CN111073025A - Method for manufacturing tissue engineering scaffold with tooth appearance - Google Patents
Method for manufacturing tissue engineering scaffold with tooth appearance Download PDFInfo
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- CN111073025A CN111073025A CN201911226907.9A CN201911226907A CN111073025A CN 111073025 A CN111073025 A CN 111073025A CN 201911226907 A CN201911226907 A CN 201911226907A CN 111073025 A CN111073025 A CN 111073025A
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- C—CHEMISTRY; METALLURGY
- C08—ORGANIC MACROMOLECULAR COMPOUNDS; THEIR PREPARATION OR CHEMICAL WORKING-UP; COMPOSITIONS BASED THEREON
- C08J—WORKING-UP; GENERAL PROCESSES OF COMPOUNDING; AFTER-TREATMENT NOT COVERED BY SUBCLASSES C08B, C08C, C08F, C08G or C08H
- C08J9/00—Working-up of macromolecular substances to porous or cellular articles or materials; After-treatment thereof
- C08J9/28—Working-up of macromolecular substances to porous or cellular articles or materials; After-treatment thereof by elimination of a liquid phase from a macromolecular composition or article, e.g. drying of coagulum
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61L—METHODS OR APPARATUS FOR STERILISING MATERIALS OR OBJECTS IN GENERAL; DISINFECTION, STERILISATION OR DEODORISATION OF AIR; CHEMICAL ASPECTS OF BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES; MATERIALS FOR BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES
- A61L27/00—Materials for grafts or prostheses or for coating grafts or prostheses
- A61L27/14—Macromolecular materials
- A61L27/20—Polysaccharides
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61L—METHODS OR APPARATUS FOR STERILISING MATERIALS OR OBJECTS IN GENERAL; DISINFECTION, STERILISATION OR DEODORISATION OF AIR; CHEMICAL ASPECTS OF BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES; MATERIALS FOR BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES
- A61L27/00—Materials for grafts or prostheses or for coating grafts or prostheses
- A61L27/14—Macromolecular materials
- A61L27/22—Polypeptides or derivatives thereof, e.g. degradation products
- A61L27/222—Gelatin
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61L—METHODS OR APPARATUS FOR STERILISING MATERIALS OR OBJECTS IN GENERAL; DISINFECTION, STERILISATION OR DEODORISATION OF AIR; CHEMICAL ASPECTS OF BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES; MATERIALS FOR BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES
- A61L27/00—Materials for grafts or prostheses or for coating grafts or prostheses
- A61L27/50—Materials characterised by their function or physical properties, e.g. injectable or lubricating compositions, shape-memory materials, surface modified materials
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61L—METHODS OR APPARATUS FOR STERILISING MATERIALS OR OBJECTS IN GENERAL; DISINFECTION, STERILISATION OR DEODORISATION OF AIR; CHEMICAL ASPECTS OF BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES; MATERIALS FOR BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES
- A61L2430/00—Materials or treatment for tissue regeneration
- A61L2430/12—Materials or treatment for tissue regeneration for dental implants or prostheses
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- C—CHEMISTRY; METALLURGY
- C08—ORGANIC MACROMOLECULAR COMPOUNDS; THEIR PREPARATION OR CHEMICAL WORKING-UP; COMPOSITIONS BASED THEREON
- C08J—WORKING-UP; GENERAL PROCESSES OF COMPOUNDING; AFTER-TREATMENT NOT COVERED BY SUBCLASSES C08B, C08C, C08F, C08G or C08H
- C08J2201/00—Foams characterised by the foaming process
- C08J2201/04—Foams characterised by the foaming process characterised by the elimination of a liquid or solid component, e.g. precipitation, leaching out, evaporation
- C08J2201/046—Elimination of a polymeric phase
- C08J2201/0462—Elimination of a polymeric phase using organic solvents
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- C—CHEMISTRY; METALLURGY
- C08—ORGANIC MACROMOLECULAR COMPOUNDS; THEIR PREPARATION OR CHEMICAL WORKING-UP; COMPOSITIONS BASED THEREON
- C08J—WORKING-UP; GENERAL PROCESSES OF COMPOUNDING; AFTER-TREATMENT NOT COVERED BY SUBCLASSES C08B, C08C, C08F, C08G or C08H
- C08J2201/00—Foams characterised by the foaming process
- C08J2201/04—Foams characterised by the foaming process characterised by the elimination of a liquid or solid component, e.g. precipitation, leaching out, evaporation
- C08J2201/048—Elimination of a frozen liquid phase
- C08J2201/0484—Elimination of a frozen liquid phase the liquid phase being aqueous
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- C—CHEMISTRY; METALLURGY
- C08—ORGANIC MACROMOLECULAR COMPOUNDS; THEIR PREPARATION OR CHEMICAL WORKING-UP; COMPOSITIONS BASED THEREON
- C08J—WORKING-UP; GENERAL PROCESSES OF COMPOUNDING; AFTER-TREATMENT NOT COVERED BY SUBCLASSES C08B, C08C, C08F, C08G or C08H
- C08J2207/00—Foams characterised by their intended use
- C08J2207/10—Medical applications, e.g. biocompatible scaffolds
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- C—CHEMISTRY; METALLURGY
- C08—ORGANIC MACROMOLECULAR COMPOUNDS; THEIR PREPARATION OR CHEMICAL WORKING-UP; COMPOSITIONS BASED THEREON
- C08J—WORKING-UP; GENERAL PROCESSES OF COMPOUNDING; AFTER-TREATMENT NOT COVERED BY SUBCLASSES C08B, C08C, C08F, C08G or C08H
- C08J2389/00—Characterised by the use of proteins; Derivatives thereof
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- C—CHEMISTRY; METALLURGY
- C08—ORGANIC MACROMOLECULAR COMPOUNDS; THEIR PREPARATION OR CHEMICAL WORKING-UP; COMPOSITIONS BASED THEREON
- C08J—WORKING-UP; GENERAL PROCESSES OF COMPOUNDING; AFTER-TREATMENT NOT COVERED BY SUBCLASSES C08B, C08C, C08F, C08G or C08H
- C08J2405/00—Characterised by the use of polysaccharides or of their derivatives not provided for in groups C08J2401/00 or C08J2403/00
- C08J2405/08—Chitin; Chondroitin sulfate; Hyaluronic acid; Derivatives thereof
Abstract
The invention discloses a method for manufacturing a tissue engineering scaffold with tooth appearance, which comprises the following steps: s1, weighing 0.1-0.25g of gelatin, 2.5-10mg of sodium hyaluronate and 0.05-0.1g of chondroitin sulfate sodium salt, adding into 2-4ml of double distilled water, and fully mixing and dissolving to form a solution; s2, weighing 10-20mg of EDC, adding into 1-2ml of double distilled water, and fully and uniformly mixing to obtain a 1% -2% EDC solution; s3, fully and uniformly mixing the solution obtained in the step S1 and the solution obtained in the step S2, and standing at room temperature until foams disappear; s4, taking the solution obtained in the step S3 to the upper jaw 4 of the double-tooth industrial silica gel model until the upper jaw is full; s5, freeze-drying the S4 by a freeze dryer at the temperature of-50 ℃ to obtain a primary freeze-dried scaffold; and S6, separating the industrial silica gel model from the scaffold to obtain the scaffold. The material of the method is an extracellular matrix component, and has good histocompatibility; the method has the advantages of high manufacturing feasibility, low failure rate, simple and easy operation steps, similar shape to the original teeth in the oral cavity, easy acceptance by patients, suitability for in vitro culture of regenerated teeth and good market prospect.
Description
Technical Field
The present invention relates to a medical tissue engineering material for tissue engineering, in particular, it relates to a method for manufacturing tissue engineering scaffold with tooth appearance.
Background
Tissue engineering refers to the application of the principles and methods of life sciences and engineering to design, build, and maintain the growth of human cells and tissues to restore the function of damaged tissues or organs. The development of tissue engineering offers the possibility of tissue or organ repair and reconstruction. Tissue engineering materials are also receiving increasing attention, and research and development of materials with good tissue compatibility have become the cornerstone for tissue engineering development.
There is now an increasing concern about dental health, and there is also a need for more effective treatment techniques in dental treatment. The technology for clinically regenerating the teeth by using the tooth germ cells (BDCs) for autograft obtained by delayed eruption of wisdom teeth and the like and the scaffolds for molding the appearance of the teeth by using tissue engineering materials and the autologous bone marrow cells to generate the related growth factors and morphogens required by tooth regeneration greatly meets the requirement of a patient on having one mouth of healthy teeth and the requirement of a dental treatment technology, and reduces the defects after the treatment of tooth loss, tooth filling and the like. The development of this technology presupposes a good cell carrier, which provides a basic site for the growth of cells.
Disclosure of Invention
Based on the above, the invention provides the manufacturing method of the tissue engineering scaffold with the tooth appearance, which has the advantages of high feasibility, low failure rate, simple and easy operation steps, similarity with the original tooth shape in the oral cavity, easy acceptance by patients and good market prospect.
The method comprises the following steps:
s1, weighing 0.1-0.25g of gelatin, 2.5-10mg of sodium hyaluronate and 0.05-0.1g of chondroitin sulfate sodium salt, adding into 2-4ml of double distilled water, and fully mixing and dissolving to form a solution;
s2, weighing 10-20mg of EDC, adding into 1-2ml of double distilled water, and fully and uniformly mixing to obtain a 1% -2% EDC solution;
s3, fully and uniformly mixing the solution obtained in the step S1 and the solution obtained in the step S2, and standing at room temperature until foams disappear;
s4, taking the solution obtained in the step S3 to the upper jaw of the double-tooth industrial silica gel model until the double-tooth industrial silica gel model is full;
s5, freeze-drying the S4 by a freeze dryer at the temperature of-50 ℃ to obtain a primary freeze-dried scaffold;
and S6, separating the double-tooth industrial silica gel model from the scaffold.
Further, the EDC in S2 has a value range of 1-2 ml.
Adding 0.9-5ml of 0.2% EDC solution into the solution obtained in S3, and reacting for 24h at room temperature; transferring the obtained solution into an upper jaw 2 industrial silica gel model until the solution is full; freezing at-20 deg.C for 24-30 hr; and then freeze-drying for 70-100 hours at the temperature of-50 ℃ in a freeze dryer to obtain the secondary freeze-dried scaffold.
Further, the freeze-drying time at S5 at-50 ℃ is in the range of 70 to 100 hours.
Compared with the prior art, the invention has the following advantages:
because the preparation conditions are adopted and the components are controlled within the ranges, the prepared upper jaw scaffold is complete in shape and white and yellowish, and a layer of colorless transparent film is arranged on the surface; the tooth shape is complete, the interocclusal fossa and sulcus point gap of the occlusal surface is clear, the shape of the cuspid ridge is regular, the buccal tongue surface is complete, the adjacent surface is quadrilateral, the neck is wider, the near-far and middle contact area is close to the edge and is deviated from the buccal side, and the near-middle sulcus is exposed.
By utilizing the preparation method, the left maxillary lateral incisor scaffold is freeze-dried twice and is white and yellowish. The scaffold has complete shape, clear edge ridge and cutting ridge and round blunt shape. The labial surface is trapezoidal, slightly longer at the proximal edge and shorter at the distal edge, and is connected with the cutting edge in an arc shape. The marginal ridge of the lingual surface is obvious, and the lingual fossa is narrow and deep. The adjacent surfaces are slightly triangular, and can distinguish the near and far adjacent surfaces.
Drawings
Fig. 1 is a scanning electron microscope image of an engineering scaffold manufactured by the method of the present invention.
Detailed Description
For the purpose of illustrating the technical content, the constructional features, the achieved objects and the effects of the invention in detail, reference will be made to the following detailed description of the embodiments in conjunction with the accompanying drawings.
Example one
Referring to FIG. 1, first, the maxilla 4 industrial silica gel model and the maxilla 2 industrial silica gel model were measured to determine the volume of maxilla 4 to be 3.5ml and the volume of maxilla 2 to be 1.5ml using pure water.
The manufacturing method of the tissue engineering scaffold with tooth appearance comprises the following steps:
s1, weighing 0.25g of gelatin, 2.5mg of sodium hyaluronate and 0.05g of chondroitin sulfate sodium salt, adding into 3.5ml of double distilled water, and fully mixing and dissolving to form a solution; s2, weighing 10mg of EDC, adding into 1ml of double distilled water, and fully and uniformly mixing to prepare 1ml of 1% EDC solution; s3, fully and uniformly mixing the solution obtained in the step S1 and the solution obtained in the step S2, and standing at room temperature until foams disappear; s4, taking the S3 mixed solution by using a pipette gun into the upper jaw 4 industrial silica gel model until the upper jaw 4 industrial silica gel model is full; s5, freeze-drying the S4 for 96 hours by using a freeze dryer at the temperature of-50 ℃ to obtain a primary freeze-dried scaffold; and S6, separating the industrial silica gel model from the scaffold.
Adding 1ml of 0.2% EDC solution into the solution obtained in S3, and reacting for 24h at room temperature; transferring the obtained solution into an upper jaw 2 industrial silica gel model until the solution is full; freezing at-20 deg.C for 24 hr; and then freeze-drying for 72 hours at the temperature of 50 ℃ below zero in a freeze dryer to obtain a secondary freeze-dried scaffold.
Example two:
the manufacturing method of the tissue engineering scaffold with tooth appearance comprises the following steps:
s1, weighing 0.25g of gelatin, 10mg of sodium hyaluronate and 0.1g of chondroitin sulfate sodium salt, adding into 5ml of double distilled water, and fully mixing and dissolving to form a solution; s2, weighing 20mg of EDC, adding into 2ml of double distilled water, and fully and uniformly mixing to obtain 2ml of 1% EDC solution; s3, fully and uniformly mixing the solution obtained in the step S1 and the solution obtained in the step S2, and standing at room temperature until foams disappear; s4, taking the S3 mixed solution by using a pipette gun into the upper jaw 4 industrial silica gel model until the upper jaw 4 industrial silica gel model is full; s5, freeze-drying the S4 for 72 hours by using a freeze dryer at the temperature of-50 ℃ to obtain a primary freeze-dried scaffold; and S6, separating the industrial silica gel model from the scaffold.
Adding 1ml of 0.2% EDC solution into the solution obtained in S3, and reacting for 24h at room temperature; transferring the obtained solution into an upper jaw 2 industrial silica gel model until the solution is full; freezing at-20 deg.C for 24 hr; and then freeze-drying for 72 hours at the temperature of 50 ℃ below zero in a freeze dryer to obtain a secondary freeze-dried scaffold.
Claims (6)
1. The manufacturing method of the tissue engineering scaffold with tooth appearance is characterized in that: the method comprises the following steps:
s1, weighing 0.1-0.25g of gelatin, 2.5-10mg of sodium hyaluronate and 0.05-0.1g of chondroitin sulfate sodium salt, adding into 2-4ml of double distilled water, and fully mixing and dissolving to form a solution;
s2, weighing 10-20mg of EDC, adding into 1-2ml of double distilled water, and fully and uniformly mixing to obtain a 1% -2% EDC solution;
s3, fully and uniformly mixing the solution obtained in the step S1 and the solution obtained in the step S2, and standing at room temperature until foams disappear;
s4, taking the solution obtained in the step S3 to the upper jaw of the double-tooth industrial silica gel model until the double-tooth industrial silica gel model is full;
s5, freeze-drying the S4 by a freeze dryer at the temperature of-50 ℃ to obtain a primary freeze-dried scaffold;
and S6, separating the double-tooth industrial silica gel model from the scaffold.
2. The method for manufacturing a tissue engineering scaffold having a dental appearance according to claim 1, wherein: the EDC solution in S2 has a value range of 1-2 ml.
3. The method for manufacturing a tissue engineering scaffold having a dental appearance according to claim 1, wherein: adding 0.9-5ml of 0.2% EDC solution into the solution obtained in S3, and reacting for 24h at room temperature; transferring the obtained solution into the upper jaw of a double-tooth industrial silica gel model until the upper jaw is full; freezing at-20 deg.C for 24-30 hr; and then freeze-drying for 70-100 hours at the temperature of-50 ℃ in a freeze dryer to obtain the secondary freeze-dried scaffold.
4. The method for manufacturing a tissue engineering scaffold having a dental appearance according to claim 1, wherein: the lyophilization time at-50 ℃ in S5 ranged from 70 to 100 hours.
5. The method of claim 1, wherein the upper jaw scaffold has a complete shape and is white and yellowish, and a colorless transparent film is formed on the surface; the teeth have complete shape, clear alveolar and sulcus gaps of the occlusal surfaces, regular shape of the apices, complete buccal and lingual surfaces, quadrangular adjacent surface, wide neck and near-far contact area
The edges are buccal and the mesial sulcus emerges.
6. The method for manufacturing a tissue engineering scaffold with tooth appearance according to claim 3, wherein the prepared freeze-dried twice left maxillary lateral incisor scaffold is white and yellowish; the scaffold has complete shape, clear edge ridge and cutting ridge and round blunt shape; the labial surface is trapezoidal, has a long near middle edge and a short far middle edge and is connected with the cutting edge in an arc shape; the marginal ridge of the lingual surface is obvious, and the lingual fossa is narrow and deep; the adjacent surfaces are slightly triangular, and can distinguish the near and far adjacent surfaces.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201911226907.9A CN111073025A (en) | 2019-12-04 | 2019-12-04 | Method for manufacturing tissue engineering scaffold with tooth appearance |
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| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201911226907.9A CN111073025A (en) | 2019-12-04 | 2019-12-04 | Method for manufacturing tissue engineering scaffold with tooth appearance |
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| CN111073025A true CN111073025A (en) | 2020-04-28 |
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| CN201911226907.9A Pending CN111073025A (en) | 2019-12-04 | 2019-12-04 | Method for manufacturing tissue engineering scaffold with tooth appearance |
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Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JP2004067630A (en) * | 2002-08-09 | 2004-03-04 | Sangaku Renkei Kiko Kyushu:Kk | Scaffold for teeth regeneration, production method therefor, and method for regenerating teeth using the same |
| CN104211982A (en) * | 2013-05-31 | 2014-12-17 | 嘉兴学院 | Manufacturing method of multi-aperture tissue engineering scaffold for cell growth |
| US20150335400A1 (en) * | 2009-06-17 | 2015-11-26 | The Trustees Of Columbia University In The City Of New York | Tooth scaffolds |
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2019
- 2019-12-04 CN CN201911226907.9A patent/CN111073025A/en active Pending
Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JP2004067630A (en) * | 2002-08-09 | 2004-03-04 | Sangaku Renkei Kiko Kyushu:Kk | Scaffold for teeth regeneration, production method therefor, and method for regenerating teeth using the same |
| US20150335400A1 (en) * | 2009-06-17 | 2015-11-26 | The Trustees Of Columbia University In The City Of New York | Tooth scaffolds |
| CN104211982A (en) * | 2013-05-31 | 2014-12-17 | 嘉兴学院 | Manufacturing method of multi-aperture tissue engineering scaffold for cell growth |
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Application publication date: 20200428 |