CN105717305A - Diagnosis method - Google Patents
Diagnosis method Download PDFInfo
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- CN105717305A CN105717305A CN201610123828.5A CN201610123828A CN105717305A CN 105717305 A CN105717305 A CN 105717305A CN 201610123828 A CN201610123828 A CN 201610123828A CN 105717305 A CN105717305 A CN 105717305A
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- enamel
- susceptibility
- concentration
- statherin
- corroded
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
- G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
- G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
- G01N33/68—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving proteins, peptides or amino acids
- G01N33/6803—General methods of protein analysis not limited to specific proteins or families of proteins
- G01N33/6806—Determination of free amino acids
- G01N33/6812—Assays for specific amino acids
Abstract
The invention discloses a method for identifying mammalians having improved tooth enamel erosion susceptibility, a kit for the method and relevant application and a method thereof.
Description
The application is the applying date be December in 2011 20 days, application number is 201180075691.6
(PCT/US2011/066078), denomination of invention is the divisional application of the application of " diagnostic method ".
Background of invention
Owing to the understanding corroded and affect being increased in dentist and patient, therefore there are the correct needs diagnosing the source corroded the qualification individual method to corroding the susceptibility having rising.Different from the dental caries owing to being exposed to the very deep sub-surface focus in antibacterial acid generation local for a long time, erosion is to be caused by following reason: due to short and be frequently exposed to low pH meals acid, the dissolving of whole enamel surface and softening, cause the loss of softening and micron order mineral, especially after brushing teeth with grinding agent toothpaste.Meals acid such as citric acid is particularly detrimental, is not only due to its acid pH, and due to its calcium sequestration capacity, it causes further dental enamel dissolution.
Acquired enamel pellicle (AEP) is attacked for adamantine acid, provides some protections natively.AEP comprises calcium ion, phosphate anion and protein and enzyme.The formation of pellicle is a complicated multi-step process, and wherein the albumen of precursor protein such as Statherin and proline rich is tied at first and merged into other albumen (mucin etc.) offer binding site.Maturation relates to the crosslinking of surface membrane protein matter and dephosphorylation and after 2 hours in the combination of later stage dental plaque.The composition of pellicle is highly dependent on saliva contents, and it changes with flow velocity, stimulation, the different time of every day, meals etc..Saliva flow rate greatly have impact on the pH of saliva, ion component and buffer capacity.
Corrode and may originate from several conditions-be frequently exposed to meals acid, gastric acid or lack enough salivas (xerostomia).The evaluation suffering from seriousness that the patient of gastric erosion damages usually by erosion and position (palate surface) is identified, suffers from those of xerostomia then by estimating saliva and measuring saliva flow rate and identify.Corrode and the intake of acidic beverages increases, there is contact between relatively low saliva flow rate and the relatively low saliva pH/ buffer capacity caused by relatively low saliva flow rate.But, some have normal saliva flow rate and the patient without gastric acid exposure sign still suffers from meals erosion damages, even if when being exposed to meals acid and being minimized, this shows the susceptibility corroded is raised, it is possible to relevant with its AEP.There are the needs of the simple and reliable method correctly to treat of the subgroup for identifying this height susceptible patient, and for identifying that patient reduces, alleviates and control the needs of the method corroded from the dentin that meals are sour with evaluation in clinical oral care active.
Invention summary
Unexpectedly, have been found that meals enamel is corroded the patients with high susceptibility goes out relative to the normal healthy controls group with similar saliva flow rate and meals, 1) significantly lower in whole saliva of buccal cavity mucin 5B, carbonic anhydrase 6 and/or higher Statherin level, and 2) total protein significantly lower in AEP, Statherin and/or calcium concentration.Utilizing the saliva based on total protein concentration, calcium analysis and Identification of Fusion Protein and the diagnostic method of pellicle, these factors can be used for identifying the patient to corroding susceptible.Owing to test sample is saliva and saliva pellicle, sample collection is to be easy to and there is the abundant information relevant to albumen and content of mineral substances.
Therefore, some embodiments of the present invention provide for identifying the method for patient that enamel corrodes the susceptibility with rising, its comprise the following steps one or two:
(i) from patient obtain the sample of whole saliva of buccal cavity and detect described saliva in mucin 5B level, carbonic anhydrase 6 level and Statherin level one or more, wherein relative to control value, relatively low mucin 5B, relatively low carbonic anhydrase and/or higher Statherin level each represent that the susceptibility that enamel is corroded raises；With
(ii) from patient obtain the sample of acquired enamel pellicle (AEP) and measure in total protein concentration described AEP, Statherin concentration and calcium concentration one or more, wherein relative to control value, the reduced levels of these materials each each represents that the susceptibility that enamel is corroded raises.
In some embodiments, the cause of disease of described erosion is meals.
The detailed description that other field of present invention application will be provided below becomes apparent.It should be understood that detailed description and specific embodiment, when representing the preferred embodiments of the invention, the purpose being intended only to illustrate, and it is not intended to limit the scope of the present invention.
Detailed Description Of The Invention
Being described below of one or more preferred embodiments is exemplary only in itself, and is intended to restriction the scope of the present invention, its application or purposes never in any form.
Therefore some embodiments of the present invention provide for identifying the method (method 1) of patient that enamel corrodes the susceptibility with rising, including measuring following one or more, for instance following any 2,3,4 or 5 kind, for instance whole 6 kinds:
I () measures the concentration of mucin 5B in the sample of the whole saliva of buccal cavity (UWMS) such as not stimulated from the whole saliva of buccal cavity of patient, wherein relative to control value, the concentration of relatively low mucin 5B represents that the susceptibility that enamel is corroded raises；
(ii) in the sample of the whole saliva of buccal cavity (UWMS) such as not stimulated from the whole saliva of buccal cavity of patient, measure the concentration of carbonic anhydrase 6, wherein relative to control value, the concentration of relatively low carbonic anhydrase 6 represents that the susceptibility that enamel is corroded raises；
(iii) in the sample of the whole saliva of buccal cavity (UWMS) such as not stimulated from the whole saliva of buccal cavity of patient, measure the concentration of Statherin, wherein relative to control value, the concentration of higher Statherin represents that the susceptibility that enamel is corroded raises；
(iv) in the concentration measuring total protein in acquired enamel pellicle (AEP) sample of patient, wherein relative to control value, relatively low total protein concentration represents that the susceptibility that enamel is corroded raises；
V (), measuring the concentration of Statherin in acquired enamel pellicle (AEP) sample of patient, wherein relative to control value, the concentration of relatively low Statherin represents that the susceptibility that enamel is corroded raises；With
(vi) in the concentration measuring calcium in acquired enamel pellicle (AEP) sample of patient, wherein relative to control value, the concentration of relatively low calcium represents that the susceptibility that enamel is corroded raises.
Such as, the invention provides
1.1 methods 1, wherein patient's fasting or taboo before collecting sample is drunk water at least one hour, for instance at least two hours.
1.2 methods 1 or method 1.1, wherein when patients go out in following standard one or more time, described patient is accredited as the susceptibility that adamantine erosion has rising:
1.3 methods 1.2, are wherein accredited as the patient of the susceptibility that adamantine erosion has rising and meet in method 1.2 at least two of judging standard.
In 1.4 preceding methods any one, described method includes mucin 5B, carbonic anhydrase and the Statherin of measuring in whole saliva of buccal cavity.
In 1.5 preceding methods any one, wherein by hydroxyapatite or dentium nitor mass are placed in patient oral cavity, it is fixed on palate mouth pallet/clamping plate, wears 0.5-2 hour such as about 1 hour, and after removing, from block, collect AEP from the oral cavity of patient, gather the AEP sample of patient.
In 1.6 preceding methods any one, farther include to treat the step of the patient being accredited as the susceptibility that adamantine erosion is had rising, including giving patient or instructing patient to give effective therapeutic agent or product to key, to control enamel or Dentinal erosion, described therapeutic agent or product include following one or more:
A () fluorine processes thing, selected from fluorine-containing collutory and the toothpaste containing high concentration fluorine；
The toothpaste of (b) phosphoric acid calcium or calcium grinding agent such as calcium pyrophosphate, Tri-Compress or calcium carbonate；(c) oral care product, for instance comprise collutory or the dentifrice of the L-arginine being free or salt form；
(d) dentin adhesive or to the varnish exposing Dentinal region；
E () comprises the oral care product of stannous ion source；
F () comprises the oral care product of calcium ion.
Some embodiments of the present invention further provide a kind of for diagnosing the test kit that the susceptibility that enamel is corroded raises, and it includes for measuring the instrument of one or more concentration in following albumen: mucin 5B, carbonic anhydrase 6 and Statherin；Such as include the antibody for described albumen and operation instructions.
Present invention also offers and preparing for diagnosing the purposes in the test kit that the susceptibility that enamel is corroded raises for one or more antibody in mucin 5B, carbonic anhydrase 6 and Statherin.
In some embodiments, present invention provide for identifying the mammiferous method that enamel corrodes the susceptibility with rising, comprising: a. obtains the sample of whole saliva of buccal cavity from mammiferous oral cavity；B. one or more concentration in following biomarker is measured: i) mucin 5B；Ii) carbonic anhydrase 6；And iii) Statherin；And the concentration of described mucin B, the concentration of carbonic anhydrase 6 and/or the concentration of Statherin are compared with the respective control value of described biomarker；Wherein said control value is taken from not suffering from the concentration of biomarker described in the whole saliva of buccal cavity sample in the mammiferous oral cavity that enamel corrodes；And the wherein control value relative to mucin 5B, relatively low mucin 5B concentration represents that the susceptibility that enamel is corroded raises；Wherein relative to the control value of carbonic anhydrase 6, relatively low carbonic anhydrase 6 concentration represents that the susceptibility that enamel is corroded raises；Wherein relative to the control value of Statherin, higher Statherin concentration represents that the susceptibility that enamel is corroded raises.In some embodiments, the cause of disease that described enamel corrodes is meals.
In some embodiments, described whole saliva of buccal cavity is the whole saliva of buccal cavity not stimulated.In some embodiments, described whole saliva of buccal cavity is through stimulating.
In some embodiments, when mammal show in following standard one or more time, described mammal is accredited as the susceptibility that adamantine erosion has rising:
In some embodiments, based at least two shown in following standard, described mammal is accredited as the susceptibility that adamantine erosion has rising:
Some of the present invention embodiment, provides following methods, it farther includes to treat the mammiferous step being accredited as the susceptibility that adamantine erosion has rising, and wherein said step includes the agent preventing or reducing dentin or enamel erosion or the product that give described mammiferous oral cavity effective dose.
In some embodiments, described prevention or minimizing dentin or enamel are corroded agent or product are selected from: fluorine-containing collutory；Toothpaste containing high concentration fluorine；The toothpaste of phosphoric acid calcium or calcium grinding agent such as calcium pyrophosphate, Tri-Compress or calcium carbonate；Comprise the oral care composition of the L-arginine being free or salt form；Dentin adhesive or varnish；Comprise the oral care composition of stannous ion source；And the combination of two or more.
Some embodiments provide a kind of for diagnosing the test kit that the susceptibility in mammal, enamel corroded raises, and it includes the instrument for measuring one or more concentration in mucin 5B, carbonic anhydrase 6 and Statherin in the whole saliva of buccal cavity sample adopted from mammiferous oral cavity and operation instructions.
In some embodiments, described mammal is selected from people, felid and Canis animals.In some embodiments, described mammal is behaved.
Some embodiments of the present invention provide to be prepared for diagnosing the purposes in the test kit that the susceptibility in mammal, enamel corroded raises for one or more antibody in mucin 5B, carbonic anhydrase 6 and Statherin.
Other embodiment of the present invention provides the mammiferous method for identifying the susceptibility that adamantine erosion has rising, comprising: a. obtains the sample of acquired enamel pellicle (AEP) from mammiferous oral cavity；B. one or more the concentration in following biomarker is measured: i) total protein；Ii) Statherin；And iii) calcium；And the concentration of total protein, the concentration of Statherin and/or the concentration of calcium are compared with the respective control value of described biomarker；Wherein said control value is taken from not suffering from the concentration of biomarker described in the AEP sample in mammiferous oral cavity that corrodes；And wherein relative to the control value of total protein, relatively low total protein concentration represents that the susceptibility that enamel is corroded raises；Wherein relative to the control value of Statherin, relatively low Statherin concentration represents that the susceptibility that enamel is corroded raises；Wherein relative to the control value of calcium, relatively low calcium concentration represents that the susceptibility that enamel is corroded raises.
In some embodiments, when showing in following standard or two when mammal, described mammal is accredited as the susceptibility that adamantine erosion has rising:
Some embodiments provide for diagnosing the test kit in mammal, the susceptibility corroded raised, and it comprises for measuring through 1 hour or the instrument of AEP calcium in sample, Statherin and/or total protein that formed more for a long time.
The scope used in full is writing a Chinese character in simplified form of each value for describing in described scope and each value.Any value in described scope is selected as the end points of this scope.Additionally, all references cited herein is incorporated integrally into herein with it by reference.
When the definition in present disclosure is inconsistent with the definition in the list of references quoted, it is as the criterion with present disclosure.
Unless otherwise indicated, otherwise other place of this paper and description expresses all percentage ratios and quantity are understood to mean that percentage by weight.The weight provided is based on the effective weight of material.
Embodiment
Embodiment 1
Carry out clinical trial to identify the mark of the susceptibility corroded for enamel.Inclusion criteria includes (i) saliva flow rate > 0.2ml/min does not stimulate, and (ii) is without gastric acid erosion damage or the parafunctional sign of saliva.
Checking two groups of patients (each n=30), one group of sign not having meals to corrode (" comparison ") and sign (" erosion ") that another group has meals to corrode, two groups have similar diet and normal saliva flow rate.Hydroxyapatite or dentium nitor mass are placed in patient oral cavity, are fixed on palate mouth pallet/clamping plate, wear 1 hour, and after removing from the oral cavity of patient, from block, collect AEP, gather AEP sample.AEP also gathers from the incisor of patient.
Mucin 5B in saliva, carbonic anhydrase 6 and Statherin : by the whole saliva of buccal cavity (UWMS) not stimulated by analyzing for the immunoblotting of MUC5b, carbonic anhydrase 6 and Statherin.Semi-quantitative analysis is undertaken by comparing the antibody staining intensity for the MUC5b of purification, carbonic anhydrase 6 and Statherin reference material.The MUC5b (p < 0.05) and carbonic anhydrase 6 (p < 0.05) ratio that corrode the whole saliva of patient compare few about 25%.On the contrary, there is the Statherin (p < 0.001) of significantly more (about twice) in same sample.
The AEP situ total protein content formed through 1 hour : utilize BCA algoscopy, compare the total protein concentration in the AEP collected by Sialostrip paper.On incisor, the total protein in AEP through being formed more than 2 hours comparison patient and corrodes zero difference between patient on natural.On the contrary, the hydroxyapatite maintained in clamping plate through 1 hour or the AEP of dentium nitor mass overlying lid show significant difference (respectively p < 0.01 and p < 0.05, n=18 in each group).This shows that the speed that AEP is formed in erosion group is slower.
Original position exposes the Statherin after 1 hour in AEP : in AEP, the analysis of specific protein shows difference only small (although can't detect albumen for a lot of experimenters) in mucin or carbonic anhydrase 6 are horizontal but Statherin is then had to obvious minimizing.
Original position exposes the calcium after 1 hour in AEP : the ion analysis (ICP/MS) of the AEP formed on 1 hour hydroxyapatite dish demonstrates calcium and the phosphorus of pronounced amount.Additionally, the level of matched group calcium is significantly higher than erosion group.Utilizing identical elution technique, from the hydroxyapatite dish that non-saliva processes, minute quantity calcium or phosphorus are eluted, it was shown that calcium is relevant to the sialoprotein of absorption with phosphorus.
To sum up, saliva and dental enamel table membrane sample are collected from the health volunteer and erosion experimenter of big group.The utilization of human enamel or hydroxyapatite does not produce any contradiction and produces similar result in all mensuration used.The utilization of ICP/MS needs to resolve further, although have shown that difference between two groups.Profile measurement shows protective effect and the group difference of pellicle.Can be calculated by the speed minimizing that pellicle is formed corroding the susceptibility raised.Although being not detected by the difference of surface membrane protein amount from the tooth (incisor) of patient self, but be maintained at the enamel in clamping plate and on hydroxy-apatite stone the difference of surface membrane protein amount be obvious.The speed that pellicle is formed can be depending on the concentration of pellicle component in saliva, but is also possible to be subject to the impact of the degraded of microorganism or other ion factor.
From above embodiment, use any one in following parameter or all diagnostic methods of developing to identify the individuality that the susceptibility that dentin is corroded raises, for instance,
。
Claims (4)
1., for diagnosing the test kit that the susceptibility in mammal, enamel corroded raises, it includes for measuring in the whole saliva of buccal cavity sample picking up from mammiferous oral cavity the instrument of one or more concentration in mucin 5B, carbonic anhydrase 6 and Statherin.
2. the test kit of claim 1, the cause of disease that wherein said enamel corrodes is meals.
3. the test kit of claim 1 or 2, wherein said includes the antibody for it for measuring the instrument of one or more concentration in mucin 5B, carbonic anhydrase 6 and Statherin.
4., for diagnosing the test kit that the susceptibility in mammal, enamel corroded raises, it comprises for measuring through 1 hour or the instrument of AEP calcium in sample, Statherin and/or total protein that formed more for a long time.
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| CN201610123828.5A CN105717305B (en) | 2011-12-20 | 2011-12-20 | Diagnostic method |
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| Application Number | Priority Date | Filing Date | Title |
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| CN201180075691.6A CN103998936B (en) | 2011-12-20 | 2011-12-20 | Diagnostic method |
| CN201610123828.5A CN105717305B (en) | 2011-12-20 | 2011-12-20 | Diagnostic method |
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| CN201180075691.6A Division CN103998936B (en) | 2011-12-20 | 2011-12-20 | Diagnostic method |
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| CN105717305B CN105717305B (en) | 2019-04-16 |
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Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1694682A (en) * | 2002-12-13 | 2005-11-09 | 宝洁公司 | Method of protecting teeth against erosion |
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Patent Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1694682A (en) * | 2002-12-13 | 2005-11-09 | 宝洁公司 | Method of protecting teeth against erosion |
Non-Patent Citations (6)
| Title |
|---|
| ALMSTÅHL A 等: "Lactoferrin, amylase and mucin MUC5B and their relation to the oral microflora in hyposalivation of different origins", 《ORAL MICROBIOL IMMUNOL》 * |
| ANDERSON T. HARA 等: "Biological Factors", 《DENTAL EROSION》 * |
| FRANCISCO MANUEL LEMOS AMADO 等: "Analysis of the human saliva proteome", 《EXPERT REV. PROTEOMICS》 * |
| J. LI 等: "Statherin is an in vivo pellicle constituent:identification and immuno-quantification", 《ARCHIVES OF ORAL BIOLOGY》 * |
| JYRKI KIVELä 等: "Salivary carbonic anhydrase isoenzyme VI", 《JOURNAL OF PHYSIOLOGY》 * |
| W. L. SIQUEIRA 等: "Evidence of Intact Histatins in the in vivo Acquired Enamel Pellicle", 《J DENT RES》 * |
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| CN105717305B (en) | 2019-04-16 |
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Granted publication date: 20190416 Termination date: 20211220 |