阅读说明：本技术 一种直流电辅助下加快促进脱矿牙本质再矿化的方法 (A kind of lower method for accelerating to promote demineralization dentine remineralization of direct current auxiliary ) 是由 周洲 许涛 于金华 李泽汉 李娜 葛兴云 于 2019-09-25 设计创作，主要内容包括：本发明公开了一种直流电辅助下加快促进脱矿牙本质再矿化的方法,属于生物材料工程技术领域。旨在提供一种加快脱矿牙本质再矿化的方法,其技术方案要点如下,S1.将脱矿的牙本质片进行磷酸化；S2.将磷酸化的脱矿牙本质片固定于一个双通管的中间,双通管一部分放入酪蛋白磷酸肽-纳米无定形磷酸钙乳剂,另一部分固定于盛有模拟唾液的溶液盒内；S3.用导线导出电泳仪正负极；S4.将接有电泳仪负极的电极放入到双通管的一部分,进入到酪蛋白磷酸肽-纳米无定形磷酸钙乳剂中,连有电泳仪正极的电极放入到盛有模拟唾液的双通管另一部分；S5.接通电泳仪电源导电,调整电流电压；S6.将S5获得的牙本质片样本清洗干净,得到所述再矿化脱矿牙本质片。(The invention discloses a kind of direct currents to assist the lower method for accelerating to promote demineralization dentine remineralization, belongs to biomaterial field of engineering technology.It is intended to provide a kind of method for accelerating demineralization dentine remineralization, drip irrigation device is as follows, and the dentine piece of demineralization is carried out phosphorylation by S1.；S2., the demineralization dentine piece of phosphorylation is fixed on to the centre of a double-way pipe, double-way pipe a part is put into casein phosphopeptide-nm amorphous calcium phosphate emulsion, and another part is fixed in the solution box for filling simulation saliva；S3. electrophoresis apparatus positive and negative anodes are exported with conducting wire；S4., the electrode for being connected to electrophoresis apparatus cathode is put into a part of double-way pipe, is entered in casein phosphopeptide-nm amorphous calcium phosphate emulsion, the electrode for being connected with electrophoresis apparatus anode is put into the double-way pipe another part for filling simulation saliva；S5. it is conductive to connect electrophoresis apparatus power supply, adjusts Current Voltage；S6. the S5 dentine piece sample obtained is cleaned up, obtains the remineralization demineralization dentine piece.)

1. a kind of lower method for accelerating to promote demineralization dentine remineralization of direct current auxiliary, which is characterized in that including operating as follows Step:

S1. phosphorylation is carried out to demineralization dentine using sodium tripolyphosphate, soaking time is 24 hours；

S2., the demineralization dentine piece of phosphorylation is fixed on to the centre of a double-way pipe, double-way pipe a part is put into casein phosphorus Sour peptide-nm amorphous calcium phosphate emulsion, emulsion and dentin surface completely attach to, and another part, which is fixed on, fills simulation saliva Solution box in, simulate saliva and the another side of dentine piece and also completely attach to；

S3. electrophoresis apparatus positive and negative anodes are exported with conducting wire；

S4., the electrode for being connected to electrophoresis apparatus cathode is put into a part of double-way pipe, enters casein phosphopeptide-nanometer without fixed In shape calcium phosphate emulsion, the electrode for being connected with electrophoresis apparatus anode is put into the double-way pipe another part for filling simulation saliva；

S5. it is conductive to connect electrophoresis apparatus power supply, adjusts Current Voltage, with 1 hour for a mineralising period, replaces casein phosphoric acid Peptide-nm amorphous calcium phosphate emulsion and simulation saliva；

S6. the S5 dentine piece sample obtained is cleaned up, obtains the remineralization demineralization dentine piece.

2. the lower method for accelerating to promote demineralization dentine remineralization of direct current auxiliary according to claim 1, feature exist In the concentration of the sodium tripolyphosphate is 20g/L.

3. the lower method for accelerating to promote demineralization dentine remineralization of direct current auxiliary according to claim 1, feature exist In casein phosphopeptide-nm amorphous calcium phosphate quality in the casein phosphopeptide-nm amorphous calcium phosphate emulsion Percentage is 10%.

4. the lower method for accelerating to promote demineralization dentine remineralization of direct current auxiliary according to claim 1, feature exist In the electrode is platinized platinum.

5. the lower method for accelerating to promote demineralization dentine remineralization of direct current auxiliary according to claim 1, feature exist In the electric current is 1 ~ 4mA, and the voltage is 8 ~ 12V.

6. the lower method for accelerating to promote demineralization dentine remineralization of direct current auxiliary according to claim 4, feature exist In being separately connected two electrodes with crocodile clip in the S3.

7. the lower method for accelerating to promote demineralization dentine remineralization of direct current auxiliary according to claim 1, feature exist In the dentine piece of the demineralization carries out ultrasonic pretreatment before phosphorylation.

8. the lower method for accelerating to promote demineralization dentine remineralization of direct current auxiliary according to claim 1, feature exist In also containing graphene oxide in the casein phosphopeptide-nm amorphous calcium phosphate emulsion.

9. the lower method for accelerating to promote demineralization dentine remineralization of direct current auxiliary according to claim 8, feature exist In cladding silicon carbide inside the graphene oxide.

10. the lower method for accelerating to promote demineralization dentine remineralization of direct current auxiliary according to claim 9, feature exist In the graphene oxide mass fraction be 5%-8%.

Technical field

The present invention relates to biomaterial field of engineering technology, specifically, being related to a kind of lower quickening promotion of direct current auxiliary The method of demineralization dentine remineralization.

Background technique

Bionical remineralization is the important research field in minimally invasive dentistry, the tooth of demineralization is repaired by bionical remineralization, most The healing potential for playing to limits tooth body itself can solve sensitivity of tooth, the prevention of tooth decay and self-regeneration etc. and ask Topic.Remineralization of teeth can be divided into the remineralization of enamel and the remineralization of dentine.The Remineralizing technique of enamel relative at It is ripe, also achieve relatively good remineralization efficacy.And dentine remineralization, because of its structural particularity, often remineralization effect is not true It is fixed, and remineralization speed is extremely slow.

Main structure of the dentine as tooth body wants low compared to enamel mineralization degree, in addition to 70% inorganic matter, also About 20% organic substrate and 10% water, organic substrate is mainly Type I collagen fiber in dentine, and collagenous fibres are dentine Fundamental structural unit, really to realize dentine remineralization, it is necessary to hydroxyapatite crystal be made to enter collagenous fibres table Between face and collagenous fibres.This is compared to Enamel remineralizations and directly completes seed crystal on the basis of remaining enamel crystal Extension and growth difficulty want greatly more.Currently usually using calcium phosphate complex, bioactivity glass in dentine remineralization The non-collagens analogs such as ion, agar gel, polyamide-amide dendritic (PAMAM) guide the bionical of dentine Remineralization, often speed is slower for effect, and generally can only form layer of surface mineralizer in dentin surface, is difficult to complete tooth Remineralization in essential fiber.This also largely constrains the application of the bionical remineralization of dentine.

Summary of the invention

In view of the status of current dentine remineralization, the present invention is directed to invent a kind of direct current auxiliary for hospital clinical The lower method for accelerating to promote demineralization dentine remineralization, the defect in addition to that can overcome current dentine remineralization accelerate tooth sheet The remineralization of matter, moreover it is possible to while realizing the interior remineralization with outside fiber of the fiber of dentine, to really realize the imitative of dentine Raw remineralization.

Above-mentioned technical purpose of the invention has the technical scheme that

A kind of lower method for accelerating to promote demineralization dentine remineralization of direct current auxiliary, including following operating procedure:

S1. the dentine piece of demineralization is subjected to phosphorylation；

S2., the demineralization dentine piece of phosphorylation is fixed on to the centre of a double-way pipe, double-way pipe a part is put into casein phosphorus Sour peptide-nm amorphous calcium phosphate emulsion, emulsion and dentin surface completely attach to, and another part, which is fixed on, fills simulation saliva Solution box in, simulate saliva and the another side of dentine piece and also completely attach to；

S3. electrophoresis apparatus positive and negative anodes are exported with conducting wire；

S4., the electrode for being connected to electrophoresis apparatus cathode is put into a part of double-way pipe, enters casein phosphopeptide-nanometer without fixed In shape calcium phosphate emulsion, the electrode for being connected with electrophoresis apparatus anode is put into the double-way pipe another part for filling simulation saliva；

S5. it is conductive to connect electrophoresis apparatus power supply, adjusts Current Voltage, with 1 hour for a mineralising period, replaces casein phosphoric acid Peptide-nm amorphous calcium phosphate emulsion and simulation saliva；

S6. the S5 dentine piece sample obtained is cleaned up, obtains the remineralization demineralization dentine piece.Using will be through TPP The demineralization dentine piece of processing is fixed on the centre of a double-way pipe, and double-way pipe top half is put into 10% casein phosphopeptide-and receives Rice amorphous calcium phosphate (CPP-ACP) emulsion, emulsion and dentin surface completely attach to, and are partially fixed under double-way pipe and fill mould In the solution box of quasi- saliva, the mode that the another side of simulation saliva and dentine piece also completely attaches to can simulate tooth in mouth The true environment of chamber, convenient for clinical popularization and use.

Further, phosphorylation is carried out to demineralization dentine using sodium tripolyphosphate in S1, soaking time is 24 hours. Because of the maximum difficult point of the bionical remineralization of dentine, it is necessary to make hydroxyapatite crystal and meanwhile enter collagenous fibres surface and Between collagenous fibres, i.e., realize that this realizes the performances such as machinery to dentine remineralization with the outer remineralization of fiber in fiber simultaneously Restore most important.By the phosphorylation dentin collagen fiber of sodium tripolyphosphate, as subsequent dentine remineralization is provided The template of remineralization, is easily advanced into the collagenous fibres of dentine convenient for hydroxyapatite crystal, reaches in fiber and fine The purpose of the outer remineralization simultaneously of dimension, improves the hardness and compression strength of remineralization dentine.

Further, the concentration of sodium tripolyphosphate is 20g/L.

Further, casein phosphopeptide-nm amorphous phosphorus in casein phosphopeptide-nm amorphous calcium phosphate emulsion The mass percent of sour calcium is 10%.

Further, electrode is platinized platinum.

Further, electric current is 1 ~ 4mA, and voltage is 8 ~ 12V.Direct current is introduced into dentine remineralization, by straight Galvanic electricity accelerates the ordered movement of remineralization molecule, to have the function that quickening and promote remineralization.It is provided by electrophoresis apparatus straight Galvanic electricity, size of current precise and safety is controllable, and size of current and voltage swing can only keep constant, this hair in actual operation Bright selection constant current, it is of the invention as long as voltage control below safe voltage, is clinical conversion service after being preferably Size of current is mainly set as 1mA, 2mA, 4mA, this is human-body safety electric current in the function and effect and practical application for take into account electric current Limitation.

Further, in S3, two electrodes are separately connected with crocodile clip.

Further, the dentine piece of demineralization carries out ultrasonic pretreatment before phosphorylation.

Further, also contain graphene oxide in casein phosphopeptide-nm amorphous calcium phosphate emulsion.

Further, silicon carbide is coated inside graphene oxide.

Further, the mass fraction of graphene oxide is 5%-8%.

In conclusion the invention has the following advantages:

1, the dentine remineralization of previous mode is made slow progress, and generally wants several weeks that could obtain apparent remineralization effect, The present invention assists remineralization using direct current, and the direct current of some strength can accelerate molecular motion velocities, thus several small When it is interior can obtain preferable remineralization effect, substantially increase the efficiency of remineralization.

2, the present invention carries out collagenous fibres phosphorylation to the dentine piece of demineralization first by sodium tripolyphosphate, is remineralization Dentine piece nucleation site is provided, when powering on, the casein phosphopeptide with anion-nm amorphous calcium phosphate will It is mobile to anode under direct current electro ultrafiltration, at this time because of the marked nucleation site of demineralization dentine piece, casein phosphopeptide-nanometer It will be a kind of orderly combination that amorphous calcium phosphate, which enters in dentine piece, may be implemented to be completed at the same time outside the fiber of dentine With remineralization in fiber, this is vital to the recovery of mechanical performance after later period dentine remineralization.

3, method security provided by the invention is high, easy to be economical, can realize for the later period to clinical conversion and application；

4. the addition of graphene oxide enters graphene oxide jointly with casein phosphopeptide-nm amorphous calcium phosphate In dentine piece, electronics can quickly be adsorbed using the porous and its big specific surface area of graphene oxide, simultaneously Conduction is carried out to electronics using graphene oxide, the conversion ratio of electrochemical deposition in the application is improved, is more caused Close dentine；And graphene oxide has good antibiotic property, can be used in various external wounds healings, prevents from infecting；

5. graphene oxide cladding silicon carbide can effectively improve the toughness and intensity of graphene oxide, later period tooth is further increased The restoring degree of the mechanical performance of essence.

Detailed description of the invention

Fig. 1 is the scanning electron microscope (SEM) photograph of demineralization dentine piece；

Fig. 2 is the scanning electron microscope (SEM) photograph for the remineralization dentine piece being powered 2 hours under 4mA direct current auxiliary；

Fig. 3 is the remineralization dentine piece that graphene oxide covering material is not added under 4mA direct current auxiliary and is powered 6 hours Scanning electron microscope (SEM) photograph；

Fig. 4 is the scanning electron microscope (SEM) photograph for the remineralization dentine piece being powered 6 hours under 4mA direct current auxiliary；

Fig. 5 is the scanning electron microscope (SEM) photograph that no direct current assists lower 6 hours remineralization dentine pieces of control group remineralization；

Fig. 6 is demineralization dentine piece 6 hours scanning electron microscope (SEM) photographs of remineralization without TPP processing under 4mA direct current auxiliary；

Fig. 7 is the X diffraction pattern of dentine piece；

Fig. 8 is that different size direct current assists the dentine piece microhardness result of variations after lower remineralization.

Specific embodiment

The present invention is described further in the following with reference to the drawings and specific embodiments.

A kind of lower method for accelerating to promote demineralization dentine remineralization of direct current auxiliary, comprising the following steps:

S1. demineralization dentine sample is prepared, is cleaned by ultrasonic spare.

S2. the dentine piece of demineralization is put into sodium tripolyphosphate (TPP) solution of 20g/L and is impregnated 24 hours, make demineralization Dentine piece complete phosphorylation, provide nucleation site for the remineralization of dentine.

S3. it is more preferable simulation tooth in the true environment in oral cavity, the demineralization dentine piece handled through TPP is fixed on one The centre of a double-way pipe, double-way pipe top half are put into 10% casein phosphopeptide-nm amorphous calcium phosphate (CPP-ACP) cream Agent and 5% graphene oxide coat silicon carbide, and emulsion and dentin surface completely attach to, be partially fixed under double-way pipe and fill mould In the solution box of quasi- saliva.Simulation saliva and the another side of dentine piece also completely attach to.

Remineralization process:

(1) electrophoresis apparatus positive and negative anodes are exported with conducting wire, is separately connected two platinum plate electrodes using crocodile clip.Two platinum plate electrode accesses It can provide direct current in circuit.

(2) platinum plate electrode for being connected to electrophoresis apparatus cathode is put into the top half of double-way pipe, enters 10% casein phosphorus In sour peptide-nm amorphous calcium phosphate emulsion；The platinum plate electrode for being connected with electrophoresis apparatus anode is put into the bilateral for filling simulation saliva Pipe lower half portion.Guarantee that current loop can be formed after voltage instrument powers on.

(3) it is conductive to connect electrophoresis apparatus power supply, adjusts suitable Current Voltage, general electric current is set as 1mA, 2mA, 4mA, such as Fruit selectes constant current, and voltage generally has fluctuation, overall to be no more than 12V.With 1 hour for a mineralising period, casein phosphorus is replaced Sour peptide-nm amorphous calcium phosphate emulsion and simulation saliva, dentine piece sample is finally cleaned up, it is de- to get remineralization Mine dentine piece sample.

Demineralization dentine piece is first placed into sodium tripolyphosphate (TPP) solution of 20g/L and is impregnated 24 hours, be in order to So that the dentine piece of demineralization is completed phosphorylation, provides nucleation site for the remineralization of dentine.

Demineralization dentine sample specific the preparation method is as follows:

(1) the complete third molar for collecting Outpatient Service Stomatology surgical extraction is impregnated with 0.005% thymol solution disinfection, in 4 It is saved in DEG C refrigerator；

(2) surface enamel is removed with high speed turbine, the dentine piece sample 68 of 4 × 4 × 1mm is prepared with cutting machine at a slow speed Piece is successively polished smooth with the sand paper of 180 mesh, 400 mesh, 800 mesh, 1200 mesh, and it is clear to be put into deionized water ultrasound in ultrasonic cleaning It washes 5 minutes, stays 4 to disregard as positive controls；

(3) remaining dentine piece sample embrocates 60s repeatedly with 35% phosphoric acid gel to simulate dentine demineralization；The tooth of demineralization Essential piece sample is cleaned in supersonic cleaning machine by deionized water, drying for standby.Stay 4 demineralization dentine pieces as yin simultaneously Property control group.

By the dentine sample packet of demineralization in following embodiment, it is divided into are as follows:

(1) 1mA direct current assists remineralization group: mineralising 2 hours, 4 hours, 6 hours each 4.

(2) 2mA direct current assists remineralization group: mineralising 2 hours, 4 hours, 6 hours each 4.

(3) 4mA direct current assists remineralization group: mineralising 2 hours, 4 hours, 6 hours each 4.

(4) 4mA direct current auxiliary is added graphene oxide and coats carbofrax material remineralization group: mineralising 2 hours, 4 hours, 6 hours each 4.

(5) assist remineralization group without direct current: mineralising 2 hours, 4 hours, 6 hours each 4.

(6) assist remineralization group without TPP processing 4mA direct current: mineralising 2 hours, 4 hours, 6 hours each 4.

(7) negative control group (demineralization dentine piece) 4.

(8) positive controls (non-demineralization dentine piece) 4.