CN112245662A - Preparation method of Hep-HA composite porous material and application of Hep-HA composite porous material in construction of root canal built-in support - Google Patents
Preparation method of Hep-HA composite porous material and application of Hep-HA composite porous material in construction of root canal built-in support Download PDFInfo
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- A—HUMAN NECESSITIES
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- 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
- A61L27/56—Porous materials, e.g. foams or sponges
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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
- A61L27/26—Mixtures of macromolecular compounds
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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
- A61L27/54—Biologically active materials, e.g. therapeutic substances
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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
- A61L2300/00—Biologically active materials used in bandages, wound dressings, absorbent pads or medical devices
- A61L2300/20—Biologically active materials used in bandages, wound dressings, absorbent pads or medical devices containing or releasing organic materials
- A61L2300/23—Carbohydrates
- A61L2300/236—Glycosaminoglycans, e.g. heparin, hyaluronic acid, chondroitin
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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
- A61L2300/00—Biologically active materials used in bandages, wound dressings, absorbent pads or medical devices
- A61L2300/40—Biologically active materials used in bandages, wound dressings, absorbent pads or medical devices characterised by a specific therapeutic activity or mode of action
- A61L2300/412—Tissue-regenerating or healing or proliferative agents
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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
- A61L2300/00—Biologically active materials used in bandages, wound dressings, absorbent pads or medical devices
- A61L2300/40—Biologically active materials used in bandages, wound dressings, absorbent pads or medical devices characterised by a specific therapeutic activity or mode of action
- A61L2300/412—Tissue-regenerating or healing or proliferative agents
- A61L2300/414—Growth factors
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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
- A61L2300/00—Biologically active materials used in bandages, wound dressings, absorbent pads or medical devices
- A61L2300/40—Biologically active materials used in bandages, wound dressings, absorbent pads or medical devices characterised by a specific therapeutic activity or mode of action
- A61L2300/42—Anti-thrombotic agents, anticoagulants, anti-platelet agents
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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
- A61L2430/00—Materials or treatment for tissue regeneration
- A61L2430/12—Materials or treatment for tissue regeneration for dental implants or prostheses
Abstract
The invention belongs to the technical field of biomaterial preparation, and particularly relates to a preparation method of a Hep-HA composite porous material and application thereof in construction of a root canal internal stent, wherein heparin is placed in 2- (N-morpholino) ethanesulfonic acid (MES) buffer solution, and 1- (3-dimethylaminopropyl) -3-Ethylcarbodiimide (EDC) and N-hydroxysuccinimide (NHS) are added; adding hyaluronic acid into the activated heparin solution; the Hep-HA composite porous material is prepared after uniform stirring, dialysis purification, freeze forming and freeze drying, the preformed bracket which can be placed into the dental canal is prepared by adopting the Hep-HA composite porous material, the form and the structure of the preformed bracket which can be placed into the dental canal and prepared by adopting the method can be regulated, the yield is higher, the process is simple, and the operation is convenient.
Description
Technical Field
The invention belongs to the technical field of preparation of biological materials, and particularly relates to a preparation method of a Hep-HA composite porous material and application of the Hep-HA composite porous material in construction of a root canal built-in support.
Background
At present, dental pulp regeneration is one of the hot problems in the field of dental pulp research, and dental pulp tissue regeneration needs to be completed by the participation of seed cells, growth factors and scaffolds, however, due to the limitation of the special anatomical structure of the root canal and the particularity of the root canal environment, the progress of the research on dental pulp regeneration is seriously hindered, and therefore, the construction of the root canal built-in scaffold which is suitable for the root canal form and is convenient to apply is one of the great challenges in the research on dental pulp regeneration.
Research shows that the composite porous material has a three-dimensional microporous structure and is widely applied in the fields of tissue engineering, drug carriers, biomedicine and the like, and thus, the composite porous material is concerned by a plurality of tissue engineering researchers. Among them, the pre-formed scaffold conveniently placed in the root canal is expected to promote the regeneration of dental pulp tissue, and this research has become a research focus in this field. Hyaluronic Acid (HA) is a degradable high molecular biomaterial, belongs to linear glycosaminoglycan, is a main component of extracellular matrix in human body, regulates cell proliferation, differentiation and movement in natural tissues, and HAs non-immunogenicity, non-antigenicity, good biocompatibility and biological activity; HA-based biomaterials and bioscaffolds do not trigger allergy or inflammation and are hydrophilic. Heparin (Hep) is an anticoagulant, which is a mucopolysaccharide sulfate formed by alternately connecting two kinds of polysaccharides, and has an anticoagulant effect both in vivo and in vitro, and on the other hand, Heparin can bind to growth factors to exert a controlled-release growth factor effect. The composite porous scaffold based on the natural polymer materials HAs better application prospects in the fields of textile, clinic, veterinary medicine, health, agriculture and the like, and is expected to construct a scaffold adaptive to the anatomical characteristics of the root canal due to the plasticity of the shape and the structure, and at present, no report about the preparation of the root canal built-in preformed scaffold by applying Hep-HA is reported in the field of dental pulp tissue regeneration.
Disclosure of Invention
In order to overcome the defects of the prior art, the primary object of the present invention is to provide a method for preparing a Hep-HA composite porous material, wherein the prepared product HAs a composite porous structure, comprises two components of hyaluronic acid and heparin (which can be combined with growth factors), and can be used as a material for manufacturing dental pulp regeneration scaffolds.
The second purpose of the invention is to provide the Hep-HA composite porous material prepared by the preparation method.
The third purpose of the invention is to provide a preformed bracket which can be placed in a root canal, is prepared by the Hep-HA composite porous material, is convenient to place in the root canal and is expected to be used as a dental pulp regeneration bracket.
It is a fourth object of the present invention to provide a method for preparing a preformed bracket that can be placed in a root canal of a tooth as described above.
In order to achieve the purpose, the invention adopts the technical scheme that:
the invention provides a preparation method of a Hep-HA composite porous material, which comprises the following steps:
s1, placing heparin (Hep) in 2- (N-morpholino) ethanesulfonic acid (MES) buffer solution, adding 1- (3-dimethylaminopropyl) -3-Ethylcarbodiimide (EDC) and N-hydroxysuccinimide (NHS), and activating Hep;
s2, adding Hyaluronic Acid (HA) into the mixed solution obtained in the step S1, and uniformly stirring;
s3, and the mixture liquid obtained in the step S2 is dialyzed, frozen, formed and dried to obtain the Hep-HA composite porous material.
Preferably, the MES buffer has a pH of 4 to 5 and a molar concentration of 0.05mol/L to 0.1 mol/L. Further, the pH of the MES buffer solution is 4.5, and the molar concentration is 0.05 mol/L.
Preferably, the molar ratio of Hep, EDC and NHS is 1 (1-3) to (1-3).
Preferably, the mass ratio of the hyaluronic acid to the heparin is (1-10): (1-10). Further, the mass ratio of the hyaluronic acid to the heparin is 1: (1-5).
Preferably, the concentration of Hep is 5mg/mL-50 mg/mL.
Preferably, the dialysis is purification by dialysis with distilled water for 3-5 days, and the molecular weight cut-off is 25000-50000 Da. Further, the dialysis is a dialysis purification with distilled water for 3 days, and the molecular weight cut-off is 50000 Da.
Preferably, the freeze forming is freezing for 24-48h under (-15 deg.C) - (-30 deg.C) conditions. Further, the freezing and forming are carried out for 48h at the temperature of-20 ℃.
Preferably, the drying is vacuum freeze drying for 72-96h, the freezing temperature is less than-60 ℃, and the vacuum degree is less than 10 Pa. Further, the drying is vacuum freeze drying for 72 hours, the temperature is-80 ℃, and the vacuum degree is ultimate vacuum 1 Pa.
The invention also provides the Hep-HA composite porous material prepared by the preparation method.
The invention also provides a preformed bracket convenient to be placed in a tooth root canal, which comprises a round bar-shaped bracket made of the Hep-HA composite porous material and a gutta-percha point inserted in the round bar-shaped bracket.
Preferably, the taper of the gutta-percha point is 0.02-0.04(15# -40 #).
Preferably, the diameters of the two ends of the circular strip-shaped bracket are equal or unequal. Furthermore, the diameter range of the two ends of the round bar-shaped bracket is 0.5mm-3mm, and the length range is 5mm-20 mm.
The invention also provides a preparation method of the preformed bracket capable of being placed in the root canal, which comprises the following steps:
s1, preparing a porous wax mold for manufacturing the round bar-shaped bracket;
s2, shearing the Hep-HA composite porous material, dissolving the cut Hep-HA composite porous material in water, pouring the cut Hep-HA composite porous material into a porous wax mold, and inserting the gutta-percha point into the pores of the wax mold in the middle;
s3, taking out the wax mould after the freeze forming and freeze drying of the wax mould of the step S2 to obtain the preformed bracket which can be placed in the root canal.
Preferably, the preparation method of the porous wax mold comprises the following steps: melting the wax block into liquid wax, pouring the liquid wax into a container, then putting the cut round table-shaped battens into the container filled with the liquid wax, taking out the battens after the liquid wax is solidified, and obtaining the porous wax mould.
Specifically, the preparation method of the porous wax mold comprises the following steps: melting wax blocks into liquid wax, pouring the liquid wax into a container, wherein the depth of the container is more than 25mm, vertically penetrating and fixing the cut round table-shaped battens (the height is Hmm, the diameter of the upper bottom surface is amm, the diameter of the lower bottom surface is bmm, wherein a is less than or equal to b) on a hard paper sheet (the round table-shaped battens are vertically penetrated and fixed on the hard paper sheet to fix the round table-shaped battens and prepare and obtain a large number of porous molds with uniform length and consistent positions), controlling the distance from the upper bottom surface to the hard paper sheet to be cmm, putting the end of the upper bottom surface into the container filled with the liquid wax, keeping the depth of the round table-shaped battens penetrating into the liquid wax to be Hmm, taking out the battens after the liquid wax is solidified, and the surface of the solidified wax can see pores with uniform, thus obtaining the porous wax mould with the height of hmm, the diameter of the upper bottom surface of amm and the diameter of the lower bottom surface of [ (b-a) H/H + a ] mm. Further, H >20 mm; h is more than or equal to 5mm and less than or equal to 20 mm; a is more than or equal to 0.5mm and less than or equal to 2 mm; b is greater than 2 mm; c >0 mm.
Preferably, the concentration of the cut Hep-HA composite porous material dissolved in water is (2.5-10) mg/(1-4) mL.
Preferably, the Hep-HA composite porous material needs to be uniformly stirred for 3-5 hours at room temperature when being sheared and dissolved in water, and the rotating speed is 1000-2000 rpm.
Preferably, the freeze forming is freezing for 24-48h under (-15 ℃) - (30 ℃).
Preferably, the freeze drying is vacuum freeze drying, the freezing temperature is less than-60 ℃, the vacuum degree is less than 10Pa, and the time is 72-96 h.
Compared with the prior art, the invention has the beneficial effects that:
the invention puts heparin into 2- (N-morpholino) ethanesulfonic acid (MES) buffer solution, and adds 1- (3-dimethylaminopropyl) -3-Ethylcarbodiimide (EDC) and N-hydroxysuccinimide (NHS); adding hyaluronic acid into the activated heparin solution; the Hep-HA composite porous material is prepared by uniformly stirring, dialyzing, purifying, freezing, forming and freeze-drying, is composite porous, comprises two components of hyaluronic acid and heparin (which can be combined with growth factors), and can be used as a material for manufacturing dental pulp regeneration scaffolds; therefore, the Hep-HA composite porous material is cut into pieces, dissolved in water, poured into a porous wax mold, and inserted into the pores of the wax mold in the middle of the gutta-percha point, and the preformed bracket capable of being placed into the dental canal is prepared after freezing forming and freeze drying.
Drawings
FIG. 1 is an SEM image of a Hep-HA composite porous material;
FIG. 2 is a visual inspection of a Hep-HA (1: 1 by mass) preformed stent prepared in example 2;
FIG. 3 is a diagram of different lengths of Hep-HA preformed scaffolds prepared in example 3;
FIG. 4 is a drawing of a different taper Hep-HA preformed stent prepared in example 4;
fig. 5 is a diagram of a model of a preformed bracket that can be conveniently placed into a canal.
Detailed Description
The following further describes the embodiments of the present invention. It should be noted that the description of the embodiments is provided to help understanding of the present invention, but the present invention is not limited thereto. In addition, the technical features involved in the embodiments of the present invention described below may be combined with each other as long as they do not conflict with each other.
The experimental procedures in the following examples were carried out by conventional methods unless otherwise specified, and the test materials used in the following examples were commercially available by conventional methods unless otherwise specified.
The hyaluronic acid in the following examples was purchased from Shandong-West Asia chemical industry Co., Ltd, heparin was purchased from Shanghai-sourced leaf Biotech, and the water used in all the experiments was deionized water.
Example 1 Hep-HA composite porous materials prepared with heparin and hyaluronic acid at different mass ratios
(1) Respectively placing HA250mg, 1250mg and 2500mg in 50mL of 0.05mol/L MES buffer solution with pH 4.5, adding EDC237.66mg and NHS142.68mg, and activating heparin; wherein n (HA), n (EDC), N (NHS) 1:2: 2.
(2) Adding 250mg of hyaluronic acid [ m (HA): m (hep): 1:1/5/10] into the mixed solution, and magnetically and uniformly stirring for 5 hours at room temperature at the rotating speed of 1500 rpm;
(3) putting the mixed solution obtained in the step (2) into a dialysis bag, and dialyzing and purifying for 3 days by using distilled water (the cut-off molecular weight is 50000 Da);
(4) putting the substance (heparin-hyaluronic acid) in the dialysis bag obtained in the step (3) into a refrigerator with the temperature of-20 ℃ for freezing for 48 hours; then putting the mixture into a vacuum freeze dryer for freeze drying for 72h (the vacuum is ultimate vacuum of 1Pa, and the temperature is-80 ℃); and (3) freeze-drying to obtain the cotton-shaped grafted Hep-HA composite porous material, and weighing. And characterizing the shape and structure of the product (Hep-HA composite porous scaffold material) by using an SEM (scanning electron microscope) under the conditions of normal temperature and normal pressure.
As can be seen from table 1, the mass ratio of hyaluronic acid to heparin added affects the final yield, when the mass ratio of hyaluronic acid to heparin is from 1:1 to 1: 5 hours, the increase in production indicates a hyaluronic acid to heparin mass ratio of 1:1 in the presence of unreacted hyaluronic acid; when the mass ratio of the hyaluronic acid to the heparin is from 1: 5 to 1: when 10 hours, the yield is not changed greatly, which indicates that the mass ratio of the hyaluronic acid to the heparin is 1: at time 5, the hyaluronic acid reaction is substantially complete.
SEM observation of the Hep-HA composite porous material prepared in the above way (as shown in figure 1) shows that the prepared material is composite porous, and the material is prepared into a preformed bracket which can be placed in a root canal, thereby being beneficial to dental pulp regeneration.
Table 1 raw material amounts and product data for example 1
EXAMPLE 2 different concentrations of heparin-hyaluronic acid preformed stents (Hep-HA preformed stents)
(1) Manufacturing a mold: melting a red wax block into a liquid state, pouring the liquid state into a paper cup, wherein the depth of the paper cup is more than 25mm, then vertically penetrating and fixing a cut round table-shaped batten (the height is 22mm, the diameter of the upper bottom surface is 1mm, and the diameter of the lower bottom surface is 3mm) on a hard paper sheet, controlling the distance from the upper bottom surface to the hard paper sheet to be 5mm, putting the upper bottom surface end of the round table-shaped batten into a container filled with liquid wax, immersing the batten into the liquid wax for 11mm, taking out the batten after the liquid wax is solidified, and obtaining a porous mould with the solidified wax surface having uniform size, namely obtaining a porous mould with the height of 11mm, the diameter of the upper bottom surface being 1mm, and the diameter of the lower bottom surface being 2 mm;
(2) the mass ratio of Hep to HA prepared in example 1 was respectively weighed as 1:1, cutting 100mg, 50mg and 25mg of the Hep-HA composite porous material, dissolving in 10mL of deionized water, magnetically stirring for 3 hours at room temperature, and rotating at 1500 rpm;
(3) placing the solution obtained in the step (2) into the die in the step (1), inserting the gutta-percha point with the taper of No. 25 and 0.02 into a hole formed by the circular truncated cone-shaped wood strip in the die, and then closing the die by using aluminum foil paper;
(4) the mold is firstly put into a refrigerator with the temperature of 20 ℃ below zero for freezing for 48h, then the mold is put into a vacuum freeze dryer for freeze drying for 72h (the vacuum degree is 1Pa at the limit vacuum, and the temperature is 80 ℃ below zero), and finally the Hep-HA is taken out of the mold, thus obtaining the Hep-HA preformed stent which can be placed into the root canal of the tooth (as shown in figure 2). The water content of the obtained preformed stent was measured, and the results are shown in table 2.
As shown in Table 2, the moisture content of the Hep-HA preformed scaffold prepared by the invention is as high as 95%, and the magnitude of the moisture content is independent of the concentration of Hep-HA.
Table 2 raw material amounts and product data for example 2
Example 3 preparation of Hep-HA Pre-formed scaffolds of varying lengths
(1) Melting the red wax block into a liquid state, pouring the liquid state into a paper cup, wherein the depth of the paper cup is more than 25mm, then pricking cut round table-shaped battens (the height is 22mm, the diameter of the upper bottom surface is 1mm, and the diameter of the lower bottom surface is 3mm) on a hard paper sheet, putting the stiff paper sheet into a container filled with liquid wax, wherein the depths of the battens immersed into the liquid wax are respectively 9mm, 11mm and 13mm, and taking out the battens after the liquid wax is solidified, thus obtaining the porous mold with the lengths of 9mm, 11mm and 13mm, the diameter of the upper bottom surface is 1mm, and the diameter of the lower bottom surface is respectively 1.82mm, 2.00mm and 2.18 mm;
(2) the mass ratio of Hep to HA prepared in example 1 was weighed as 1:1, shearing 100mg of Hep-HA composite porous material, dissolving in 10mL of deionized water, magnetically stirring at room temperature for 3h, and rotating at 1500 rpm;
(3) placing the solution obtained in the step (2) in the mould in the step (1), inserting the gutta-percha point with the taper of No. 25 and 0.02 into the gap, and then sealing the mould by using aluminum foil paper;
(4) the mould is firstly put into a refrigerator with the temperature of minus 20 ℃ for freezing for 48h, then the mould is put into a vacuum freeze dryer for freeze drying for 72h, and finally the Hep-HA is taken out of the mould, thus obtaining the preformed stents with different lengths which can be placed into the root canal (as shown in figure 3).
As shown in Table 3, the length of the pre-formed Hep-HA scaffold prepared by the method of the present invention is controllable.
Table 3 raw material amounts and product data for example 3
EXAMPLE 4 preparation of different taper Hep-HA preformed stents
(1) Melting the red wax block into a liquid state, pouring the liquid state into a paper cup, wherein the size of the paper cup is larger than 25mm, then pricking cut round table-shaped battens (the height is 22mm, the diameter of the upper bottom surface is 1mm, and the diameters of the lower bottom surfaces are 1.5mm, 2.1mm and 3mm respectively) on a hard paper sheet, putting the stiff paper sheet into a container containing liquid wax, immersing the battens into the liquid wax to a depth of 7mm, and taking out the battens after the liquid wax is solidified, thus obtaining a porous mold with the height of 7mm, the diameter of the upper bottom surface is 1mm, and the diameter of the lower bottom surface is 2 mm;
(2) the mass ratio of Hep to HA prepared in example 1 was weighed as 1:1, shearing 100mg of Hep-HA composite porous material, dissolving in 10mL of deionized water, magnetically stirring at room temperature for 3h, and rotating at 1500 rpm;
(3) placing the solution obtained in the step (2) in the mould in the step (1), inserting the gutta-percha point with the taper of No. 25 and 0.02 into the gap, and then sealing the mould by using aluminum foil paper;
(4) firstly, putting the mould into a refrigerator with the temperature of-20 ℃ for freezing for 48 h; then putting the mould into a vacuum freeze dryer for freeze drying for 72 hours; finally, the Hep-HA is taken out of the mould, so that the Hep-HA preformed bracket with different tapers which can be placed in the root canal can be obtained (as shown in figure 4).
As shown in Table 4, the degree of taper of the preformed stent made of Hep-HA according to the present invention was controllable.
Table 4 raw material amounts and product data for example 4
The Hep-HA preformed scaffold prepared in the above example can be conveniently placed in a root canal (as shown in fig. 5) and can be applied to dental pulp tissue regeneration, and the construction method of the scaffold is not only applicable to Hep-HA as listed in the examples, but also applicable to other composite porous materials besides the examples. Other template materials, solvent materials, dialysis times, agitation rates and times, freezing and lyophilization times, different scaffold sizes and morphologies, and the like, other than those used in the examples, are within the contemplation of the present invention.
The embodiments of the present invention have been described in detail, but the present invention is not limited to the described embodiments. It will be apparent to those skilled in the art that various changes, modifications, substitutions and alterations can be made in these embodiments without departing from the principles and spirit of the invention, and the scope of protection is still within the scope of the invention.
Claims (9)
1. A preparation method of a Hep-HA composite porous material is characterized by comprising the following steps:
s1, placing Hep in MES buffer solution, and adding EDC and NHS;
s2, adding HA into the mixed solution obtained in the step S1, and uniformly stirring;
s3, and the mixture liquid obtained in the step S2 is dialyzed, frozen, formed and dried to obtain the Hep-HA composite porous material.
2. The method for preparing a Hep-HA composite porous material according to claim 1, wherein the MES buffer HAs a pH of 4-5 and a molarity of 0.05mol/L-0.1 mol/L.
3. The method for preparing a Hep-HA composite porous material as defined in claim 1, wherein the molar ratio of Hep, EDC and NHS is 1 (1-3) to (1-3).
4. The method for preparing a Hep-HA composite porous material according to claim 1, wherein the mass ratio of HA to Hep is (1-10): (1-10).
5. A Hep-HA composite porous material prepared by the preparation method of any one of claims 1-4.
6. A preformed bracket which can be placed in a root canal, comprising a round bar-shaped bracket made of the Hep-HA composite porous material according to claim 5 and a gutta-percha point inserted in the round bar-shaped bracket.
7. The preformed bracket implantable in a root canal of claim 6, wherein the taper of the gutta percha point is 0.02 to 0.04.
8. The pre-shaped bracket capable of being placed in a root canal of claim 7, wherein the diameter of the two ends of the round bar-shaped bracket is equal or unequal.
9. A method of preparing a preformed bracket that can be placed in a root canal of a tooth according to any of claims 6 to 8, comprising the steps of:
s1, preparing a porous wax mold for manufacturing the round bar-shaped bracket;
s2, cutting the Hep-HA composite porous scaffold material of claim 5 into pieces, dissolving the cut pieces in water, pouring the pieces into a porous wax mold, and inserting the gutta-percha points into the pores of the wax mold;
s3, taking out the wax mould after the freeze forming and freeze drying of the wax mould of the step S2 to obtain the preformed bracket which can be placed in the root canal.
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| CN116569916B (en) * | 2023-07-13 | 2023-11-28 | 九天览月生物科技(天津)有限公司 | Frozen cells, cell frozen solution, frozen method and application |
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