CN102068318B - Manufacturing method of biological tooth root bracket material - Google Patents
Manufacturing method of biological tooth root bracket material Download PDFInfo
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- CN102068318B CN102068318B CN2010105976407A CN201010597640A CN102068318B CN 102068318 B CN102068318 B CN 102068318B CN 2010105976407 A CN2010105976407 A CN 2010105976407A CN 201010597640 A CN201010597640 A CN 201010597640A CN 102068318 B CN102068318 B CN 102068318B
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Abstract
The invention discloses a manufacturing method of a biological tooth root bracket material. The method comprises the following steps of: flushing teeth with phosphate buffered saline (PBS) in a sterile state for three times; removing dental crown parts by grinding with a carborundum disc, and keeping tooth root parts; pulling pulp tissues out of the tooth roots and repeatedly flushing the inside of root canals with physiological saline; gradually expanding the inside of teeth with a dental expander and repeatedly flushing the inside of the root canals with 5 to 17 percent of ethylene diamine tetraacetic acid (EDTA) solution; grinding outer side tissues of the tooth roots by grinding with a dental turbine and constructing a biological tooth root profile as required so as to obtain a bracket with a specific shape; preserving the teeth in the PBS and treating in an ultrasonic oscillator for 3 to 8 times; performing stepped demineralization on a dentine with EDTA with different concentrations in a magnetic stirrer with the rotating speed of between 100 and 500 revolutions per minute so as to obtain a dentine substrate; and preserving the dentine substrate in double-antibody-containing PBS. The biological tooth root bracket material has a tooth root profile, the thickness of 1mm to 3mm, the length of 1cm to 2.5cm and certain tooth inducing capability, comprises a large number of proteins and factors related to the manufacturing of teeth, can release the proteins and factors continuously, can better solve the problem of induction micro-environment, and contributes to application to tooth socket intra-alveolar orthotopic transplantation.
Description
Technical field
The invention discloses a kind of construction method of biological root of the tooth timbering material.
Background technology
At present, the absence of tooth prevalence due to a variety of causes increases with age growth, is about 80% to person in middle and old age's prevalence.Absence of tooth and dysplasia dentalis seriously hinder functions such as the chewing of patient, language; And cause patient's appearance deformity, cognitive defect, mental maladjustment; Thereby greatly endangered patient's physical and mental health, therefore, seeking the efficacious therapy approach is urgent and difficult task.
Present absence of tooth all adopts the reparation of artificial material prosthesis, exists defectives such as physiological function recovers limited, service life short, damage normal structure, lacks the reparation of real physiological significance.It is the focus and emphasis of treating anodontia at present that missing tooth is repaired by implantation body, but implantation body and periodontal tissue can only realize synostosis, can't obtain newborn shortcomings such as periodontal membrane.How effectively on present restorative procedure basis, obtaining newborn cementum, periodontal membrane and alveolar bone, is the key of treatment anodontia.
Timbering material and hemopoietic inductive microenviroment problem are the major issues of carrying out root of the tooth regenerating tissues engineering.In recent years, go deep into along with what various timbering materials were studied, adopting biological organic material to be applied to the root of the tooth organizational project as timbering material has become the research focus, and mesentery, calcining Os Bovis seu Bubali, dentinal matrix etc. all have application report.Research to Dentinal histology and protein science shows: in dentin, exist multiplely for tooth development and tooth the sustain damage important protein and the factor in the repair process of back, very strong expression or higher content are wherein much all arranged.Guo etc. are object of study with the dentin of rat; Success has induced cementum, periodontal membrane and neurovascular regeneration (Weihua Guo around the dentin; Yong He, Xiaojun Zhang, et al; The use of dentin matrix scaffold and dental follicle cells for dentin regeneration, Biomaterials 2009; 30:6708-6723).The research of Guo etc. shows: with respect to the other biological timbering material; Dentinal matrix can better be induced to differentiate into cementoblast like cell, osteoblast-like cells and fibroblast-like cell; Form the similar natural periodontal membrane appearance connective tissue and the structure of periodontal membrane/cementum complex; Help the reconstruction of biological root of the tooth, this is one of key character that is different from the other biological timbering material.But prepared dentinal matrix is the mouse dentin in this research; Wait the dentin of other species to exist scarcely together with the mankind; That constructs is organized as dentin, can't be designed to the timbering material of dental root shaped; And biological root of the tooth material is at the important biomolecule active substance that can not destroy in the dentinal matrix, with guarantee that these bioactive substances can continue from material, be discharged in the surrounding time; Need keep as far as possible root of the tooth shape, keep this material to have certain mechanical strength and fully demineralization, be beneficial to the soft tissue of dental pulp and periodontal tissue and the common structure of sclerous tissues.Simultaneously, the ectopic Dentinal regeneration of using in the research of Guo is not discussed for the regeneration situation in the alveolar bone accordingly.In sum, the reconstruction of this research and the biological root of the tooth timbering material truly of being unrealized.
Summary of the invention
The objective of the invention is to overcome existing above-mentioned deficiency in the prior art, a kind of construction method of biological root of the tooth timbering material is provided.Construction method easy master of the present invention, safe and feasible, with low cost.
To achieve these goals, the invention provides following technical scheme:
A kind of construction method of biological root of the tooth timbering material may further comprise the steps:
(1) donor is fresh tooth washes with the PBS buffer under aseptic condition;
(2) step (1) gained tooth is ground except that the corona part on the turbine at a slow speed at the dental that water injector is arranged; The retained dental root part; The said dental rotating speed of turbine at a slow speed is 1000-10000 rev/min, and gained tooth root branch is stored in the culture bottle or other vessel that contains the PBS buffer;
(3) extract the pulp tissue in step (2) the gained tooth root of the tooth with the dental barbed broach, wash the tooth canal repeatedly inner 3-8 time, each 10-30 milliliter with normal saline;
(4) use dental to enlarge machine step (3) gained tooth inside is enlarged gradually, remove tissues such as pulp tissue and predentin, use the EDTA solution of 5%-17% to wash the root pipe repeatedly inner 3-8 time, each 10-30 milliliter in this process;
(5) use the dental turbine to remove the part dental tissue, to remove tissues such as periodontal tissue and cementum according to biological root of the tooth profile mill.The material thickness 1-3mm that finally obtains, length 1-2.5cm;
(6) tooth of step (5) being handled well is stored in the PBS buffer, in ultrasonator, handles 3-8 time, handles 10-30min at every turn;
(7) in rotating speed is 100-500 rev/min magnetic stirring apparatus, use the EDTA of variable concentrations that dentin is carried out the gradient demineralization, obtain dentinal matrix.
The dentinal matrix that step (7) is handled well is stored in and contains in two anti-PBS buffer subsequent usely, saidly contains that penicillin concn is 50-500unit/mL among two anti-PBS, and streptomycin concentration is 50-500unit/mL.
In the construction method of above-mentioned biological root of the tooth timbering material; Use dental to enlarge machine described in the step (4) method that step (3) gained tooth inside enlarges is gradually enlarged tooth canal inside at first using No. 15 dentistry handss with common NiTi file; No. 20 common NiTi file enlarges tooth canal inside; From the trumpet to the large size, use the Sx dental to enlarge pin then and enlarge tooth root canal orifice part; Promptly enlarge pin and enlarge tooth root canal orifice part with S1 number, S2 number, F1 number, F2 dental successively, enlarge pin expansion tooth root canal orifice part with the F3 dental in case of necessity.The said use dental of step (5) turbine with the method that the outer side grinding of step (4) gained root of the tooth removes is: use corundum car pin, remove part dental tissue, the material thickness 1-3mm that finally obtains, length 1-2.5cm according to biological root of the tooth profile mill.
In the construction method of above-mentioned biological root of the tooth timbering material, the method for gradient demineralization is described in the step (7):
At first use the EDTA solution demineralization of 17%-25% to handle 5-15min, use the EDTA solution demineralization of 7%-15% to handle 5-15min then, use 5% EDTA solution demineralization processing 5-15min at last.
Dentin owing to itself have certain mechanical strength, can be used as a kind of novel regenerated timbering material of realization root of the tooth as a kind of biomaterial; And dentin itself derives from tooth source sexual cell, has the important protein and the factor in some tooth development processes, again can be as a kind of cell tooth to the induced differentiation microenvironment.Simultaneously, because various clinical needs and teeth extracted be except being used for cell culture, the sclerous tissues of tooth part is nearly all by as MEDICAL WASTE TREATMENT, and dentin is again the key component of tooth, and this has just well solved source problem.But; Utilize dentinal matrix as biological root of the tooth timbering material; Need keep the shape of root of the tooth and fully demineralization as far as possible, but can not destroy the important biomolecule active substance in the dentinal matrix, what guarantee that these bioactive substances can continue is discharged into from material in the surrounding; In addition, also to keep this material to have certain mechanical strength.Thereby the present invention adopts from the trumpet to the large size, uses the dental enlarger with the inner expansion of tooth canal gradually, and adopts the method for gradient demineralization that tooth is carried out the demineralization processing.Through said method; In the important biomolecule active substance that does not destroy in the dentinal matrix; Kept as far as possible root of the tooth shape, keep this material to have certain mechanical strength and fully demineralization; Help finally constructing the root of the tooth timbering material of biologically active in the soft tissue of dental pulp and periodontal tissue and the common structure of sclerous tissues.
The evaluation of the dentinal matrix timbering material that the present invention makes up: 1. with the dentinal matrix of handling well as in the culture medium, 37 ℃ hatch 3 days after; Culture medium is limpid, no muddy; Prove that this material can use, sterilization reaches a standard, and this culture medium is analyzed; Find that the inside contains and possess into the tooth relevant albumen and the factor in a large number, like TGF-β 1, DMP-1, DSPP etc.2. the surface sweeping electron microscopic observation finds that material surface dentinal tubule is unobstructed, and dentin collagen fully exposes between pipe, and cell is at the material surface well-grown.3. cytoactive detects, and finds that this material helps the propagation and the growth of cell.
The construction method of biological root of the tooth timbering material of the present invention can obtain a large amount of people's dentinal matrixs from depleted people's healthy tooth, set up the tissue engineering technique biologic bracket material, makes full use of efficient resource.According to the method for the invention, the biological root of the tooth timbering material and the hemopoietic inductive microenviroment of preparation can induce seed cell to be divided into cementum, periodontal membrane, dentin and pulp tissue after compound cells is handled, and realize the reconstruction of the biological root of the tooth of real meaning.Compare with existing some other timbering material, the structure of this material easy master simple to operate, safe and feasible, with low cost, and can provide seed cell to induce the microenvironment of differentiation.In addition, employed tooth is because of a variety of causes and is pulled out depleted tooth in this method, and this has practiced thrift medical resource greatly.Therefore, the preparation of this biology root of the tooth timbering material and hemopoietic inductive microenviroment has wide practical use.
Biological root of the tooth timbering material of the present invention at first possesses the profile of root of the tooth appearance, more near the profile of natural teeth; Secondly; Contain in the root of the tooth timbering material of the present invention preparation and possess into the tooth relevant albumen and the factor in a large number, like TGF-β 1, DMP-1, DSPP, alveolar bone inside has certain inductivity for the one-tenth tooth ability of cell, material; Timbering material itself also has certain tooth to inducibility; Both complement each other, and can better solve the problem of hemopoietic inductive microenviroment, help this biology root of the tooth timbering material is carried out transplanting in the alveolar bone.In a word, the reconstruction that realizes biological root of the tooth that is prepared as of dentinal matrix provides new approaches, and is significant to the treatment absence of tooth, and the research and the clinical practice of dentinal matrix then needed a large amount of this timbering materials of dentinal matrix.
Description of drawings:
Fig. 1 is people's labial teeth dentinal matrix timbering material that the embodiment of the invention 1 makes up;
Fig. 2 is the electron-microscope scanning figure of people's labial teeth dentinal matrix timbering material of the embodiment of the invention 1 structure;
Fig. 3 is the cell growth figure of people's labial teeth dentinal matrix timbering material of the embodiment of the invention 1 structure;
Fig. 4 is the active influence of people's labial teeth dentinal matrix pair cell that the embodiment of the invention 1 makes up;
The excretory relevant albumen of people's labial teeth dentinal matrix and the factor that Fig. 5,6 embodiment of the invention 1 make up;
Fig. 7 is people's labial teeth dentinal matrix timbering material that the embodiment of the invention 2 makes up;
Fig. 8 is the electron-microscope scanning figure of people's labial teeth dentinal matrix timbering material of the embodiment of the invention 2 structures;
Fig. 9 is the cell growth figure of people's labial teeth dentinal matrix timbering material of the embodiment of the invention 2 structures; Figure 10 is people's labial teeth dentinal matrix timbering material that the embodiment of the invention 3 makes up;
Figure 11 is the electron-microscope scanning figure of people's labial teeth dentinal matrix timbering material of the embodiment of the invention 3 structures;
Figure 12 is the cell growth figure of people's labial teeth dentinal matrix timbering material of the embodiment of the invention 3 structures.
The specific embodiment
Below in conjunction with the Test Example and the specific embodiment the present invention is made further detailed description.But should this be interpreted as that the scope of the above-mentioned theme of the present invention only limits to following embodiment, allly all belong to scope of the present invention based on the technology that content of the present invention realized.Operation is all carried out in the Strict aseptic environment in the instance, and is not contaminated for guaranteeing cell, in the PBS of all uses, DMEM, all is added with 100U/ml penicillin and 100U/ml streptomycin.
The method for preparing of the biological root of the tooth timbering material that present embodiment is enumerated may further comprise the steps:
(1) gets four of healthy labial teeths,, be stored in the culture bottle or other vessel that contains PBS, seal the film phonograph seal bottleneck and be stored in 4 ℃ of refrigerators with PBS liquid flushing three times;
(2) on the turbine carborundum disc is installed at a slow speed at the dental that water injector is arranged; The adjustment rotating speed is 3000 rev/mins; Use the carborundum disc mill to remove the corona part of step (1) gained tooth; The retained dental root part also is stored in the culture bottle or other vessel that contains PBS, seals the film phonograph seal bottleneck and is stored in 4 ℃ of refrigerators;
(3) use the complete pulp tissue of extracting in step (2) the gained tooth root of the tooth of dental barbed broach, and use normal saline to wash the root pipe repeatedly inner 3 times, about at every turn 20mL;
(4) use dental to enlarge machine step (3) gained tooth is carried out the tooth inter-process; At first use No. 15 dentistry handss to enlarge tooth canal inside with common NiTi file; It is inner that No. 20 common NiTis files enlarge tooth, from the trumpet to the large size, uses the Sx dental to enlarge pin then and enlarge tooth root canal orifice part, promptly enlarges pin expansion tooth root canal orifice part with S1 number, S2 number, F1 number, F2 dental successively; And use 10% EDTA solution flushing root pipe repeatedly inner 3 times, each 10mL;
(5) use the dental turbine that the tooth of step (4) gained is removed the part dental tissue according to labial teeth root of the tooth profile mill, to remove tissues such as periodontal tissue and cementum.The material thickness 1mm that finally obtains, length 1.5cm;
(6) tooth of step (5) being handled well is stored in the PBS buffer, and in ultrasonator, handles 4 times, handles 10min at every turn;
(7) in rotating speed is 100 rev/mins magnetic stirring apparatus; Use the EDTA of variable concentrations that dentin is carried out the gradient demineralization; At first use 15% EDTA solution demineralization processing 5min, use 10% EDTA solution demineralization processing 5min then, use 5% EDTA solution demineralization processing 5min at last;
The dentinal matrix that step (7) is handled well is stored in and contains that two anti-(penicillin concn is 100unit/ml; Streptomycin concentration is 100unit/ml) the PBS buffer in, during use the dentinal matrix of above-mentioned made used according to clinical requirement and practical application to get final product.
The evaluation of the dentinal matrix timbering material that present embodiment makes up:
The dentinal matrix that
handles well as in the DMEM culture medium, 37 ℃ hatch 3 days after; Check finds that culture medium is limpid, it is muddy not have; Prove that this material can use, sterilization reaches a standard.Collect the culture medium of above-mentioned soak-out material, and this culture medium is analyzed, adopt the ELISA test kit to detect the GAP-associated protein GAP and the factor.The result finds that culture medium the inside contains and possess into the tooth relevant albumen and the factor in a large number, like TGF-β 1, DMP-1, DSPP etc., and does not soak the no above-mentioned GAP-associated protein GAP and the factor in the culture medium of material.
2. present embodiment is constructed dentinal matrix timbering material places under the surface sweeping electron microscopic observation and observes, and finds that material surface dentinal tubule is unobstructed, and dentin collagen fully exposes between pipe, and cell is at the material surface well-grown.
3. cytoactive detects, and adopts the CCK-8 experimental technique, promptly collects to have used this material to carry out cultured cells and do not use this material, be with normal cultured base cultured cells; Be prepared into cell suspension; Be seeded in 96 orifice plates, every plate 100 μ l add 10 μ lCCK-8 solution simultaneously; Hatch after 2 hours for 37 ℃ and use ELIASA to detect absorbance, detect cytoactive at the 450nm place.Find that this material helps the propagation and the growth of cell.
The method for preparing of the biological root of the tooth timbering material that present embodiment is enumerated may further comprise the steps:
(1) gets four of premolarss,, be stored in the culture bottle or other vessel that contains PBS, seal the film phonograph seal bottleneck and be stored in 4 ℃ of refrigerators with PBS liquid flushing three times.
(2) having under the situation of water injector; On the turbine carborundum disc is installed at a slow speed at dental; 6500 rev/mins of adjustment rotating speeds; Use the carborundum disc mill to remove step (1) corona part, the retained dental root part also is stored in the culture bottle or other vessel that contains PBS, seals the film phonograph seal bottleneck and is stored in 4 ℃ of refrigerators.
(3) use dental barbed broach is complete extracts pulp tissue in step (2) the gained tooth root of the tooth, and uses normal saline to wash the root pipe repeatedly inner 5 times, about at every turn 15ml.
(4) use dental to enlarge machine and carry out the tooth inter-process; At first use No. 15 dentistry handss to enlarge tooth canal inside with common NiTi file; It is inner that No. 20 common NiTis files enlarge tooth, from the trumpet to the large size, uses the Sx dental to enlarge pin then and enlarge tooth root canal orifice part, promptly enlarges pin expansion tooth root canal orifice part with S1 number, S2 number, F1 number, F2 dental successively; And use 10% EDTA solution flushing root pipe repeatedly inner 5 times, each 20mL;
(5) use the dental turbine that the tooth of step (4) gained is removed part root of the tooth tissue according to premolars root of the tooth profile mill, to remove tissues such as periodontal tissue and cementum.The material thickness 1.5mm that finally obtains, length 1.5cm;
(6) tooth of step (5) being handled well is stored in the PBS buffer, and in ultrasonator, handles 4 times, handles 20min at every turn.
(7) in rotating speed is 200 rev/mins magnetic stirring apparatus; Use the EDTA of variable concentrations to carry out the gradient demineralization; 5min is handled in 20% EDTA solution demineralization, uses 12% EDTA solution demineralization processing 5min then, uses 5% EDTA solution demineralization processing 5min at last.
The dentinal matrix that step (7) is handled well is stored in and contains that two anti-(penicillin concn is 100unit/ml; Streptomycin concentration is 100unit/ml) PBS in, during use the dentinal matrix of above-mentioned made used according to clinical requirement and practical application to get final product.
The evaluation of the dentinal matrix timbering material that present embodiment makes up:
The dentinal matrix that
handles well as in the DMEM culture medium, 37 ℃ hatch 3 days after; Check finds that culture medium is limpid, it is muddy not have; Prove that this material can use, sterilization reaches a standard.
2. present embodiment is constructed dentinal matrix timbering material places under the surface sweeping electron microscopic observation and observes, and finds that material surface dentinal tubule is unobstructed, and dentin collagen fully exposes between pipe, and cell is at the material surface well-grown.
The method for preparing of the biological root of the tooth timbering material that present embodiment is enumerated may further comprise the steps:
(1) gets health and grind one's teeth in sleep four,, be stored in the culture bottle or other vessel that contains PBS, seal the film phonograph seal bottleneck and be stored in 4 ℃ of refrigerators with PBS liquid flushing three times;
(2) on the turbine carborundum disc is installed at a slow speed at the dental that water injector is arranged; The adjustment rotating speed is 4000 rev/mins; Use the carborundum disc mill to remove the corona part of step (1) gained tooth; The retained dental root part also is stored in the culture bottle or other vessel that contains PBS, seals the film phonograph seal bottleneck and is stored in 4 ℃ of refrigerators;
(3) use the complete pulp tissue of extracting in step (2) the gained tooth root of the tooth of dental barbed broach, and use normal saline to wash the root pipe repeatedly inner 7 times, about at every turn 20mL;
(4) use dental to enlarge machine step (3) gained tooth is carried out the tooth inter-process; At first use No. 15 dentistry handss to enlarge tooth canal inside with common NiTi file; It is inner that No. 20 common NiTis files enlarge tooth, from the trumpet to the large size, uses the Sx dental to enlarge pin then and enlarge tooth root canal orifice part, promptly enlarges pin expansion tooth root canal orifice part with S1 number, S2 number, F1 number, F2 dental successively; And use 10% EDTA solution flushing root pipe repeatedly inner 7 times, each 20mL;
(5) use the dental turbine that the root of the tooth of step (4) gained is removed the part dental tissue according to the root of the tooth profile mill of grinding one's teeth in sleep, to remove tissues such as periodontal tissue and cementum.The material thickness 1mm that finally obtains, length 1cm;
(6) tooth of step (5) being handled well is stored in the PBS buffer, and in ultrasonator, handles 4 times, handles 20min at every turn;
(7) in rotating speed is 300 rev/mins magnetic stirring apparatus; Use the EDTA of variable concentrations that dentin is carried out the gradient demineralization; At first use 20% EDTA solution demineralization processing 10min; Use 10% EDTA solution demineralization processing 10min then, use 5% EDTA solution demineralization processing 10min at last;
The dentinal matrix that step (7) is handled well is stored in and contains that two anti-(penicillin concn is 100unit/ml; Streptomycin concentration is 100unit/ml) the PBS buffer in, during use the dentinal matrix of above-mentioned made used can the dentinal matrix of above-mentioned made be used according to clinical requirement and practical application according to clinical requirement and practical application to get final product.
1. the inferior essential substrate that will handle well as in the culture medium, 37 ℃ hatch 3 days after, culture medium is limpid, no muddy, proves that this material can use, sterilization reaches a standard.2. the surface sweeping electron microscopic observation finds that material surface dentinal tubule is unobstructed, and dentin collagen fully exposes between pipe, and cell is at the material surface well-grown.
Claims (3)
1. the construction method of a biological root of the tooth timbering material is characterized in that may further comprise the steps:
(1) donor is fresh tooth washes with the PBS buffer under aseptic condition;
(2) step (1) gained tooth is ground except that the corona part on the turbine at a slow speed at the dental that water injector is arranged; The retained dental root part; The said dental rotating speed of turbine at a slow speed is 1000-10000 rev/min, and gained tooth root branch is stored in the culture bottle or other vessel that contains the PBS buffer;
(3) extract the pulp tissue in step (2) the gained tooth root of the tooth with the dental barbed broach, wash the tooth canal repeatedly inner 3-8 time, each 10-30 milliliter with normal saline;
(4) use dental to enlarge machine step (3) gained tooth inside is enlarged gradually, remove pulp tissue and predentin tissue, use the EDTA solution of 5%-17% to wash the root pipe repeatedly inner 3-8 time, each 10-30 milliliter in this process;
(5) use the dental turbine to remove portion of tissue,, obtain the timbering material that thickness is 1-3mm, length 1-2.5cm to remove periodontal tissue and cementum tissue according to biological root of the tooth profile mill;
(6) timbering material of step (5) being handled well is stored in the PBS buffer, in ultrasonator, handles 3-8 time, handles 10-30min at every turn;
(7) in rotating speed is 100-500 rev/min magnetic stirring apparatus, use the EDTA of variable concentrations that dentin is carried out the gradient demineralization, obtain biological root of the tooth timbering material.
2. the construction method of biological root of the tooth timbering material according to claim 1 is characterized in that:
Use dental expansion machine to do described in the step (4) with the method that step (3) gained tooth inside enlarges gradually; At first use No. 15 dentistry handss to enlarge tooth canal inside with common NiTi file; No. 20 common NiTi file enlarges tooth canal inside; From the trumpet to the large size, use the Sx dental to enlarge pin then and enlarge tooth root canal orifice part, promptly enlarge pin expansion tooth root canal orifice part with S1 number, S2 number, F1 number, F2 dental successively.
3. the construction method of biological root of the tooth timbering material according to claim 1 and 2; It is characterized in that: the method for gradient demineralization does described in the step (7); At first use the EDTA solution demineralization of 17%-25% to handle 5-15min; Use the EDTA solution demineralization of 7%-15% to handle 5-15min then, use 5% EDTA solution demineralization processing 5-15min at last.
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| CN103340755B (en) * | 2013-06-20 | 2014-11-26 | 北京大学 | Dental prosthetic material and preparation method thereof |
| CN106823001B (en) * | 2017-04-12 | 2019-11-05 | 吉林大学 | A kind of biologic bracket material for dental root regeneration, preparation method and applications |
| CN107648670A (en) * | 2017-11-02 | 2018-02-02 | 南京医科大学附属口腔医院 | A kind of preparation method of dentin-pulp complex |
| CN107674111A (en) * | 2017-11-02 | 2018-02-09 | 南京医科大学附属口腔医院 | A kind of sequencing preparation method of dentin non-collagenous protein |
| CN110327221A (en) * | 2019-04-16 | 2019-10-15 | 中国人民解放军第四军医大学 | A kind of dentin collagen fiber demineralization material and the preparation method and application thereof |
| CN114767937B (en) * | 2022-04-18 | 2024-02-20 | 武汉大学 | Tooth bone material for external demineralization of collagen |
| CN115300147B (en) * | 2022-08-24 | 2023-05-09 | 四川大学 | Construction method of 3D printing personalized biological tooth root bracket |
| CN116099051B (en) * | 2022-11-16 | 2023-11-14 | 首都医科大学附属北京口腔医院 | Method for constructing tissue engineering vascularized dental pulp by acellular matrix and application |
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