CN108728388A - Porphyromonas gingivalis, resisting porphyromonas gingivalis specific IgY preparation and Compound formula of toothpaste - Google Patents
Porphyromonas gingivalis, resisting porphyromonas gingivalis specific IgY preparation and Compound formula of toothpaste Download PDFInfo
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N1/00—Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
- C12N1/20—Bacteria; Culture media therefor
- C12N1/205—Bacterial isolates
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12R—INDEXING SCHEME ASSOCIATED WITH SUBCLASSES C12C - C12Q, RELATING TO MICROORGANISMS
- C12R2001/00—Microorganisms ; Processes using microorganisms
- C12R2001/01—Bacteria or Actinomycetales ; using bacteria or Actinomycetales
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K39/00—Medicinal preparations containing antigens or antibodies
- A61K39/395—Antibodies; Immunoglobulins; Immune serum, e.g. antilymphocytic serum
- A61K39/40—Antibodies; Immunoglobulins; Immune serum, e.g. antilymphocytic serum bacterial
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K8/00—Cosmetics or similar toiletry preparations
- A61K8/18—Cosmetics or similar toiletry preparations characterised by the composition
- A61K8/19—Cosmetics or similar toiletry preparations characterised by the composition containing inorganic ingredients
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K8/00—Cosmetics or similar toiletry preparations
- A61K8/18—Cosmetics or similar toiletry preparations characterised by the composition
- A61K8/19—Cosmetics or similar toiletry preparations characterised by the composition containing inorganic ingredients
- A61K8/20—Halogens; Compounds thereof
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K8/00—Cosmetics or similar toiletry preparations
- A61K8/18—Cosmetics or similar toiletry preparations characterised by the composition
- A61K8/30—Cosmetics or similar toiletry preparations characterised by the composition containing organic compounds
- A61K8/33—Cosmetics or similar toiletry preparations characterised by the composition containing organic compounds containing oxygen
- A61K8/35—Ketones, e.g. benzophenone
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K8/00—Cosmetics or similar toiletry preparations
- A61K8/18—Cosmetics or similar toiletry preparations characterised by the composition
- A61K8/30—Cosmetics or similar toiletry preparations characterised by the composition containing organic compounds
- A61K8/64—Proteins; Peptides; Derivatives or degradation products thereof
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P1/00—Drugs for disorders of the alimentary tract or the digestive system
- A61P1/02—Stomatological preparations, e.g. drugs for caries, aphtae, periodontitis
-
- 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
- A61P31/04—Antibacterial agents
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P37/00—Drugs for immunological or allergic disorders
- A61P37/08—Antiallergic agents
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61Q—SPECIFIC USE OF COSMETICS OR SIMILAR TOILETRY PREPARATIONS
- A61Q11/00—Preparations for care of the teeth, of the oral cavity or of dentures; Dentifrices, e.g. toothpastes; Mouth rinses
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61Q—SPECIFIC USE OF COSMETICS OR SIMILAR TOILETRY PREPARATIONS
- A61Q17/00—Barrier preparations; Preparations brought into direct contact with the skin for affording protection against external influences, e.g. sunlight, X-rays or other harmful rays, corrosive materials, bacteria or insect stings
- A61Q17/005—Antimicrobial preparations
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K2800/00—Properties of cosmetic compositions or active ingredients thereof or formulation aids used therein and process related aspects
- A61K2800/80—Process related aspects concerning the preparation of the cosmetic composition or the storage or application thereof
- A61K2800/86—Products or compounds obtained by genetic engineering
Abstract
The present invention relates to a kind of porphyromonas gingivalis, porphyromonas gingivalis is porphyromonas gingivalis (Porphyromonas gingivalis) Ma01, is deposited in China typical culture collection center, and deposit number is CCTCC NO：M 2017790.Further relate to a kind of resisting porphyromonas gingivalis specific IgY preparation.Further relate to a kind of preparation method of resisting porphyromonas gingivalis specific IgY preparation.Further relate to a kind of Compound formula of toothpaste containing resisting porphyromonas gingivalis specific IgY preparation.It the advantage is that there is significant antibacterial and bactericidal effect, halitosis to reduce the generation of the mouth diseases such as periodontitis porphyromonas gingivalis；It carries out compounding Compound formula of toothpaste obtained with strontium salt, Paeonol, there are significant anti-allergic effects.
Description
Technical field
The present invention relates to microorganisms technical field more particularly to a kind of porphyromonas gingivalis, resisting porphyromonas gingivalis
Specific IgY preparation and Compound formula of toothpaste.
Background technology
In China, gum, periodontal disease incidence account for 70% of population or more, and porphyromonas gingivalis is chronic gum edge
The main pathogenic bacteria of the periodontosis such as inflammation, chronic periodontitis, aggressive periodontitis.In the hair of patients with periodontal disease simultaneous halitosis
It is raw, currently without the effective ways for the treatment of halitosis.Common treatment halitosis method is to be treated using antimicrobial DP finish, but use
Antimicrobial DP finish usually has many drawbacks.
Prevention and treatment periodontosis and its halitosis are set about being crucial institute from main pathogenic bacteria porphyromonas gingivalis
?.
The prior art is used, and is had and is carried out treatment periodontosis and halitosis using specific IgY.But experiments verify that it is rear without or
The effect of without significantly inhibiting periodontosis and its halitosis.
Invention content
The purpose of the present invention is being directed to deficiency in the prior art, a kind of porphyromonas gingivalis, resisting porphyromonas are provided
Monad specific IgY preparation and preparation method thereof and toothpaste.
To achieve the above object, the technical solution adopted by the present invention is that：
The present invention provides a kind of porphyromonas gingivalis, which is porphyromonas gingivalis
(Porphyromonas gingivalis) Ma01 is deposited on December 14th, 2017 in China typical culture collection
The heart, deposit number are CCTCC NO：M 2017790, preservation address are the Wuhan Wuhan Universitys of China.
The present invention also provides a kind of resisting porphyromonas gingivalis specific IgY preparations, for inhibiting porphyromonas gingivalis
Ma01。
Preferably, the immune bird inlay of antigen is combined by porphyromonas gingivalis Ma01 thalline and bacterial chip and made
It is standby.
The present invention also provides a kind of preparation methods of resisting porphyromonas gingivalis specific IgY preparation, include the following steps：
A. porphyromonas gingivalis incubation step
A.1 bacterial species：Porphyromonas gingivalis (Porphyromonas gingivalis)
A.2 porphyromonas gingivalis Ma01 bacterial strains are taken, are separately cultured respectively through BHI-S culture mediums and amplification cultivation, through mirror
Surely it is confirmed as porphyromonas gingivalis, obtains porphyromonas gingivalis thalline and bacterial chip；
B. immunogene preparation process：
B.1 take porphyromonas gingivalis thalline and bacterial chip that bacterium solution antigen is made；
B.2 isometric bacterium solution antigen and isometric good fortune formula Freund's complete adjuvant is compound, stir breast using high speed agitator
Change, is finally made Water-In-Oil good fortune formula Freund's complete adjuvant porphyromonas gingivalis complex immunogen；
B.3 isometric bacterium solution antigen and isometric good fortune formula Freund's incomplete adjuvant is compound, it is stirred using high speed agitator
Emulsification, is finally made Water-In-Oil good fortune formula Freund's incomplete adjuvant porphyromonas gingivalis complex immunogen；
C. step is immunized
Good fortune formula Freund's complete adjuvant porphyromonas gingivalis complex immunogen is given to production using syringe through subcutaneous and intramuscular injection
Then egg hen uses good fortune formula Freund's incomplete adjuvant porphyromonas gingivalis complex immunogen through subcutaneous and intramuscular injection week about
To bird inlay；After booster immunization is secondary, it is compound that good fortune formula Freund's incomplete adjuvant porphyromonas gingivalis was hereafter reused every ten weeks
Immunogene is front and back for eight times immune through subcutaneously and intramuscular injection is to bird inlay booster immunization five times；
D. resisting porphyromonas gingivalis specific IgY extract preparation process
Immune latter 28th day immune hen egg immune for the first time is collected to be smashed with 75% alcohol disinfecting egg shell
Egg eggshell, leaves and takes yolk；After 10 times of deionized water dilution yolk, with glacial acetic acid and anhydrous sodium acetate adjust pH to 5.30~
Supernatant is taken after 5.50,4 DEG C of sedimentations, adds 2% chitosan and sodium alginate, 2% calcium chloride and sodium polyphosphate flocculation are heavy
Drop, then supernatant is taken to remove extra deionized water with gel filtration system ultrafiltration, egg yolk liquid is concentrated 20 times or so, with 1N hydrogen
Sodium oxide molybdena adjusts pH to 7.0~8.0, obtains resisting porphyromonas gingivalis specific IgY extract, -20 DEG C of preservations of juxtaposition；
E. resisting porphyromonas gingivalis specific IgY extract purification step
Add amine sulfate to 55% saturation degree resisting porphyromonas gingivalis specific IgY extract, is sufficiently stirred, room temperature
It is static to carry out high speed centrifugation after a certain period of time, take precipitation, precipitation is diluted to substance accumulated amount using distilled water, then add amine sulfate
It to 33% saturation degree, is sufficiently stirred, room temperature is static after a certain period of time, carries out high speed centrifugation, takes precipitation, and precipitation is used distillation
It is packed into bag filter after water dilution dissolving, with distilled water through hollow fiber filter desalination, that is, it is special to obtain resisting porphyromonas gingivalis
Anisotropic IgY preparations.
Preferably, it is immunized in step in the C., subcutaneous and intramuscular injection good fortune formula Freund's complete adjuvant porphyromonas gingivalis
The injection volume of complex immunogen is 0.5ml~3.0ml.
Preferably, it is immunized in step in the C., subcutaneous and intramuscular injection good fortune formula Freund's incomplete adjuvant porphyromonas unit cell
The injection volume of bacterium complex immunogen is 0.5ml~3.0ml.
Preferably, in the E. resisting porphyromonas gingivalis specific IgY extract purification step, room temperature static 2 is small
Shi Hou, carries out high speed centrifugation 10 minutes.
The present invention also provides a kind of Compound formula of toothpaste, for inhibiting porphyromonas gingivalis Ma01, including：
Resisting porphyromonas gingivalis specific IgY preparation, the resisting porphyromonas gingivalis specific IgY preparation account for toothpaste
The mass percent of gross mass is 0.5%~5.0%.
Preferably, further include：
Antiallergic ingredient, the antiallergic ingredient includes strontium salt and Paeonol；
Base ingredients, the base ingredients include sorbierite, glycerine, polyethylene glycol, rubbing agent, saccharin sodium, thickener, table
Face activating agent, pigment, essence and water.
Preferably, the rubbing agent is any one in silica, calcium carbonate, carbonic acid fine particle calcium carbonate, aluminium hydroxide or two
Kind or more combination.
Preferably, the surfactant is lauryl sodium sulfate, N- sarcosinates, cocamidopropyl propyl amide beet
Any one in alkali, poloxamer or two or more combinations.
Preferably, the thickener be CMC, HEC, carragheen, xanthans, in guar gum any one or it is two or more
Combination.
Preferably, the pigment is the combination of any one or two kinds in CI77891, CI42090.
Preferably, the essence is the combination of Mint Essence, any one or two kinds in spearmint essence.
Preferably, the water is deionized water or sterile water.
Preferably, each component account for toothpaste gross mass mass percent it is as follows：
Strontium salt：1.0%~3.0%
Paeonol：0.2%~1.0%
Sorbierite：20.0%~70.0%
Glycerine：0.5%~30.0%
Polyethylene glycol：0.5%~10%
Rubbing agent：5.0%~50.0%
Saccharin sodium：0.1%~0.3%
Thickener：0.1%~2.0%
Surfactant：0.5%~8.0%
Pigment：0.001%~1.0%
Essence：0.5%~2.0%
Watering balance is to 100.0%.
Preferably, the strontium salt is strontium chloride, and the mass percent that the strontium salt accounts for toothpaste gross mass is 2.0%.
Preferably, the mass percent that the Paeonol accounts for toothpaste gross mass is 0.5%.
The present invention is had the following technical effect that compared with prior art using above technical scheme：
The present invention's has resisting porphyromonas gingivalis specific IgY preparation prepared by porphyromonas gingivalis, to gum porphin
There is quinoline monad significant antibacterial and bactericidal effect, halitosis to reduce the generation of the mouth diseases such as periodontitis；With strontium salt, pellet
Skin phenol carries out compounding Compound formula of toothpaste obtained, has significant anti-allergic effects.
Specific implementation mode
Below in conjunction with the embodiment of the present invention, technical scheme in the embodiment of the invention is clearly and completely described,
Obviously, described embodiments are only a part of the embodiments of the present invention, instead of all the embodiments.Based in the present invention
Embodiment, the every other embodiment that those of ordinary skill in the art are obtained under the premise of not making creative work,
It shall fall within the protection scope of the present invention.
It should be noted that in the absence of conflict, the feature in embodiment and embodiment in the present invention can phase
Mutually combination.
The present invention is further explained in the light of specific embodiments, but not as limiting to the invention.
Embodiment 1
The present embodiment is related to the porphyromonas gingivalis Ma01 of the present invention.
Using the sulphur ethyl alcohol culture medium containing sheep blood, in 85% nitrogen, 10% hydrogen and 5% carbon dioxide anaerobic environment
Culture three days, takes 1*105~1*1062ml BHI fluid nutrient mediums are added in 100 μ l of cfu/ml bacterium solutions, continue to cultivate 48h, select leather
Blue Albert'stain Albert microscopy is the bacterial strain of the positive.
One plant of isolated best bacterium of activity is named as Ma01.It is identified by Physiology and biochemistry, while using PCR
The bacterium rRNA sequences are expanded, are compared, as a result show the same of the sequence and ncbi database Porphyromonas gingivalis
Source property is up to 99%, therefore the dientification of bacteria is porphyromonas gingivalis, is named as porphyromonas gingivalis Ma01
(Porphyromonas gingivalis Ma01), and be deposited in China typical culture collection on December 14th, 2017
The heart, deposit number are CCTCC NO：M 2017790, preservation address are wuchang, wuhan Luo Jia Shan road Wuhan University.
Embodiment 2
The present embodiment is related to the resisting porphyromonas gingivalis specific IgY preparation and preparation method thereof of the present invention.
The porphyromonas gingivalis Ma01 obtained using embodiment 1 prepares resisting porphyromonas gingivalis specific IgY system
Agent is combined into the immune bird inlay of antigen by the thalline and bacterial chip of porphyromonas gingivalis Ma01 and is prepared.
The preparation method of resisting porphyromonas gingivalis specific IgY preparation, includes the following steps：
A. porphyromonas gingivalis incubation step
A.1 bacterial species：Porphyromonas gingivalis (Porphyromonas gingivalis)
A.2 porphyromonas gingivalis Ma01 bacterial strains are taken, are separately cultured respectively through BHI-S culture mediums and amplification cultivation, through mirror
Surely it is confirmed as porphyromonas gingivalis, obtains porphyromonas gingivalis thalline and bacterial chip；
B. immunogene preparation process：
B.1 take porphyromonas gingivalis thalline and bacterial chip that bacterium solution antigen is made；
B.2 isometric bacterium solution antigen and isometric good fortune formula Freund's complete adjuvant is compound, stir breast using high speed agitator
Change, is finally made Water-In-Oil good fortune formula Freund's complete adjuvant porphyromonas gingivalis complex immunogen, every milliliter of good fortune formula Freund's complete adjuvant gum
Detection of Porphyromonas complex immunogen contains bacterium 1*109~9*109；
B.3 isometric bacterium solution antigen and isometric good fortune formula Freund's incomplete adjuvant is compound, it is stirred using high speed agitator
Emulsification, is finally made Water-In-Oil good fortune formula Freund's incomplete adjuvant porphyromonas gingivalis complex immunogen, every milliliter of good fortune formula is not exclusively helped
Agent porphyromonas gingivalis complex immunogen contains bacterium 1*109~9*109；
C. step is immunized
Using syringe by good fortune formula Freund's complete adjuvant porphyromonas gingivalis complex immunogen 0.5ml~3.0ml through subcutaneously and
Then intramuscular injection uses good fortune formula Freund's incomplete adjuvant porphyromonas gingivalis complex immunogen week about to bird inlay
0.5ml~3.0ml is through subcutaneous and intramuscular injection to bird inlay；After booster immunization is secondary, hereafter good fortune formula was reused every ten weeks
Freund's incomplete adjuvant porphyromonas gingivalis complex immunogen 0.5ml~3.0ml reinforces through subcutaneous and intramuscular injection to bird inlay
It is five times immune, it is front and back for eight times immune；
D. resisting porphyromonas gingivalis specific IgY extract preparation process
Immune latter 28th day immune hen egg immune for the first time is collected to be smashed with 75% alcohol disinfecting egg shell
Egg eggshell, leaves and takes yolk；After 10 times of deionized water dilution yolk, with glacial acetic acid and anhydrous sodium acetate adjust pH to 5.30~
Supernatant is taken after 5.50,4 DEG C of sedimentations, adds 2% chitosan and sodium alginate, 2% calcium chloride and sodium polyphosphate flocculation are heavy
Drop, then supernatant is taken to remove extra deionized water with gel filtration system ultrafiltration, egg yolk liquid is concentrated 20 times or so, with 1N hydrogen
Sodium oxide molybdena adjusts pH to 7.0~8.0, obtains resisting porphyromonas gingivalis specific IgY extract, -20 DEG C of preservations of juxtaposition；
E. resisting porphyromonas gingivalis specific IgY extract purification step
Add amine sulfate to 55% saturation degree resisting porphyromonas gingivalis specific IgY extract, is sufficiently stirred, room temperature
Static 2 hours, high speed centrifugation 10 minutes took precipitation, precipitation was diluted to substance accumulated amount using distilled water, then add amine sulfate extremely
33% saturation degree, is sufficiently stirred, and static 2 hours of room temperature, high speed centrifugation 10 minutes takes precipitation, and precipitation is dilute using distilled water
It is packed into bag filter after releasing dissolving, with distilled water through hollow fiber filter desalination, that is, obtains resisting porphyromonas gingivalis specificity
IgY preparations.
Resisting porphyromonas gingivalis specific IgY preparation sterling is measured through ELISA method, and resisting porphyromonas gingivalis is special
Property IgY preparation sterlings in the potency of specific IgY antibody in every milligram of albumen be 1：1000/mg albumen or anti-gum porphin
In quinoline monad specific IgY preparation sterling, the specific activity of the specific IgY antibody in every milligram of albumen is 1：1000.?
In IgY preparation sterlings, the purity of IgY antibody is measured as 92.5% through irreducibility SDS-PAGE.
Embodiment 3
The present embodiment is related to the Compound formula of toothpaste of the present invention.
The resisting porphyromonas gingivalis specific IgY preparation and antiallergic ingredient, base ingredients being prepared using embodiment 2
Prepare Compound formula of toothpaste.
Antiallergic ingredient includes strontium salt and Paeonol, and base ingredients include sorbierite, glycerine, polyethylene glycol, rubbing agent, saccharin
Sodium, thickener, surfactant, pigment, essence and water.
Wherein, strontium salt is strontium chloride.
Wherein, rubbing agent be silica, calcium carbonate, carbonic acid fine particle calcium carbonate, in aluminium hydroxide any one or it is two or more
Combination.
Wherein, any one in thickener CMC, HEC, carragheen, xanthans, guar gum or two or more groups
It closes.
Wherein, surfactant is lauryl sodium sulfate, N- sarcosinates, Cocoamidopropyl betaine, pool
Any one in Luo Shamu or two or more combinations.
Wherein, any one in pigment CI77891, CI42090 or two kinds of combination.
Wherein, essence is the combination of Mint Essence, any one or two kinds in spearmint essence.
Wherein, water is sterile water or deionized water.
In Compound formula of toothpaste, the mass percent that each component accounts for toothpaste gross mass is as follows：
Resisting porphyromonas gingivalis specific IgY preparation：0.5%~5.0%
Strontium salt：1.0%~3.0%
Paeonol：0.2%~1.0%
Sorbierite：20.0%~70.0%
Glycerine：0.5%~30.0%
Polyethylene glycol：0.5%~10%
Rubbing agent：5.0%~50.0%
Saccharin sodium：0.1%~0.3%
Thickener：0.1%~2.0%
Surfactant：0.5%~8.0%
Pigment：0.001%~1.0%
Essence：0.5%~2.0%
Watering balance is to 100.0%.
The production method of Compound formula of toothpaste is：At normal temperatures, cream processed is added in the sorbierite weighed, glycerine and polyethylene glycol
It stirs in machine, then successively puts into thickener, surfactant, strontium salt, saccharin sodium, pigment in Manufacturing medicine extract machine, Paeonol is abundant
It is dissolved in the water, is added in Manufacturing medicine extract machine, strong stirring is uniform；Resisting porphyromonas gingivalis specific IgY preparation is added to system
In cream machine, stir evenly；Silica is added, is stirred evenly, stirring is then vacuumized；Manufacturing medicine extract machine finally is added in essence
In, it stirs evenly, deaerates, lotion is made.
Embodiment 4
The present embodiment is a preferred embodiment of the Compound formula of toothpaste of embodiment 3.
The compounding ointment of the present embodiment, each component and mass percent are as follows：
Resisting porphyromonas gingivalis specific IgY preparation：1%
Strontium chloride：1.0%
Paeonol：0.2%
Sorbierite：40.0%
Glycerine：5%
Polyethylene glycol：5%
Silica：25%
Saccharin sodium：0.3%
Carragheen：1%
Lauryl sodium sulfate：3%
CI77891：0.005%
Mint Essence：0.5%
Deionized water：Surplus.
The production method of Compound formula of toothpaste is：At normal temperatures, cream processed is added in the sorbierite weighed, glycerine and polyethylene glycol
It stirs in machine, then successively puts into carragheen, lauryl sodium sulfate, strontium chloride, saccharin sodium, CI77891 in Manufacturing medicine extract machine, it will
Paeonol is completely dissolved in water, is added in Manufacturing medicine extract machine, strong stirring is uniform；By resisting porphyromonas gingivalis specific IgY system
Agent is added in Manufacturing medicine extract machine, is stirred evenly；Silica is added, is stirred evenly, stirring is then vacuumized；Finally by minty note
Finishing enters in Manufacturing medicine extract machine, stirs evenly, and lotion is made in degassing.
Embodiment 5
The present embodiment is a preferred embodiment of the Compound formula of toothpaste of embodiment 3.
The compounding ointment of the present embodiment, each component and mass percent are as follows：
Resisting porphyromonas gingivalis specific IgY preparation：2.5%
Strontium chloride：2.0%
Paeonol：0.4%
Sorbierite：35%
Glycerine：5%
Polyethylene glycol：5%
Aluminium hydroxide：35%
Saccharin sodium：0.2%
Guar gum：1.5%
N- sarcosinates：2.5%
CI42090：0.01%
Spearmint essence：1.0%
Deionized water：Surplus.
The production method of Compound formula of toothpaste is：At normal temperatures, cream processed is added in the sorbierite weighed, glycerine and polyethylene glycol
It stirs in machine, then successively puts into guar gum, N- sarcosinates, strontium chloride, saccharin sodium, CI42090 in Manufacturing medicine extract machine, it will
Paeonol is completely dissolved in water, is added in Manufacturing medicine extract machine, strong stirring is uniform；By resisting porphyromonas gingivalis specific IgY system
Agent is added in Manufacturing medicine extract machine, is stirred evenly；Silica is added, is stirred evenly, stirring is then vacuumized；Finally by spearmint
Essence is added in Manufacturing medicine extract machine, stirs evenly, and deaerates, lotion is made.
Embodiment 6
The present embodiment is a preferred embodiment of the Compound formula of toothpaste of embodiment 3.
The compounding ointment of the present embodiment, each component and mass percent are as follows：
Resisting porphyromonas gingivalis specific IgY preparation：4%
Strontium chloride：3.0%
Paeonol：0.6%
Sorbierite：45.0%
Glycerine：5%
Polyethylene glycol：10%
Calcium carbonate：20%
Saccharin sodium：0.3%
CMC：1.5%
Poloxamer：4%
CI42090：0.002%
Mint Essence：1.5%
Deionized water：Surplus.
The production method of Compound formula of toothpaste is：At normal temperatures, cream processed is added in the sorbierite weighed, glycerine and polyethylene glycol
It stirs in machine, then successively puts into CMC, poloxamer, strontium chloride, saccharin sodium, CI42090 in Manufacturing medicine extract machine, Paeonol is abundant
It is dissolved in the water, is added in Manufacturing medicine extract machine, strong stirring is uniform；Resisting porphyromonas gingivalis specific IgY preparation is added to system
In cream machine, stir evenly；Silica is added, is stirred evenly, stirring is then vacuumized；Cream processed finally is added in Mint Essence
It in machine, stirs evenly, deaerates, lotion is made.
Embodiment 7
The present embodiment is related to the halitosis compliance test result of Compound formula of toothpaste.
Artificial saliva is prepared first, and the ingredient and concentration of artificial saliva are as follows：
1163mg/l KCl、535mg/l NaH2PO4、43mg/l MgCl2、210mg/l CaCl2、354mg/l KH2PO4、
875mg/l NaHPO4、233mg/l NH4Cl, 41mg/l kilnitamin, 1.0mg/l creatinines, 273mg/l urea, 0.8mg/l
Mixed vitamin and 2000mg/l carboxymethyl celluloses.
Artificial saliva after preparation is handled 10 minutes at 100 DEG C, transparent liquid is obtained by filtration, it is spare after sterilizing 15 minutes.
0.9ml artificial salivas are dispensed into multiple 2.5ml centrifuge tubes, after being incubated 1 hour, 0.2ml blood plasma is added, then
100 μ l 3*10 are added6Cfu/ml porphyromonas gingivalis.Being incubated under the conditions of 37 DEG C of amphimicrobians makes it generate stink, uses
Halimeter apparatus measures VSCs (volatile sulfur compounds, including hydrogen sulfide, methyl mercaptan, diethyl sulfide etc.) is horizontal.
Three experimental groups, respectively Compound formula of toothpaste group, blank control group and positive controls are set, and each experimental group carries out
It at least tests three times, each experimental group takes experimental result average value as evaluation result.Experiment condition is in 37 DEG C of amphimicrobians
Under the conditions of be incubated, in 8 hours each 4 hours it is horizontal with Halimeter apparatus measures VSCs.
In Compound formula of toothpaste group, the compounding tooth of 20 μ l is added in the centrifuge tube containing artificial saliva after each incubation
Cream；In blank control group, the distilled water of 20 μ l is added in the centrifuge tube containing artificial saliva after each incubation；In sun
In property control group, the 25 μ g/ml metronidazoles of 20 μ l are added in the centrifuge tube containing artificial saliva after each incubation.
Experimental result is as follows：
| 4 hours VSCs levels, mg/m3 | 8 hours VSCs levels, mg/m3 | |
| Compound formula of toothpaste group | 54 | 32 |
| Blank control group | 124 | 251 |
| Positive controls | 67 | 56 |
From above-mentioned experimental result it is found that the Compound formula of toothpaste of the present invention can effectively contain the mouth that porphyromonas gingivalis generates
Smelly problem, effect are better than metronidazole.Compared with metronidazole, resisting porphyromonas gingivalis specific IgY preparation not will produce resistance to
Pharmacological property, and be not carcinogen.
Embodiment 8
The present embodiment is related to the real-time antiallergic effect verification of Compound formula of toothpaste.
It takes Compound formula of toothpaste prepared by the embodiment of the present invention 4~6 to be tested, while contrast groups 1 and control group 2 is set, it is right
It is all made of antiallergy toothpaste prepared by existing known formulations according to group 1 and control group 2, control group 1 is 0.5% strontium chloride, 0.1% pellet
Skin phenol and toothpaste base-material, control group 2 are 4% strontium chloride, 2.5% Paeonol and toothpaste base-material.
It choosing 200 person-times and carries out blind time experiment with odontalgia hemodia patient, every group of 40 people, brushing time is after 1 minute,
Being gargled using cold water is stimulated with air-flow.
Criterion is：It is effective, no any feeling of pain (having remarkable result)；Effectively, there is sense of discomfort but substantially painless
(effective)；Generally, there is feeling of pain but can still endure and (have general effect)；In vain, there is feeling of pain and be difficult to endure (without apparent effect
Fruit).
Its experiment effect is as shown in the table：
| n | It is effective | Effectively | Generally | In vain | Total effective rate | |
| 4 groups of embodiment | 40 | 37(92.5) | 2(5.0) | 1(2.5) | 0(0.0) | 40(100.0) |
| 5 groups of embodiment | 40 | 35(87.5) | 2(5.0) | 3(7.5) | 0(0.0) | 40(100.0) |
| 6 groups of embodiment | 40 | 36(90.0) | 4(10.0) | 0(0.0) | 0(0.0) | 40(100.0) |
| Control group 1 | 40 | 30(75.0) | 4(10.0) | 4(10.0) | 2(5.0) | 38(95.0) |
| Control group 2 | 40 | 36(90.0) | 4(10.0) | 0(0.0) | 0(0.0) | 40(100.0) |
By the experimental result of upper table it is found that the Compound formula of toothpaste of the present invention has significant analgesia desensitization treatment effect, in reality
When dental sensitivity treatment on it is efficient up to 100%.
Embodiment 9
The present embodiment is related to the long-term antiallergic effect verification of Compound formula of toothpaste.
It takes Compound formula of toothpaste prepared by the embodiment of the present invention 4~6 to be tested, while contrast groups 3 and control group 4 is set, it is right
It is all made of antiallergy toothpaste prepared by existing known formulations according to group 3 and control group 4, control group 3 is 0.5% strontium chloride, 0.1% pellet
Skin phenol and toothpaste base-material, control group 4 are 4% strontium chloride, 2.5% Paeonol and toothpaste base-material.
It chooses 200 person-times and carries out blind experiment with odontalgia hemodia patient, every group of 40 people brush teeth, every time sooner or later daily
Brushing time 3 minutes after persistently using 3 months, carries out probe acute test.
Probe acute is tested：The test of sensitivity of tooth index, probe contact are carried out using electron pressure responsive probe
The compacting of the cheek surface exposure of chosen tooth, is placed on enamel essence intersection, sets the strength of probe contact as 50g.
Criterion is：0, probe is contacted without any feeling of pain (having remarkable result)；1, probe contact slightly discomfort but base
This is painless (effective)；2, probe has feeling of pain but can still endure and (have general effect) when contacting；3, probe has when contacting
Severe pain is felt and is difficult to endure (no positive effect).
Subjective assessment：0~2=improves；3=is without change.
Its experiment effect is as shown in the table：
| n | 0 | 1 | 2 | 3 | Total effective rate | |
| 4 groups of embodiment | 40 | 31(77.5) | 2(5.0) | 6(15.0) | 1(2.5) | 39(97.5) |
| 5 groups of embodiment | 40 | 30(75.0) | 6(15.0) | 3(7.5) | 2(5.0) | 38(95.0) |
| 6 groups of embodiment | 40 | 32(80.0) | 4(10.0) | 3(7.5) | 1(2.5) | 39(97.0) |
| Control group 3 | 40 | 15(37.5) | 4(10.0) | 8(20.0) | 13(32.5) | 27(67.5) |
| Control group 4 | 40 | 30(75.0) | 4(10.0) | 4(10.0) | 2(5.0) | 38(95.0) |
By the experimental result of upper table it is found that the Compound formula of toothpaste of the present invention has significant analgesia desensitization treatment effect, in length
Improvement rate on the dental sensitivity treatment of phase is substantially better than control group 3.
By embodiment 6~7 it is found that the present invention the Compound formula of toothpaste containing resisting porphyromonas gingivalis specific IgY preparation,
Not only there is significant analgesia desensitization treatment effect, but also there is excellent halitosis effect.
Embodiment 10
The present embodiment is related to the mass percent of the resisting porphyromonas gingivalis specific IgY preparation in Compound formula of toothpaste.
In Compound formula of toothpaste, the mass percent of resisting porphyromonas gingivalis specific IgY preparation is respectively 0%, 0.1%,
0.2%, 0.3%, 0.4%, 0.5%, 0.75%, 1.0%, 1.5%, 2%, 2.5%, 3.0%, 3.5%, 4.0%,
5.0%, 6.0%, 7.0%, 8.0%, 10%, 15%, 20%, 25.0%, strontium chloride 0.5%, Paeonol 0.1%,
Remaining is toothpaste base-material.
The Compound formula of toothpaste of above-mentioned different component is subjected to test experience as described in Example 7, experimental result such as following table
It is shown：
As seen from the above table, when the mass percent of resisting porphyromonas gingivalis specific IgY preparation is less than 0.5%,
Halitosis effect is poor；When its mass percent is 0.5%~5.0%, halitosis effect gradually increases；When its quality percentage
When than being 5.0%~10.0%, halitosis effect is identical, the trend not significantly increased；When its mass percent is more than 15%
When, halitosis effect has slight increase.In conclusion the mass percent of resisting porphyromonas gingivalis specific IgY preparation is
When 0.5%~5.0%, halitosis effect is preferable, and cost is relatively low.
The foregoing is merely preferred embodiments of the present invention, are not intended to limit embodiments of the present invention and protection model
It encloses, to those skilled in the art, should can appreciate that and all be replaced with being equal made by present specification
It changes and obviously changes obtained scheme, should all be included within the scope of the present invention.
Claims (10)
1. a kind of porphyromonas gingivalis, which is characterized in that the porphyromonas gingivalis is porphyromonas gingivalis
(Porphyromonas gingivalis) Ma01, is deposited in China typical culture collection center, deposit number CCTCC
NO：M 2017790.
2. a kind of resisting porphyromonas gingivalis specific IgY preparation, for inhibiting porphyromonas unit cell as described in claim 1
Bacterium Ma01.
3. resisting porphyromonas gingivalis specific IgY preparation according to claim 2, which is characterized in that by porphyromonas
Monad Ma01 thalline and bacterial chip are combined into antigen and bird inlay are immunized and prepares.
4. a kind of Compound formula of toothpaste, for inhibiting porphyromonas gingivalis as described in claim 1, which is characterized in that including：
Resisting porphyromonas gingivalis specific IgY preparation as described in claim 2~3 is any, the resisting porphyromonas unit cell
The mass percent that bacterium specific IgY preparation accounts for toothpaste gross mass is 0.5%~5.0%.
5. Compound formula of toothpaste according to claim 4, which is characterized in that further include：
Antiallergic ingredient, the antiallergic ingredient includes strontium salt and Paeonol；
Base ingredients, the base ingredients include sorbierite, glycerine, polyethylene glycol, rubbing agent, saccharin sodium, thickener, surface work
Property agent, pigment, essence and water.
6. Compound formula of toothpaste according to claim 5, which is characterized in that the thickener is CMC, HEC, carragheen, xanthan
Any one in glue, guar gum or two or more combinations.
7. Compound formula of toothpaste according to claim 5, which is characterized in that the surfactant be lauryl sodium sulfate,
Any one in N- sarcosinates, Cocoamidopropyl betaine, poloxamer or two or more combinations.
8. Compound formula of toothpaste according to claim 5, which is characterized in that the pigment is appointing in CI77891, CI42090
The combination for one or two of anticipating.
9. Compound formula of toothpaste according to claim 5, which is characterized in that the essence is in Mint Essence, spearmint essence
Any one or two kinds combination.
10. Compound formula of toothpaste according to claim 5, which is characterized in that each component accounts for the mass percent of toothpaste gross mass
It is as follows：
Strontium salt：1.0%~3.0%
Paeonol：0.2%~1.0%
Sorbierite：20.0%~70.0%
Glycerine：0.5%~30.0%
Polyethylene glycol：0.5%~10%
Rubbing agent：5.0%~50.0%
Saccharin sodium：0.1%~0.3%
Thickener：0.1%~2.0%
Surfactant：0.5%~8.0%
Pigment：0.001%~1.0%
Essence：0.5%~2.0%
Watering balance is to 100.0%.
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| CN111588647A (en) * | 2020-05-20 | 2020-08-28 | 谭骏 | Natural small-molecule mouthwash and toothpaste formula for inhibiting porphyromonas gingivalis |
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