polymerizable resins containing at least one of antimicrobially active quaternary ammonium and phosphonium compounds at least one polymerizable group, wherein according to the various embodiments, the antimicrobially active quaternary ammonium and phosphonium compounds are present in compositions, articles and coatings in amounts of from about 0.1 weight percent to about 10 weight percent, the amount selected to achieve balanced biofilm attenuating activity, antibacterial activity/microbial cytotoxicity and mechanical properties of the compositions, articles and coatings. Thus, in some embodiments, the antimicrobially active quaternary ammonium and phosphonium compounds are present in amounts from about 0.1 weight percent to about 10 weight percent, and in some embodiments up to 50 weight percent or more, including 0.1, 0.2, 0.3, 0.4, 0.5, 0.6, 0.7, 0.8, 0.9, 1.0, 2.0, 3.0, 4.0, 5.0, 6.0, 7.0, 8.0, 9.0, 10.0, 20.0, 30.0, 40.0 and 50.0 and fractional increments there between.

[0168] In accordance with the various embodiments, compositions may be formulated with or incorporated or dispersed in resins known in various arts for forming or coating articles of manufacture. And in accordance with the compositions hereof, resins for composites may be selected from, by way of non-limiting examples, HEMA and HPMA, which are typical monomethacrylate resins; BisGMA, TEGDMA, UDMA are typical conventional dimethacrylate resins, which are polymerizable/curable by heat, light and redox initiation processes. -CQ and LTPO are typical photoinitaiors. Tertiary aromatic amines, such as EDAB, may be included as an accelerator for CO-based photoinitiator. Other additives such as inhibitors, UV stabilizers or fluorescent agents may also be used. In addition, a variety of particles, polymeric, inorganic, organic particles may be incorporated to reinforce the mechanical properties, rheological properties, and sometime biological functionalities.

[0169] The following abbreviations may be used: BisGMA: 2,2-bis(4-(3-methacryloyloxy-2-hydroxypropoxy)-phenyl)propane HEMA: 2-hydroxyethyl methacrylate HPMA: 2-hydroxypropyl methacrylate TEGDMA: tri ethylene glycol dimethacrylate UDMA: di(methacryloxyethyptrimethyl-1,6-hexaethylenediurethane BHT: butylhydroxytoluene CQ; cannphorquinone LTPO: lucirin TP0/2,4,6-trimethylbenzoyldiphenylphosphine oxide EDAB: 4-Ethyl dimethylaminobenzonate AMAHP: 3-(acryloyloxy)-2-hydroxypropyl methacrylate EGAMA: ethyleneglycol acrylate methacrylate TCDC: 4,8-bis(hydroxymethyl)-tricyclo[5,2,1,02=6] CDI: 1,1-carbonyl-diimidazole SR295: pentaerythritol tetraacrylate.

Experimental Examples

[0170] Influence of Composite Material on the Development, 3D Architecture and Mechanical Stability of S. mutans Biofilms

[0171] Goal: Examine how the biofilm formation is affected by the test composite in terms of biomass, and how its mechanical stability is changed.

[0172] Biofilm growth protocol is shown in FIG. 5.

[0173] Test Groups:

[0174] HA disc

[0175] Conventional Dental Composite/IJ8-095 (control) sterilized by autoclaving

[0176] Experimental Antibacterial Composite/IJ8-083 (test) sterilized by autoclaving

[0177] Analyses

[0178] Inhibition of biofilm formation

[0179] Intact biofilm 3D architecture

[0180] Intact biofilm biomass (dry-weight)

[0181] pH changes of supernatant

[0182] Variation of antibiofilm effect

[0183] Mechanical stability by applying shear stress

[0184] Biofilm removal profile

[0185] Sheared biofilm 3D architecture

[0186] Analysis of EPS-matrix

[0187] EPS-matrix in 2D Cartesian coordinate system (XY, YZ, and XZ planes)

[0188] Analysis of EPS-matrix via topological skeleton method

[0189] Results

[0190] Inhibition of biofilm formation

[0191] Intact biofilm 3D architecture

[0192] FIG. 6 shows the 3D architecture of 67 h-old biofilms formed on each surface.

[0193] Biofilm formation was clearly disrupted by the test composite. Confocal images show that biofilm formation and accumulation were significantly compromised by the test composite.

[0194] Use of saliva coating evidenced no impact on the antibacterial effect of the test composite.

[0195] The composites were sterilized by using 70% EtOH+UV. However, the test composite was much less effective than the autoclaved test composite, and prone to contamination. Therefore, autoclaved composites were used.

[0196] Intact Biofilm Biomass

[0197] FIG. 7 shows the quantitative data of biomass from each surface.

[0198] At 67 h, biomass from the test composite was 2.3 times less than the biomass from control composite, which agrees very well with the confocal imaging data.

[0199] Inhibition of biofilm formation was maintained even after the initial biofilm formation period (29 h), indicating lasting effect for prolonged period.

[0200] pH Changes

[0201] FIG. 8 shows that the pH of the supernatant surrounding test composites was significantly higher than the pH of supernatant of control composite. It indicates that biofilm formation and accumulation were affected during the whole experimental period. However, pH deviation was largely due to some variation of antibiofilm effect.

[0202] Variation of the antibiofilm effect can be visualized, and a new finding about potential long-term effect of the material (see later section).

[0203] FIG. 9 shows images of the supernatant during biofilm growth

[0204] Above images are the 24-well plates containing supernatant during biofilm growth period.

[0205] Usually, the supernatant became turbid when the bacterial growth is active in the first 29 to 43 h, then it became clear again (after 53 h) once the biofilm growth became stable.

[0206] Between 29-43 h (active bacterial growth transitioning to biofilm phase), all the supernatant from control composite were turbid. Then after 53 h, all the supernatant of control composite became clear, as biofilm growth establishes.

[0207] In contrast, all the supernatant from test composite (except one) were mostly clear between 29-43 h, indicating antibacterial activity. However, some variability was observed on the effects after 43 h, indicating variability of the antibacterial release profile among the different test samples.

[0208] One supernatant from the test composite (box with green dotted line) never became turbid by the end of biofilm growth (67 h), indicating strong antibacterial activity and no biofilm growth on the surface.

[0209] Additional Information:

[0210] Further analyses were conducted to determine if the used test composite would be effective. Surprisingly, re-used test composites were still interfering with the initial biofilm formation and accumulation, which suggests a long term effect even after re-use.

[0211] Mechanical Stability

[0212] Biofilm Removal Profile

[0213] FIG. 10 shows the remaining biomass from each composite surface after applying shear stress (n>=12).

[0214] Biomass removal patterns were similar, while the amount of biomass from the test composite was significantly lower than the one from the control composite.

[0215] At 0.804 N/m.sup.2, biofilm removal from the test composite already reached a detection limit (˜0.0003 g), while the percentage of biomass removal from the control composite was still only ˜50%. There was no significant further removal from the control composite at 1.785 N/m.sup.2.

[0216] Sheared 3-D Biofilm Architecture

[0217] FIG. 11 shows the representative confocal image of 67 h biofilms after exposure to shear stress of 0.804 N/m.sup.2.

[0218] Although biofilms on the control composite were flattened under application of shear force of 0.804 N/m.sup.2, numerous bacterial microcolonies still attached to the control composite.

[0219] Strikingly, most of the bacterial biomass and EPS-matrix on the test composite were clearly removed, while a few tiny aggregates remained.

[0220] Quite surprisingly, the results show that dental composites comprising the compositions according to the invention can disrupt both the initial biofilm formation and its further development. Although biofilms are not completely inhibited on the test composite, the biofilm accumulated can be easily removed and detached by low external shear forces.

[0221] Analysis of EPS-Matrix

[0222] FIG. 12 shows EPS-matrix in 2-D Cartesian coordinate system (XY, YZ, and XZ planes)

[0223] To understand why the biofilms on the test composite are easily removed, the structural morphology of EPS-matrix was assessed. FIG. 12 shows the representative projection images of intact 67-h biofilms in XY, YZ, and XZ planes.

[0224] EPS-matrix on the control composite was thick and relatively evenly distributed over the entire surface. Also, the EPS-matrix is structurally more organized, which appeared to be connected to each other forming a network that likely provides a strong and stable architecture.

[0225] In contrast, the EPS-matrix on the test composite was much thinner compared to the matrix on the control composite. Further, the shape of the matrix appeared scattered and unorganized. It may indicate lack of structural stability (in sharp contrast to control composite) of the scattered EPS-matrix formed on the test composite.

[0226] Additional analyses were conducted to verify whether there was significant differences in the geometrical pattern of the EPS formed on control vs test composite surfaces.

[0227] Analysis of EPS-matrix via Mathematical morphology

[0228] To further analyze the structure of EPS-matrix, the topological skeleton method was applied which is based on theoretical analysis and processing of geometrical structures. The skeleton usually emphasizes geometrical and topological properties of the shape, such as its connectivity, topology, length, direction, and width. Thus, it can provide basic information regarding how the EPS-matrix is developed and organized.

[0229] FIG. 13 demonstrates that the projected image of skeletonized EPS-matrix on the control composite is clearly a well-structured surrounding EPS-matrix that is connected by thick filaments, while the inside structure is densely filled with thin filaments. Clearly, the assembly of the entire EPS-matrix is highly organized, which may explain the mechanical resistance of biofilm to external shear forces.

[0230] In contrast to the control composite, the EPS-matrix on the test composite was devoid of thick filaments, but rather thin and short filaments without any pattern were observed. At 40 μm height, the EPS-matrix was already disconnected and its density was reducing with increased height. The projection image shows poorly developed overall EPS-matrix which may not be able to withstand external shear forces.

[0231] Collectively, the test composite may impede the formation of a typical EPS-matrix with densely packed thick and thin filaments that provides strong resistance to mechanical stress.

[0232] As used herein, the singular forms “a,” “an,” and “the” are intended to include the plural forms as well, unless the context clearly indicates otherwise.

[0233] As used herein the term “biofilm” refers to an extracellular polymeric substance produced by and including microbes and having three-dimensional structural characteristics. Biofilms, whether on a surface or in a suspension, provide a matrix that can support the retention and growth of one or more of discrete microbial species and mixed species populations selected from bacteria, fungi, protozoa, algae, and others. In some embodiments, biofilms comprise co-aggregating organisms.

[0234] The term “coating” as used herein refers to a topically applied or superficial layer or surface of an underlying material that constitutes a material covering an article such as a medical device, a dental composite or apparatus, a container such as for food or industrial goods, and the like.

[0235] As used herein, the term “microbe” refers to a microorganism and is intended to encompass both an individual organism, and hetero and homogenous populations comprising any number of the organisms. As used herein, the term “microorganism” refers to any of a variety of species or microorganism, including but not limited to, archaea, bacteria, fungi, protozoans, mycoplasma, and parasitic organisms, wherein the term “fungi” is used in reference to eukaryotic organisms such as the molds and yeasts, including dimorphic fungi, and the terms “bacteria” and “bacterium” refers to the various examples as specifically disclosed in the tables and description herein, broadly including prokaryotic organisms within the phyla in the kingdom Procaryotae, the microorganisms including Actinomyces, Chlamydia, Streptomyce, and all cocci, bacilli, spirochetes, spheroplasts, protoplasts, all Gram-negative and Gram-positive “Gram-negative” and “Gram-positive” refer to staining patterns with the Gram-staining process, and all non-pathogenic bacteria and pathogenic bacteria. In particular, the term “pathogen” refers to a biological organism that causes or to which can be at least partially attributed any of a variety of disease states in a host, and include, but are not limited to, archaea, bacteria, fungi, protozoans, mycoplasma, parasites, and viruses.

[0236] As used herein, the term “antimicrobial agent” refers to composition that decreases, prevents or inhibits the growth of bacterial and/or fungal organisms. In some specific examples of antimicrobial agents, antibiotics are those substances that inhibit the growth of microorganisms, ideally without damage to the host. In various different examples, antibiotics may affect one or more of a microbial cell's activity resulting in cell death, including but not limited to inhibition or alteration of one or more of membrane function and nucleic acid, protein, and cellular component/cell wall synthesis. Antibiotics can include, but are not limited to, macrolides (e.g., erythromycin), penicillins (e.g., nafcillin), cephalosporins (e.g., cefazolin), carbapenems (e.g., imipenem), monobactam (e.g., aztreonam), other beta-lactam antibiotics, beta-lactam inhibitors (e.g., sulbactam), oxalines (e.g., linezolid), aminoglycosides (e.g., gentamicin), chloramphenicol, 15 sufonamides (e.g., sulfamethoxazole), glycopeptides (e.g., vancomycin), quinolones (e.g., ciprofloxacin), tetracyclines (e.g., minocycline), fusidic acid, trimethoprim, metronidazole, clindamycin, mupirocin, rifamycins (e.g., rifampin), streptogramins (e.g., quinupristin and dalfopristin) lipoprotein (e.g., daptomycin), polyenes (e.g., amphotericin B), azoles (e.g., fluconazole), and echinocandins (e.g., caspofungin acetate). Examples of specific antibiotics include, but are not limited to, amifloxacin, amphotericin B, and nystatin, azithromycin, aztreonam, cefazolin, ciprofloxacin, clarithromycin, clavulanic acid, clinafloxacin, clindamycin, enoxacin, erythromycin, fleroxacin, fluconazole, gatifloxacin, gemifloxacin, gentamicin, imipenem, itraconazole, ketoconazole, linezolid, lomefloxacin, metronidazole, minocycline, moxifloxacin, mupirocin, nafcillin, nalidixic acid, norfloxacin, ofloxacin, pefloxacin, rifampin, sparfloxacin, sulbactam, sulfamethoxazole, teicoplanin, temafloxacin, tosufloxacin, trimethoprim, vancomycin.

[0237] As used herein, the term “medical devices” includes any material or device that is used on, in, or through a subject's or patient's body, for example, in the course of medical treatment to address, to minimize or prevent an illness or injury. Medical devices include, but are not limited to, such items as CPAP, Ventilation equipment, Central lines, Kwires and screws for fracture fixation, and orthopedic reduction or distraction and other medical implants, catheters, intravascular catheters, dialysis shunts, wound drainage tubes, skin sutures, vascular grafts, implantable meshes, intraocular devices, heart valves, graft materials, needles, transdermal and transmucosal patches, sponges, and personal care and hygiene products selected from but not limited to tampons, sponges, intrauterine devices, diaphragms, condoms, gloves, drapes and films, wound dressings, tapes and dressings, and the like.

[0238] Dental devices include, but are not limited to Dental Composite, Dental Adhesive, Dental Cement, Dental Sealant, Dental Liner, Dental Varnish, Denture, Root Canal Sealer, Implant Cement, Orthodontic Cement, Self-disinfected Dental Impression Material, Wearable or removable dental plaque treatment device (Antibacterial Night Guard). According to such embodiments, the compositions can be used in Resin Composite-based CAD/CAM Blocks; for Temporary Crown-bridge Composite; for -Pediatric Crown; for Esthetic Orthodontic Aligner; for Esthetic Polymer based Orthodontic Bracket (and maybe coating for metal/ceramic bracket); and in some particular embodiments, the compositions can be used in Coating for Dental Implant Abutment. And according to other such embodiments, the compositions may be provided in suspension or coated on micro or nanoparticles for use in mouthwashes, dental strips, dental films and gels, toothpaste and other dental care items.

[0239] The general inventive concepts herein are described with occasional reference to the exemplary embodiments of the invention. Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art encompassing the general inventive concepts. The terminology set forth in this detailed description is for describing particular embodiments only and is not intended to be limiting of the general inventive concepts.

[0240] Unless otherwise indicated, all numbers expressing quantities, properties, and so forth as used in the specification and claims are to be understood as being modified in all instances by the term “about.” Accordingly, unless otherwise indicated, the numerical properties set forth in the specification and claims are approximations that may vary depending on the suitable properties desired in embodiments of the present invention. Notwithstanding that the numerical ranges and parameters setting forth the broad scope of the general inventive concepts are approximations, the numerical values set forth in the specific examples are reported as precisely as possible. Any numerical values, however, inherently contain certain errors necessarily resulting from error found in their respective measurements.

[0241] While various inventive aspects, concepts and features of the general inventive concepts are described and illustrated herein in the context of various exemplary embodiments, these various aspects, concepts and features may be used in many alternative embodiments, either individually or in various combinations and sub-combinations thereof. Unless expressly excluded herein all such combinations and sub-combinations are intended to be within the scope of the general inventive concepts. Still further, while various alternative embodiments as to the various aspects, concepts and features of the inventions (such as alternative materials, structures, configurations, methods, devices and components, alternatives as to form, fit and function, and so on) may be described herein, such descriptions are not intended to be a complete or exhaustive list of available alternative embodiments, whether presently known or later developed.

[0242] Those skilled in the art may readily adopt one or more of the inventive aspects, concepts or features into additional embodiments and uses within the scope of the general inventive concepts even if such embodiments are not expressly disclosed herein. Additionally, even though some features, concepts or aspects of the inventions may be described herein as being a preferred arrangement or method, such description is not intended to suggest that such feature is required or necessary unless expressly so stated. Still further, exemplary or representative values and ranges may be included to assist in understanding the present disclosure; however, such values and ranges are not to be construed in a limiting sense and are intended to be critical values or ranges only if so expressly stated.