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Molecular biological model for eliminating oral pigment biofilm with protease and preparation method thereof

A molecular biology and biofilm technology, which is applied in biochemical equipment and methods, microbial determination/inspection, biological testing, etc. Effect

Inactive Publication Date: 2012-04-11
姚江武
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003] The disadvantage of the prior art is that it is impossible to monitor the formation and removal process of the pigmented film on the tooth surface or restoration surface in real time, and the removal of the pigmented layer through physical and chemical methods, although it has a certain effect on removing the pigment, but also Causes varying degrees of mechanical wear and demineralization of the enamel layer on the tooth surface, resulting in irreparable damage

Method used

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  • Molecular biological model for eliminating oral pigment biofilm with protease and preparation method thereof
  • Molecular biological model for eliminating oral pigment biofilm with protease and preparation method thereof
  • Molecular biological model for eliminating oral pigment biofilm with protease and preparation method thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0031] Embodiment 1, QCM-D chip

[0032] 1. Self-assembled protein film: pass through 1ml of 10mM 11-mercapto-11 alkanoic acid solution to activate the chip for 10 minutes; then add 1 mL of 0.2M EDC and 0.1M NHS mixed solution with a volume ratio of 1:1 to activate the QCM-D chip for 10 minutes . Inject 1mL of protein solution until the adsorption reaches a steady state. The protein is human saliva, and then inject 1mL of 1M ethanolamine hydrochloric acid solution with pH 8.5, and keep it closed for 10min. Ethanolamine blocks nonspecific binding sites on protein surfaces. Ethanolamine was drained and a solution of 20 mM hydrochloric acid pH 2.0 was passed through to remove non-specifically bound proteins. So far, a biofilm assembled by protein membranes (such as saliva) has formed on the surface of the activated chip.

[0033] 2. Formation of pigment-saliva biofilm: Before the pigment is passed, the buffer is continuously passed. Under the conditions of 25°C, pH 7.0, and i...

Embodiment 2

[0039] Embodiment 2, QCM-D chip

[0040] 1. Self-assembled protein film: pass through 1ml of 8mM 11-mercapto-11 alkanoic acid solution to activate the chip for 12min; then pass through 1mL of a mixed solution of 0.2M EDC and 0.1M NHS with a volume ratio of 2:1 to activate the QCM-D chip 10min. Inject 1mL of protein solution until the adsorption reaches a steady state, the protein is human saliva, and then inject 1mL of 1.2M ethanolamine hydrochloric acid solution with pH 8.0, and keep it closed for 10min. Ethanolamine blocks nonspecific binding sites on protein surfaces. Ethanolamine was drained and a solution of 20 mM hydrochloric acid pH 2.0 was passed through to remove non-specifically bound proteins. So far, a biofilm assembled by protein membranes (such as saliva) has formed on the surface of the activated chip.

[0041] 2. Formation of pigment-saliva biofilm: Before the pigment is passed, the buffer is continuously passed. Under the conditions of 25°C, pH 7.0, and io...

Embodiment 3

[0046] Embodiment 3 SPR chip

[0047] 1. Self-assembled protein film: pass through 1ml of 12mM 11-mercapto-11alkanoic acid solution to activate the chip for 8 minutes; then pass through 1mL of a mixed solution of 0.3M EDC and 0.15M NHS with a volume ratio of 1:2 to activate the SPR chip for 10 minutes. Inject 1mL of protein solution until the adsorption reaches a steady state. The protein is artificially synthesized saliva, and then inject 1mL of 1M ethanolamine hydrochloric acid solution with pH 8.5, and keep blocking for 10min. Ethanolamine was drained and a solution of 20 mM hydrochloric acid pH 2.0 was passed through to remove non-specifically bound proteins. So far, a biofilm assembled by protein membranes (such as saliva) has formed on the surface of the activated chip.

[0048] 2. Formation of pigment-saliva biofilm: Before the pigment is passed, the buffer is continuously passed. Under the conditions of 25°C, pH 7.0, and ionic strength of 10mM PBST, inject the theafl...

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Abstract

The invention discloses a molecular biological model for eliminating oral pigment biofilm with protease and a preparation method thereof. According to the invention, a self-assembly protein film is formed on the surface of a QCM-D (quartz crystal microbalance with dissipation monitoring) or SPR (surface plasmon resonance) chip to build an in-vitro model of an oral protein biofilm; then a common edible pigment compound is added, and the adsorption and desorption reaction between the pigment and protein on a biochip is in-situ, real-time and dynamically monitored on QCM and SPR, thus a colored biofilm model on the surfaces of teeth and a repairing material is built, the mechanism of interaction between the pigment and protein is researched, and the mechanism of coloring, caused by the pigment, of the surfaces of natural teeth and artificial material is explored; and the adsorbed protein / pigment film is further catalytically degraded by protease (such as papain), the efficiency of the biological protease in degrading the pigment under different experiment conditions is researched, and the bioactivity of the biological protease under different experiment conditions is explored, thus an optimal experiment condition for safely and efficiently hydrolyzing the pigment biofilm with the biological protease is screened.

Description

technical field [0001] The invention relates to an oral pigment removal model, in particular to a molecular biological model for protease removal of oral pigment biofilm and a preparation method thereof. Background technique [0002] In previous studies on the removal of oral pigment, the method was to select natural teeth with surface coloring or form an exogenous coloring layer on the surface of isolated teeth, and then bleach them with high-concentration peroxide or apply whitening toothpaste, denture cleaner, etc. Physical and chemical methods to remove the stained layer on the surface of teeth and restorations. [0003] The disadvantage of the prior art is that it is impossible to monitor the formation and removal process of the pigmented film on the tooth surface or restoration surface in real time, and the removal of the pigmented layer through physical and chemical methods, although it has a certain effect on removing the pigment, but also Different degrees of mecha...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/37G01N33/68
Inventor 姚江武
Owner 姚江武
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