CN102860932B - Gingivitis and gingivitis ozostomia preventing mouthwash prepared by anti-porphyromonas gingivalis and IgY antibody with fusobacterium nucleatum specificity - Google Patents
Gingivitis and gingivitis ozostomia preventing mouthwash prepared by anti-porphyromonas gingivalis and IgY antibody with fusobacterium nucleatum specificity Download PDFInfo
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Abstract
The invention discloses IgY mouthwash prepared by anti-porphyromonas gingivalis and an IgY antibody with fusobacterium nucleatum specificity, which is used for preventing gingivitis and gingivitis ozostomia. The IgY mouthwash comprises an IgY preparation for resisting periodontal pathogens and basic mouthwash materials, wherein the IgY preparation for resisting periodontal pathogens is prepared by raw materials according to the following mass percent accounting for the total mass of toothpaste: 0.01%-0.5% of anti-porphyromonas gingivalis and IgY antibody preparation with fusobacterium nucleatum specificity, 0.01-2.0% of chlorhexidine, 1.0-30.0% of glycerol, and 0.1-2.0% of polyvinylpyrrolidone, wherein a protective preparation of the anti-porphyromonas gingivalis and the IgY antibody preparation with fusobacterium nucleatum specificity is prepared by the chlorhexidine, the glycerol, and the polyvinylpyrrolidone. The IgY mouthwash which is disclosed by the invention and is used for treating and preventing gingivitis and ozostomia has an obvious effect on treating and preventing the gingivitis and the ozostomia.
Description
Technical field
The present invention relates to oral-cavity article technical field, the prevention that particularly prepared by the anti-Fusobacterium nucleatum of a kind of use and porphyromonas gingivalis specific IgY antibody is by the collutory of these two kinds of microbial gingivitis and halitosis thereof.
Background technology
China's gum, periodontal disease sickness rate account for the more than 70% of population, porphyromonas gingivalis (Porphyromonas gingivalis) is the main suspicious pathogenic bacterium of the periodontopathy such as marginal gingivitis, chronic periodontitis, aggressive periodontitis, its pili can be combined with the glyceraldehyde-3-phosphate dehydrogenase on mouth epithelial cells surface, invade gingival epithelium cell and Gingival Fibroblasts, escape from immune is removed.Porphyromonas gingivalis produces dipeptides acyl aminopeptidase degrade connective tissues, promotes the active increase of MMP-2 and MMP-1.The arginine of porphyromonas gingivalis and Methionin specific cysteine protease (RgpA, RgpB, Kgp) can be hydrolyzed L-12 and cause INN γ to produce minimizing.Part porphyromonas unicellular hair can induce Expression of Macrophages L-1 α, L-1 β, L-6, TNF α to cause alveolar bone to destroy.In the generation of patients with periodontal disease simultaneous halitosis, its halitosis mainly contains Fusobacterium nucleatum and causes.At present there is no effectively to treat the method for halitosis, use Antibiotics pharmacological agent halitosis to have a lot of drawbacks.
Setting about from main cause of disease bacterium porphyromonas gingivalis and Fusobacterium nucleatum for prevention and treatment periodontopathy and halitosis thereof is key point.
At present be generally for the collutory preventing and treat gingivitis the collutory that contains medicinal herb components.Although the patent of this class has a lot, all without clear and definite curative effect.
Existing bibliographical information shows to use clinically specific IgY antibody preventing dental caries and treatment infant rotavirus diarrhoea, and prevention pulmonary fibrosis patients charrin's disease, alleviates the microbial stomachache effect of helicobacter pylorus remarkable, and have no side effect and habituation.
In the IgY of numerous species of our research, find, some periodontopathy the experiment proved that the trend and the curative effect that are not with or without obvious inhibition periodontopathy and halitosis thereof because of the IgY antibody that bacterial immunity hen produces.Briefly, not every IgY has obvious therapeutic action.
Because the suspicious cause of disease bacterium of periodontopathy and halitosis thereof has 11 kinds more than, but main suspicious cause of disease bacterium comprises porphyromonas gingivalis, Fusobacterium nucleatum, companion actinomycetes actinobacillus, the treponema dentium of dwelling, Fu Shi bacterioide, capnocytophaga gingivalis etc.When IgY corresponding to exploration preparation is used for the treatment of periodontopathy and halitosis thereof, we infer that porphyromonas gingivalis, Fusobacterium nucleatum, companion actinomycetes actinobacillus, the treponema dentium of dwelling, Fu Shi bacterioide, capnocytophaga gingivalis all likely cause periodontopathy and halitosis thereof; Because our prior unpredictable which bacterium in the middle of this is the principal element that causes periodontopathy and halitosis thereof, which bacterium is to cause that periodontopathy and halitosis thereof are secondary causes.
We think must have the test of suitable anti-periodontopathy and halitosis thereof to determine which bacterium is the principal element that causes periodontopathy and halitosis thereof.Thereby guidance system is for medicine or the health product of anti-periodontopathy and halitosis thereof.
The prompting guidance technology personnel that current prior art is not correlated with find that resisting porphyromonas gingivalis, Fusobacterium nucleatum composite IgY are the keys that solves treatment periodontopathy and halitosis thereof.
Technician cannot be in advance under the prerequisite of experiment of not doing anti-periodontopathy and halitosis thereof, learns that in advance porphyromonas gingivalis, Fusobacterium nucleatum are the principal elements that causes periodontopathy and halitosis thereof.
Technician cannot be in advance under the prerequisite of experiment of not doing anti-periodontopathy and halitosis thereof, learns that in advance resisting porphyromonas gingivalis, Fusobacterium nucleatum composite IgY are than the better effects if of independent resisting porphyromonas gingivalis IgY, independent anti-Fusobacterium nucleatum IgY.
Summary of the invention
Technical problem to be solved by this invention is to provide for the not good problem of existing periodontopathy medicine collutory result for the treatment of prevention prepared by the anti-Fusobacterium nucleatum of a kind of use and the porphyromonas gingivalis specific IgY antibody collutory by these two kinds of microbial gingivitis and halitosis thereof.
Technical problem to be solved by this invention can be achieved through the following technical solutions:
A kind of prevention of gingivitis of preparing with resisting porphyromonas gingivalis and Fusobacterium nucleatum specific IgY antibody and the collutory of halitosis thereof, formed by anti-Periodontal Pathogens IgY preparation and collutory base-material, described anti-Periodontal Pathogens IgY preparation is by being made up of the materials in percentage by mass that accounts for collutory total mass: resisting porphyromonas gingivalis and Fusobacterium nucleatum mix specific IgY antibody preparation 0.01～0.5%, Tubulicid 0.01-2.0%, glycerine 1.0-30.0%, polyvinylpyrrolidone 0.1-2.0%, wherein Tubulicid, glycerine, polyvinylpyrrolidone forms resisting porphyromonas gingivalis and the compound specific IgY antibody preparation of Fusobacterium nucleatum protective material.
In a preferred embodiment of the invention, described collutory base-material is made up of the materials in percentage by mass that accounts for collutory total mass:
Glycerine 1.0-10.0%, sorbyl alcohol 10-20%, propylene glycol 1.0-5.0%, Xylitol 1.0-5.0%, polysorbas20 1.0-3.0%, tween 80 1.0-3.0%, CrmopherRH400.5-3.0%, PVM/MA polymkeric substance 0.6-2.0%, Sodium Benzoate 0.5-1.5%, aspartame 0.1-0.5%, asccharin 0.1-0.3%, essence 0.01-0.8%, food dye is appropriate, deionized water surplus.
The present invention, for the collutory of prevention of periodontal disease and halitosis thereof, has a significant effect to prevention of periodontal disease and halitosis.
Embodiment
Describe implementation of the present invention in detail below in conjunction with embodiment.The related bacterial classification of embodiment is all provided by Medical College, Shanghai Communication Univ. below.
Embodiment 1:
The preparation of the compound specific IgY antibody preparation of resisting porphyromonas gingivalis and Fusobacterium nucleatum:
A. porphyromonas gingivalis and Fusobacterium nucleatum culturing step
A.1 bacterial species: porphyromonas gingivalis (Porphyromonas gingivalis) and Fusobacterium nucleatum (Fusobacterium nucleatum);
A.2 get porphyromonas gingivalis (P.g) ATCC33277 bacterial strain and Fusobacterium nucleatum (Fn) ATCC10953 bacterial strain, respectively through the unicellular bacterium of porphyromonas, the Fusobacterium nucleatum of BHI-S substratum separation and Culture and amplification cultivation, and confirm as porphyromonas gingivalis, Fusobacterium nucleatum and obtain porphyromonas gingivalis thalline and bacterial chip, Fusobacterium nucleatum thalline and bacterial chip through identifying;
B. immunogen preparation process:
B.1 get porphyromonas gingivalis thalline and bacterial chip 10～70 weight parts, Fusobacterium nucleatum thalline and bacterial chip 30～90 weight parts are mixed into bacterium liquid antigen;
B.2 isopyknic bacterium liquid antigen is mixed with isopyknic Freund's complete adjuvant, use high speed agitator stirring and emulsifying, finally makes water-in-oil Freund's complete adjuvant porphyromonas gingivalis and Fusobacterium nucleatum mixed immunity is former; Every milliliter of former bacterium 1,000,000,000-8,000,000,000 of containing of Freund's complete adjuvant mixed immunity;
B.3 isopyknic mixed bacteria liquid is mixed with isopyknic freund 's incomplete adjuvant, use high speed agitator stirring and emulsifying, finally make water-in-oil freund 's incomplete adjuvant porphyromonas gingivalis and Fusobacterium nucleatum mixed immunity is former, during every milliliter of freund 's incomplete adjuvant mixed immunity is former, contain bacterium 1,000,000,000-8,000,000,000;
C. immune step:
Use syringe by former Freund's complete adjuvant mixed immunity subcutaneous and intramuscular injection to bird inlay, every bird inlay injection 0.5ml-3.0ml immunogen, is then used the subcutaneous and intramuscular injection of the former 0.5ml-3.0ml of freund 's incomplete adjuvant mixed immunity to bird inlay week about.After booster immunization secondary, after this re-used the subcutaneous and intramuscular injection of the former 0.5ml-3.0ml of freund 's incomplete adjuvant mixed immunity every ten weeks to bird inlay booster immunization five times, front and back immunity eight times altogether;
D. resisting porphyromonas gingivalis and Fusobacterium nucleatum mix the preparation of specific IgY extract:
Collect the immunity immunity immune hen egg of latter the 28th day for the first time, the alcohol of use 75% carries out egg surface sterilization, open eggshell and get yolk, remove membrane of yolk and get egg yolk liquid, by egg yolk liquid and the distilled water ratio adding distil water of 1:10 by volume, fully stir, and use Glacial acetic acid to regulate potential of hydrogen to pH5.30-5.50, then place-20 ℃ of cold storage takes out and thaws for 3 days afterwards, get supernatant liquor, by concentrated 20 times of egg yolk liquid, with 1N NaOH adjusting PH to 7.0-8.0, put-20 ℃ of preservations by gel-filtration system.
E.IgY purification step:
Resisting porphyromonas gingivalis and Fusobacterium nucleatum are mixed to the saturation ratio that specific IgY extract adds sulfuric acid amine to 55%, fully stir, room temperature leaves standstill 2 hours, then high speed centrifugation 10 minutes, get precipitation, to precipitate and use distilled water diluting to substance accumulated amount, add again the saturation ratio of sulfuric acid amine to 33%, fully stir, room temperature leaves standstill 2 hours, then high speed centrifugation 10 minutes, get precipitation, after dissolving with distilled water diluting, pack dialysis tubing into, with distilled water through hollow fiber filter desalination, obtain resisting porphyromonas gingivalis and Fusobacterium nucleatum and mix the direct sterling of specific IgY antibody.
Resisting porphyromonas gingivalis and Fusobacterium nucleatum mix specific IgY antibody preparation sterling and measure through ELISA method, tiring as 1:32000/mg albumen of specific IgY antibody in resisting porphyromonas gingivalis and Fusobacterium nucleatum mixing specific IgY sterling in every milligram of albumen, or claim resisting porphyromonas gingivalis and Fusobacterium nucleatum to mix in specific IgY sterling, the specific activity of the specific IgY antibody in every milligram of albumen is 1:32000.In IgY antibody sterling, the purity of IgY antibody is determined as 89.3% through irreducibility SDS-PAGE.
Embodiment 2:
Resisting porphyromonas gingivalis and Fusobacterium nucleatum mix the preparation of specific IgY antibody preparation:
A. porphyromonas gingivalis and Fusobacterium nucleatum culturing step
A.1 bacterial species: porphyromonas gingivalis (Porphyromonas gingivalis) and Fusobacterium nucleatum (Fusobacterium nucleatum);
A.2 get porphyromonas gingivalis (P.g) ATCC33277 bacterial strain and Fusobacterium nucleatum (Fn) ATCC10953 bacterial strain, respectively through the unicellular bacterium of porphyromonas, the Fusobacterium nucleatum of BHI-S substratum separation and Culture and amplification cultivation, and confirm as porphyromonas gingivalis, Fusobacterium nucleatum and obtain porphyromonas gingivalis thalline and bacterial chip, Fusobacterium nucleatum thalline and bacterial chip through identifying;
B. immunogen preparation process:
B.1 get porphyromonas gingivalis thalline and bacterial chip 10～70 weight parts, Fusobacterium nucleatum thalline and bacterial chip 30～90 weight parts are mixed into bacterium liquid antigen;
B.2 isopyknic bacterium liquid antigen is mixed with isopyknic Freund's complete adjuvant, use high speed agitator stirring and emulsifying, finally makes water-in-oil Freund's complete adjuvant porphyromonas gingivalis and Fusobacterium nucleatum mixed immunity is former; Every milliliter of former bacterium 1,000,000,000-8,000,000,000 of containing of Freund's complete adjuvant mixed immunity;
B.3 isopyknic mixed bacteria liquid is mixed with isopyknic freund 's incomplete adjuvant, use high speed agitator stirring and emulsifying, finally make water-in-oil freund 's incomplete adjuvant porphyromonas gingivalis and Fusobacterium nucleatum mixed immunity is former, during every milliliter of freund 's incomplete adjuvant mixed immunity is former, contain bacterium 1,000,000,000-8,000,000,000;
C. immune step:
Use syringe by former Freund's complete adjuvant mixed immunity subcutaneous and intramuscular injection to bird inlay, every bird inlay injection 0.5ml-3.0ml immunogen, is then used the subcutaneous and intramuscular injection of the former 0.5ml-3.0ml of freund 's incomplete adjuvant mixed immunity to bird inlay week about.After booster immunization secondary, after this re-used the subcutaneous and intramuscular injection of the former 0.5ml-3.0ml of freund 's incomplete adjuvant mixed immunity every ten weeks to bird inlay booster immunization five times, front and back immunity eight times altogether;
D. resisting porphyromonas gingivalis and Fusobacterium nucleatum mix the preparation of specific IgY extract:
Collect the immunity immunity immune hen egg of latter the 28th day for the first time, with 75% alcohol disinfecting egg shell, smash eggshell, leave and take yolk.With after 10 times of dilution yolk of deionized water, with Glacial acetic acid and anhydrous sodium acetate adjusting PH to 5.30-5.50., after 4 ℃ of sedimentations, get supernatant liquor, add 2% chitosan and sodium alginate, 2% calcium chloride and sodium polyphosphate flocculating settling, then get the ultrafiltration of supernatant liquor gel-filtration system and remove unnecessary deionized water, by concentrated egg yolk liquid 20 times of left and right, with 1N sodium hydroxide adjusting PH to 7.0-8.0, put-20 ℃ of preservations.
E.IgY purification step:
Resisting porphyromonas gingivalis and Fusobacterium nucleatum are mixed to the saturation ratio that specific IgY extract adds sulfuric acid amine to 55%, fully stir, room temperature leaves standstill 2 hours, then high speed centrifugation 10 minutes, get precipitation, to precipitate and use distilled water diluting to substance accumulated amount, add again the saturation ratio of sulfuric acid amine to 33%, fully stir, room temperature leaves standstill 2 hours, then high speed centrifugation 10 minutes, get precipitation, after dissolving with distilled water diluting, pack dialysis tubing into, with distilled water through hollow fiber filter desalination, obtain resisting porphyromonas gingivalis and Fusobacterium nucleatum and mix the direct sterling of specific IgY antibody.
Resisting porphyromonas gingivalis and Fusobacterium nucleatum mix specific IgY antibody preparation sterling and measure through ELISA method, tiring as 1:32000/mg albumen of specific IgY antibody in resisting porphyromonas gingivalis and Fusobacterium nucleatum mixing specific IgY sterling in every milligram of albumen, or claim resisting porphyromonas gingivalis and Fusobacterium nucleatum to mix in specific IgY sterling, the specific activity of the specific IgY antibody in every milligram of albumen is 1:32000.In IgY antibody sterling, the purity of IgY antibody is determined as 89.3% through irreducibility SDS-PAGE.
Embodiment 3:
Get separately porphyromonas gingivalis thalline and bacterial chip as bacterium liquid; Prepare separately resisting porphyromonas gingivalis specific IgY extract.All the other are prepared with reference to embodiment 1.
Embodiment 4:
Get separately Fusobacterium nucleatum bacterial chip as bacterium liquid; The anti-Fusobacterium nucleatum specific IgY extract of preparation separately.All the other are prepared with reference to embodiment 1.
Experimental example 1:
The immunogen of two kinds of bacteria combination such as porphyromonas gingivalis, Fusobacterium nucleatum is by method immunity bird inlay of the present invention, get immune latter 30 days for the first time immune egg, and prepare by method of the present invention the extract that resisting porphyromonas gingivalis and Fusobacterium nucleatum mix specific IgY, application ELISA method is measured resisting porphyromonas gingivalis and Fusobacterium nucleatum and is mixed specific IgY in conjunction with the biological value of synantigen (in table 1) not:
Tiring of table 1 resisting porphyromonas gingivalis and anti-Fusobacterium nucleatum IgY antibody extract
Can be clear that according to above-mentioned experimental result:
(1). porphyromonas gingivalis, two kinds of bacterium thalline+bacterial chip immunogen immune bird inlays of Fusobacterium nucleatum, can produce the specific IgY antibody of anti-two kinds of bacteriums respectively;
. the specific IgY antibody to various bacteriums of generation tire not with immune bird inlay time every kind of bacterium dosage of using change, that is: be not that tiring of bacteriogenic IgY antibody that immunizing dose is large is also high, this is relevant to the immunogenicity of antigen;
(3). the hybrid antigen of resisting porphyromonas gingivalis and Fusobacterium nucleatum composite IgY antibody and porphyromonas gingivalis, Fusobacterium nucleatum can react, its additive effect of having tired;
(4). from above-mentioned experiment, a kind of resisting porphyromonas gingivalis and Fusobacterium nucleatum mix specific IgY and can prevent by above-mentioned bacterial periodontopathy, halitosis etc.;
(5). also can mix specific IgY with a kind of resisting porphyromonas gingivalis and Fusobacterium nucleatum simultaneously and prevent by any bacterial periodontopathy, halitosis etc. in above-mentioned bacterium;
(6). can also expand to thus by having bacterial periodontopathy, the halitosis etc. of common antigenic determinant with above-mentioned bacterium, can use resisting porphyromonas gingivalis of the present invention and Fusobacterium nucleatum to mix specific IgY prevention.
Experimental example 2:
Porphyromonas gingivalis, the former immune bird inlay of method of the present invention of pressing of Fusobacterium nucleatum mixed immunity, get the egg of the immune hen of immune latter 60 days for the first time, and prepare resisting porphyromonas gingivalis and Fusobacterium nucleatum mixing specific IgY antibody preparation sterling by method of the present invention, application ELISA method measures resisting porphyromonas gingivalis and Fusobacterium nucleatum mixes the not biological value of synantigen (in table 2) of specific IgY combination:
Tiring of table 2 resisting porphyromonas gingivalis and anti-Fusobacterium nucleatum IgY antibody sterling
Can be clear that according to above-mentioned experimental result:
(1). prepare sterling that resisting porphyromonas gingivalis and Fusobacterium nucleatum mix specific IgY than preparing by method of the present invention specific binding porphyromonas gingivalis and the Fusobacterium nucleatum hybrid antigen that resisting porphyromonas gingivalis and Fusobacterium nucleatum mix the extract of specific IgY in experimental example 1 by method of the present invention, and the bacterium such as porphyromonas gingivalis, Fusobacterium nucleatum separately the biological value of antigen all want high, be to increase in proportion substantially;
(2). the former immune bird inlay of porphyromonas gingivalis and Fusobacterium nucleatum mixed immunity, can produce the specific IgY antibody of anti-two kinds of bacteriums respectively;
. the specific IgY antibody to various bacteriums of generation tire not with immune bird inlay time every kind of bacterium dosage of using change, that is: be not that bacterium that immunizing dose is large produces tiring of IgY antibody high yet, this is relevant to the immunogenicity of antigen;
(4). the hybrid antigen of two kinds of bacterium such as the compound specific IgY of resisting porphyromonas gingivalis and Fusobacterium nucleatum and porphyromonas gingivalis, Fusobacterium nucleatum can react, and tiring of total specific IgY antibody also increased, its additive effect of having tired;
(5). from above-mentioned experiment, the compound specific IgY of a kind of resisting porphyromonas gingivalis and Fusobacterium nucleatum can prevent by above-mentioned bacterial periodontopathy and halitosis thereof etc.;
(6). also can prevent by any bacterial periodontopathy, halitosis etc. in above-mentioned bacterium with a kind of resisting porphyromonas gingivalis and the compound specific IgY of Fusobacterium nucleatum simultaneously;
(7). can also expand to thus by having bacterial periodontopathy, the halitosis etc. of common antigenic determinant with above-mentioned bacterium, can use the compound specific IgY prevention of resisting porphyromonas gingivalis of the present invention and Fusobacterium nucleatum.
Experimental example 3:
The activity of resist gingivitis and halitosis IgY collutory thereof is tired
With the IgY collutory of the resist gingivitis after protective material and halitosis thereof ((instant RT) and do not add protectant resist gingivitis and halitosis thereof IgY collutory (result of tiring of (RT immediately) is as table 3:
Table 3
Obviously the IgY collutory with the resist gingivitis after protective material and halitosis thereof that this patent provides can effectively be protected the activity of resist gingivitis IgY in collutory.
Experimental example 4:
Bacteriostatic test
Porphyromonas gingivalis anaerobism is cultivated 48h, after identifying, several colonies typicals is suspended in to BHI substratum, and adjusting bacterial concentration is (2 × 10
8) CPU/L～(1 × 10
9) CPU/L, IgY is diluted with aseptic BHI substratum, adjust the unicellular bacterium IgY of resisting porphyromonas final concentration and be 5.0,1.0,0.1g/L.
Get the unicellular bacterium 100 μ l of porphyromonas and be placed in 96 well culture plates, then add the unicellular bacterium IgY100 of each concentration group resisting porphyromonas μ l, every group of 3 holes, blank hole does not add bacterium liquid and only adds BHI substratum.96 well culture plates are cultivated to 24h, 72h in 37 ℃, anaerobism.
A pH-value determination pH, gets anaerobism and cultivates 24h, 72h and obtain 96 well culture plates, surveys A by microplate reader
490, read absorbance (A).
Statistical procedures is used t check.
Result
Resisting porphyromonas gingivalis IgY extracts
The purity of the resisting porphyromonas gingivalis IgY of the different batches that application water dilution method extracts reaches 31.4～34.6%, reaches 1:800 in conjunction with tiring of porphyromonas gingivalis.
The unicellular bacterium IgY of resisting porphyromonas purifying, the IgY purity of 55% saturated ammonium sulphate of different batches reaches 58.3%～61.2%, the 33% saturated ammonium sulphate further IgY purity of precipitation reaches 87.6%～89.1%, and the tiring of the unicellular bacterium of combination porphyromonas of 33% saturated ammonium sulphate purifying reaches 1:3200.
The unicellular bacterium IgY of resisting porphyromonas bacteriostasis rate is in table 4.
The inhibiting rate of the resisting porphyromonas gingivalis IgY of table 4. purifying to porphyromonas gingivalis
As can be seen from Table 4, resisting porphyromonas gingivalis IgY has remarkable effect to suppressing the unicellular bacteria growing rate of porphyromonas.
The inhibiting rate of the anti-Fusobacterium nucleatum IgY of purifying to Fusobacterium nucleatum
Tool nucleic acid fusobacterium anaerobism is cultivated 48h, after identifying, several colonies typicals is suspended in to BHI substratum, and adjusting bacterial concentration is (2 × 10
8) CPU/L～(1 × 10
9) CPU/L, IgY is diluted with aseptic BHI substratum, adjust anti-tool nucleic acid fusobacterium IgY final concentration and be 2.0,1.0,0.1g/L.
Get tool nucleic acid fusobacterium 100 μ l and be placed in 96 well culture plates, then add the anti-tool nucleic acid of each concentration group fusobacterium IgY100 μ l, every group of 3 holes, blank hole does not add bacterium liquid and only adds BHI substratum.96 well culture plates are cultivated to 24h, 72h in 37 ℃, anaerobism.
A pH-value determination pH, gets anaerobism and cultivates 24h, 72h and obtain 96 well culture plates, surveys A by microplate reader
490, read absorbance (A).
Statistical procedures is used t check.
Result
Anti-Fusobacterium nucleatum IgY extracts
The purity of the anti-tool nucleic acid fusobacterium IgY of the different batches that application water dilution method extracts reaches 31.4～34.6%, reaches 1:25600 in conjunction with tiring of Fusobacterium nucleatum.
The unicellular bacterium IgY of resisting porphyromonas purifying, the IgY purity of 55% saturated ammonium sulphate of different batches reaches 58.3%～61.2%, the 33% saturated ammonium sulphate further IgY purity of precipitation reaches 87.6%～89.1%, and the tiring of combination tool nucleic acid fusobacterium of 33% saturated ammonium sulphate purifying reaches 1:51200.
The anti-Fusobacterium nucleatum IgY of purifying to the inhibiting rate of Fusobacterium nucleatum in table 5
Table 5
As can be seen from Table 5, anti-Fusobacterium nucleatum IgY has remarkable effect to suppressing Fusobacterium nucleatum growth rate.
Claims (2)
1. a prevention of gingivitis of preparing with resisting porphyromonas gingivalis and Fusobacterium nucleatum specific IgY antibody and the IgY collutory of halitosis thereof, it is characterized in that, formed by anti-Periodontal Pathogens IgY preparation and collutory base-material, described anti-Periodontal Pathogens IgY preparation is by being made up of the materials in percentage by mass that accounts for collutory total mass: the compound specific IgY antibody preparation 0.01～0.5% of resisting porphyromonas gingivalis and Fusobacterium nucleatum, Tubulicid 0.01-2.0%, glycerine 1.0-30.0%, polyvinylpyrrolidone 0.1-2.0%, wherein Tubulicid, glycerine, polyvinylpyrrolidone forms resisting porphyromonas gingivalis and the compound specific IgY antibody preparation of Fusobacterium nucleatum protective material.
2. the IgY collutory of prevention of gingivitis as claimed in claim 1 and halitosis thereof, its spy is, described collutory base-material is made up of the materials in percentage by mass that accounts for collutory total mass:
Glycerine 1.0-10.0%, sorbyl alcohol 10-20%, propylene glycol 1.0-5.0%, Xylitol 1.0-5.0%, polysorbas20 1.0-3.0%, tween 80 1.0-3.0%, CrmopherRH400.5-3.0%, PVM/MA polymkeric substance 0.6-2.0%, Sodium Benzoate 0.5-1.5%, aspartame 0.1-0.5%, asccharin 0.1-0.3%, essence 0.01-0.8%, food dye is appropriate, deionized water surplus.
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| CN108728388A (en) * | 2018-07-24 | 2018-11-02 | 上海美加净日化有限公司 | Porphyromonas gingivalis, resisting porphyromonas gingivalis specific IgY preparation and Compound formula of toothpaste |
| CN110128541A (en) * | 2019-04-16 | 2019-08-16 | 广东工业大学 | A kind of preparation method and application of Yolk antibody that preventing and treating chronic periodontitis |
| CN110357955B (en) * | 2019-06-24 | 2023-05-12 | 桂林三金药业股份有限公司 | Composite specificity egg yolk antibody and preparation method and application thereof |
| CN114052124A (en) * | 2021-11-26 | 2022-02-18 | 北京中科昊盈生物技术有限公司 | Composition, preparation and application thereof |
Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1482141A (en) * | 2003-08-15 | 2004-03-17 | 雅臣药业集团（远东）有限公司 | Method for preparing specific composite IgY against periodontal disease and group system thereof |
| CN1504179A (en) * | 2002-10-17 | 2004-06-16 | 上海白猫股份有限公司 | Non-fluorin anti-caries safety toothpaste of double cavity tube and its process |
-
2012
- 2012-10-18 CN CN201210396508.9A patent/CN102860932B/en active Active
Patent Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1504179A (en) * | 2002-10-17 | 2004-06-16 | 上海白猫股份有限公司 | Non-fluorin anti-caries safety toothpaste of double cavity tube and its process |
| CN1482141A (en) * | 2003-08-15 | 2004-03-17 | 雅臣药业集团（远东）有限公司 | Method for preparing specific composite IgY against periodontal disease and group system thereof |
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