CN102188754B - Nanometer pore hydroxyl calcium phosphate/aquogel materials - Google Patents
Nanometer pore hydroxyl calcium phosphate/aquogel materials Download PDFInfo
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- CN102188754B CN102188754B CN2011101047139A CN201110104713A CN102188754B CN 102188754 B CN102188754 B CN 102188754B CN 2011101047139 A CN2011101047139 A CN 2011101047139A CN 201110104713 A CN201110104713 A CN 201110104713A CN 102188754 B CN102188754 B CN 102188754B
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Abstract
The invention belongs to the field of biological medicines, and particularly relates to hydroxyl calcium phosphate/aquogel materials which are used as repair and implanting materials for bones or teeth. In order to solve the problems that high bonding interfaces are not formed between tooth implants and dental bones and the like during tooth or bone repair in the prior art, the invention provides nanometer pore hydroxyl calcium phosphate/aquogel materials, which are characterized in that an aquogel surface is modified by polypeptides, and hydroxyl calcium phosphate is deposited on aquogel by the bionic mineralization process. The nanometer pore hydroxyl calcium phosphate/aquogel materials are materials which are highly close to or consistent with materials of human bodies in aspects of macroscopic forms and microstructures and are used for orthopedics and dentistry, are high in biocompatibility, and can be used as repair materials, filling materials and biological support materials for the teeth or the bones.
Description
Technical field
The invention belongs to biomedicine field, be specifically related to a kind of as bone or the reparation of tooth and the calcium hydroxy phosphate/hydrogel material of embedded material.
Background technology
According to interrelated data statistics, China's population morbidity of dental caries is 37%, and on average everyone has 2.47 of dental caries, and the filler that is used for repairing dental caries is in great demand, and estimates that annual requirement is 1 ton of left and right.But the gear division packing material that uses at present all can not promote the quick reparation of dental tissue.
For the dental caries that drops, artificial tooth root is installed normally.Artificial tooth root claims again tooth implant, is by in the jawbone of surgical operation with its implant into body agomphosis position, repairs at an upper portion thereof the device of artificial tooth after healing.Tooth implant is different by its material, can be divided into substantially five types, i.e. (1) Metal and Alloy material type: comprise gold, 316L rustless steel (ferrum-chromium-nickel alloy), casting vitallium, titanium and alloy thereof etc.(2) ceramic material class: comprise bio-inert ceramic, bioactive ceramics, biological degradability pottery etc.(3) carbon materials class: comprise vitreous carbon, Low Temperature Isotropic Carbon etc.(4) macromolecular material class: comprise esters of acrylic acid, politef class, polysulfones etc.(5) composite class, namely above-mentioned two or more material is compound, as ceramic coated etc. in the metal surface.The material metal class that at present tooth implant is commonly used is mainly pure titanium or titanium alloy, the titanium light specific gravity, intensity is high, and is nonmagnetic, contractility is little, stable chemical nature, have splendid corrosion resistance and with the compatibility of on every side biological tissue.Although titanium alloy has biocompatibility preferably, there is no to form very strong combination interface between implant and dentale.
In addition, in surgical operation, the reparation of bone and regeneration are general problems, in order to obtain desirable osseous tissue substitution material, people have carried out a lot of researchs, normally adopt the natural high polymers such as calcium hydroxy phosphate and collagen protein, gelatin, chitosan or the synthetic high polymers such as polyvinyl alcohol, polyacrylamide to prepare composite.
But these composites all also have weak point, the material of preparation and natural bone structural similarity, and improving its biocompatibility is the emphasis of bone tissue engineer research.
Our research discovery, mineralizing of organism is collaborative completing under the joint effect of many factors.Take the bone mineral to turn to example, osteocyte is under the guide of somatomedin, at two kinds of albumen: (1) water-fast skelemin (resembling collagen protein is the skeleton of people's bone); (2) water-solubility protein (as, bone contains osteopontin (osteopontin, OPN), tooth includes with phosphoprotein (phosphophoryn)) co-controlling under complete in order.Traditional theoretical conception has been ignored the effect of water-solubility protein.We find that also water-solubility protein is not all to be evenly distributed in the liquid phase that mineralizes, and wherein most of and skelemin interacts and is adsorbed on skelemin.The out-phase water-solubility protein that is adsorbed adsorbs by ionic bond the ion that mineralizes, and resembles Ca/Mg ion etc., thereby has effectively improved the degree of supersaturation of calcium ion, carbanion and phosphate anion.Because this is the process of a dynamics Controlling, a large amount of hydrones also are involved in wherein, thereby form liquid phase mineral precursor.Liquid phase mineral precursor is progressively dehydration under skelemin is controlled subsequently, and the heterogeneous nucleation principle (epitaxy) by classics changes into crystal.
Summary of the invention
In view of this, the mimic biology of the present invention process that mineralizes, by with water-solubility protein, the biodegradable organization bracket being carried out body and surface modification, and support mineralizes by the bionical process that mineralizes (being similar to the condition that human teeth and bone form), a kind of nanometer pore hydroxyl calcium phosphate/aquogel materials is provided, this material be macroscopic form aspect microstructure all with close or consistent orthopaedics and the dental materials of human body self material height, these materials can with plant tooth, bone-grafting material, timbering material and active packing material.
In order to achieve the above object, the present invention adopts following technical proposals:
Nanometer pore hydroxyl calcium phosphate/aquogel materials provided by the invention, its feature is, with polypeptide, described hydrogel surface is carried out modification, and by the bionical process that mineralizes, calcium hydroxy phosphate is deposited on hydrogel, the preparation method of described nanometer pore hydroxyl calcium phosphate/aquogel materials comprises the steps:
(1) the preparation mass concentration is the hydrophilic macromolecule aqueous solution of 1-5%, the preparation mass concentration is the glutaraldehyde water solution of 3-8%, compound concentration is the polypeptid solution of 10-300 μ g/ml, described hydrophilic macromolecule is selected from collagen, gelatin, polyvinyl alcohol or alginic acid or the compositions of at least two kinds wherein
(2) preparation hydrogel substrate: the ratio that the prepared three kinds of aqueous solutions of step (1) are 1:1:1 according to volume ratio is mixed, stirred at normal temperatures 0.5-10 minute, then mixture is placed in (on sheet glass) on carrier, prepare the hydrogel substrate by phase detachment technique, prepared hydrogel substrate soaks 6-10 hour to remove free glutaraldehyde with the glycine solution of 0.1-0.3M
Above-mentioned phase detachment technique is several phase detachment techniques such as heat-induced gel commonly used, emulsifying/lyophilization, Solid-Liquid Separation.
(3) preparation TRIS buffer: with the Tri(Hydroxymethyl) Amino Methane Hydrochloride of 5-8 gram, 0.5-1.5 gram Tris and 6-10 gram sodium chloride join in 100 ml deionized water, the pH value of this buffer is 7.0-8.0, Tri(Hydroxymethyl) Amino Methane Hydrochloride is used for regulating the pH value of this buffer, PH the best is 7.2-7.4
(4) take respectively calcium chloride and dipotassium hydrogen phosphate, and dissolve in respectively in the buffer that step (3) makes, be mixed with respectively calcium chloride solution and the dipotassium hydrogen phosphate solution of 0.002-0.006mol/L, be the ratio mix and blend mutually of 1:1:X (X=0-6) according to volume ratio with the polypeptid solution that makes in calcium chloride solution and dipotassium hydrogen phosphate solution and step (1)
(5) the prepared hydrogel substrate that is carried on carrier of step (2) is placed in the prepared mixed solution of step (4), after standing 24-48 hour, priority water and washing with alcohol, drying namely makes described nanometer pore hydroxyl calcium phosphate/aquogel materials.
Preferably, the invention provides a kind of nanometer pore hydroxyl calcium phosphate/aquogel materials, its feature is that the preparation method of described nanometer pore hydroxyl calcium phosphate/aquogel materials comprises the steps:
(1) the preparation mass concentration is the hydrophilic macromolecule aqueous solution of 1-3%, and the preparation mass concentration is the glutaraldehyde water solution of 4-6%, and compound concentration is the polypeptid solution of 150-250 μ g/ml,
(2) preparation hydrogel substrate: the ratio that the prepared three kinds of aqueous solutions of step (1) are 1:1:1 according to volume ratio is mixed mutually, stirred at normal temperatures 0.5-10 minute, then mixture is placed on carrier, prepare the hydrogel substrate by phase detachment technique, prepared hydrogel substrate soaks 6-10 hour to remove free glutaraldehyde with the glycine solution of 0.1-0.3M
(3) preparation TRIS buffer: with the Tri(Hydroxymethyl) Amino Methane Hydrochloride of 6-7 gram, 0.6-1.2 gram Tris and 8-9 gram sodium chloride join in 100 ml deionized water, and the pH value of this buffer is 7.2-7.4,
(4) take respectively calcium chloride and dipotassium hydrogen phosphate, and dissolve in respectively in the buffer that step (3) makes, be mixed with respectively calcium chloride solution and the dipotassium hydrogen phosphate solution of 0.002-0.006mol/L, the ratio that the polypeptid solution that makes in calcium chloride solution and dipotassium hydrogen phosphate solution and step (1) is 1:1:X (X=0-6) according to volume ratio is mixed mutually, stirred at normal temperatures 0.5-10 minute
(5) the prepared hydrogel substrate that is carried on carrier of step (2) is placed in the prepared mixed solution of step (4), after standing 24-36 hour, priority water and washing with alcohol, drying namely makes described nanometer pore hydroxyl calcium phosphate/aquogel materials.
Further preferred, the invention provides a kind of nanometer pore hydroxyl calcium phosphate/aquogel materials, it is characterized in that, the preparation method of described nanometer pore hydroxyl calcium phosphate/aquogel materials comprises the steps:
(1) the preparation mass concentration is 1% hydrophilic macromolecule aqueous solution, and the preparation mass concentration is 5% glutaraldehyde water solution, and compound concentration is the polypeptid solution of 200 μ g/ml,
(2) preparation hydrogel substrate: the ratio that the prepared three kinds of aqueous solutions of step (1) are 1:1:1 according to volume ratio is mixed mutually, stirred at normal temperatures 0.5-10 minute, then mixture is placed on carrier, prepare the hydrogel substrate by phase detachment technique, prepared hydrogel substrate soaks 6-10 hour to remove free glutaraldehyde with the glycine solution of 0.1-0.3M
(3) preparation Tris (Tris) buffer: with the Tri(Hydroxymethyl) Amino Methane Hydrochloride (Tris-HCl) of 6.61 grams, 0.97 gram Tris (Tris) and 8.77 gram sodium chloride join in 100 ml deionized water, the pH value of this buffer is 7.2-7.4
(4) take respectively calcium chloride and dipotassium hydrogen phosphate, and dissolve in respectively in the buffer that step (3) makes, be mixed with respectively calcium chloride solution and the dipotassium hydrogen phosphate solution of 0.002-0.006mol/L, the ratio that the polypeptid solution that makes in calcium chloride solution and dipotassium hydrogen phosphate solution and step (1) is 1:1:X (X=0-6) according to volume ratio is mixed mutually, stirred at normal temperatures 0.5-10 minute
(5) the prepared hydrogel substrate that is carried on carrier of step (2) is placed in the prepared mixed solution of step (4), after standing 24 hours, priority water and washing with alcohol, drying namely makes described nanometer pore hydroxyl calcium phosphate/aquogel materials.
The present invention also provides a kind of nanometer pore hydroxyl calcium phosphate/aquogel materials, its feature is, with polypeptide, described hydrogel surface is carried out modification, and by the bionical process that mineralizes, calcium hydroxy phosphate is deposited on hydrogel, the preparation method of described nanometer pore hydroxyl calcium phosphate/aquogel materials comprises the steps:
(1) the preparation mass concentration is the hydrophilic macromolecule aqueous solution of 1-5%, and compound concentration is the polypeptid solution of 10-300 μ g/ml, and described hydrophilic macromolecule is selected from collagen, gelatin, polyvinyl alcohol or alginic acid or the compositions of at least two kinds wherein,
(2) preparation hydrogel substrate: the ratio that the prepared two kinds of aqueous solutions of step (1) are 1:1 according to volume ratio is mixed, stirred at normal temperatures 1-10 minute, and then mixture was placed on carrier, under room temperature dry 24-48 hour, then vacuum drying at the temperature of 80 ℃-100 ℃
(3) preparation TRIS buffer: with the Tri(Hydroxymethyl) Amino Methane Hydrochloride of 5-8 gram, 0.5-1.5 gram Tris and 6-10 gram sodium chloride join in 100 ml deionized water, the pH value of this buffer is 7.0-8.0, preferably, the pH value of this buffer is 7.2-7.4
(4) take respectively calcium chloride and dipotassium hydrogen phosphate, and dissolve in respectively in the buffer that step (3) makes, be mixed with respectively calcium chloride solution and the dipotassium hydrogen phosphate solution of 0.002-0.006mol/L, be the ratio mix and blend mutually of 1:1:X (X=0-6) according to volume ratio with the polypeptid solution that makes in calcium chloride solution and dipotassium hydrogen phosphate solution and step (1)
(5) the prepared hydrogel substrate that is carried on carrier of step (2) is placed in the prepared mixed solution of step (4), after standing 24-48 hour, priority water and washing with alcohol, drying namely makes described nanometer pore hydroxyl calcium phosphate/aquogel materials.
Hydrogel, the gel take water as disperse medium.Have introduce a part of hydrophobic group in the water soluble polymer of cross-linked structure and form can water-swellable cross linked polymer.Be a kind of macromolecule network system, character is soft, can keep certain shape, can absorb a large amount of water.Every water solublity or hydrophilic macromolecule by certain chemical crosslinking or physical crosslinking, can form hydrogel.These macromolecules can be divided into natural and synthetic two large classes by its source.Natural hydrophilic macromolecule comprises polysaccharide (starch, cellulose, alginic acid, hyaluronic acid, chitosan etc.) and polypeptide class (collagen, polylysine, poly-L-glutamic acid etc.).Synthetic hydrophilic high mol comprises polyvinyl alcohol, acrylic acid and derivatives class thereof (polyacrylamide, poly-N-is poly-for acrylamide etc. for polyacrylic acid, polymethylacrylic acid).
Above-mentioned hydrogel is preferably collagen, gelatin, polyvinyl alcohol, alginic acid or the compositions of at least two kinds wherein; Described polypeptide is the osteopontin (osteopontin, OPN) that contains of bone, phosphoprotein (phosphophoryn) or its fragment that tooth contains; Or, described polypeptide can be rich in acidic-group, can be for being rich in the polypeptide of aspartic acid, glutamic acid, phosphorylation or Sulfated serine/tyrosine, wherein, to account for the percentage by weight of polypeptide be 10-90% to the summation of described aspartic acid, glutamic acid, phosphorylation or Sulfated serine/tyrosine; Or described polypeptide is poly-aspartic-acid, polyglutamic acid, poly-Sulfated serine/tyrosine.
Be the further quick reparation of quickening tooth or bone, being added with mass percent in above-mentioned hydrogel material is the micromolecular compound of the promotion osteoblast breeding of 0.0001-5%.Promote the micromolecular compound osteoblast breeding or that promote bone to form to comprise that international publication number is the micromolecular compound of the promoted osteocyte breeding of announcing in the international application of WO2010017472A1, also comprises the compound of following structure:
R wherein
1, R
3, R
4, R
6, R
8, R
11, R
12Or R
13In at least one be hydrogen atom, and R wherein
1, R
3, R
4, R
6, R
8, R
11, R
12Or R
13In at least one comprise the atom of non-hydrogen atom.
Promote the micromolecular compound osteoblast breeding or that promote bone to form also to comprise other the existing known micromolecular compound such as diosgenin, amygdaloside.
the present invention also provides a kind of tooth implant, comprise titanium alloy-based, its feature is, describedly be provided with above-mentioned nanometer pore hydroxyl calcium phosphate/aquogel materials provided by the invention on titanium alloy-based, in its preparation method, step (1) is constant, the process of step (2) preparation hydrogel substrate comprises: the ratio that the prepared three kinds of aqueous solutions of step (1) are 1:1:1 according to volume ratio is mixed mutually, stirred at normal temperatures 0.5-10 minute, then mixture is sprayed to titanium alloy-based on, rapid draing under room temperature was placed in 90 ℃ to 100 ℃ heat treatments 5 to 15 minutes again after (as passing through vacuum drying), prepared hydrogel is two-dimentional, it is the continuous or discrete thin film of one deck, the width of the discontinuous part of described discontinuous thin film is 1 μ m-3mm, prepared hydrogel substrate soaks 6-10 hour to remove free glutaraldehyde with the glycine solution of 0.1-0.3M, step (3) is constant to step (5).
the present invention also provides a kind of tooth implant, comprise titanium alloy-based, describedly be provided with nanometer pore hydroxyl calcium phosphate/aquogel materials provided by the invention on titanium alloy-based, in its preparation method, step (1) is constant, the process of step (2) preparation hydrogel substrate comprises: the ratio that the prepared two kinds of aqueous solutions of step (1) are 1:1 according to volume ratio is mixed, stirred at normal temperatures 1-10 minute, then mixture is sprayed to titanium alloy-based on, be placed in again 90 ℃ to 100 ℃ heat treatments under room temperature after rapid draing 5 to 15 minutes, prepared hydrogel is the continuous or discrete thin film of one deck, the width of the discontinuous part of described discontinuous thin film is 1 μ m-3mm, step (3) is constant to step (5).
the present invention also provides a kind of tooth implant, comprises titanium alloy-basedly, and its feature is, describedly is provided with above-mentioned nanometer pore hydroxyl calcium phosphate/aquogel materials provided by the invention on titanium alloy-based, in its preparation method, step (1) is constant, the process of step (2) preparation hydrogel substrate comprises: the ratio that the prepared three kinds of aqueous solutions of step (1) are 1:1:1 according to volume ratio is mixed mutually, stirred at normal temperatures 0.5-10 minute, process titanium alloy-based surface with chemical method, then with mixture freezing on titanium alloy-based under-8 ℃ to-10 ℃,-10 ℃ to-60 ℃ lower freezing vacuum sublimation dryings 6-8 hour, prepared hydrogel is three-dimensional framework, its thickness is 0.01mm to 5mm, prepared hydrogel substrate soaks 6-10 hour to remove free glutaraldehyde with the glycine solution of 0.1-0.3M, step (3) is constant to step (5).
The present invention also provides a kind of tooth implant, comprises titanium alloy-basedly, and its feature is, describedly is provided with above-mentioned nanometer pore hydroxyl calcium phosphate/aquogel materials provided by the invention on titanium alloy-based; In its preparation method, step (1) is constant, the process of step (2) preparation hydrogel substrate comprises: the ratio that the prepared two kinds of aqueous solutions of step (1) are 1:1 according to volume ratio is mixed, stirred at normal temperatures 1-10 minute, process titanium alloy-based surface with chemical method, then with mixture freezing on titanium alloy-based under-8 ℃ to-10 ℃ ,-10 ℃ to-60 ℃ lower freezing vacuum sublimation dryings 6-8 hour, the thickness of prepared hydrogel was 0.01mm to 5mm; Step (3) is constant to step (5).
The present invention also provides a kind of calcium-supplementing nutritive product, and its feature is that described nutriment mainly contains above-mentioned nanometer pore hydroxyl calcium phosphate/aquogel materials provided by the invention.
The present invention also provides a kind of orthopaedics or the biological filling/timbering material of gear division, and its feature is that described packing material mainly contains above-mentioned nanometer pore hydroxyl calcium phosphate/aquogel materials provided by the invention.
Compared with prior art, nanometer pore hydroxyl calcium phosphate/aquogel materials provided by the invention, wherein, hydrogel be micron to millimetre-sized poroid two dimension or three-dimensional material, the three-dimensional material hole is interconnected, the aperture is: 1 micron to 3 millimeters; The calcium hydroxy phosphate that is deposited on hydrogel is the poroid calcium hydroxy phosphate of three-dimensional communication, and the aperture is 50-1000nm, and hole wall is thick is the 30-80 nanometer.This material be macroscopic form aspect microstructure all with close or consistent orthopaedics and the dental materials of human body self material height, biocompatibility is stronger.These materials can be used for the repair materials of tooth or bone, packing material, biologic bracket material.
Compared with prior art, calcium-supplementing nutritive product provided by the invention, because its calcium hydroxy phosphate is the poroid calcium hydroxy phosphate of three-dimensional communication, the aperture is 50-1000nm, hole wall is thick is the 30-80 nanometer, so nutrition more easily is absorbed by the body.
Compared with prior art, tooth implant provided by the invention is provided with two dimension or three-dimensional nanometer pore hydroxyl calcium phosphate/aquogel materials provided by the invention on titanium alloy-based due to it, so, dental tissue can be repaired by myna fast, and, formed stronger combination interface between implant and dentale.
Compared with prior art, the biological filling/timbering material of orthopaedics provided by the invention or gear division, biocompatibility is stronger.
Description of drawings
Fig. 1 is the schematic diagram of turkey tendon of mineralizing: (A) the turkey tendon by along its length laterally solution cut open, (B) be cross section Electronic Speculum figure under nonactive polypeptide condition, (C) be longitudinal section Electronic Speculum figure under nonactive polypeptide condition, (D) be the cross section Electronic Speculum figure under the active polypeptide condition of the present invention, (E) be the longitudinal section Electronic Speculum figure under the active polypeptide condition of the present invention
Fig. 2 (A) is the scanning electron microscope (SEM) photograph of the collagen fabric net that mineralizes of the present invention, (B) is the transmission electron microscope picture of the collagen fabric net that mineralizes of the present invention;
Fig. 3 (A) is deposited on the electron-microscope scanning figure on the hydrogel material of polypeptide modification for continuous micron adsorbed property of porous hydroxyl calcium phosphate of the present invention, (B) for the continuous calcium hydroxy phosphate of densification of the present invention is deposited on electron-microscope scanning figure on the hydrogel material of polypeptide modification, (C) be deposited on electron-microscope scanning figure on the hydrogel material of polypeptide modification for the continuous netted calcium hydroxy phosphate of nanoporous of the present invention;
Fig. 4 is the process schematic representation for preparing the poroid calcium hydroxy phosphate/hydrogel material of three-dimensional manometer with freezing method of the present invention, (A) solid free forming schematic diagram, (B) multiinjector deposit manufacture system schematic, (C) three-dimensional framework schematic diagram;
Fig. 5 (A) is arranged at structural representation on tooth implant titanium alloy-based for of the present invention with hydrogel material, (B) is the post-depositional hydrogel material structural representation of calcium hydroxy phosphate of the present invention.
Description of reference numerals
101, tendon elastin laminin protection cortex: mineral liquid phase precursor cannot infiltrate
102, tendon cross section: mineral liquid phase precursor infiltrates thus
103, tendon longitudinal section: will use electronic microscope photos after mineralizing
201, calcium hydroxy phosphate crystal
501, screw
502, crown lid
503, titanium alloy-based
504, two dimension or the three-dimensional framework of hydrogel material formation
505, the post-depositional hydrogel skeleton of calcium hydroxy phosphate
The specific embodiment
Hydrogel material used in embodiment is as gelatin, collagen, alginic acid etc.; The polypeptide such as osteopontin (osteopontin, OPN), phosphoprotein (phosphophoryn) are the product commonly used of selling on market;
Tri(Hydroxymethyl) Amino Methane Hydrochloride (Tris-HCl), Tris (Tris), calcium chloride, dipotassium hydrogen phosphate, glutaraldehyde, sodium chloride used in embodiment are analytical pure.
The prepared sample of embodiment is analyzed with X-ray diffraction (XRD)/transmission electron microscope/scanning electron microscope (SEM is called for short scanning electron microscope).The described carrier of step in embodiment 1-16 (2) can be sheet glass.
As shown in Figure 1: (A) the turkey tendon by along its length laterally solution cut open.The periphery of tendon is wrapped up by elastin laminin 101, and mineral liquid phase precursor cannot infiltrate thus, and can only infiltrate from cross section 102.(B) and (C) be the Electronic Speculum figure under the condition that there is no active polypeptide, as shown in (B), do not having under the condition of nonactive polypeptide, owing to not having the liquid phase precursor to generate, mineral can only be on cross section random deposition, and can not effectively infiltrate longitudinal section (C).On the contrary, at active polypeptide, when existing as osteopontin, liquid phase mineral precursor not only can cover cross section with meticulous matter attitude, as shown in (D), forms and the similar mineral parcel of people's bone collagen fabric, and can effectively infiltrate the longitudinal section, as shown in (E), form the collagen fabric of fine and close mineral parcel, rather than fragmentary random deposition.This has not only greatly improved the density of mineral, and due to the extremely strong interaction force that has formed between mineral and collagen protein and greatly improved the intensity of material monolithic, thereby all closer to the bone material of organism self, can be used as artificial sclerotin and transplant in human body on Macrocosm and microcosm.
As shown in Figure 2, (A) scanning electron microscope: in calcium hydroxy phosphate was wrapped in collagen fabric, the support that mineralizes was the collagen fabric net.The Electronic Speculum result shows that the nanometer pore hydroxyl calcium phosphate/aquogel materials that the present invention prepares is the same with the pattern of people's bone.(B) transmission electron microscope picture (Dark field TEM): transmission electron microscope results shows that the nanometer pore hydroxyl calcium phosphate/aquogel materials that the present invention prepares has identical microstructure with people's bone, and namely the center crystallographic axis of calcium hydroxy phosphate crystal (C axle) is parallel with the major axis of collagen fabric.
As shown in Figure 3, (A) continuous micron adsorbed property of porous hydroxyl calcium phosphate is deposited on the hydrogel material of polypeptide modification.(B) fine and close continuous calcium hydroxy phosphate is deposited on polypeptide modification or the hydrogel material without the polypeptide modification.(C) the netted calcium hydroxy phosphate of continuous nanoporous is deposited on the hydrogel material of polypeptide modification.By hydrogel material is carried out the polypeptide modification, the present invention has obtained the calcium hydroxy phosphate of different densities and form.Because people's bone density of different parts is different, the present invention can synthesize the calcium hydroxy phosphate of proper density and pattern to be used for filling or to be implanted in people's bone of different densities.The holey calcium hydroxy phosphate can be used as the bone cell growth skeleton, and does not need additionally to add somatomedin.The calcium hydroxy phosphate of different densities and form is determined by many factors, as, with the kind of hydrogel, whether hydrogel is modified, by which kind of polypeptide modification, the albumen that adds in the solution that mineralizes, polypeptide or the high molecular sequential structure of imitative albumen with and concentration etc. relation is all arranged.As, poly-aspartate can produce the calcium hydroxy phosphate (atresia) of densification with the hydrogel without modification, as shown in Fig. 3 (B).
As shown in Figure 4: (A) solid free forming.Solid free forming (solid free-form, SFF) technology is also referred to as rapid prototyping input method (Rapid Protyping).Its ultimate principle is that extruder or spraying machine are directly set up three-dimensional physical model layer by layer under the computer digit model is controlled.These mathematical models are from the CAD modeling, three-dimensional reconstruction CT/MRI image or three-dimensional body digitization system.In extruding/course of injection, on computer-controlled nozzle extruded material long filament or liquid droplets deposition of material to a platform, storehouse goes out a three dimensional object from level to level.(B) multiinjector deposit manufacture system schematic.Hydrogel material in water-soluble or solvent is deposit to can be frozen at once on platform, and three-dimensional framework will be stablized and temporarily can be not destroyed.Three-dimensional framework is frozen drying subsequently, thereby obtains stable three-dimensional framework (C).In this support manufacture process, promote albumen, the bone cell growth factor of mineral precursor, the micromolecular compound that promotes that osteoblast is bred or the formation of promotion bone and anti-infective etc. to add to very easily in timbering material, form a regenerating tissues Engineering System.
As shown in Figure 5, a kind of tooth implant provided by the invention comprises titanium alloy-based 503 as the load-bearing matrix, and screw thread 501 is carved with on its top, and can screw on crown lid 502(crown lid can change afterwards in case of necessity); As shown in figure (A), the outer surface of its underpart, at first the hydrogel material that adopts spraying method or freezing method setting shown in Figure 4 not to mineralize, form two dimension or three-dimensional framework 504, by the bionical process that mineralizes of the present invention, calcium hydroxy phosphate is deposited on again subsequently and forms skeleton 505 on hydrogel material, as shown in figure (B).Calcium hydroxy phosphate has strengthened the hydrogel skeleton, has kept simultaneously the connection cavernous structure of three dimensional hydrogel skeleton.Like this osteoblast can be within the shortest time bioelectric interface between repairing transplant body and dentale, tooth implant is firmly got off and can not lose for the fixing adjacent teeth of transplant.
Embodiment 1
Nanometer pore hydroxyl calcium phosphate/aquogel materials provided by the invention, preparation method comprises the steps:
(1) the preparation mass concentration is 1% hydrophilic macromolecule aqueous solution, and the preparation mass concentration is 3% glutaraldehyde water solution, and compound concentration is the polypeptid solution of 10 μ g/ml,
Above-mentioned hydrophilic macromolecule is collagen; Aforementioned polypeptides is the osteopontin (osteopontin, OPN) that bone contains; Be added with mass percent in described hydrophilic high molecular material and be the micromolecular compound of 0.0001% promotion osteoblast breeding.
(2) preparation hydrogel substrate: the ratio that the prepared three kinds of aqueous solutions of step (1) are 1:1:1 according to volume ratio is mixed, stirred at normal temperatures 0.5 minute, then mixture is placed on carrier, prepare the hydrogel substrate by phase detachment technique, prepared hydrogel substrate soaks 6 hours to remove free glutaraldehyde with the glycine solution of 0.1M
(3) preparation TRIS buffer: with the Tri(Hydroxymethyl) Amino Methane Hydrochloride of 5 grams, 0.5 gram Tris and 6 gram sodium chloride join in 100 ml deionized water, and the pH value of this buffer is 7.0-8.0,
(4) take respectively calcium chloride and dipotassium hydrogen phosphate, and dissolve in respectively in the buffer that step (3) makes, be mixed with respectively calcium chloride solution and the dipotassium hydrogen phosphate solution of 0.002mol/L, be that mix and blend is (namely mutually for the ratio of 1:1:0 with the polypeptid solution that makes in calcium chloride solution and dipotassium hydrogen phosphate solution and step (1) according to volume ratio, be the ratio mix and blend mutually of 1:1 according to volume ratio with calcium chloride solution and dipotassium hydrogen phosphate solution)
(5) the prepared hydrogel substrate that is carried on carrier of step (2) is placed in the prepared mixed solution of step (4), after standing 24 hours, priority water and washing with alcohol, drying namely makes described nanometer pore hydroxyl calcium phosphate/aquogel materials.
Embodiment 2
Nanometer pore hydroxyl calcium phosphate/aquogel materials provided by the invention, preparation method comprises the steps:
(1) the preparation mass concentration is 5% hydrophilic macromolecule aqueous solution, and the preparation mass concentration is 8% glutaraldehyde water solution, and compound concentration is the polypeptid solution of 300 μ g/ml,
Above-mentioned hydrophilic macromolecule is gelatin; Aforementioned polypeptides is phosphoprotein (phosphophoryn) or its fragment that tooth contains; Be added with mass percent in above-mentioned hydrophilic high molecular material and be the micromolecular compound of 5% promotion osteoblast breeding.
(2) preparation hydrogel substrate: the ratio that the prepared three kinds of aqueous solutions of step (1) are 1:1:1 according to volume ratio is mixed, stirred at normal temperatures 10 minutes, then mixture is placed on carrier, prepare the hydrogel substrate by phase detachment technique, prepared hydrogel substrate soaks 10 hours to remove free glutaraldehyde with the glycine solution of 0.3M
(3) preparation TRIS buffer: with the Tri(Hydroxymethyl) Amino Methane Hydrochloride of 8 grams, 1.5 gram Tris and 10 gram sodium chloride join in 100 ml deionized water, and the pH value of this buffer is 7.0-8.0,
(4) take respectively calcium chloride and dipotassium hydrogen phosphate, and dissolve in respectively in the buffer that step (3) makes, be mixed with respectively calcium chloride solution and the dipotassium hydrogen phosphate solution of 0.006mol/L, be the ratio mix and blend mutually of 1:1:6 according to volume ratio with the polypeptid solution that makes in calcium chloride solution and dipotassium hydrogen phosphate solution and step (1)
(5) the prepared hydrogel substrate that is carried on carrier of step (2) is placed in the prepared mixed solution of step (4), after standing 48 hours, priority water and washing with alcohol, drying namely makes described nanometer pore hydroxyl calcium phosphate/aquogel materials.
Embodiment 3
Nanometer pore hydroxyl calcium phosphate/aquogel materials provided by the invention, preparation method comprises the steps:
(1) the preparation mass concentration is 3% hydrophilic macromolecule aqueous solution, and the preparation mass concentration is 6% glutaraldehyde water solution, and compound concentration is the polypeptid solution of 150 μ g/ml,
Above-mentioned hydrophilic macromolecule is the compositions of collagen and gelatin, and the part by weight of collagen and gelatin is 1:3; Aforementioned polypeptides is the polypeptide that is rich in aspartic acid, and wherein, the percentage by weight that the summation of described aspartic acid accounts for polypeptide is 10%; Be added with mass percent in described hydrophilic high molecular material and be the micromolecular compound of 2.5% promotion osteoblast breeding.
(2) preparation hydrogel substrate: the ratio that the prepared three kinds of aqueous solutions of step (1) are 1:1:1 according to volume ratio is mixed, stirred at normal temperatures 6 minutes, then mixture is placed on carrier, prepare the hydrogel substrate by phase detachment technique, prepared hydrogel substrate soaks 8 hours to remove free glutaraldehyde with the glycine solution of 0.2M
(3) preparation TRIS buffer: with the Tri(Hydroxymethyl) Amino Methane Hydrochloride of 6 grams, 1 gram Tris and 8 gram sodium chloride join in 100 ml deionized water, and the pH value of this buffer is 7.0-8.0,
(4) take respectively calcium chloride and dipotassium hydrogen phosphate, and dissolve in respectively in the buffer that step (3) makes, be mixed with respectively calcium chloride solution and the dipotassium hydrogen phosphate solution of 0.004mol/L, be the ratio mix and blend mutually of 1:1:3 according to volume ratio with the polypeptid solution that makes in calcium chloride solution and dipotassium hydrogen phosphate solution and step (1)
(5) the prepared hydrogel substrate that is carried on carrier of step (2) is placed in the prepared mixed solution of step (4), after standing 36 hours, priority water and washing with alcohol, drying namely makes described nanometer pore hydroxyl calcium phosphate/aquogel materials.
Nanometer pore hydroxyl calcium phosphate/aquogel materials provided by the invention, preparation method comprises the steps:
(1) the preparation mass concentration is 1% hydrophilic macromolecule aqueous solution, and the preparation mass concentration is 8% glutaraldehyde water solution, and compound concentration is the polypeptid solution of 300 μ g/ml,
Above-mentioned hydrophilic macromolecule is polyvinyl alcohol; Aforementioned polypeptides is the fragment of the osteopontin (osteopontin, OPN) that contains of bone; Be added with mass percent in above-mentioned hydrophilic high molecular material and be the micromolecular compound of 0.001 promotion osteoblast breeding.
(2) preparation hydrogel substrate: the ratio that the prepared three kinds of aqueous solutions of step (1) are 1:1:1 according to volume ratio is mixed, stirred at normal temperatures 9 minutes, then mixture is placed on carrier, prepare the hydrogel substrate by phase detachment technique, prepared hydrogel substrate soaks 10 hours to remove free glutaraldehyde with the glycine solution of 0.3M
(3) preparation TRIS buffer: with the Tri(Hydroxymethyl) Amino Methane Hydrochloride of 5 grams, 1.5 gram Tris and 10 gram sodium chloride join in 100 ml deionized water, and the pH value of this buffer is 7.0-8.0,
(4) take respectively calcium chloride and dipotassium hydrogen phosphate, and dissolve in respectively in the buffer that step (3) makes, be mixed with respectively calcium chloride solution and the dipotassium hydrogen phosphate solution of 0.006mol/L, be the ratio mix and blend mutually of 1:1:1 according to volume ratio with the polypeptid solution that makes in calcium chloride solution and dipotassium hydrogen phosphate solution and step (1)
(5) the prepared hydrogel substrate that is carried on carrier of step (2) is placed in the prepared mixed solution of step (4), after standing 48 hours, priority water and washing with alcohol, drying namely makes described nanometer pore hydroxyl calcium phosphate/aquogel materials.
Embodiment 5
Nanometer pore hydroxyl calcium phosphate/aquogel materials provided by the invention, preparation method comprises the steps:
(1) the preparation mass concentration is 1% hydrophilic macromolecule aqueous solution, and the preparation mass concentration is 4% glutaraldehyde water solution, and compound concentration is the polypeptid solution of 150 μ g/ml,
Above-mentioned hydrophilic macromolecule is alginic acid; Aforementioned polypeptides is the polypeptide that is rich in glutamic acid, and wherein, the percentage by weight that the summation of described glutamic acid accounts for polypeptide is 10%; Be added with mass percent in above-mentioned hydrophilic high molecular material and be the micromolecular compound of 0.01% promotion osteoblast breeding.
(2) preparation hydrogel substrate: the ratio that the prepared three kinds of aqueous solutions of step (1) are 1:1:1 according to volume ratio is mixed mutually, stirred at normal temperatures 0.5 minute, then mixture is placed on carrier, prepare the hydrogel substrate by phase detachment technique, prepared hydrogel substrate soaks 6 hours to remove free glutaraldehyde with the glycine solution of 0.1M
(3) preparation TRIS buffer: with the Tri(Hydroxymethyl) Amino Methane Hydrochloride of 6 grams, 0.6 gram Tris and 8 gram sodium chloride join in 100 ml deionized water, and the pH value of this buffer is 7.2-7.4,
(4) take respectively calcium chloride and dipotassium hydrogen phosphate, and dissolve in respectively in the buffer that step (3) makes, be mixed with respectively calcium chloride solution and the dipotassium hydrogen phosphate solution of 0.002mol/L, the ratio that the polypeptid solution that makes in calcium chloride solution and dipotassium hydrogen phosphate solution and step (1) is 1:1:0 according to volume ratio is mixed (namely mutually, be the ratio mix and blend mutually of 1:1 according to volume ratio with calcium chloride solution and dipotassium hydrogen phosphate solution), stirred at normal temperatures 0.5 minute
(5) the prepared hydrogel substrate that is carried on carrier of step (2) is placed in the prepared mixed solution of step (4), after standing 24 hours, priority water and washing with alcohol, drying namely makes described nanometer pore hydroxyl calcium phosphate/aquogel materials.
Nanometer pore hydroxyl calcium phosphate/aquogel materials provided by the invention, preparation method comprises the steps:
(1) the preparation mass concentration is 3% hydrophilic macromolecule aqueous solution, and the preparation mass concentration is 6% glutaraldehyde water solution, and compound concentration is the polypeptid solution of 250 μ g/ml,
Above-mentioned hydrophilic macromolecule is the compositions of gelatin and alginic acid, and the part by weight of gelatin and alginic acid is 1:2; Aforementioned polypeptides is the polypeptide that is rich in the serine of phosphorylation, and wherein, the percentage by weight that the summation of described serine accounts for polypeptide is 90%; Be added with mass percent in above-mentioned hydrophilic high molecular material and be the micromolecular compound of 0.01% promotion osteoblast breeding.
(2) preparation hydrogel substrate: the ratio that the prepared three kinds of aqueous solutions of step (1) are 1:1:1 according to volume ratio is mixed mutually, stirred at normal temperatures 10 minutes, then mixture is placed on carrier, prepare the hydrogel substrate by phase detachment technique, prepared hydrogel substrate soaks 10 hours to remove free glutaraldehyde with the glycine solution of 0.3M
(3) preparation TRIS buffer: with the Tri(Hydroxymethyl) Amino Methane Hydrochloride of 7 grams, 1.2 gram Tris and 9 gram sodium chloride join in 100 ml deionized water, and the pH value of this buffer is 7.2-7.4,
(4) take respectively calcium chloride and dipotassium hydrogen phosphate, and dissolve in respectively in the buffer that step (3) makes, be mixed with respectively calcium chloride solution and the dipotassium hydrogen phosphate solution of 0.006mol/L, the ratio that the polypeptid solution that makes in calcium chloride solution and dipotassium hydrogen phosphate solution and step (1) is 1:1:6 according to volume ratio is mixed mutually, stirred at normal temperatures 10 minutes
(5) the prepared hydrogel substrate that is carried on carrier of step (2) is placed in the prepared mixed solution of step (4), after standing 36 hours, priority water and washing with alcohol, drying namely makes described nanometer pore hydroxyl calcium phosphate/aquogel materials.
Embodiment 7
Nanometer pore hydroxyl calcium phosphate/aquogel materials provided by the invention, preparation method comprises the steps:
(1) the preparation mass concentration is 2% hydrophilic macromolecule aqueous solution, and the preparation mass concentration is 5% glutaraldehyde water solution, and compound concentration is the polypeptid solution of 200 μ g/ml,
Above-mentioned hydrophilic macromolecule is alginic acid; Aforementioned polypeptides is the polypeptide that is rich in Sulfated serine, and wherein, the percentage by weight that the summation of described Sulfated serine accounts for polypeptide is 50%; Be added with mass percent in above-mentioned hydrophilic high molecular material and be the micromolecular compound of 0.0005% promotion osteoblast breeding.
(2) preparation hydrogel substrate: the ratio that the prepared three kinds of aqueous solutions of step (1) are 1:1:1 according to volume ratio is mixed mutually, stirred at normal temperatures 5 minutes, then mixture is placed on carrier, prepare the hydrogel substrate by phase detachment technique, prepared hydrogel substrate soaks 8 hours to remove free glutaraldehyde with the glycine solution of 0.2M
(3) preparation TRIS buffer: with the Tri(Hydroxymethyl) Amino Methane Hydrochloride of 6.5 grams, 1.0 gram Tris and 8.5 gram sodium chloride join in 100 ml deionized water, and the pH value of this buffer is 7.2-7.4,
(4) take respectively calcium chloride and dipotassium hydrogen phosphate, and dissolve in respectively in the buffer that step (3) makes, be mixed with respectively calcium chloride solution and the dipotassium hydrogen phosphate solution of 0.004mol/L, the ratio that the polypeptid solution that makes in calcium chloride solution and dipotassium hydrogen phosphate solution and step (1) is 1:1:3 according to volume ratio is mixed mutually, stirred at normal temperatures 5 minutes
(5) the prepared hydrogel substrate that is carried on carrier of step (2) is placed in the prepared mixed solution of step (4), after standing 30 hours, priority water and washing with alcohol, drying namely makes described nanometer pore hydroxyl calcium phosphate/aquogel materials.
Embodiment 8
Nanometer pore hydroxyl calcium phosphate/aquogel materials provided by the invention, preparation method comprises the steps:
(1) the preparation mass concentration is 1% hydrophilic macromolecule aqueous solution, and the preparation mass concentration is 6% glutaraldehyde water solution, and compound concentration is the polypeptid solution of 250 μ g/ml,
Above-mentioned hydrophilic macromolecule is the compositions of collagen, gelatin and alginic acid, and the part by weight of collagen, gelatin and alginic acid is 1:1:3; Aforementioned polypeptides is the polypeptide that is rich in sulphation tyrosine, and wherein, the percentage by weight that the summation of described Sulfated tyrosine accounts for polypeptide is 30%; Be added with mass percent in above-mentioned hydrophilic high molecular material and be the micromolecular compound of 0.001% promotion osteoblast breeding.
(2) preparation hydrogel substrate: the ratio that the prepared three kinds of aqueous solutions of step (1) are 1:1:1 according to volume ratio is mixed mutually, stirred at normal temperatures 0.5 minute, then mixture is placed on carrier, prepare the hydrogel substrate by phase detachment technique, prepared hydrogel substrate soaks 6-10 hour to remove free glutaraldehyde with the glycine solution of 0.3M
(3) preparation TRIS buffer: with the Tri(Hydroxymethyl) Amino Methane Hydrochloride of 7 grams, 0.6 gram Tris and 8 gram sodium chloride join in 100 ml deionized water, and the pH value of this buffer is 7.2-7.4,
(4) take respectively calcium chloride and dipotassium hydrogen phosphate, and dissolve in respectively in the buffer that step (3) makes, be mixed with respectively calcium chloride solution and the dipotassium hydrogen phosphate solution of 0.002mol/L, the ratio that the polypeptid solution that makes in calcium chloride solution and dipotassium hydrogen phosphate solution and step (1) is 1:1:1 according to volume ratio is mixed mutually, stirred at normal temperatures 10 minutes
(5) the prepared hydrogel substrate that is carried on carrier of step (2) is placed in the prepared mixed solution of step (4), after standing 36 hours, priority water and washing with alcohol, drying namely makes described nanometer pore hydroxyl calcium phosphate/aquogel materials.
Embodiment 9
Nanometer pore hydroxyl calcium phosphate/aquogel materials provided by the invention, preparation method comprises the steps:
(1) the preparation mass concentration is 3% hydrophilic macromolecule aqueous solution, and the preparation mass concentration is 4% glutaraldehyde water solution, and compound concentration is the polypeptid solution of 150 μ g/ml,
Above-mentioned hydrophilic macromolecule is the compositions of collagen, gelatin, and the part by weight of collagen, gelatin is 3:1; Aforementioned polypeptides is the polypeptide that is rich in aspartic acid and glutamic acid, and wherein, the percentage by weight that the summation of described aspartic acid and glutamic acid accounts for polypeptide is 60%; Be added with mass percent in above-mentioned hydrophilic high molecular material and be the micromolecular compound of 0.005% promotion osteoblast breeding.
(2) preparation hydrogel substrate: the ratio that the prepared three kinds of aqueous solutions of step (1) are 1:1:1 according to volume ratio is mixed mutually, stirred at normal temperatures 10 minutes, then mixture is placed on carrier, prepare the hydrogel substrate by phase detachment technique, prepared hydrogel substrate soaks 10 hours to remove free glutaraldehyde with the glycine solution of 0.3M
(3) preparation TRIS buffer: with the Tri(Hydroxymethyl) Amino Methane Hydrochloride of 7 grams, 1.2 gram Tris and 8 gram sodium chloride join in 100 ml deionized water, and the pH value of this buffer is 7.2-7.4,
(4) take respectively calcium chloride and dipotassium hydrogen phosphate, and dissolve in respectively in the buffer that step (3) makes, be mixed with respectively calcium chloride solution and the dipotassium hydrogen phosphate solution of 0.006mol/L, the ratio that the polypeptid solution that makes in calcium chloride solution and dipotassium hydrogen phosphate solution and step (1) is 1:1:1 according to volume ratio is mixed mutually, stirred at normal temperatures 10 minutes
(5) the prepared hydrogel substrate that is carried on carrier of step (2) is placed in the prepared mixed solution of step (4), after standing 24 hours, priority water and washing with alcohol, drying namely makes described nanometer pore hydroxyl calcium phosphate/aquogel materials.
Embodiment 10
Nanometer pore hydroxyl calcium phosphate/aquogel materials provided by the invention, preparation method comprises the steps:
(1) the preparation mass concentration is 1% hydrophilic macromolecule aqueous solution, and the preparation mass concentration is 5% glutaraldehyde water solution, and compound concentration is the polypeptid solution of 200 μ g/ml,
Above-mentioned hydrophilic macromolecule is the compositions of gelatin and alginic acid, and the part by weight of gelatin and alginic acid is 1:1; Aforementioned polypeptides is the polypeptide that is rich in phosphorylation or Sulfated serine/tyrosine, and wherein, the percentage by weight that the summation of described phosphorylation or Sulfated serine/tyrosine accounts for polypeptide is 70%; Be added with mass percent in above-mentioned hydrophilic high molecular material and be the micromolecular compound of 0.05% promotion osteoblast breeding.
(2) preparation hydrogel substrate: the ratio that the prepared three kinds of aqueous solutions of step (1) are 1:1:1 according to volume ratio is mixed mutually, stirred at normal temperatures 0.5 minute, then mixture is placed on carrier, prepare the hydrogel substrate by phase detachment technique, prepared hydrogel substrate soaks 6 hours to remove free glutaraldehyde with the glycine solution of 0.1M
(3) preparation TRIS buffer: with the Tri(Hydroxymethyl) Amino Methane Hydrochloride (Tris-HCl) of 6.61 grams, 0.97 gram Tris and 8.77 gram sodium chloride join in 100 ml deionized water, the pH value of this buffer is 7.2-7.4
(4) take respectively calcium chloride and dipotassium hydrogen phosphate, and dissolve in respectively in the buffer that step (3) makes, be mixed with respectively calcium chloride solution and the dipotassium hydrogen phosphate solution of 0.002mol/L, the ratio that the polypeptid solution that makes in calcium chloride solution and dipotassium hydrogen phosphate solution and step (1) is 1:1:0 according to volume ratio is mixed (namely mutually, be the ratio mix and blend mutually of 1:1 according to volume ratio with calcium chloride solution and dipotassium hydrogen phosphate solution), stirred at normal temperatures 0.5 minute
(5) the prepared hydrogel substrate that is carried on carrier of step (2) is placed in the prepared mixed solution of step (4), after standing 24 hours, priority water and washing with alcohol, drying namely makes described nanometer pore hydroxyl calcium phosphate/aquogel materials.
Embodiment 11
Nanometer pore hydroxyl calcium phosphate/aquogel materials provided by the invention, preparation method comprises the steps:
(1) the preparation mass concentration is 1% hydrophilic macromolecule aqueous solution, and the preparation mass concentration is 5% glutaraldehyde water solution, and compound concentration is the polypeptid solution of 200 μ g/ml,
Above-mentioned hydrophilic macromolecule is the compositions of collagen, gelatin and alginic acid, and the part by weight of collagen, gelatin and alginic acid is 1:3:1; Aforementioned polypeptides is poly-aspartic-acid, is added with mass percent in above-mentioned hydrophilic high molecular material and is the micromolecular compound of 0.05% promotion osteoblast breeding.
(2) preparation hydrogel substrate: the ratio that the prepared three kinds of aqueous solutions of step (1) are 1:1:1 according to volume ratio is mixed mutually, stirred at normal temperatures 10 minutes, then mixture is placed on carrier, prepare the hydrogel substrate by phase detachment technique, prepared hydrogel substrate soaks 10 hours to remove free glutaraldehyde with the glycine solution of 0.3M
(3) preparation TRIS buffer: with the Tri(Hydroxymethyl) Amino Methane Hydrochloride (Tris-HCl) of 6.61 grams, 0.97 gram Tris and 8.77 gram sodium chloride join in 100 ml deionized water, the pH value of this buffer is 7.2-7.4
(4) take respectively calcium chloride and dipotassium hydrogen phosphate, and dissolve in respectively in the buffer that step (3) makes, be mixed with respectively calcium chloride solution and the dipotassium hydrogen phosphate solution of 0.006mol/L, the ratio that the polypeptid solution that makes in calcium chloride solution and dipotassium hydrogen phosphate solution and step (1) is 1:1:6 according to volume ratio is mixed mutually, stirred at normal temperatures 10 minutes
(5) the prepared hydrogel substrate that is carried on carrier of step (2) is placed in the prepared mixed solution of step (4), after standing 24 hours, priority water and washing with alcohol, drying namely makes described nanometer pore hydroxyl calcium phosphate/aquogel materials.
Embodiment 12
Nanometer pore hydroxyl calcium phosphate/aquogel materials provided by the invention, preparation method comprises the steps:
(1) the preparation mass concentration is 1% hydrophilic macromolecule aqueous solution, and the preparation mass concentration is 5% glutaraldehyde water solution, and compound concentration is the polypeptid solution of 200 μ g/ml,
Above-mentioned hydrophilic macromolecule is polyvinyl alcohol; Aforementioned polypeptides is polyglutamic acid; Be added with mass percent in above-mentioned hydrophilic high molecular material and be the micromolecular compound of 0.5% promotion osteoblast breeding.
(2) preparation hydrogel substrate: the ratio that the prepared three kinds of aqueous solutions of step (1) are 1:1:1 according to volume ratio is mixed mutually, stirred at normal temperatures 5 minutes, then mixture is placed on carrier, prepare the hydrogel substrate by phase detachment technique, prepared hydrogel substrate soaks 8 hours to remove free glutaraldehyde with the glycine solution of 0.2M
(3) preparation TRIS buffer: with the Tri(Hydroxymethyl) Amino Methane Hydrochloride (Tris-HCl) of 6.61 grams, 0.97 gram Tris and 8.77 gram sodium chloride join in 100 ml deionized water, the pH value of this buffer is 7.2-7.4
(4) take respectively calcium chloride and dipotassium hydrogen phosphate, and dissolve in respectively in the buffer that step (3) makes, be mixed with respectively calcium chloride solution and the dipotassium hydrogen phosphate solution of 0.004mol/L, the ratio that the polypeptid solution that makes in calcium chloride solution and dipotassium hydrogen phosphate solution and step (1) is 1:1:3 according to volume ratio is mixed mutually, stirred at normal temperatures 5 minutes
(5) the prepared hydrogel substrate that is carried on carrier of step (2) is placed in the prepared mixed solution of step (4), after standing 24 hours, priority water and washing with alcohol, drying namely makes described nanometer pore hydroxyl calcium phosphate/aquogel materials.
Embodiment 13
Nanometer pore hydroxyl calcium phosphate/aquogel materials provided by the invention, with polypeptide, described hydrogel surface is carried out modification, and by the bionical process that mineralizes, calcium hydroxy phosphate is deposited on hydrogel, the preparation method of described nanometer pore hydroxyl calcium phosphate/aquogel materials comprises the steps:
(1) the preparation mass concentration is 1% hydrophilic macromolecule aqueous solution, and compound concentration is the polypeptid solution of 10 μ g/ml,
Above-mentioned hydrophilic macromolecule is collagen; Aforementioned polypeptides is the osteopontin (osteopontin, OPN) that bone contains; Be added with mass percent in described hydrophilic high molecular material and be the micromolecular compound of 0.0001% promotion osteoblast breeding.
(2) preparation hydrogel substrate: the ratio that the prepared two kinds of aqueous solutions of step (1) are 1:1 according to volume ratio is mixed, stirred at normal temperatures 1 minute, and then mixture was placed on carrier, under room temperature, drying is 24 hours, then vacuum drying at the temperature of 80 ℃
(3) preparation TRIS buffer: with the Tri(Hydroxymethyl) Amino Methane Hydrochloride of 5 grams, 0.5 gram Tris and 6 gram sodium chloride join in 100 ml deionized water, and the pH value of this buffer is 7.0-8.0,
(4) take respectively calcium chloride and dipotassium hydrogen phosphate, and dissolve in respectively in the buffer that step (3) makes, be mixed with respectively calcium chloride solution and the dipotassium hydrogen phosphate solution of 0.002mol/L, be that mix and blend is (namely mutually for the ratio of 1:1:0 with the polypeptid solution that makes in calcium chloride solution and dipotassium hydrogen phosphate solution and step (1) according to volume ratio, be the ratio mix and blend mutually of 1:1 according to volume ratio with calcium chloride solution and dipotassium hydrogen phosphate solution)
(5) the prepared hydrogel substrate that is carried on carrier of step (2) is placed in the prepared mixed solution of step (4), after standing 24 hours, priority water and washing with alcohol, drying namely makes described nanometer pore hydroxyl calcium phosphate/aquogel materials.
Embodiment 14
Nanometer pore hydroxyl calcium phosphate/aquogel materials provided by the invention, with polypeptide, described hydrogel surface is carried out modification, and by the bionical process that mineralizes, calcium hydroxy phosphate is deposited on hydrogel, the preparation method of described nanometer pore hydroxyl calcium phosphate/aquogel materials comprises the steps:
(1) the preparation mass concentration is 5% hydrophilic macromolecule aqueous solution, and compound concentration is the polypeptid solution of 300 μ g/ml,
Above-mentioned hydrophilic macromolecule is gelatin; Aforementioned polypeptides is phosphoprotein (phosphophoryn) or its fragment that tooth contains; Be added with mass percent in above-mentioned hydrophilic high molecular material and be the micromolecular compound of 5% promotion osteoblast breeding.
(2) preparation hydrogel substrate: the ratio that the prepared two kinds of aqueous solutions of step (1) are 1:1 according to volume ratio is mixed, stirred at normal temperatures 10 minutes, and then mixture was placed on carrier, under room temperature, drying is 48 hours, then vacuum drying at the temperature of 100 ℃
(3) preparation TRIS buffer: with the Tri(Hydroxymethyl) Amino Methane Hydrochloride of 8 grams, 1.5 gram Tris and 10 gram sodium chloride join in 100 ml deionized water, and the pH value of this buffer is 7.2-7.4,
(4) take respectively calcium chloride and dipotassium hydrogen phosphate, and dissolve in respectively in the buffer that step (3) makes, be mixed with respectively calcium chloride solution and the dipotassium hydrogen phosphate solution of 0.006mol/L, be the ratio mix and blend mutually of 1:1:6 according to volume ratio with the polypeptid solution that makes in calcium chloride solution and dipotassium hydrogen phosphate solution and step (1)
(5) the prepared hydrogel substrate that is carried on carrier of step (2) is placed in the prepared mixed solution of step (4), after standing 48 hours, priority water and washing with alcohol, drying namely makes described nanometer pore hydroxyl calcium phosphate/aquogel materials.
Embodiment 15
Nanometer pore hydroxyl calcium phosphate/aquogel materials provided by the invention, with polypeptide, described hydrogel surface is carried out modification, and by the bionical process that mineralizes, calcium hydroxy phosphate is deposited on hydrogel, the preparation method of described nanometer pore hydroxyl calcium phosphate/aquogel materials comprises the steps:
(1) the preparation mass concentration is 3% hydrophilic macromolecule aqueous solution, and compound concentration is the polypeptid solution of 150 μ g/ml,
Above-mentioned hydrophilic macromolecule is the compositions of collagen and gelatin, and the part by weight of collagen and gelatin is 1:3; Aforementioned polypeptides is the polypeptide that is rich in aspartic acid, and wherein, the percentage by weight that the summation of described aspartic acid accounts for polypeptide is 10%; Be added with mass percent in described hydrophilic high molecular material and be the micromolecular compound of 2.5% promotion osteoblast breeding.
(2) preparation hydrogel substrate: the ratio that the prepared two kinds of aqueous solutions of step (1) are 1:1 according to volume ratio is mixed, stirred at normal temperatures 6 minutes, and then mixture was placed on carrier, under room temperature, drying is 36 hours, then vacuum drying at the temperature of 90 ℃
(3) preparation TRIS buffer: with the Tri(Hydroxymethyl) Amino Methane Hydrochloride of 6.5 grams, 1 gram Tris and 8 gram sodium chloride join in 100 ml deionized water, and the pH value of this buffer is 7.0-8.0,
(4) take respectively calcium chloride and dipotassium hydrogen phosphate, and dissolve in respectively in the buffer that step (3) makes, be mixed with respectively calcium chloride solution and the dipotassium hydrogen phosphate solution of 0.004mol/L, be the ratio mix and blend mutually of 1:1:3 according to volume ratio with the polypeptid solution that makes in calcium chloride solution and dipotassium hydrogen phosphate solution and step (1)
(5) the prepared hydrogel substrate that is carried on carrier of step (2) is placed in the prepared mixed solution of step (4), after standing 36 hours, priority water and washing with alcohol, drying namely makes described nanometer pore hydroxyl calcium phosphate/aquogel materials.
Embodiment 16
Nanometer pore hydroxyl calcium phosphate/aquogel materials provided by the invention, with polypeptide, described hydrogel surface is carried out modification, and by the bionical process that mineralizes, calcium hydroxy phosphate is deposited on hydrogel, the preparation method of described nanometer pore hydroxyl calcium phosphate/aquogel materials comprises the steps:
(1) the preparation mass concentration is 5% hydrophilic macromolecule aqueous solution, and compound concentration is the polypeptid solution of 100 μ g/ml,
Above-mentioned hydrophilic macromolecule is polyvinyl alcohol; Aforementioned polypeptides is the fragment of the osteopontin (osteopontin, OPN) that contains of bone; Be added with mass percent in above-mentioned hydrophilic high molecular material and be the micromolecular compound of 0.001 promotion osteoblast breeding.
(2) preparation hydrogel substrate: the ratio that the prepared two kinds of aqueous solutions of step (1) are 1:1 according to volume ratio is mixed, stirred at normal temperatures 5 minutes, and then mixture was placed on carrier, under room temperature, drying is 24 hours, then vacuum drying at the temperature of 100 ℃
(3) preparation TRIS buffer: with the Tri(Hydroxymethyl) Amino Methane Hydrochloride of 8 grams, 1.5 gram Tris and 6 gram sodium chloride join in 100 ml deionized water, and the pH value of this buffer is 7.0-8.0,
(4) take respectively calcium chloride and dipotassium hydrogen phosphate, and dissolve in respectively in the buffer that step (3) makes, be mixed with respectively calcium chloride solution and the dipotassium hydrogen phosphate solution of 0.002mol/L, be the ratio mix and blend mutually of 1:1:1 according to volume ratio with the polypeptid solution that makes in calcium chloride solution and dipotassium hydrogen phosphate solution and step (1)
(5) the prepared hydrogel substrate that is carried on carrier of step (2) is placed in the prepared mixed solution of step (4), after standing 48 hours, priority water and washing with alcohol, drying namely makes described nanometer pore hydroxyl calcium phosphate/aquogel materials.
Embodiment 17
tooth implant provided by the invention, comprise titanium alloy-based, preparation method is is the ratio phase mix and blend of 1:1:1 according to volume ratio with the prepared three kinds of aqueous solutions of step (1) in embodiment 1, then mixture is sprayed to titanium alloy-based on, rapid draing under room temperature, as by being placed in again 90 ℃ of heat treatments after vacuum drying 5 minutes, prepared hydrogel is the continuous or discrete thin film of one deck, the width of the discontinuous part of described discontinuous thin film is 1 μ m-100 μ m, other content of preparation method is constant, identical with the method in embodiment 1, namely make the tooth implant that is provided with nanometer pore hydroxyl calcium phosphate/aquogel materials of the present invention.
Embodiment 18
tooth implant provided by the invention, comprise titanium alloy-based, preparation method is is the ratio phase mix and blend of 1:1:1 according to volume ratio with the prepared three kinds of aqueous solutions of step (1) in embodiment 2, then mixture is sprayed to titanium alloy-based on, be placed in again 100 ℃ of heat treatments after vacuum rapid draing 15 minutes under room temperature, prepared hydrogel is the continuous or discrete thin film of one deck, the width of the discontinuous part of described discontinuous thin film is 1mm-3mm, other content of preparation method is constant, identical with the method in embodiment 2, namely make the tooth implant that is provided with nanometer pore hydroxyl calcium phosphate/aquogel materials of the present invention.
Embodiment 19
tooth implant provided by the invention, comprise titanium alloy-based, preparation method is is the ratio phase mix and blend of 1:1:1 according to volume ratio with the prepared three kinds of aqueous solutions of step (1) in embodiment 6, then mixture is sprayed to titanium alloy-based on, be placed in again 95 ℃ of heat treatments after vacuum rapid draing 10 minutes under room temperature, prepared hydrogel is the continuous or discrete thin film of one deck, the width of the discontinuous part of described discontinuous thin film is 500 μ m-1mm, other content of preparation method is constant, identical with the method in embodiment 6, namely make the tooth implant that is provided with nanometer pore hydroxyl calcium phosphate/aquogel materials of the present invention.
tooth implant provided by the invention, comprise titanium alloy-based, preparation method is is the ratio phase mix and blend of 1:1 according to volume ratio with the prepared two kinds of aqueous solutions of step (1) in embodiment 13, then mixture is sprayed to titanium alloy-based on, be placed in again 90 ℃ of heat treatments after vacuum rapid draing 5 minutes under room temperature, prepared hydrogel is the continuous or discrete thin film of one deck, the width of the discontinuous part of described discontinuous thin film is 100 μ m-500 μ m, other content of preparation method is constant, identical with the method in embodiment 13, namely make the tooth implant that is provided with nanometer pore hydroxyl calcium phosphate/aquogel materials of the present invention.
Embodiment 21
tooth implant provided by the invention, comprise titanium alloy-based, preparation method is is the ratio phase mix and blend of 1:1 according to volume ratio with the prepared two kinds of aqueous solutions of step (1) in embodiment 14, then mixture is sprayed to titanium alloy-based on, be placed in again 100 ℃ of heat treatments after vacuum rapid draing 15 minutes under room temperature, prepared hydrogel is the continuous or discrete thin film of one deck, the width of the discontinuous part of described discontinuous thin film is 1mm-3mm, other content of preparation method is constant, identical with the method in embodiment 14, namely make the tooth implant that is provided with nanometer pore hydroxyl calcium phosphate/aquogel materials of the present invention.
Embodiment 22
tooth implant provided by the invention, comprise titanium alloy-based, preparation method is is the ratio phase mix and blend of 1:1 according to volume ratio with the prepared two kinds of aqueous solutions of step (1) in embodiment 15, then mixture is sprayed to titanium alloy-based on, be placed in again 95 ℃ of heat treatments after vacuum rapid draing 10 minutes under room temperature, prepared hydrogel is the continuous or discrete thin film of one deck, the width of the discontinuous part of described discontinuous thin film is 2mm, other content of preparation method is constant, identical with the method in embodiment 15, namely make the tooth implant that is provided with nanometer pore hydroxyl calcium phosphate/aquogel materials of the present invention.
Embodiment 23
tooth implant provided by the invention, comprise titanium alloy-based, preparation method is is the ratio phase mix and blend of 1:1:1 according to volume ratio with the prepared three kinds of aqueous solutions of the step in embodiment 8 (1), process titanium alloy-based surface with chemical method, then with mixture freezing on titanium alloy-based under-8 ℃,-10 ℃ of lower freezing vacuum sublimation dryings 6 hours, the thickness of prepared hydrogel is 0.01mm-0.1mm, other content of preparation method is constant, identical with the method in embodiment 8, namely make the tooth implant that is provided with nanometer pore hydroxyl calcium phosphate/aquogel materials of the present invention.
Embodiment 24
tooth implant provided by the invention, comprise titanium alloy-based, preparation method is is the ratio phase mix and blend of 1:1:1 according to volume ratio with the prepared three kinds of aqueous solutions of the step in embodiment 9 (1), process titanium alloy-based surface with chemical method, then with mixture freezing on titanium alloy-based under-10 ℃,-60 ℃ of lower freezing vacuum sublimation dryings 8 hours, the thickness of prepared hydrogel is 4mm-5mm, other content of preparation method is constant, identical with the method in embodiment 9, namely make the tooth implant that is provided with nanometer pore hydroxyl calcium phosphate/aquogel materials of the present invention.
Embodiment 25
tooth implant provided by the invention, comprise titanium alloy-based, preparation method is is the ratio phase mix and blend of 1:1:1 according to volume ratio with the prepared three kinds of aqueous solutions of the step in embodiment 11 (1), process titanium alloy-based surface with chemical method, then with mixture freezing on titanium alloy-based under-9 ℃,-35 ℃ of lower freezing vacuum sublimation dryings 7 hours, the thickness of prepared hydrogel is 2mm-3mm, other content of preparation method is constant, identical with the method in embodiment 11, namely make the tooth implant that is provided with nanometer pore hydroxyl calcium phosphate/aquogel materials of the present invention.
Embodiment 26
tooth implant provided by the invention, comprise titanium alloy-based, preparation method is is the ratio phase mix and blend of 1:1 according to volume ratio with the prepared two kinds of aqueous solutions of the step in embodiment 13 (1), process titanium alloy-based surface with chemical method, then with mixture at-8 ℃ of lower freezing vacuum sublimation dryings on titanium alloy-based, under-10 ℃ freezing 6 hours, the thickness of prepared hydrogel is 0.01mm-0.1mm, other content of preparation method is constant, identical with the method in embodiment 13, namely make the tooth implant that is provided with nanometer pore hydroxyl calcium phosphate/aquogel materials of the present invention.
Embodiment 27
tooth implant provided by the invention, comprise titanium alloy-based, its preparation method is is the ratio phase mix and blend of 1:1 according to volume ratio with the prepared two kinds of aqueous solutions of the step in embodiment 14 (1), process titanium alloy-based surface with chemical method, then with mixture freezing on titanium alloy-based under-10 ℃,-60 ℃ of lower freezing vacuum sublimation dryings 8 hours, the thickness of prepared hydrogel is 1mm-5mm, other content of preparation method is constant, identical with the method in embodiment 14, namely make the tooth implant that is provided with nanometer pore hydroxyl calcium phosphate/aquogel materials of the present invention.
Embodiment 28
tooth implant provided by the invention, comprise titanium alloy-based, its preparation method is is the ratio phase mix and blend of 1:1 according to volume ratio with the prepared two kinds of aqueous solutions of the step in embodiment 15 (1), process titanium alloy-based surface with chemical method, then with mixture freezing on titanium alloy-based under-9 ℃,-35 ℃ of lower freezing vacuum sublimation dryings 7 hours, the thickness of prepared hydrogel is 0.5mm-1mm, other content of preparation method is constant, identical with the method in embodiment 15, namely make the tooth implant that is provided with nanometer pore hydroxyl calcium phosphate/aquogel materials of the present invention.
Two, method of testing
Utilize scanning electron microscope to measure hydrogel material aperture, poroid calcium hydroxy phosphate aperture and wall thickness;
The detection data of table 1, the prepared nanometer pore hydroxyl calcium phosphate/aquogel materials of embodiment
Can be found out by above-mentioned experimental data, nanometer pore hydroxyl calcium phosphate/aquogel materials provided by the invention, wherein, hydrogel be micron to millimetre-sized poroid two dimension or three-dimensional material, the three-dimensional material hole is interconnected, the aperture is: 1 micron to 3 millimeters; The calcium hydroxy phosphate that is deposited on hydrogel is the poroid calcium hydroxy phosphate of three-dimensional communication, and the aperture is the 50-1000 nanometer, and hole wall is thick is the 30-80 nanometer.
The above is only preferred embodiment of the present invention, is not for limiting protection scope of the present invention.Every equalization that content is done according to the present invention changes and modifies, and all is encompassed in the scope of the claims of the present invention.
Claims (12)
1. nanometer pore hydroxyl calcium phosphate/aquogel materials, it is characterized in that, with polypeptide, described hydrogel surface is carried out modification, and by the bionical process that mineralizes, calcium hydroxy phosphate is deposited on hydrogel, the preparation method of described nanometer pore hydroxyl calcium phosphate/aquogel materials comprises the steps:
(1) the preparation mass concentration is the hydrophilic macromolecule aqueous solution of 1-5%, the preparation mass concentration is the glutaraldehyde water solution of 3-8%, compound concentration is the polypeptid solution of 10-300 μ g/ml, described hydrophilic macromolecule is selected from collagen, gelatin, polyvinyl alcohol or alginic acid or the compositions of at least two kinds wherein
(2) preparation hydrogel substrate: the ratio that the prepared three kinds of aqueous solutions of step (1) are 1:1:1 according to volume ratio is mixed, stirred at normal temperatures 0.5-10 minute, then mixture is placed on carrier, prepare the hydrogel substrate by phase detachment technique, prepared hydrogel substrate soaks 6-10 hour to remove free glutaraldehyde with the glycine solution of 0.1-0.3M
(3) preparation TRIS buffer: with the Tri(Hydroxymethyl) Amino Methane Hydrochloride of 5-8 gram, 0.5-1.5 gram Tris and 6-10 gram sodium chloride join in 100 ml deionized water, and the pH value of this buffer is 7.0-8.0,
(4) take respectively calcium chloride and dipotassium hydrogen phosphate, and dissolve in respectively in the buffer that step (3) makes, be mixed with respectively calcium chloride solution and the dipotassium hydrogen phosphate solution of 0.002-0.006mol/L, be the ratio mix and blend mutually of 1:1:X (X=0-6) according to volume ratio with the polypeptid solution that makes in calcium chloride solution and dipotassium hydrogen phosphate solution and step (1)
(5) the prepared hydrogel substrate that is carried on carrier of step (2) is placed in the prepared mixed solution of step (4), after standing 24-48 hour, priority water and washing with alcohol, drying namely makes described nanometer pore hydroxyl calcium phosphate/aquogel materials.
2. a nanometer pore hydroxyl calcium phosphate/aquogel materials as claimed in claim 1, is characterized in that, the preparation method of described nanometer pore hydroxyl calcium phosphate/aquogel materials comprises the steps:
(1) the preparation mass concentration is the hydrophilic macromolecule aqueous solution of 1-3%, and the preparation mass concentration is the glutaraldehyde water solution of 4-6%, and compound concentration is the polypeptid solution of 150-250 μ g/ml,
(2) preparation hydrogel substrate: the ratio that the prepared three kinds of aqueous solutions of step (1) are 1:1:1 according to volume ratio is mixed mutually, stirred at normal temperatures 0.5-10 minute, then mixture is placed on carrier, prepare the hydrogel substrate by phase detachment technique, prepared hydrogel substrate soaks 6-10 hour to remove free glutaraldehyde with the glycine solution of 0.1-0.3M
(3) preparation TRIS buffer: with the Tri(Hydroxymethyl) Amino Methane Hydrochloride of 6-7 gram, 0.6-1.2 gram Tris and 8-9 gram sodium chloride join in 100 ml deionized water, and the pH value of this buffer is 7.2-7.4,
(4) take respectively calcium chloride and dipotassium hydrogen phosphate, and dissolve in respectively in the buffer that step (3) makes, be mixed with respectively calcium chloride solution and the dipotassium hydrogen phosphate solution of 0.002-0.006mol/L, the ratio that the polypeptid solution that makes in calcium chloride solution and dipotassium hydrogen phosphate solution and step (1) is 1:1:X (X=0-6) according to volume ratio is mixed mutually, stirred at normal temperatures 0.5-10 minute
(5) the prepared hydrogel substrate that is carried on carrier of step (2) is placed in the prepared mixed solution of step (4), after standing 24-36 hour, priority water and washing with alcohol, drying namely makes described nanometer pore hydroxyl calcium phosphate/aquogel materials.
3. a nanometer pore hydroxyl calcium phosphate/aquogel materials as claimed in claim 1, is characterized in that, the preparation method of described nanometer pore hydroxyl calcium phosphate/aquogel materials comprises the steps:
(1) the preparation mass concentration is 1% hydrophilic macromolecule aqueous solution, and the preparation mass concentration is 5% glutaraldehyde water solution, and compound concentration is the polypeptid solution of 200 μ g/ml,
(2) preparation hydrogel substrate: the ratio that the prepared three kinds of aqueous solutions of step (1) are 1:1:1 according to volume ratio is mixed mutually, stirred at normal temperatures 0.5-10 minute, then mixture is placed on carrier, prepare the hydrogel substrate by phase detachment technique, prepared hydrogel substrate soaks 6-10 hour to remove free glutaraldehyde with the glycine solution of 0.1-0.3M
(3) preparation TRIS buffer: with the Tri(Hydroxymethyl) Amino Methane Hydrochloride (Tris-HCl) of 6.61 grams, 0.97 gram Tris and 8.77 gram sodium chloride join in 100 ml deionized water, the pH value of this buffer is 7.2-7.4
(4) take respectively calcium chloride and dipotassium hydrogen phosphate, and dissolve in respectively in the buffer that step (3) makes, be mixed with respectively calcium chloride solution and the dipotassium hydrogen phosphate solution of 0.002-0.006mol/L, the ratio that the polypeptid solution that makes in calcium chloride solution and dipotassium hydrogen phosphate solution and step (1) is 1:1:X (X=0-6) according to volume ratio is mixed mutually, stirred at normal temperatures 0.5-10 minute
(5) the prepared hydrogel substrate that is carried on carrier of step (2) is placed in the prepared mixed solution of step (4), after standing 24 hours, priority water and washing with alcohol, drying namely makes described nanometer pore hydroxyl calcium phosphate/aquogel materials.
4. nanometer pore hydroxyl calcium phosphate/aquogel materials, it is characterized in that, with polypeptide, described hydrogel surface is carried out modification, and by the bionical process that mineralizes, calcium hydroxy phosphate is deposited on hydrogel, the preparation method of described nanometer pore hydroxyl calcium phosphate/aquogel materials comprises the steps:
(1) the preparation mass concentration is the hydrophilic macromolecule aqueous solution of 1-5%, and compound concentration is the polypeptid solution of 10-300 μ g/ml, and described hydrophilic macromolecule is selected from collagen, gelatin, polyvinyl alcohol or alginic acid or the compositions of at least two kinds wherein,
(2) preparation hydrogel substrate: the ratio that the prepared two kinds of aqueous solutions of step (1) are 1:1 according to volume ratio is mixed, stirred at normal temperatures 1-10 minute, and then mixture was placed on carrier, under room temperature dry 24-48 hour, then vacuum drying at the temperature of 80 ℃-100 ℃
(3) preparation TRIS buffer: with the Tri(Hydroxymethyl) Amino Methane Hydrochloride of 5-8 gram, 0.5-1.5 gram Tris and 6-10 gram sodium chloride join in 100 ml deionized water, and the pH value of this buffer is 7.0-8.0,
(4) take respectively calcium chloride and dipotassium hydrogen phosphate, and dissolve in respectively in the buffer that step (3) makes, be mixed with respectively calcium chloride solution and the dipotassium hydrogen phosphate solution of 0.002-0.006mol/L, be the ratio mix and blend mutually of 1:1:X (X=0-6) according to volume ratio with the polypeptid solution that makes in calcium chloride solution and dipotassium hydrogen phosphate solution and step (1)
(5) the prepared hydrogel substrate that is carried on carrier of step (2) is placed in the prepared mixed solution of step (4), after standing 24-48 hour, priority water and washing with alcohol, drying namely makes described nanometer pore hydroxyl calcium phosphate/aquogel materials.
5. as claim 1,2,3 or 4 described nanometer pore hydroxyl calcium phosphate/aquogel materials, it is characterized in that, described polypeptide is the osteopontin (osteopontin, OPN) that contains of bone, phosphoprotein (phosphophoryn) or its fragment that tooth contains; Or, described polypeptide is the polypeptide that is rich in aspartic acid, glutamic acid, phosphorylation or Sulfated serine/tyrosine, wherein, to account for the percentage by weight of polypeptide be 10-90% to the summation of described aspartic acid, glutamic acid, phosphorylation or Sulfated serine/tyrosine; Or described polypeptide is poly-aspartic-acid, polyglutamic acid, poly-Sulfated serine/tyrosine.
6. as claim 1,2,3 or 4 described nanometer pore hydroxyl calcium phosphate/aquogel materials, it is characterized in that, being added with mass percent in described hydrogel material is the micromolecular compound of the promotion osteoblast breeding of 0.0001-5%, and described micromolecular compound is selected from diosgenin or amygdaloside.
7. tooth implant, comprise titanium alloy-based, it is characterized in that, describedly be provided with the described nanometer pore hydroxyl calcium phosphate/aquogel materials of one of claims 1 to 3 on titanium alloy-based, in its preparation method, step (1) is constant, the process of step (2) preparation hydrogel substrate comprises: the ratio that the prepared three kinds of aqueous solutions of step (1) are 1:1:1 according to volume ratio is mixed mutually, stirred at normal temperatures 0.5-10 minute, then mixture is sprayed to titanium alloy-based on, be placed in again 90 ℃ to 100 ℃ heat treatments under room temperature after rapid draing 5 to 15 minutes, prepared hydrogel is the continuous or discrete thin film of one deck, the width of the discontinuous part of described discontinuous thin film is 1 μ m-3mm, prepared hydrogel substrate soaks 6-10 hour to remove free glutaraldehyde with the glycine solution of 0.1-0.3M, step (3) is constant to step (5).
8. tooth implant, comprise titanium alloy-based, it is characterized in that, describedly be provided with nanometer pore hydroxyl calcium phosphate/aquogel materials claimed in claim 4 on titanium alloy-based, in its preparation method, step (1) is constant, the process of step (2) preparation hydrogel substrate comprises: the ratio that the prepared two kinds of aqueous solutions of step (1) are 1:1 according to volume ratio is mixed, stirred at normal temperatures 1-10 minute, then mixture is sprayed to titanium alloy-based on, be placed in again 90 ℃ to 100 ℃ heat treatments under room temperature after rapid draing 5 to 15 minutes, prepared hydrogel is the continuous or discrete thin film of one deck, the width of the discontinuous part of described discontinuous thin film is 1 μ m-3mm, step (3) is constant to step (5).
9. tooth implant comprises titanium alloy-basedly, it is characterized in that, describedly is provided with the described nanometer pore hydroxyl calcium phosphate/aquogel materials of one of claims 1 to 3 on titanium alloy-based, in its preparation method, step (1) is constant, the process of step (2) preparation hydrogel substrate comprises: the ratio that the prepared three kinds of aqueous solutions of step (1) are 1:1:1 according to volume ratio is mixed mutually, stirred at normal temperatures 0.5-10 minute, process titanium alloy-based surface with chemical method, then with mixture freezing on titanium alloy-based under-8 ℃ to-10 ℃,-10 ℃ to-60 ℃ lower freezing vacuum sublimation dryings 6-8 hour, the thickness of prepared hydrogel is 0.01mm to 5mm, prepared hydrogel substrate soaks 6-10 hour to remove free glutaraldehyde with the glycine solution of 0.1-0.3M, step (3) is constant to step (5).
10. tooth implant comprises titanium alloy-basedly, it is characterized in that, describedly is provided with nanometer pore hydroxyl calcium phosphate/aquogel materials claimed in claim 4 on titanium alloy-based; In its preparation method, step (1) is constant, the process of step (2) preparation hydrogel substrate comprises: the ratio that the prepared two kinds of aqueous solutions of step (1) are 1:1 according to volume ratio is mixed, stirred at normal temperatures 1-10 minute, process titanium alloy-based surface with chemical method, then with mixture freezing on titanium alloy-based under-8 ℃ to-10 ℃ ,-10 ℃ to-60 ℃ lower freezing vacuum sublimation dryings 6-8 hour, the thickness of prepared hydrogel was 0.01mm to 5mm; Step (3) is constant to step (5).
11. calcium-supplementing nutritive product is characterized in that, described nutriment mainly contains the described nanometer pore hydroxyl calcium phosphate/aquogel materials of one of claim 1 to 6.
12. the biological filling/timbering material of an orthopaedics or gear division is characterized in that described material mainly contains the described nanometer pore hydroxyl calcium phosphate/aquogel materials of one of claim 1 to 6.
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| CN104548211B (en) * | 2014-12-19 | 2016-11-23 | 戴立军 | A kind of orthopaedics or gear division packing material, a kind of tooth implant and a kind of degradable artificial bone |
| CN106725743A (en) * | 2017-01-11 | 2017-05-31 | 首都医科大学附属北京天坛医院 | A kind of huge and very significant meningioma overall cutting method |
| CN107469137B (en) * | 2017-09-11 | 2020-04-07 | 曲阜师范大学 | Injectable hemostatic hydrogel material and preparation method and application thereof |
| CN107497375B (en) * | 2017-10-16 | 2019-11-08 | 中国石油大学(华东) | Calcium phosphate nanoparticles and ion complementary peptide composite water gel and preparation method thereof |
| CN108558285A (en) * | 2018-04-24 | 2018-09-21 | 常州思宇知识产权运营有限公司 | A kind of cracking resistance heat-insulating finishing mortar |
| CN109758615B (en) * | 2019-02-13 | 2021-05-25 | 四川大学 | Double-sided composite hydrogel and preparation method thereof |
| WO2022265585A1 (en) * | 2021-06-18 | 2022-12-22 | Chulalongkorn University | Method of fabricating an implantable construct and an implantable construct derived from the same |
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