CN102188754A - Nanometer pore hydroxyl calcium phosphate/aquogel materials - Google Patents
Nanometer pore hydroxyl calcium phosphate/aquogel materials Download PDFInfo
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- CN102188754A CN102188754A CN2011101047139A CN201110104713A CN102188754A CN 102188754 A CN102188754 A CN 102188754A CN 2011101047139 A CN2011101047139 A CN 2011101047139A CN 201110104713 A CN201110104713 A CN 201110104713A CN 102188754 A CN102188754 A CN 102188754A
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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 dental caries sickness rate 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 about 1 ton.But the tooth section packing material that uses all can not promote the quick reparation of dental tissue at present.
For the dental caries that drops, artificial tooth root is installed normally.Artificial tooth root claims tooth implant again, is by in the jawbone of surgical operation with its implant into body agomphosis position, repairs the device of artificial tooth after the healing at an upper portion thereof.Tooth implant can be divided into five types, i.e. (1) Metal and Alloy material type substantially by its material difference: 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, promptly above-mentioned two or more material is compound, ceramic coated etc. as the metal surface.At present the material metal class that tooth implant is commonly used mainly is pure titanium or titanium alloy, the titanium light specific gravity, the intensity height, nonmagnetic, contractility is little, chemical property is stable, have splendid corrosion resistance and with the compatibility of on every side biological tissue.Though titanium alloy has better biocompatibility, there is not to form very strong combination interface between implant and dentale.
In addition, in surgical operation, the reparation of bone is a general problem with regeneration, in order to obtain ideal osseous tissue substitution material, people have carried out a lot of researchs, normally adopt natural high polymers such as calcium hydroxy phosphate and collagen protein, gelatin, chitosan or synthetic high polymers such as polyvinyl alcohol, polyacrylamide to prepare composite.
But these composites all also have weak point, the material of preparation and nature bone structural similarity, and improving its biocompatibility is the emphasis of bone tissue engineer research.
We discover, mineralizing of organism is collaborative finishing under the common influence of multiple factor.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 (contains as, bone and finishes in order under the common control of osteopontin (osteopontin, OPN), tooth includes with phosphoprotein (phosphophoryn)).Traditional theoretical conception has then 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 the skelemin.The out-phase water-solubility protein that is adsorbed adsorbs the ion that mineralizes by ionic bond, resemble Ca/Mg ion etc., thereby improved the degree of supersaturation of local particle effectively, improved the degree of supersaturation of calcium ion, carbanion and/or phosphate anion calcium carbonate/calcium phosphate (CaCO3/CaP).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 control 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 the biodegradable organization bracket being carried out body and surface modification with water-solubility protein, and by the bionical process that mineralizes (be similar to human teeth and bone form condition) support mineralizes, provide a kind of nanometer poroid calcium hydroxy phosphate/hydrogel material, this material be macroscopical 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:
Poroid calcium hydroxy phosphate/the hydrogel material of nanometer 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 the hydrogel, the preparation method of the poroid calcium hydroxy phosphate/hydrogel material of described nanometer comprises the steps:
(1) the preparation mass concentration is the hydrogel aqueous solution of 1-5%, and the preparation mass concentration is the glutaraldehyde water solution of 3-8%, and compound concentration is the polypeptid solution of 10-300ug/mL,
(2) prepare the hydrogel substrate: the mixed that with the prepared three kinds of aqueous solutions of step (1) is 1: 1: 1 according to volume ratio, stirred at normal temperatures 0.5-10 minute, then mixture is placed (on sheet glass) on the 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 thermic gelation 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 to regulate the pH value of this buffer, PH the best is 7.2-7.4
(4) take by weighing calcium chloride and dipotassium hydrogen phosphate respectively, and dissolve in respectively in the buffer that step (3) makes, be mixed with calcium chloride solution and the dipotassium hydrogen phosphate solution of 0.002-0.006mol/L respectively, is 1: 1 with the polypeptid solution that makes in calcium chloride solution and dipotassium hydrogen phosphate solution and the step (1) according to volume ratio: the ratio of X (X=0-6) is mixed stirring mutually
(5) the prepared hydrogel substrate that is carried on the carrier of step (2) is placed the prepared mixed solution of step (4), leave standstill 24-48 hour after, successively water and washing with alcohol, drying promptly makes the poroid calcium hydroxy phosphate/hydrogel material of described nanometer.
Preferably, the invention provides the poroid calcium hydroxy phosphate/hydrogel material of a kind of nanometer, its feature is that the preparation method of the poroid calcium hydroxy phosphate/hydrogel material of described nanometer comprises the steps:
(1) the preparation mass concentration is the hydrogel 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: is that 1: 1: 1 ratio is mixed mutually with the prepared three kinds of aqueous solutions of step (1) according to volume ratio, stirred at normal temperatures 0.5-10 minute, then mixture is placed on the 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 by weighing calcium chloride and dipotassium hydrogen phosphate respectively, and dissolve in respectively in the buffer that step (3) makes, be mixed with calcium chloride solution and the dipotassium hydrogen phosphate solution of 0.002-0.006mol/L respectively, is 1: 1 with the polypeptid solution that makes in calcium chloride solution and dipotassium hydrogen phosphate solution and the step (1) according to volume ratio: the ratio of X (X=0-6) is mixed mutually, stirred at normal temperatures 0.5-10 minute
(5) the prepared hydrogel substrate that is carried on the carrier of step (2) is placed the prepared mixed solution of step (4), leave standstill 24-36 hour after, successively water and washing with alcohol, drying promptly makes the poroid calcium hydroxy phosphate/hydrogel material of described nanometer.
Further preferred, the invention provides the poroid calcium hydroxy phosphate/hydrogel material of a kind of nanometer, it is characterized in that the preparation method of the poroid calcium hydroxy phosphate/hydrogel material of described nanometer comprises the steps:
(1) the preparation mass concentration is 1% hydrogel aqueous solution, and the preparation mass concentration is 5% glutaraldehyde water solution, and compound concentration is the polypeptid solution of 200ug/ml,
(2) preparation hydrogel substrate: is that 1: 1: 1 ratio is mixed mutually with the prepared three kinds of aqueous solutions of step (1) according to volume ratio, stirred at normal temperatures 0.5-10 minute, then mixture is placed on the 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 by weighing calcium chloride and dipotassium hydrogen phosphate respectively, and dissolve in respectively in the buffer that step (3) makes, be mixed with calcium chloride solution and the dipotassium hydrogen phosphate solution of 0.002-0.006mol/L respectively, is 1: 1 with the polypeptid solution that makes in calcium chloride solution and dipotassium hydrogen phosphate solution and the step (1) according to volume ratio: the ratio of X (X=0-6) is mixed mutually, stirred at normal temperatures 0.5-10 minute
(5) the prepared hydrogel substrate that is carried on the carrier of step (2) is placed the prepared mixed solution of step (4), leave standstill 24 hours after, successively water and washing with alcohol, drying promptly makes the poroid calcium hydroxy phosphate/hydrogel material of described nanometer.
The present invention also provides a kind of nanometer poroid calcium hydroxy phosphate/hydrogel material, 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 the hydrogel, the preparation method of the poroid calcium hydroxy phosphate/hydrogel material of described nanometer comprises the steps:
(1) the preparation mass concentration is the hydrogel aqueous solution of 1-5%, and compound concentration is the polypeptid solution of 10-300ug/mL,
(2) prepare the hydrogel substrate: the mixed that with the prepared two kinds of aqueous solutions of step (1) is 1: 1 according to volume ratio, stirred at normal temperatures 1-10 minute, and then mixture was placed on the carrier, under the room temperature dry 24-48 hour, vacuum drying under 80 ℃-100 ℃ temperature then
(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 by weighing calcium chloride and dipotassium hydrogen phosphate respectively, and dissolve in respectively in the buffer that step (3) makes, be mixed with calcium chloride solution and the dipotassium hydrogen phosphate solution of 0.002-0.006mol/L respectively, is 1: 1 with the polypeptid solution that makes in calcium chloride solution and dipotassium hydrogen phosphate solution and the step (1) according to volume ratio: the ratio of X (X=0-6) is mixed stirring mutually
(5) the prepared hydrogel substrate that is carried on the carrier of step (2) is placed the prepared mixed solution of step (4), leave standstill 24-48 hour after, successively water and washing with alcohol, drying promptly makes the poroid calcium hydroxy phosphate/hydrogel material of described nanometer.
Hydrogel is the gel of disperse medium with water.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, the character softness 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 big classes by its source.Natural hydrophilic macromolecule comprises polysaccharide (starch, cellulose, alginic acid, hyaluronic acid, chitosan etc.) and polypeptide class (collagen, poly-L-lysine, 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 at least two kinds compositions wherein; Described polypeptide be the osteopontin that contains of bone (osteopontin, OPN), the tooth phosphoprotein (phosphophoryn) or its fragment that contain; 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 a 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 the 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 chemical 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 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, the described titanium alloy-based poroid calcium hydroxy phosphate/hydrogel material of above-mentioned nanometer provided by the invention that is provided with, in its preparation method, step (1) is constant, the process of step (2) preparation hydrogel substrate comprise with mixture spray to titanium alloy-based on, rapid draing under the room temperature placed 90 ℃ to 100 ℃ heat treatments 5 to 15 minutes again after (as passing through vacuum drying), prepared hydrogel is two-dimentional, be the continuous or discrete thin film of one deck, the width of the discontinuous part of described discontinuous thin film is 1um-3mm, and other content in this process is constant; 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 the described titanium alloy-based poroid calcium hydroxy phosphate/hydrogel material of above-mentioned nanometer provided by the invention that is provided with; In its preparation method, step (1) is constant, the process of step (2) preparation hydrogel substrate comprises with chemical method handles titanium alloy-based surface, then that mixture is down freezing on titanium alloy-based at-8 ℃ to-10 ℃,-10 ℃ to-60 ℃ following freezing vacuum sublimation dryings 6-8 hour, prepared hydrogel is a three-dimensional framework, and its thickness is 0.01mm to 5mm, and other content in this process is constant; 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 the poroid calcium hydroxy phosphate/hydrogel material of above-mentioned nanometer provided by the invention.
The present invention also provides a kind of orthopaedics or tooth section biological filling/timbering material, and its feature is that described packing material mainly contains the poroid calcium hydroxy phosphate/hydrogel material of above-mentioned nanometer provided by the invention.
Compared with prior art, the poroid calcium hydroxy phosphate/hydrogel material of nanometer provided by the invention, wherein, hydrogel is extremely millimetre-sized poroid two dimension or a three-dimensional material of micron, and the three-dimensional material hole is interconnected, and the aperture is: 1 micron to 3 millimeters; The calcium hydroxy phosphate that is deposited on the hydrogel is the poroid calcium hydroxy phosphate of three-dimensional communication, and the aperture is 50-1000nm, and hole wall is thick to be the 30-80 nanometer.This material be macroscopical 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 to be the 30-80 nanometer, so nutrition is easier to be absorbed by the body.
Compared with prior art, tooth implant provided by the invention is because its titanium alloy-based two dimension or poroid calcium hydroxy phosphate/hydrogel material of three-dimensional nanometer provided by the invention of being provided with, 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 tooth section, biocompatibility is stronger.
Description of drawings
Fig. 1 is the sketch map of turkey tendon of mineralizing: (A) the turkey tendon is laterally separated along its length and is 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 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 successive micron adsorbed property of porous hydroxyl calcium phosphate of the present invention, (B), (C) be deposited on electron-microscope scanning figure on the hydrogel material of polypeptide modification for the netted calcium hydroxy phosphate of successive nanoporous of the present invention for the successive calcium hydroxy phosphate of densification of the present invention is deposited on electron-microscope scanning figure on the hydrogel material of polypeptide modification;
Fig. 4 prepares the process schematic representation of the poroid calcium hydroxy phosphate/hydrogel material of three-dimensional manometer for of the present invention with freezing method, (A) solid free forming sketch map, (B) multiinjector deposition manufacturing system sketch map, (C) three-dimensional framework sketch map;
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, the two dimension or the three-dimensional framework of hydrogel material formation
505, the post-depositional hydrogel skeleton of calcium hydroxy phosphate
The specific embodiment
Used hydrogel material among the embodiment is as gelatin, collagen, alginic acid etc.; Osteopontin (osteopontin, OPN), phosphoprotein polypeptide such as (phosphophoryn) is the product of selling on the market commonly used;
Tri(Hydroxymethyl) Amino Methane Hydrochloride (Tris-HCl), Tris (Tris), calcium chloride, dipotassium hydrogen phosphate, glutaraldehyde, sodium chloride used among the 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 among the embodiment 1-16 (2) can be a sheet glass.
As shown in Figure 1: (A) the turkey tendon is laterally separated along its length and is 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) for the Electronic Speculum figure under the condition that does not have active polypeptide, shown in (B), do not having under the condition of nonactive polypeptide owing to there is not 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, shown in (D), forms and the similar mineral parcel of people's bone collagen fabric, and can effectively infiltrate the longitudinal section, shown in (E), form the collagen fabric of fine and close mineral parcel, rather than odd random deposition.This has not only greatly improved the density of mineral, and because the extremely strong interaction force that has formed between mineral and collagen protein and greatly improved the intensity of material monolithic, thereby on macroscopic view and microcosmic, all more approach the bone material of organism self, can be used as artificial sclerotin and transplant in human body.
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.Electronic Speculum result shows that the poroid calcium hydroxy phosphate/hydrogel material of nanometer 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 poroid calcium hydroxy phosphate/hydrogel material of nanometer that the present invention prepares has identical microstructure with people's bone, and promptly the crystalline center of calcium hydroxy phosphate crystallographic axis (C axle) is parallel with the major axis of collagen fabric.
As shown in Figure 3, (A) successive micron adsorbed property of porous hydroxyl calcium phosphate is deposited on the hydrogel material of polypeptide modification.(B) fine and close successive calcium hydroxy phosphate is deposited on polypeptide modification or the hydrogel material without the polypeptide modification.(C) the netted calcium hydroxy phosphate of successive 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 calcium hydroxy phosphate that the present invention can synthesize proper density and pattern is filled or is implanted in people's bone of different densities being used to.The holey calcium hydroxy phosphate can be used as osteocyte growth skeleton, and does not need additionally to add somatomedin.The calcium hydroxy phosphate of different densities and form is determined by multiple factor, as, with the kind of hydrogel, whether hydrogel is modified, by which kind of polypeptide modification, the albumen that is added 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 fine and close calcium hydroxy phosphate (atresia) with the hydrogel of no modification, shown in Fig. 3 (B).
As shown in Figure 4: (A) solid free forming.(solid free-form, SFF) technology is also referred to as rapid prototyping input method (Rapid Protyping) to the solid free forming.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, deposit on the platform by computer-controlled nozzle extruded material long filament or liquid droplets material, storehouse goes out a three dimensional object from level to level.(B) multiinjector deposition manufacturing system sketch map.Hydrogel material in the water-soluble or solvent is in that deposit to can be frozen at once on the 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 osteocyte somatomedin 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 the 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, can screw on crown lid 502 (the crown lid can be changed afterwards in case of necessity); Shown in figure (A), the outer surface of its underpart, the hydrogel material that at first 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 the hydrogel material, shown in figure (B).Calcium hydroxy phosphate has strengthened the hydrogel skeleton, has kept the connection cavernous structure of three dimensional hydrogel skeleton simultaneously.Like this osteoblast can be in the shortest time bioelectric interface between repairing transplant body and dentale, tooth implant is firmly got off and can not lose the adjacent teeth that is used for fixing transplant.
Embodiment 1
Poroid calcium hydroxy phosphate/the hydrogel material of nanometer provided by the invention, preparation method comprises the steps:
(1) the preparation mass concentration is 1% hydrogel aqueous solution, and the preparation mass concentration is 3% glutaraldehyde water solution, and compound concentration is the polypeptid solution of 10ug/mL,
Above-mentioned hydrogel is a collagen; Aforementioned polypeptides be the osteopontin that contains of bone (osteopontin, OPN); Be added with mass percent in the described hydrogel material and be the micromolecular compound of 0.0001% promotion osteoblast breeding.
(2) prepare the hydrogel substrate: the mixed that with the prepared three kinds of aqueous solutions of step (1) is 1: 1: 1 according to volume ratio, stirred at normal temperatures 0.5 minute, then mixture is placed on the 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 by weighing calcium chloride and dipotassium hydrogen phosphate respectively, and dissolve in respectively in the buffer that step (3) makes, be mixed with calcium chloride solution and the dipotassium hydrogen phosphate solution of 0.002mol/L respectively, is that 1: 1: 0 ratio is mixed stirring (promptly mutually with the polypeptid solution that makes in calcium chloride solution and dipotassium hydrogen phosphate solution and the step (1) according to volume ratio, is that 1: 1 ratio is mixed stirring mutually with calcium chloride solution and dipotassium hydrogen phosphate solution according to volume ratio)
(5) the prepared hydrogel substrate that is carried on the carrier of step (2) is placed the prepared mixed solution of step (4), leave standstill 24 hours after, successively water and washing with alcohol, drying promptly makes the poroid calcium hydroxy phosphate/hydrogel material of described nanometer.
Poroid calcium hydroxy phosphate/the hydrogel material of nanometer provided by the invention, preparation method comprises the steps:
(1) the preparation mass concentration is 5% hydrogel aqueous solution, and the preparation mass concentration is 8% glutaraldehyde water solution, and compound concentration is the polypeptid solution of 300ug/mL,
Above-mentioned hydrogel is a gelatin; Aforementioned polypeptides is phosphoprotein (phosphophoryn) or its fragment that tooth contains; Be added with mass percent in the above-mentioned hydrogel material and be the micromolecular compound of 5% promotion osteoblast breeding.
(2) prepare the hydrogel substrate: the mixed that with the prepared three kinds of aqueous solutions of step (1) is 1: 1: 1 according to volume ratio, stirred at normal temperatures 10 minutes, then mixture is placed on the 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 by weighing calcium chloride and dipotassium hydrogen phosphate respectively, and dissolve in respectively in the buffer that step (3) makes, be mixed with calcium chloride solution and the dipotassium hydrogen phosphate solution of 0.006mol/L respectively, is that 1: 1: 6 ratio is mixed stirring mutually with the polypeptid solution that makes in calcium chloride solution and dipotassium hydrogen phosphate solution and the step (1) according to volume ratio
(5) the prepared hydrogel substrate that is carried on the carrier of step (2) is placed the prepared mixed solution of step (4), leave standstill 48 hours after, successively water and washing with alcohol, drying promptly makes the poroid calcium hydroxy phosphate/hydrogel material of described nanometer.
Embodiment 3
Poroid calcium hydroxy phosphate/the hydrogel material of nanometer provided by the invention, preparation method comprises the steps:
(1) the preparation mass concentration is 3% hydrogel aqueous solution, and the preparation mass concentration is 6% glutaraldehyde water solution, and compound concentration is the polypeptid solution of 150ug/mL,
Above-mentioned hydrogel 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 the described hydrogel material and be the micromolecular compound of 2.5% promotion osteoblast breeding.
(2) prepare the hydrogel substrate: the mixed that with the prepared three kinds of aqueous solutions of step (1) is 1: 1: 1 according to volume ratio, stirred at normal temperatures 6 minutes, then mixture is placed on the 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 by weighing calcium chloride and dipotassium hydrogen phosphate respectively, and dissolve in respectively in the buffer that step (3) makes, be mixed with calcium chloride solution and the dipotassium hydrogen phosphate solution of 0.004mol/L respectively, is that 1: 1: 3 ratio is mixed stirring mutually with the polypeptid solution that makes in calcium chloride solution and dipotassium hydrogen phosphate solution and the step (1) according to volume ratio
(5) the prepared hydrogel substrate that is carried on the carrier of step (2) is placed the prepared mixed solution of step (4), leave standstill 36 hours after, successively water and washing with alcohol, drying promptly makes the poroid calcium hydroxy phosphate/hydrogel material of described nanometer.
Poroid calcium hydroxy phosphate/the hydrogel material of nanometer provided by the invention, preparation method comprises the steps:
(1) the preparation mass concentration is 1% hydrogel aqueous solution, and the preparation mass concentration is 8% glutaraldehyde water solution, and compound concentration is the polypeptid solution of 300ug/mL,
Above-mentioned hydrogel is a polyvinyl alcohol; Aforementioned polypeptides is osteopontin (osteopontin, fragment OPN) that bone contains; Be added with mass percent in the above-mentioned hydrogel material and be the micromolecular compound of 0.001 promotion osteoblast breeding.
(2) prepare the hydrogel substrate: the mixed that with the prepared three kinds of aqueous solutions of step (1) is 1: 1: 1 according to volume ratio, stirred at normal temperatures 9 minutes, then mixture is placed on the 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 by weighing calcium chloride and dipotassium hydrogen phosphate respectively, and dissolve in respectively in the buffer that step (3) makes, be mixed with calcium chloride solution and the dipotassium hydrogen phosphate solution of 0.006mol/L respectively, is that 1: 1: 1 ratio is mixed stirring mutually with the polypeptid solution that makes in calcium chloride solution and dipotassium hydrogen phosphate solution and the step (1) according to volume ratio
(5) the prepared hydrogel substrate that is carried on the carrier of step (2) is placed the prepared mixed solution of step (4), leave standstill 48 hours after, successively water and washing with alcohol, drying promptly makes the poroid calcium hydroxy phosphate/hydrogel material of described nanometer.
Embodiment 5
Poroid calcium hydroxy phosphate/the hydrogel material of nanometer provided by the invention, preparation method comprises the steps:
(1) the preparation mass concentration is 1% hydrogel 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 hydrogel is an 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 the above-mentioned hydrogel material and be the micromolecular compound of 0.01% promotion osteoblast breeding.
(2) preparation hydrogel substrate: is that 1: 1: 1 ratio is mixed mutually with the prepared three kinds of aqueous solutions of step (1) according to volume ratio, stirred at normal temperatures 0.5 minute, then mixture is placed on the 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 by weighing calcium chloride and dipotassium hydrogen phosphate respectively, and dissolve in respectively in the buffer that step (3) makes, be mixed with calcium chloride solution and the dipotassium hydrogen phosphate solution of 0.002mol/L respectively, is that 1: 1: 0 ratio is mixed (promptly mutually with the polypeptid solution that makes in calcium chloride solution and dipotassium hydrogen phosphate solution and the step (1) according to volume ratio, is that 1: 1 ratio is mixed stirring mutually with calcium chloride solution and dipotassium hydrogen phosphate solution according to volume ratio), stirred at normal temperatures 0.5 minute
(5) the prepared hydrogel substrate that is carried on the carrier of step (2) is placed the prepared mixed solution of step (4), leave standstill 24 hours after, successively water and washing with alcohol, drying promptly makes the poroid calcium hydroxy phosphate/hydrogel material of described nanometer.
Poroid calcium hydroxy phosphate/the hydrogel material of nanometer provided by the invention, preparation method comprises the steps:
(1) the preparation mass concentration is 3% hydrogel 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 hydrogel 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 the above-mentioned hydrogel material and be the micromolecular compound of 0.01% promotion osteoblast breeding.
(2) preparation hydrogel substrate: is that 1: 1: 1 ratio is mixed mutually with the prepared three kinds of aqueous solutions of step (1) according to volume ratio, stirred at normal temperatures 10 minutes, then mixture is placed on the 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 by weighing calcium chloride and dipotassium hydrogen phosphate respectively, and dissolve in respectively in the buffer that step (3) makes, be mixed with calcium chloride solution and the dipotassium hydrogen phosphate solution of 0.006mol/L respectively, is that 1: 1: 6 ratio is mixed mutually with the polypeptid solution that makes in calcium chloride solution and dipotassium hydrogen phosphate solution and the step (1) according to volume ratio, stirred at normal temperatures 10 minutes
(5) the prepared hydrogel substrate that is carried on the carrier of step (2) is placed the prepared mixed solution of step (4), leave standstill 36 hours after, successively water and washing with alcohol, drying promptly makes the poroid calcium hydroxy phosphate/hydrogel material of described nanometer.
Embodiment 7
Poroid calcium hydroxy phosphate/the hydrogel material of nanometer provided by the invention, preparation method comprises the steps:
(1) the preparation mass concentration is 2% hydrogel 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 hydrogel is an 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 the above-mentioned hydrogel material and be the micromolecular compound of 0.0005% promotion osteoblast breeding.
(2) preparation hydrogel substrate: is that 1: 1: 1 ratio is mixed mutually with the prepared three kinds of aqueous solutions of step (1) according to volume ratio, stirred at normal temperatures 5 minutes, then mixture is placed on the 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 by weighing calcium chloride and dipotassium hydrogen phosphate respectively, and dissolve in respectively in the buffer that step (3) makes, be mixed with calcium chloride solution and the dipotassium hydrogen phosphate solution of 0.004mol/L respectively, is that 1: 1: 3 ratio is mixed mutually with the polypeptid solution that makes in calcium chloride solution and dipotassium hydrogen phosphate solution and the step (1) according to volume ratio, stirred at normal temperatures 5 minutes
(5) the prepared hydrogel substrate that is carried on the carrier of step (2) is placed the prepared mixed solution of step (4), leave standstill 30 hours after, successively water and washing with alcohol, drying promptly makes the poroid calcium hydroxy phosphate/hydrogel material of described nanometer.
Embodiment 8
Poroid calcium hydroxy phosphate/the hydrogel material of nanometer provided by the invention, preparation method comprises the steps:
(1) the preparation mass concentration is 1% hydrogel 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 hydrogel 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 the above-mentioned hydrogel material and be the micromolecular compound of 0.001% promotion osteoblast breeding.
(2) preparation hydrogel substrate: is that 1: 1: 1 ratio is mixed mutually with the prepared three kinds of aqueous solutions of step (1) according to volume ratio, stirred at normal temperatures 0.5 minute, then mixture is placed on the 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 by weighing calcium chloride and dipotassium hydrogen phosphate respectively, and dissolve in respectively in the buffer that step (3) makes, be mixed with calcium chloride solution and the dipotassium hydrogen phosphate solution of 0.002mol/L respectively, is that 1: 1: 1 ratio is mixed mutually with the polypeptid solution that makes in calcium chloride solution and dipotassium hydrogen phosphate solution and the step (1) according to volume ratio, stirred at normal temperatures 10 minutes
(5) the prepared hydrogel substrate that is carried on the carrier of step (2) is placed the prepared mixed solution of step (4), leave standstill 36 hours after, successively water and washing with alcohol, drying promptly makes the poroid calcium hydroxy phosphate/hydrogel material of described nanometer.
Embodiment 9
Poroid calcium hydroxy phosphate/the hydrogel material of nanometer provided by the invention, preparation method comprises the steps:
(1) the preparation mass concentration is 3% hydrogel 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 hydrogel 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 the above-mentioned hydrogel material and be the micromolecular compound of 0.005% promotion osteoblast breeding.
(2) preparation hydrogel substrate: is that 1: 1: 1 ratio is mixed mutually with the prepared three kinds of aqueous solutions of step (1) according to volume ratio, stirred at normal temperatures 10 minutes, then mixture is placed on the 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 by weighing calcium chloride and dipotassium hydrogen phosphate respectively, and dissolve in respectively in the buffer that step (3) makes, be mixed with calcium chloride solution and the dipotassium hydrogen phosphate solution of 0.006mol/L respectively, is that 1: 1: 1 ratio is mixed mutually with the polypeptid solution that makes in calcium chloride solution and dipotassium hydrogen phosphate solution and the step (1) according to volume ratio, stirred at normal temperatures 10 minutes
(5) the prepared hydrogel substrate that is carried on the carrier of step (2) is placed the prepared mixed solution of step (4), leave standstill 24 hours after, successively water and washing with alcohol, drying promptly makes the poroid calcium hydroxy phosphate/hydrogel material of described nanometer.
Embodiment 10
Poroid calcium hydroxy phosphate/the hydrogel material of nanometer provided by the invention, preparation method comprises the steps:
(1) the preparation mass concentration is 1% hydrogel aqueous solution, and the preparation mass concentration is 5% glutaraldehyde water solution, and compound concentration is the polypeptid solution of 200ug/ml,
Above-mentioned hydrogel 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 the above-mentioned hydrogel material and be the micromolecular compound of 0.05% promotion osteoblast breeding.
(2) preparation hydrogel substrate: is that 1: 1: 1 ratio is mixed mutually with the prepared three kinds of aqueous solutions of step (1) according to volume ratio, stirred at normal temperatures 0.5 minute, then mixture is placed on the 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, and the pH value of this buffer is 7.2-7.4,
(4) take by weighing calcium chloride and dipotassium hydrogen phosphate respectively, and dissolve in respectively in the buffer that step (3) makes, be mixed with calcium chloride solution and the dipotassium hydrogen phosphate solution of 0.002mol/L respectively, is that 1: 1: 0 ratio is mixed (promptly mutually with the polypeptid solution that makes in calcium chloride solution and dipotassium hydrogen phosphate solution and the step (1) according to volume ratio, is that 1: 1 ratio is mixed stirring mutually with calcium chloride solution and dipotassium hydrogen phosphate solution according to volume ratio), stirred at normal temperatures 0.5 minute
(5) the prepared hydrogel substrate that is carried on the carrier of step (2) is placed the prepared mixed solution of step (4), leave standstill 24 hours after, successively water and washing with alcohol, drying promptly makes the poroid calcium hydroxy phosphate/hydrogel material of described nanometer.
Embodiment 11
Poroid calcium hydroxy phosphate/the hydrogel material of nanometer provided by the invention, preparation method comprises the steps:
(1) the preparation mass concentration is 1% hydrogel aqueous solution, and the preparation mass concentration is 5% glutaraldehyde water solution, and compound concentration is the polypeptid solution of 200ug/ml,
Above-mentioned hydrogel 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 a poly-aspartic-acid, is added with mass percent in the above-mentioned hydrogel material and is the micromolecular compound of 0.05% promotion osteoblast breeding.
(2) preparation hydrogel substrate: is that 1: 1: 1 ratio is mixed mutually with the prepared three kinds of aqueous solutions of step (1) according to volume ratio, stirred at normal temperatures 10 minutes, then mixture is placed on the 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, and the pH value of this buffer is 7.2-7.4,
(4) take by weighing calcium chloride and dipotassium hydrogen phosphate respectively, and dissolve in respectively in the buffer that step (3) makes, be mixed with calcium chloride solution and the dipotassium hydrogen phosphate solution of 0.006mol/L respectively, is that 1: 1: 6 ratio is mixed mutually with the polypeptid solution that makes in calcium chloride solution and dipotassium hydrogen phosphate solution and the step (1) according to volume ratio, stirred at normal temperatures 10 minutes
(5) the prepared hydrogel substrate that is carried on the carrier of step (2) is placed the prepared mixed solution of step (4), leave standstill 24 hours after, successively water and washing with alcohol, drying promptly makes the poroid calcium hydroxy phosphate/hydrogel material of described nanometer.
Embodiment 12
Poroid calcium hydroxy phosphate/the hydrogel material of nanometer provided by the invention, preparation method comprises the steps:
(1) the preparation mass concentration is 1% hydrogel aqueous solution, and the preparation mass concentration is 5% glutaraldehyde water solution, and compound concentration is the polypeptid solution of 200ug/ml,
Above-mentioned hydrogel is a polyvinyl alcohol; Aforementioned polypeptides is a polyglutamic acid; Be added with mass percent in the above-mentioned hydrogel material and be the micromolecular compound of 0.5% promotion osteoblast breeding.
(2) preparation hydrogel substrate: is that 1: 1: 1 ratio is mixed mutually with the prepared three kinds of aqueous solutions of step (1) according to volume ratio, stirred at normal temperatures 5 minutes, then mixture is placed on the 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, and the pH value of this buffer is 7.2-7.4,
(4) take by weighing calcium chloride and dipotassium hydrogen phosphate respectively, and dissolve in respectively in the buffer that step (3) makes, be mixed with calcium chloride solution and the dipotassium hydrogen phosphate solution of 0.004mol/L respectively, is that 1: 1: 3 ratio is mixed mutually with the polypeptid solution that makes in calcium chloride solution and dipotassium hydrogen phosphate solution and the step (1) according to volume ratio, stirred at normal temperatures 5 minutes
(5) the prepared hydrogel substrate that is carried on the carrier of step (2) is placed the prepared mixed solution of step (4), leave standstill 24 hours after, successively water and washing with alcohol, drying promptly makes the poroid calcium hydroxy phosphate/hydrogel material of described nanometer.
Embodiment 13
Poroid calcium hydroxy phosphate/the hydrogel material of nanometer 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 the hydrogel, the preparation method of the poroid calcium hydroxy phosphate/hydrogel material of described nanometer comprises the steps:
(1) the preparation mass concentration is 1% hydrogel aqueous solution, and compound concentration is the polypeptid solution of 10ug/mL,
Above-mentioned hydrogel is a collagen; Aforementioned polypeptides be the osteopontin that contains of bone (osteopontin, OPN); Be added with mass percent in the described hydrogel material and be the micromolecular compound of 0.0001% promotion osteoblast breeding.
(2) preparation hydrogel substrate: is 1: 1 mixed with the prepared two kinds of aqueous solutions of step (1) according to volume ratio, stirs at normal temperatures 1 minute, then mixture placed on the carrier, and under the room temperature dry 24 hours, vacuum drying under 80 ℃ temperature then,
(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 by weighing calcium chloride and dipotassium hydrogen phosphate respectively, and dissolve in respectively in the buffer that step (3) makes, be mixed with calcium chloride solution and the dipotassium hydrogen phosphate solution of 0.002mol/L respectively, is that 1: 1: 0 ratio is mixed stirring (promptly mutually with the polypeptid solution that makes in calcium chloride solution and dipotassium hydrogen phosphate solution and the step (1) according to volume ratio, is that 1: 1 ratio is mixed stirring mutually with calcium chloride solution and dipotassium hydrogen phosphate solution according to volume ratio)
(5) the prepared hydrogel substrate that is carried on the carrier of step (2) is placed the prepared mixed solution of step (4), leave standstill 24 hours after, successively water and washing with alcohol, drying promptly makes the poroid calcium hydroxy phosphate/hydrogel material of described nanometer.
Embodiment 14
Poroid calcium hydroxy phosphate/the hydrogel material of nanometer 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 the hydrogel, the preparation method of the poroid calcium hydroxy phosphate/hydrogel material of described nanometer comprises the steps:
(1) the preparation mass concentration is 5% hydrogel aqueous solution, and compound concentration is the polypeptid solution of 300ug/mL,
Above-mentioned hydrogel is a gelatin; Aforementioned polypeptides is phosphoprotein (phosphophoryn) or its fragment that tooth contains; Be added with mass percent in the above-mentioned hydrogel material and be the micromolecular compound of 5% promotion osteoblast breeding.
(2) prepare the hydrogel substrate: the mixed that with the prepared two kinds of aqueous solutions of step (1) is 1: 1 according to volume ratio, stirred at normal temperatures 10 minutes, and then mixture was placed on the carrier, drying is 48 hours under the room temperature, vacuum drying under 100 ℃ temperature then
(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 by weighing calcium chloride and dipotassium hydrogen phosphate respectively, and dissolve in respectively in the buffer that step (3) makes, be mixed with calcium chloride solution and the dipotassium hydrogen phosphate solution of 0.006mol/L respectively, is that 1: 1: 6 ratio is mixed stirring mutually with the polypeptid solution that makes in calcium chloride solution and dipotassium hydrogen phosphate solution and the step (1) according to volume ratio
(5) the prepared hydrogel substrate that is carried on the carrier of step (2) is placed the prepared mixed solution of step (4), leave standstill 48 hours after, successively water and washing with alcohol, drying promptly makes the poroid calcium hydroxy phosphate/hydrogel material of described nanometer.
Embodiment 15
Poroid calcium hydroxy phosphate/the hydrogel material of nanometer 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 the hydrogel, the preparation method of the poroid calcium hydroxy phosphate/hydrogel material of described nanometer comprises the steps:
(1) the preparation mass concentration is 3% hydrogel aqueous solution, and compound concentration is the polypeptid solution of 150ug/mL,
Above-mentioned hydrogel 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 the described hydrogel material and be the micromolecular compound of 2.5% promotion osteoblast breeding.
(2) preparation hydrogel substrate: is 1: 1 mixed with the prepared two kinds of aqueous solutions of step (1) according to volume ratio, stirs at normal temperatures 6 minutes, then mixture placed on the carrier, and under the room temperature dry 36 hours, vacuum drying under 90 ℃ temperature then,
(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 by weighing calcium chloride and dipotassium hydrogen phosphate respectively, and dissolve in respectively in the buffer that step (3) makes, be mixed with calcium chloride solution and the dipotassium hydrogen phosphate solution of 0.004mol/L respectively, is that 1: 1: 3 ratio is mixed stirring mutually with the polypeptid solution that makes in calcium chloride solution and dipotassium hydrogen phosphate solution and the step (1) according to volume ratio
(5) the prepared hydrogel substrate that is carried on the carrier of step (2) is placed the prepared mixed solution of step (4), leave standstill 36 hours after, successively water and washing with alcohol, drying promptly makes the poroid calcium hydroxy phosphate/hydrogel material of described nanometer.
Embodiment 16
Poroid calcium hydroxy phosphate/the hydrogel material of nanometer 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 the hydrogel, the preparation method of the poroid calcium hydroxy phosphate/hydrogel material of described nanometer comprises the steps:
(1) the preparation mass concentration is 5% hydrogel aqueous solution, and compound concentration is the polypeptid solution of 100ug/mL,
Above-mentioned hydrogel is a polyvinyl alcohol; Aforementioned polypeptides is osteopontin (osteopontin, fragment OPN) that bone contains; Be added with mass percent in the above-mentioned hydrogel material and be the micromolecular compound of 0.001 promotion osteoblast breeding.
(2) prepare the hydrogel substrate: the mixed that with the prepared two kinds of aqueous solutions of step (1) is 1: 1 according to volume ratio, stirred at normal temperatures 5 minutes, and then mixture was placed on the carrier, drying is 24 hours under the room temperature, vacuum drying under 100 ℃ temperature then
(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 by weighing calcium chloride and dipotassium hydrogen phosphate respectively, and dissolve in respectively in the buffer that step (3) makes, be mixed with calcium chloride solution and the dipotassium hydrogen phosphate solution of 0.002mol/L respectively, is that 1: 1: 1 ratio is mixed stirring mutually with the polypeptid solution that makes in calcium chloride solution and dipotassium hydrogen phosphate solution and the step (1) according to volume ratio
(5) the prepared hydrogel substrate that is carried on the carrier of step (2) is placed the prepared mixed solution of step (4), leave standstill 48 hours after, successively water and washing with alcohol, drying promptly makes the poroid calcium hydroxy phosphate/hydrogel material of described nanometer.
Embodiment 17
Tooth implant provided by the invention, comprise titanium alloy-based, preparation method is for being that 1: 1: 1 ratio is mixed stirring mutually with the prepared three kinds of aqueous solutions of step (1) among the embodiment 1 according to volume ratio, then mixture is sprayed to titanium alloy-based on, rapid draing under the room temperature, as by placing 90 ℃ of heat treatments behind the vacuum drying again 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 1um-100um, other content of preparation method is constant, identical with the method among the embodiment 1, promptly make the tooth implant that is provided with the poroid calcium hydroxy phosphate/hydrogel material of nanometer of the present invention.
Embodiment 18
Tooth implant provided by the invention, comprise titanium alloy-based, preparation method is for being that 1: 1: 1 ratio is mixed stirring mutually with the prepared three kinds of aqueous solutions of step (1) among the embodiment 2 according to volume ratio, then mixture is sprayed to titanium alloy-based on, placed 100 ℃ of heat treatments after the vacuum rapid draing again 15 minutes under the 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 among the embodiment 2, promptly make the tooth implant that is provided with the poroid calcium hydroxy phosphate/hydrogel material of nanometer of the present invention.
Embodiment 19
Tooth implant provided by the invention, comprise titanium alloy-based, preparation method is for being that 1: 1: 1 ratio is mixed stirring mutually with the prepared three kinds of aqueous solutions of step (1) among the embodiment 6 according to volume ratio, then mixture is sprayed to titanium alloy-based on, placed 95 ℃ of heat treatments after the vacuum rapid draing again 10 minutes under the 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 500um-1mm, other content of preparation method is constant, identical with the method among the embodiment 6, promptly make the tooth implant that is provided with the poroid calcium hydroxy phosphate/hydrogel material of nanometer of the present invention.
Embodiment 20
Tooth implant provided by the invention, comprise titanium alloy-based, preparation method is for being that 1: 1 ratio is mixed stirring mutually with the prepared two kinds of aqueous solutions of step (1) among the embodiment 13 according to volume ratio, then mixture is sprayed to titanium alloy-based on, placed 90 ℃ of heat treatments after the vacuum rapid draing again 5 minutes under the 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 100um-500um, other content of preparation method is constant, identical with the method among the embodiment 13, promptly make the tooth implant that is provided with the poroid calcium hydroxy phosphate/hydrogel material of nanometer of the present invention.
Embodiment 21
Tooth implant provided by the invention, comprise titanium alloy-based, preparation method is for being that 1: 1 ratio is mixed stirring mutually with the prepared two kinds of aqueous solutions of step (1) among the embodiment 14 according to volume ratio, then mixture is sprayed to titanium alloy-based on, placed 100 ℃ of heat treatments after the vacuum rapid draing again 15 minutes under the 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 among the embodiment 14, promptly make the tooth implant that is provided with the poroid calcium hydroxy phosphate/hydrogel material of nanometer of the present invention.
Embodiment 22
Tooth implant provided by the invention, comprise titanium alloy-based, preparation method is for being that 1: 1 ratio is mixed stirring mutually with the prepared two kinds of aqueous solutions of step (1) among the embodiment 15 according to volume ratio, then mixture is sprayed to titanium alloy-based on, placed 95 ℃ of heat treatments after the vacuum rapid draing again 10 minutes under the 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 among the embodiment 15, promptly make the tooth implant that is provided with the poroid calcium hydroxy phosphate/hydrogel material of nanometer of the present invention.
Embodiment 23
Tooth implant provided by the invention, comprise titanium alloy-based, preparation method is for being that 1: 1: 1 ratio is mixed stirring mutually with the prepared three kinds of aqueous solutions of the step among the embodiment 8 (1) according to volume ratio, handle titanium alloy-based surface with chemical method, then that mixture is down freezing on titanium alloy-based at-8 ℃,-10 ℃ of following 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 among the embodiment 8, promptly make the tooth implant that is provided with the poroid calcium hydroxy phosphate/hydrogel material of nanometer of the present invention.
Embodiment 24
Tooth implant provided by the invention, comprise titanium alloy-based, preparation method is for being that 1: 1: 1 ratio is mixed stirring mutually with the prepared three kinds of aqueous solutions of the step among the embodiment 9 (1) according to volume ratio, handle titanium alloy-based surface with chemical method, then that mixture is down freezing on titanium alloy-based at-10 ℃,-60 ℃ of following 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 among the embodiment 9, promptly make the tooth implant that is provided with the poroid calcium hydroxy phosphate/hydrogel material of nanometer of the present invention.
Embodiment 25
Tooth implant provided by the invention, comprise titanium alloy-based, preparation method is for being that 1: 1: 1 ratio is mixed stirring mutually with the prepared three kinds of aqueous solutions of the step among the embodiment 11 (1) according to volume ratio, handle titanium alloy-based surface with chemical method, then that mixture is down freezing on titanium alloy-based at-9 ℃,-35 ℃ of following 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 among the embodiment 11, promptly make the tooth implant that is provided with the poroid calcium hydroxy phosphate/hydrogel material of nanometer of the present invention.
Embodiment 26
Tooth implant provided by the invention, comprise titanium alloy-based, preparation method is for being that 1: 1 ratio is mixed stirring mutually with the prepared two kinds of aqueous solutions of the step among the embodiment 13 (1) according to volume ratio, handle titanium alloy-based surface with chemical method, then with mixture at-8 ℃ of following freezing vacuum sublimation dryings on titanium alloy-based, descended freezing 6 hours at-10 ℃, the thickness of prepared hydrogel is 0.01mm-0.1mm, other content of preparation method is constant, identical with the method among the embodiment 13, promptly make the tooth implant that is provided with the poroid calcium hydroxy phosphate/hydrogel material of nanometer of the present invention.
Embodiment 27
Tooth implant provided by the invention, comprise titanium alloy-based, its preparation method is for being that 1: 1 ratio is mixed stirring mutually with the prepared two kinds of aqueous solutions of the step among the embodiment 14 (1) according to volume ratio, handle titanium alloy-based surface with chemical method, then that mixture is down freezing on titanium alloy-based at-10 ℃,-60 ℃ of following 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 among the embodiment 14, promptly make the tooth implant that is provided with the poroid calcium hydroxy phosphate/hydrogel material of nanometer of the present invention.
Embodiment 28
Tooth implant provided by the invention, comprise titanium alloy-based, its preparation method is for being that 1: 1 ratio is mixed stirring mutually with the prepared two kinds of aqueous solutions of the step among the embodiment 15 (1) according to volume ratio, handle titanium alloy-based surface with chemical method, then that mixture is down freezing on titanium alloy-based at-9 ℃,-35 ℃ of following 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 among the embodiment 15, promptly make the tooth implant that is provided with the poroid calcium hydroxy phosphate/hydrogel material of nanometer of the present invention.
Two, method of testing
Utilize scanning electron microscope to measure the hydrogel material aperture, poroid calcium hydroxy phosphate aperture and wall thickness;
The detection data of table 1, the poroid calcium hydroxy phosphate/hydrogel material of the prepared nanometer of embodiment
Can find out by above-mentioned experimental data, the poroid calcium hydroxy phosphate/hydrogel material of nanometer provided by the invention, wherein, hydrogel is extremely millimetre-sized poroid two dimension or a three-dimensional material of micron, and the three-dimensional material hole is interconnected, and the aperture is: 1 micron to 3 millimeters; The calcium hydroxy phosphate that is deposited on the hydrogel is the poroid calcium hydroxy phosphate of three-dimensional communication, and the aperture is the 50-1000 nanometer, and hole wall is thick to be the 30-80 nanometer.
The above is preferred embodiment of the present invention only, is not to be used to limit 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 claim of the present invention.
Claims (10)
1. poroid calcium hydroxy phosphate/hydrogel material of nanometer, 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 the hydrogel, the preparation method of the poroid calcium hydroxy phosphate/hydrogel material of described nanometer comprises the steps:
(1) the preparation mass concentration is the hydrogel aqueous solution of 1-5%, and the preparation mass concentration is the glutaraldehyde water solution of 3-8%, and compound concentration is the polypeptid solution of 10-300ug/mL,
(2) prepare the hydrogel substrate: the mixed that with the prepared three kinds of aqueous solutions of step (1) is 1: 1: 1 according to volume ratio, stirred at normal temperatures 0.5-10 minute, then mixture is placed on the 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 by weighing calcium chloride and dipotassium hydrogen phosphate respectively, and dissolve in respectively in the buffer that step (3) makes, be mixed with calcium chloride solution and the dipotassium hydrogen phosphate solution of 0.002-0.006mol/L respectively, is 1: 1 with the polypeptid solution that makes in calcium chloride solution and dipotassium hydrogen phosphate solution and the step (1) according to volume ratio: the ratio of X (X=0-6) is mixed stirring mutually
(5) the prepared hydrogel substrate that is carried on the carrier of step (2) is placed the prepared mixed solution of step (4), leave standstill 24-48 hour after, successively water and washing with alcohol, drying promptly makes the poroid calcium hydroxy phosphate/hydrogel material of described nanometer.
2. the poroid calcium hydroxy phosphate/hydrogel material of nanometer as claimed in claim 1 is characterized in that, the preparation method of the poroid calcium hydroxy phosphate/hydrogel material of described nanometer comprises the steps:
(1) the preparation mass concentration is the hydrogel 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: is that 1: 1: 1 ratio is mixed mutually with the prepared three kinds of aqueous solutions of step (1) according to volume ratio, stirred at normal temperatures 0.5-10 minute, then mixture is placed on the 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 by weighing calcium chloride and dipotassium hydrogen phosphate respectively, and dissolve in respectively in the buffer that step (3) makes, be mixed with calcium chloride solution and the dipotassium hydrogen phosphate solution of 0.002-0.006mol/L respectively, is 1: 1 with the polypeptid solution that makes in calcium chloride solution and dipotassium hydrogen phosphate solution and the step (1) according to volume ratio: the ratio of X (X=0-6) is mixed mutually, stirred at normal temperatures 0.5-10 minute
(5) the prepared hydrogel substrate that is carried on the carrier of step (2) is placed the prepared mixed solution of step (4), leave standstill 24-36 hour after, successively water and washing with alcohol, drying promptly makes the poroid calcium hydroxy phosphate/hydrogel material of described nanometer.
3. the poroid calcium hydroxy phosphate/hydrogel material of nanometer as claimed in claim 1 is characterized in that, the preparation method of the poroid calcium hydroxy phosphate/hydrogel material of described nanometer comprises the steps:
(1) the preparation mass concentration is 1% hydrogel aqueous solution, and the preparation mass concentration is 5% glutaraldehyde water solution, and compound concentration is the polypeptid solution of 200ug/ml,
(2) preparation hydrogel substrate: is that 1: 1: 1 ratio is mixed mutually with the prepared three kinds of aqueous solutions of step (1) according to volume ratio, stirred at normal temperatures 0.5-10 minute, then mixture is placed on the 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, and the pH value of this buffer is 7.2-7.4,
(4) take by weighing calcium chloride and dipotassium hydrogen phosphate respectively, and dissolve in respectively in the buffer that step (3) makes, be mixed with calcium chloride solution and the dipotassium hydrogen phosphate solution of 0.002-0.006mol/L respectively, is 1: 1 with the polypeptid solution that makes in calcium chloride solution and dipotassium hydrogen phosphate solution and the step (1) according to volume ratio: the ratio of X (X=0-6) is mixed mutually, stirred at normal temperatures 0.5-10 minute
(5) the prepared hydrogel substrate that is carried on the carrier of step (2) is placed the prepared mixed solution of step (4), leave standstill 24 hours after, successively water and washing with alcohol, drying promptly makes the poroid calcium hydroxy phosphate/hydrogel material of described nanometer.
4. poroid calcium hydroxy phosphate/hydrogel material of nanometer, 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 the hydrogel, the preparation method of the poroid calcium hydroxy phosphate/hydrogel material of described nanometer comprises the steps:
(1) the preparation mass concentration is the hydrogel aqueous solution of 1-5%, and compound concentration is the polypeptid solution of 10-300ug/mL,
(2) prepare the hydrogel substrate: the mixed that with the prepared two kinds of aqueous solutions of step (1) is 1: 1 according to volume ratio, stirred at normal temperatures 1-10 minute, and then mixture was placed on the carrier, under the room temperature dry 24-48 hour, vacuum drying under 80 ℃-100 ℃ temperature then
(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 by weighing calcium chloride and dipotassium hydrogen phosphate respectively, and dissolve in respectively in the buffer that step (3) makes, be mixed with calcium chloride solution and the dipotassium hydrogen phosphate solution of 0.002-0.006mol/L respectively, is 1: 1 with the polypeptid solution that makes in calcium chloride solution and dipotassium hydrogen phosphate solution and the step (1) according to volume ratio: the ratio of X (X=0-6) is mixed stirring mutually
(5) the prepared hydrogel substrate that is carried on the carrier of step (2) is placed the prepared mixed solution of step (4), leave standstill 24-48 hour after, successively water and washing with alcohol, drying promptly makes the poroid calcium hydroxy phosphate/hydrogel material of described nanometer.
5. as claim 1,2, the poroid calcium hydroxy phosphate/hydrogel material of 3 or 4 described nanometers, it is characterized in that described hydrogel is collagen, gelatin, polyvinyl alcohol, alginic acid or at least two kinds compositions wherein; Described polypeptide be the osteopontin that contains of bone (osteopontin, OPN), the tooth phosphoprotein (phosphophoryn) or its fragment that contain; 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 a poly-aspartic-acid, polyglutamic acid, poly-Sulfated serine/tyrosine.
6. as claim 1,2, the poroid calcium hydroxy phosphate/hydrogel material of 3 or 4 described nanometers, it is characterized in that being added with mass percent in the described hydrogel material is the micromolecular compound of the promotion osteoblast breeding of 0.0001-5%.
7. tooth implant, comprise titanium alloy-based, it is characterized in that, the described titanium alloy-based poroid calcium hydroxy phosphate/hydrogel material of the described nanometer of one of claim 1 to 6 that is provided with, in its preparation method, step (1) is constant, the process of step (2) preparation hydrogel substrate comprise with mixture spray to titanium alloy-based on, placed 90 ℃ to 100 ℃ heat treatments under the room temperature after the rapid draing again 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 1um-3mm, and other content in this process is constant; Step (3) is constant to step (5).
8. tooth implant comprises titanium alloy-basedly, it is characterized in that the described titanium alloy-based poroid calcium hydroxy phosphate/hydrogel material of the described nanometer of one of claim 1 to 6 that is provided with; In its preparation method, step (1) is constant, the process of step (2) preparation hydrogel substrate comprises with chemical method handles titanium alloy-based surface, then that mixture is down freezing on titanium alloy-based at-8 ℃ to-10 ℃,-10 ℃ to-60 ℃ following freezing vacuum sublimation dryings 6-8 hour, the thickness of prepared hydrogel is 0.01mm to 5mm, and other content in this process is constant; Step (3) is constant to step (5).
9. calcium-supplementing nutritive product is characterized in that, described nutriment mainly contains the poroid calcium hydroxy phosphate/hydrogel material of the described nanometer of one of claim 1 to 6.
10. the biological filling/timbering material of orthopaedics or tooth section is characterized in that described material mainly contains the poroid calcium hydroxy phosphate/hydrogel material of the described nanometer of one of claim 1 to 6.
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