CN102860932A - 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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- CN102860932A CN102860932A CN2012103965089A CN201210396508A CN102860932A CN 102860932 A CN102860932 A CN 102860932A CN 2012103965089 A CN2012103965089 A CN 2012103965089A CN 201210396508 A CN201210396508 A CN 201210396508A CN 102860932 A CN102860932 A CN 102860932A
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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 the oral-cavity article technical field, particularly the prevention of the anti-Fusobacterium nucleatum of a kind of usefulness and porphyromonas gingivalis specific IgY antibody preparation is by the collutory of these two kinds of microbial gingivitiss and halitosis thereof.
Background technology
China's gingiva, periodontal sickness rate account for more than 70% of population, porphyromonas gingivalis (Porphyromonas gingivalis) is the main suspicious pathogenic bacterium of the periodontal disease 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 lysine 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 do not have effectively to treat the method for halitosis, use Antibiotics Drug therapy halitosis that a lot of drawbacks are arranged.
Setting about from main cause of disease bacterium porphyromonas gingivalis and Fusobacterium nucleatum for prevention and treatment periodontal disease and halitosis thereof is key point.
The collutory that is used at present prevention and treatment gingivitis is generally 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 and uses clinically specific IgY antibody caries prevention and treatment infant rotavirus diarrhoea, prevention pulmonary fibrosis patients charrin's disease, and it is remarkable to alleviate the microbial stomachache effect of helicobacter pylorus, and has no side effect and addiction.
Find that in the IgY of the numerous species that we study the periodontal disease that has the experiment proved that trend and the curative effect that does not have or do not have significantly to suppress periodontal disease and halitosis thereof because of the IgY antibody that the bacterial immunity hen produces.Briefly, not every IgY has obvious therapeutical effect.
Because the suspicious cause of disease bacterium of periodontal disease 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 bacteroid, capnocytophaga gingivalis etc.When the IgY that explores the preparation correspondence was used for the treatment of periodontal disease and halitosis thereof, we inferred that porphyromonas gingivalis, Fusobacterium nucleatum, companion actinomycetes Actinobacillus, the treponema dentium of dwelling, Fu Shi bacteroid, capnocytophaga gingivalis all might cause periodontal disease and halitosis thereof; Because our prior unpredictable which bacterium in the middle of this is the principal element that causes periodontal disease and halitosis thereof, which bacterium is to cause that periodontal disease and halitosis thereof are secondary causes.
We think must have the test of suitable anti-periodontal disease and halitosis thereof to determine which bacterium is the principal element that causes periodontal disease and halitosis thereof.Thereby medicine or the health product of the standby anti-periodontal disease of guidance system and halitosis thereof.
Present prior art does not have relevant prompting guidance technology personnel to find that resisting porphyromonas gingivalis, Fusobacterium nucleatum composite IgY are the keys that solves treatment periodontal disease and halitosis thereof.
The technical staff can't in advance under the prerequisite of the experiment of not doing anti-periodontal disease and halitosis thereof, learn that in advance porphyromonas gingivalis, Fusobacterium nucleatum are the principal elements that causes periodontal disease and halitosis thereof.
The technical staff can't be in advance under the prerequisite of the experiment of not doing anti-periodontal disease 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 the collutory of the prevention of the anti-Fusobacterium nucleatum of a kind of usefulness and the preparation of porphyromonas gingivalis specific IgY antibody by these two kinds of microbial gingivitiss and halitosis thereof for the existing not good problem of periodontal disease medicine collutory therapeutic effect.
Technical problem to be solved by this invention can be achieved through the following technical solutions:
A kind of with resisting porphyromonas gingivalis and the prevention of gingivitis of Fusobacterium nucleatum specific IgY antibody preparation 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 comprised of the materials in percentage by mass that accounts for the collutory gross mass: resisting porphyromonas gingivalis and Fusobacterium nucleatum mix specific IgY antibody preparation 0.01～0.5%; hibitane 0.01-2.0%; glycerol 1.0-30.0%; polyvinylpyrrolidone 0.1-2.0%, wherein hibitane; glycerol; polyvinylpyrrolidone consists of resisting porphyromonas gingivalis and the compound specific IgY antibody preparation of Fusobacterium nucleatum protective agent.
In a preferred embodiment of the invention, described collutory base material is comprised of the materials in percentage by mass that accounts for the collutory gross mass:
Glycerol 1.0-10.0%, sorbitol 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 polymer 0.6-2.0%, sodium benzoate 0.5-1.5%, aspartame 0.1-0.5%, glucide 0.1-0.3%, essence 0.01-0.8%, food coloring is an amount of, the deionized water surplus.
The present invention is used for the collutory of prevention of periodontal disease and halitosis thereof, and prevention of periodontal disease and halitosis are had a significant effect.
The specific embodiment
Describe implementation of the present invention in detail below in conjunction with embodiment.The related strain of following embodiment is all provided by Medical College, Shanghai Communication Univ..
Embodiment 1:
The preparation of the compound specific IgY antibody preparation of resisting porphyromonas gingivalis and Fusobacterium nucleatum:
A. porphyromonas gingivalis and Fusobacterium nucleatum incubation 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 the porphyromonas of BHI-S culture medium separation and Culture and amplification cultivation, Fusobacterium nucleatum, and confirm as porphyromonas gingivalis, Fusobacterium nucleatum through evaluation and obtain porphyromonas gingivalis thalline and bacterial chip, Fusobacterium nucleatum thalline and bacterial chip;
B. immunogen preparation process:
B.1 get porphyromonas gingivalis thalline and bacterial chip 10～70 weight portions, Fusobacterium nucleatum thalline and bacterial chip 30～90 weight portions are mixed into bacterium liquid antigen;
B.2 isopyknic bacterium liquid antigen is mixed with isopyknic Fu Shi Freund's complete adjuvant, use the high speed agitator stirring and emulsifying, it is former finally to make Water-In-Oil Fu Shi Freund's complete adjuvant porphyromonas gingivalis and Fusobacterium nucleatum mixed immunity; Every milliliter of Fu Shi Freund's complete adjuvant mixed immunity is former to contain antibacterial 1,000,000,000-8,000,000,000;
B.3 isopyknic mixed bacteria liquid is mixed with isopyknic freund 's incomplete adjuvant, use the high speed agitator stirring and emulsifying, finally make Water-In-Oil freund 's incomplete adjuvant porphyromonas gingivalis and the Fusobacterium nucleatum mixed immunity is former, contain antibacterial 1,000,000,000-8,000,000,000 during every milliliter of freund 's incomplete adjuvant mixed immunity is former;
C. immune step:
Use syringe with the former subcutaneous and intramuscular injection of Fu Shi Freund's complete adjuvant mixed immunity to bird inlay, then every bird inlay injection 0.5ml-3.0ml immunogen uses the subcutaneous and intramuscular injection of the former 0.5ml-3.0ml of freund 's incomplete adjuvant mixed immunity to bird inlay week about.Behind the booster immunization secondary, after this every re-using the subcutaneous and intramuscular injection of the former 0.5ml-3.0ml of freund 's incomplete adjuvant mixed immunity ten weeks to bird inlay booster immunization five times, altogether immunity eight times of front and back;
D. resisting porphyromonas gingivalis and Fusobacterium nucleatum mix the preparation of specific IgY extract:
Collect for the first time rear the 28th day immune hen egg of immunity of immunity, the ethanol of use 75% carries out the egg surface sterilization, open eggshell and get egg yolk, remove membrane of yolk and get egg yolk liquid, with 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 acid-base value to pH5.30-5.50, then-20 ℃ of cold storage of placement were taken out and are thawed in 3 days afterwards, get supernatant, with concentrated 20 times of egg yolk liquid, regulate PH to 7.0-8.0 with 1N NaOH with the gel filtration system, put-20 ℃ of preservations.
The E.IgY purification step:
Resisting porphyromonas gingivalis and Fusobacterium nucleatum are mixed the saturation that the specific IgY extract adds sulphuric acid amine to 55%, fully stir, room temperature left standstill 2 hours, then high speed centrifugation is 10 minutes, get precipitation, to precipitate and use distilled water diluting to the substance accumulated amount, the saturation that adds again sulphuric acid amine to 33%, fully stir, room temperature left standstill 2 hours, and then high speed centrifugation is 10 minutes, get precipitation, with the bag filter of packing into after the distilled water diluting dissolving,, namely get resisting porphyromonas gingivalis and Fusobacterium nucleatum and mix the direct sterling of specific IgY antibody through the hollow fiber filter desalination with distilled water.
Resisting porphyromonas gingivalis and Fusobacterium nucleatum mix specific IgY antibody preparation sterling and measure through the ELISA method, the tiring of specific IgY antibody that resisting porphyromonas gingivalis and Fusobacterium nucleatum mix in the specific IgY sterling in every milligram of albumen is 1:32000/mg albumen, perhaps claim resisting porphyromonas gingivalis and Fusobacterium nucleatum to mix in the 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 incubation 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 the porphyromonas of BHI-S culture medium separation and Culture and amplification cultivation, Fusobacterium nucleatum, and confirm as porphyromonas gingivalis, Fusobacterium nucleatum through evaluation and obtain porphyromonas gingivalis thalline and bacterial chip, Fusobacterium nucleatum thalline and bacterial chip;
B. immunogen preparation process:
B.1 get porphyromonas gingivalis thalline and bacterial chip 10～70 weight portions, Fusobacterium nucleatum thalline and bacterial chip 30～90 weight portions are mixed into bacterium liquid antigen;
B.2 isopyknic bacterium liquid antigen is mixed with isopyknic Fu Shi Freund's complete adjuvant, use the high speed agitator stirring and emulsifying, it is former finally to make Water-In-Oil Fu Shi Freund's complete adjuvant porphyromonas gingivalis and Fusobacterium nucleatum mixed immunity; Every milliliter of Fu Shi Freund's complete adjuvant mixed immunity is former to contain antibacterial 1,000,000,000-8,000,000,000;
B.3 isopyknic mixed bacteria liquid is mixed with isopyknic freund 's incomplete adjuvant, use the high speed agitator stirring and emulsifying, finally make Water-In-Oil freund 's incomplete adjuvant porphyromonas gingivalis and the Fusobacterium nucleatum mixed immunity is former, contain antibacterial 1,000,000,000-8,000,000,000 during every milliliter of freund 's incomplete adjuvant mixed immunity is former;
C. immune step:
Use syringe with the former subcutaneous and intramuscular injection of Fu Shi Freund's complete adjuvant mixed immunity to bird inlay, then every bird inlay injection 0.5ml-3.0ml immunogen uses the subcutaneous and intramuscular injection of the former 0.5ml-3.0ml of freund 's incomplete adjuvant mixed immunity to bird inlay week about.Behind the booster immunization secondary, after this every re-using the subcutaneous and intramuscular injection of the former 0.5ml-3.0ml of freund 's incomplete adjuvant mixed immunity ten weeks to bird inlay booster immunization five times, altogether immunity eight times of front and back;
D. resisting porphyromonas gingivalis and Fusobacterium nucleatum mix the preparation of specific IgY extract:
Collect for the first time rear the 28th day immune hen egg of immunity of immunity, with 75% alcohol disinfecting egg shell, smash eggshell, leave and take egg yolk.Behind 10 times of dilutions of deionized water egg yolk, regulate PH to 5.30-5.50. with glacial acetic acid and anhydrous sodium acetate, get supernatant after 4 ℃ of sedimentations, add 2% chitosan and sodium alginate, 2% calcium chloride and sodium polyphosphate flocculating sedimentation are got supernatant again and are removed unnecessary deionized water with the ultrafiltration of gel filtration system, with concentrated about 20 times of egg yolk liquid, regulate PH to 7.0-8.0 with the 1N sodium hydroxide, put-20 ℃ of preservations.
The E.IgY purification step:
Resisting porphyromonas gingivalis and Fusobacterium nucleatum are mixed the saturation that the specific IgY extract adds sulphuric acid amine to 55%, fully stir, room temperature left standstill 2 hours, then high speed centrifugation is 10 minutes, get precipitation, to precipitate and use distilled water diluting to the substance accumulated amount, the saturation that adds again sulphuric acid amine to 33%, fully stir, room temperature left standstill 2 hours, and then high speed centrifugation is 10 minutes, get precipitation, with the bag filter of packing into after the distilled water diluting dissolving,, namely get resisting porphyromonas gingivalis and Fusobacterium nucleatum and mix the direct sterling of specific IgY antibody through the hollow fiber filter desalination with distilled water.
Resisting porphyromonas gingivalis and Fusobacterium nucleatum mix specific IgY antibody preparation sterling and measure through the ELISA method, the tiring of specific IgY antibody that resisting porphyromonas gingivalis and Fusobacterium nucleatum mix in the specific IgY sterling in every milligram of albumen is 1:32000/mg albumen, perhaps claim resisting porphyromonas gingivalis and Fusobacterium nucleatum to mix in the 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 preparations are with reference to embodiment 1.
Embodiment 4:
Get separately the Fusobacterium nucleatum bacterial chip as bacterium liquid; Prepare separately anti-Fusobacterium nucleatum specific IgY extract.All the other preparations are 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 for the first time rear 30 days immune egg of immunity, and prepare the extract that resisting porphyromonas gingivalis and Fusobacterium nucleatum mix specific IgY by method of the present invention, use the ELISA method and measure resisting porphyromonas gingivalis and Fusobacterium nucleatum and mix specific IgY in conjunction with the biological value of synantigen (seeing 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:
⑴. porphyromonas gingivalis, two kinds of bacterium thalline of Fusobacterium nucleatum+bacterial chip immunogen immune bird inlay can produce the respectively specific IgY antibody of anti-two kinds of antibacterials;
. generation to the specific IgY antibody of various antibacterials tire not with immune bird inlay the time every kind of antibacterial dosage of using change, that is: be not that tiring of the large bacteriogenic IgY antibody of immunizing dose is also high, this is relevant with the immunogenicity of antigen;
⑶. the hybrid antigen of resisting porphyromonas gingivalis and Fusobacterium nucleatum composite IgY antibody and porphyromonas gingivalis, Fusobacterium nucleatum can both react, its superposition of having tired;
⑷. by above-mentioned experiment as seen, a kind of resisting porphyromonas gingivalis and Fusobacterium nucleatum mix specific IgY and can prevent by above-mentioned bacterial periodontal disease, halitosis etc.;
⑸. also can mix the specific IgY prevention by any bacterial periodontal disease in the above-mentioned antibacterial, halitosis etc. with a kind of resisting porphyromonas gingivalis and Fusobacterium nucleatum simultaneously;
⑹. can also expand to thus by with above-mentioned antibacterial the bacterial periodontal disease, halitosis etc. of common antigenic determinant being arranged, can use resisting porphyromonas gingivalis of the present invention and Fusobacterium nucleatum to mix the 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 rear 60 days immune hen of for the first time immunity, and prepare resisting porphyromonas gingivalis and Fusobacterium nucleatum by method of the present invention and mix specific IgY antibody preparation sterling, use the ELISA method and measure resisting porphyromonas gingivalis and Fusobacterium nucleatum and mix specific IgY in conjunction with the biological value of synantigen (seeing Table 2) not:
Tiring of table 2 resisting porphyromonas gingivalis and anti-Fusobacterium nucleatum IgY antibody sterling
Can be clear that according to above-mentioned experimental result:
⑴. prepare sterling that resisting porphyromonas gingivalis and Fusobacterium nucleatum mix specific IgY than preparing specific binding porphyromonas gingivalis and the Fusobacterium nucleatum hybrid antigen that resisting porphyromonas gingivalis and Fusobacterium nucleatum mix the extract of specific IgY by method of the present invention in the experimental example 1 by method of the present invention, and the antibacterials such as porphyromonas gingivalis, Fusobacterium nucleatum separately the biological value of antigen all want high, be to increase in proportion substantially;
⑵. the former immune bird inlay of porphyromonas gingivalis and Fusobacterium nucleatum mixed immunity can produce the respectively specific IgY antibody of anti-two kinds of antibacterials;
. generation to the specific IgY antibody of various antibacterials tire not with immune bird inlay the time every kind of antibacterial dosage of using change, that is: it is also high not to be that the large antibacterial of immunizing dose produces tiring of IgY antibody, this is relevant with the immunogenicity of antigen;
⑷. 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 both react, and tiring of total specific IgY antibody also increased its superposition of having tired;
⑸. by above-mentioned experiment as seen, the compound specific IgY of a kind of resisting porphyromonas gingivalis and Fusobacterium nucleatum can prevent by above-mentioned bacterial periodontal disease and halitosis thereof etc.;
⑹. also can prevent by any bacterial periodontal disease in the above-mentioned antibacterial, halitosis etc. with a kind of resisting porphyromonas gingivalis and the compound specific IgY of Fusobacterium nucleatum simultaneously;
⑺. can also expand to thus by with above-mentioned antibacterial the bacterial periodontal disease, halitosis etc. of common antigenic determinant being arranged, 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 the protective agent and halitosis thereof ((instant RT) and do not add protectant resist gingivitis and halitosis thereof the IgY collutory (tire result such as the table 3 of (immediately RT):
Table 3
The IgY collutory with the resist gingivitis after the protective agent and halitosis thereof that obvious this patent provides can effectively be protected the activity of resist gingivitis IgY in collutory.
Experimental example 4:
Bacteriostatic test
The porphyromonas gingivalis anaerobism is cultivated 48h, after identifying several colonies typicals is suspended in the BHI culture medium, and adjusting bacterial concentration is (2 * 10
8) CPU/L～(1 * 10
9) CPU/L, with aseptic BHI culture medium IgY is diluted, 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 place 96 well culture plates, add again the unicellular bacterium IgY100 of each concentration group resisting porphyromonas μ l, every group of 3 holes, the blank hole does not add bacterium liquid and only adds the BHI culture medium.96 well culture plates are cultivated 24h, 72h in 37 ℃, anaerobism.
The A pH-value determination pH is got anaerobism cultivation 24h, 72h and is got 96 well culture plates, surveys A with microplate reader
490, read absorbance (A).
Statistical procedures is used the t check.
The result
Resisting porphyromonas gingivalis IgY extracts
The purity of the resisting porphyromonas gingivalis IgY of the different batches that the 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 purification, the IgY purity of 55% saturated ammonium sulphate of different batches reaches 58.3%～61.2%, the 33% saturated ammonium sulfate further IgY purity of precipitation reaches 87.6%～89.1%, 33% saturated ammonium sulfate purification reach 1:3200 in conjunction with tiring of the unicellular bacterium of porphyromonas.
The unicellular bacterium IgY of resisting porphyromonas bacteriostasis rate sees Table 4.
The resisting porphyromonas gingivalis IgY of table 4. purification is to the suppression ratio of 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 anti-Fusobacterium nucleatum IgY of purification is to the suppression ratio of Fusobacterium nucleatum
Tool nucleic acid Fusobacterium anaerobism is cultivated 48h, after identifying several colonies typicals is suspended in the BHI culture medium, and adjusting bacterial concentration is (2 * 10
8) CPU/L～(1 * 10
9) CPU/L, with aseptic BHI culture medium IgY is diluted, 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 place 96 well culture plates, add again the anti-tool nucleic acid of each concentration group Fusobacterium IgY100 μ l, every group of 3 holes, the blank hole does not add bacterium liquid and only adds the BHI culture medium.96 well culture plates are cultivated 24h, 72h in 37 ℃, anaerobism.
The A pH-value determination pH is got anaerobism cultivation 24h, 72h and is got 96 well culture plates, surveys A with microplate reader
490, read absorbance (A).
Statistical procedures is used the t check.
The result
Anti-Fusobacterium nucleatum IgY extracts
The purity of the anti-tool nucleic acid Fusobacterium IgY of the different batches that the 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 purification, the IgY purity of 55% saturated ammonium sulphate of different batches reaches 58.3%～61.2%, the 33% saturated ammonium sulfate further IgY purity of precipitation reaches 87.6%～89.1%, 33% saturated ammonium sulfate purification reach 1:51200 in conjunction with tiring of tool nucleic acid Fusobacterium.
The anti-Fusobacterium nucleatum IgY of purification sees Table 5 to the suppression ratio of Fusobacterium nucleatum
Table 5
As can be seen from Table 5, anti-Fusobacterium nucleatum IgY has remarkable effect to suppressing the Fusobacterium nucleatum rate of growth.
Claims (2)
1. one kind with the prevention of gingivitis of resisting porphyromonas gingivalis and Fusobacterium nucleatum specific IgY antibody preparation 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 comprised of the materials in percentage by mass that accounts for the collutory gross mass: the compound specific IgY antibody preparation 0.01～0.5% of resisting porphyromonas gingivalis and Fusobacterium nucleatum; hibitane 0.01-2.0%; glycerol 1.0-30.0%; polyvinylpyrrolidone 0.1-2.0%, wherein hibitane; glycerol; polyvinylpyrrolidone consists of resisting porphyromonas gingivalis and the compound specific IgY antibody preparation of Fusobacterium nucleatum protective agent.
2. the IgY collutory of prevention of gingivitis as claimed in claim 1 and halitosis thereof, its spy is that described collutory base material is comprised of the materials in percentage by mass that accounts for the collutory gross mass:
Glycerol 1.0-10.0%, sorbitol 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 polymer 0.6-2.0%, sodium benzoate 0.5-1.5%, aspartame 0.1-0.5%, glucide 0.1-0.3%, essence 0.01-0.8%, food coloring is an amount of, the 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 |
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| 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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| CN104188816A (en) * | 2014-08-11 | 2014-12-10 | 中国人民解放军第四军医大学 | Mouth wash and preparation method thereof |
| CN106214510A (en) * | 2016-08-31 | 2016-12-14 | 广东工业大学 | A kind of anti-antozostomatic yolk antibody collutory and preparation method thereof |
| CN106467572A (en) * | 2016-08-31 | 2017-03-01 | 广东工业大学 | One species specificity composite yolk antibody and its preparation method and application |
| CN109381493A (en) * | 2017-08-09 | 2019-02-26 | 鼎赫生物科技股份有限公司 | For inhibiting or killing except porphyromonas gingivalis and/or turn streptococcic composition of sugar and application thereof |
| CN108272649A (en) * | 2018-04-02 | 2018-07-13 | 广东药科大学 | A kind of clarification gargle and preparation method thereof containing IgY |
| CN108272649B (en) * | 2018-04-02 | 2021-01-29 | 广东药科大学 | IgY-containing clear mouthwash and preparation method thereof |
| 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 |
| CN110357955A (en) * | 2019-06-24 | 2019-10-22 | 桂林三金药业股份有限公司 | A kind of compound special yolk antibody and its preparation method and application |
| 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 |
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