CN110292661A - Compound porous bone material of mineralized collagen/macromolecule and preparation method thereof - Google Patents
Compound porous bone material of mineralized collagen/macromolecule and preparation method thereof Download PDFInfo
- Publication number
- CN110292661A CN110292661A CN201910736390.1A CN201910736390A CN110292661A CN 110292661 A CN110292661 A CN 110292661A CN 201910736390 A CN201910736390 A CN 201910736390A CN 110292661 A CN110292661 A CN 110292661A
- Authority
- CN
- China
- Prior art keywords
- collagen
- solution
- calcium
- macromolecule
- aqueous solution
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Pending
Links
Classifications
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61L—METHODS OR APPARATUS FOR STERILISING MATERIALS OR OBJECTS IN GENERAL; DISINFECTION, STERILISATION OR DEODORISATION OF AIR; CHEMICAL ASPECTS OF BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES; MATERIALS FOR BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES
- A61L27/00—Materials for grafts or prostheses or for coating grafts or prostheses
- A61L27/02—Inorganic materials
- A61L27/025—Other specific inorganic materials not covered by A61L27/04 - A61L27/12
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61L—METHODS OR APPARATUS FOR STERILISING MATERIALS OR OBJECTS IN GENERAL; DISINFECTION, STERILISATION OR DEODORISATION OF AIR; CHEMICAL ASPECTS OF BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES; MATERIALS FOR BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES
- A61L27/00—Materials for grafts or prostheses or for coating grafts or prostheses
- A61L27/14—Macromolecular materials
- A61L27/18—Macromolecular materials obtained otherwise than by reactions only involving carbon-to-carbon unsaturated bonds
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61L—METHODS OR APPARATUS FOR STERILISING MATERIALS OR OBJECTS IN GENERAL; DISINFECTION, STERILISATION OR DEODORISATION OF AIR; CHEMICAL ASPECTS OF BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES; MATERIALS FOR BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES
- A61L27/00—Materials for grafts or prostheses or for coating grafts or prostheses
- A61L27/14—Macromolecular materials
- A61L27/22—Polypeptides or derivatives thereof, e.g. degradation products
- A61L27/24—Collagen
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61L—METHODS OR APPARATUS FOR STERILISING MATERIALS OR OBJECTS IN GENERAL; DISINFECTION, STERILISATION OR DEODORISATION OF AIR; CHEMICAL ASPECTS OF BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES; MATERIALS FOR BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES
- A61L27/00—Materials for grafts or prostheses or for coating grafts or prostheses
- A61L27/50—Materials characterised by their function or physical properties, e.g. injectable or lubricating compositions, shape-memory materials, surface modified materials
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61L—METHODS OR APPARATUS FOR STERILISING MATERIALS OR OBJECTS IN GENERAL; DISINFECTION, STERILISATION OR DEODORISATION OF AIR; CHEMICAL ASPECTS OF BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES; MATERIALS FOR BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES
- A61L27/00—Materials for grafts or prostheses or for coating grafts or prostheses
- A61L27/50—Materials characterised by their function or physical properties, e.g. injectable or lubricating compositions, shape-memory materials, surface modified materials
- A61L27/56—Porous materials, e.g. foams or sponges
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61L—METHODS OR APPARATUS FOR STERILISING MATERIALS OR OBJECTS IN GENERAL; DISINFECTION, STERILISATION OR DEODORISATION OF AIR; CHEMICAL ASPECTS OF BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES; MATERIALS FOR BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES
- A61L2430/00—Materials or treatment for tissue regeneration
- A61L2430/02—Materials or treatment for tissue regeneration for reconstruction of bones; weight-bearing implants
Landscapes
- Health & Medical Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Epidemiology (AREA)
- General Health & Medical Sciences (AREA)
- Medicinal Chemistry (AREA)
- Oral & Maxillofacial Surgery (AREA)
- Transplantation (AREA)
- Veterinary Medicine (AREA)
- Animal Behavior & Ethology (AREA)
- Dermatology (AREA)
- Public Health (AREA)
- Biophysics (AREA)
- Inorganic Chemistry (AREA)
- Chemical Kinetics & Catalysis (AREA)
- Dispersion Chemistry (AREA)
- Materials For Medical Uses (AREA)
- Medicines That Contain Protein Lipid Enzymes And Other Medicines (AREA)
- Medicinal Preparation (AREA)
Abstract
The present invention relates to compound porous bone materials of mineralized collagen/macromolecule and preparation method thereof；The preparation method includes the following steps: to prepare collagen solution；Add calcium ions aqueous solution；Add phosphorus-containing acid ion aqueous solution；Being adjusted to pH value is 8；Washing precipitate simultaneously filters；Freeze-drying；It grinds and mineralized collagen dry powder is made；Prepare macromolecule polymer solution；Prepared polymer/calcium-phosphorus salt/collagen/mixed solution；Freeze-drying；Supersonic extraction and drying；Sterilizing, so that the compound porous bone material of mineralized collagen-macromolecule be made.The method of the present invention has many advantages, such as that at low cost, stock utilization is high, easy to operate, and the compound porous bone material of mineralized collagen-macromolecule obtained by this method is high with homogenieity, mechanical strength is high, degradation time is long, organic solvent residual is few, porosity is high, good biocompatibility, degradation time are matched with bone remoulding.
Description
The application is entitled " compound porous bone material of mineralized collagen/macromolecule and preparation method thereof ", the applying date
For " on October 31st, 2016 ", application No. is the divisional applications of the application for a patent for invention of " 201610928746.8 ".
Technical field
The present invention relates to medical material fields, use polymer, nanometer phase calcium-phosphorus salt, collagen for original more particularly to a kind of
Expect the porous bone material for preparing with and preparation method thereof.
Background technique
According to statistics, limbs not free patient in China's has 50,000,000 people or more, wherein due to lacking bone remoulding operation and aggregate
Material has 5,000,000 people's amputation；Bone is caused every year because of factors such as all kinds of traffic accidents, work safety accident, orthopaedic diseases in the whole nation
The patient of defect or bone injury has 12,000,000 people, and the unsound number of bone has up to ten million；National tooth defect, anodontia patient are
Exceed 400,000,000.These people are badly in need of properly being treated.Clinically the bone defect due to caused by wound, tumour, infection is very common, estimates
Meter China has 12,000,000 orthopaedics cases to need bone grafting every year.
Field of medical materials is filled in bone defect, for a long time, autologous bone and allograph bone are clinical treatment bone defects
Common bone-grafting material.Although there is limited source and cause for bone area multiple complications, because of it in autologous bone transplanting
Bone genetic function is had concurrently again with self-bone grafting, all the time by " goldstandard " as bone defect healing；Allograph bone still cannot be complete
Immunological rejection after overcoming transplanting entirely has potential pathogeny to propagate dangerous, exists simultaneously the obstacle in terms of Medicine Ethics.
In order to overcome the problems, such as that the use of artificial bone has been a concern, and American-European countries is in recent years brought by its transplanting
The use of autologous bone and allograph bone compared to artificial bone just tends to apparent downward trend, such as gel, demineralization people's bone base
Matter, titanium alloy, high polymer, calcium sulfate, bioactivity glass, calcium phosphate bone cement and bioceramic, collagen-based material etc..However
Collagen-based material is that most prospect can promote in early days because collagen is main key components in natural bone tissue
Mineralising and the formation for supporting new bone, while material biodegradable can finally be replaced by new bone.The product being commercialized both at home and abroad
Mainly have chronOs, Medtronic's of Vitoss, Synthes of HEALOS, Orthovita of DePuy Spine
The BonGold etc. of Mastergraft, Allgens, collagen content is between 20 to 70%.
It gains enlightenment from the research carried out to natural bone, nanometer hydroxyapatite (nano-HA)/collagen base complex receives
Great concern.Initially, synthetic is prepared by directly mixing for nanometer hydroxyapatite and collagen, nano-hydroxy-apatite
Stone is from advanced ceramics technique；And collagen is to extract from animal tissue, but because collagen and hydroxyapatite are relatively weak
In conjunction with so that collagen degradation obtains comparatively fast in vivo, and hydroxyapatite still keeps originality shape and do not generate collaboration to Bone Defect Repari
Effect.
However bionical thinking is used, can prepare the hydroxyapatite similar with nature bone and be combined into collagen has
The compound of the hierarchical structure of unique law.The synergistic effect of the two is reported at first in nineteen ninety-five, repairs for the bionical bone for preparing
Multiple material provides thinking, and in order to overcome the obstruction that inside collagen or the continuous mineralising at interval is formed, collagen is first in acidity
Dispersed in solution, makes its co precipitation by adjusting pH value or additive being added, hydroxide radical phosphorite nanocrystalline body starts
It is formed in the suspension system of calcium salt, phosphate and collagen solution mixing, i.e. biomineralization.
Good biocompatibility, degradation rate are controllable in order to obtain simultaneously, easily molded have height porosity, suitable aperture
The frame material of size prepares good biological bone by high molecular material and the compound mode of nanocrystalline calcium phosphate collagen-based
Material.
Polylactide (PLGA) has nontoxic, reliable biological safety, is to be approved by the fda in the United States for clinical drop
Solve biomaterial.Polylactide has certain intensity, and the porous network structure of material can adapt to a certain range of pressure
Power variation, while porous organization can make blood circulation, ensure that the eubolism of bone tissue.After porous material implants,
Due to the increase of its specific surface area (surface area/volume), be conducive to tiny blood vessels, fibrous connective tissue grow into and nutrition and
The conveying of metabolite guides the migration of osteoblast；Product itself is converted into the master of new bone in the forming process of bone simultaneously
Composition is wanted, new bone formation is promoted, so that the ability of material guidance cell and osteogenic activity be made to increase.However, due to the drop of PLGA
The solution time is copolymerized in PLA section with PGA sections, and degradation time is very fast, be unfavorable for osteanagenesis, so that degradation time and bone remoulding
It mismatches.In addition, in the prior art, when manufacturing mineralized collagen dry powder, there are localized ion concentrations too large or too small, phosphoric acid
The problems such as wash time caused by radical ion and/or calcium ion remain is too long, sediment washing is time-consuming, calcium microcosmic salt utilization rate is low,
The mineralized collagen when the mechanical property of the preparation compound porous bone material of mineralized collagen-macromolecule is low, degradation time is short and prepares
Dry powder dispersion is poor, Determination of Residual Organic Solvents it is excessive be difficult to remove, porosity is low the problems such as.
Summary of the invention
To achieve the goals above, the above problem existing for this field is solved, the present invention provides one kind in first aspect
The method for preparing mineralized collagen dry powder, described method includes following steps:
(1) type i collagen fiber is configured to the collagen solution that concentration is 0.0001 to 0.001g/ml using acid solution；
(2) that calcium ions are added into the collagen solution with 0.08 to 0.12 mole of calcium ion/gram collagen amount is water-soluble
Liquid, so that calcium/collagen composite solution be made；
(3) phosphorus-containing acid ion aqueous solution is added into the calcium/collagen composite solution with the amount of Ca/P=1.2~1.8,
To which calcium/phosphorus/collagen composite solution be made；
(4) calcium/phosphorus/collagen composite solution is adjusted to pH value is 8, and stirring, precipitation and separation, water washing and precipitating are until supernatant
PH value 7.0~7.5, to obtain sediment；
(5) using purifying water washing precipitate and filtering, filter cake is obtained；
(6) it under normal pressure by filter cake in -30~-20 DEG C of progress pre-freezes, then is risen under vacuum conditions in -10~0 DEG C
China, to be freeze-dried to filter cake；
(7) the mineralized collagen dry powder that partial size is not more than 80 mesh will be ground by the filter cake of freeze-drying.
The present invention provides mineralized collagen dry powder made from first aspect present invention the method in second aspect.
The present invention provides mineralized collagen dry powder described in second aspect of the present invention in preparation mineralized collagen-in the third aspect
Application in the compound porous bone material of macromolecule.
The present invention provides a kind of method for preparing the compound porous bone material of mineralized collagen-macromolecule, institute in fourth aspect
The method of stating includes the following steps:
(a) macromolecule polymer solution that mass-volume concentration is 0.05 to 0.10g/ml is prepared, the solvent used is choosing
Any one of free 1,4- dioxane, chloroform or group of dimethyl sulfoxide composition；
(b) mineralized collagen dry powder, obtained polymer/calcium-phosphorus salt/collagen/mixed are added in Xiang Suoshu macromolecule polymer solution
The mass ratio of conjunction solution, the mineralized collagen dry powder and the high molecular polymer is 1:2~3:2；
(c) under normal pressure by polymer/calcium-phosphorus salt/collagen/mixed solution in -30~-20 DEG C of progress pre-freezes, then in vacuum
Under the conditions of distil in -10~0 DEG C；
(d) remaining organic solvent and drying after being lyophilized are removed by way of supersonic extraction；
(e) it sterilizes to the material for eliminating organic solvent, so that it is compound porous that the mineralized collagen-macromolecule is made
Bone material.
The present invention additionally provides a kind of method for preparing the compound porous bone material of mineralized collagen-macromolecule at the 5th aspect,
Described method includes following steps:
(1) type i collagen fiber is configured to the collagen solution that concentration is 0.0001 to 0.001g/ml using acid solution；
(2) that calcium ions are added into the collagen solution with 0.08 to 0.12 mole of calcium ion/gram collagen amount is water-soluble
Liquid, so that calcium/collagen composite solution be made；
(3) phosphorus-containing acid ion aqueous solution is added into the calcium/collagen composite solution with the amount of Ca/P=1.2~1.8,
To which calcium/phosphorus/collagen composite solution be made；
(4) calcium/phosphorus/collagen composite solution is adjusted to pH value is 8, and stirring, precipitation and separation, water washing and precipitating are until supernatant
PH value 7.0~7.5, to obtain sediment；
(5) using purifying water washing precipitate and filtering, filter cake is obtained；
(6) it under normal pressure by filter cake in -30~-20 DEG C of progress pre-freezes, then is risen under vacuum conditions in -10~0 DEG C
China, to be freeze-dried to filter cake；
(7) the mineralized collagen dry powder that partial size is not more than 80 mesh will be ground by the filter cake of freeze-drying；
(8) macromolecule polymer solution that mass-volume concentration is 0.05 to 0.10g/ml is prepared, the solvent used is choosing
Any one of free 1,4- dioxane, chloroform or group of dimethyl sulfoxide composition；
(9) mineralized collagen dry powder, obtained polymer/calcium-phosphorus salt/collagen/mixed are added in Xiang Suoshu macromolecule polymer solution
The mass ratio of conjunction solution, the mineralized collagen dry powder and the high molecular polymer is 1:2~3:2；
(10) under normal pressure by polymer/calcium-phosphorus salt/collagen/mixed solution in -30~-20 DEG C of progress pre-freezes, then true
It distils under empty condition in -10~0 DEG C；
(11) remaining organic solvent and drying after being lyophilized are removed by way of supersonic extraction；
(12) it sterilizes to the material for eliminating organic solvent, so that it is compound more that the mineralized collagen-macromolecule is made
Hole bone material.
The present invention provides mineralized collagen-high score made from the 4th or the 5th aspect the method for the invention at the 6th aspect
The compound porous bone material of son.
The present invention provides the compound porous aggregate of mineralized collagen-macromolecule described in sixth aspect present invention at the 7th aspect
Expect the application in manufacture bone implant.
Advantages of the present invention and brought unexpected technical effect include at least following several points:
(1) the collagen solution concentration used is 0.0001 to 0.001g/ml, under the collagen concentration, whole solution viscosity
It is more moderate, it not will cause viscosity influence excessive for localized ion concentration or too small.
(2) it uses calcium phosphorus molar feed ratio for Ca:P=1.2 to 1.8, improves the utilization efficiency of calcium salt and microcosmic salt, reduce
Dissociate in material the residual of calcium salt and microcosmic salt.
(3) pH value is used to significantly reduce for 8 precipitating terminal so that mineralized collagen precipitates quicker and abundant
Stirring and time of repose.
(4) 80 mesh mineralized collagen dry powder below is used, so that it with collagen adhesives when mixing, it is easier to disperse.
(5) it uses 40kgy irradiation dose below and carries out sterilizing processing, significantly reduce the mechanical property and drop of material
Solve the time.
(6) use the mass volume ratio of high molecular polymer and organic solvent for 0.05 to 0.10g/ml, according to as group
The porous support of weaver's journey, different histocytes have different requirements to aperture, and bone and cartilage tissue engineered, need
100 to 250 μm of aperture is preferred 200 to 400 μm of degradable stephanoporate stent material, therefore organic solvent pore-creating under the concentration
Aperture is close to theoretical value, and porosity is 70 to 95%；
(7) it uses room temperature and the swelling under good solvent is carried out to high molecular polymer, because using the good molten of polymer
Agent, therefore swelling can be completed at room temperature, reduce temperature compared with risks such as the volatilization of relative superiority or inferiority solvent and toxicity；
(8) it by way of segmentation freeze-drying, allows the material to solidify at a lower temperature, distil at slightly higher temperature,
Shorten freeze-drying time.
(9) replace PLGA as polymeric stent material using polylactic acid (PLA) such as l-lactic acid and polycaprolactone (PCL)
Material, so that degradation time is matched with bone remoulding.
(10) due to removing the residual of organic solvent after freeze-drying by the way of supersonic extraction, the time is short, high-efficient, residual
It measures small.
Specific embodiment
As described above, the present invention first aspect provide a kind of method for preparing mineralized collagen dry powder the method includes
Following steps:
(1) using acid solution by type i collagen fiber be configured to concentration be 0.0001 to 0.001g/ml (such as 0.0001,
0.0005 or 0.001g/ml) collagen solution；
(2) molten to the collagen with the amount of 0.08 to 0.12 (such as 0.08,0.10 or 0.12) mole calcium ion/gram collagen
Calcium ions aqueous solution is added in liquid, so that calcium/collagen composite solution be made；
(3) with the amount of Ca/P=1.2~1.8 (such as 1.2,1.4,1.6 or 1.8) into the calcium/collagen composite solution
Phosphorus-containing acid ion aqueous solution is added, so that calcium/phosphorus/collagen composite solution be made；
(4) calcium/phosphorus/collagen composite solution is adjusted to pH value is 8, and stirring, precipitation and separation, water washing and precipitating are until supernatant
PH value in 7.0~7.5 (such as 7.0,7.1,7.2,7.3,7.4 or 7.5), to obtain sediment；
(5) using purifying water washing precipitate and filtering, filter cake is obtained；
(6) under normal pressure by filter cake in -30~-20 DEG C (such as -30, -25 or -20 DEG C) progress pre-freezes, then in vacuum item
It distils under part in -10~0 DEG C (such as -10, -5 or 0 DEG C), to be freeze-dried to filter cake；
(7) partial size will be ground by the filter cake of freeze-drying no more than 80 mesh (such as no more than 80,90,100 or 120
Mesh) mineralized collagen dry powder.
In some preferred embodiments, the acid solution in step (1) is selected from by aqueous hydrochloric acid solution, aqueous solution of nitric acid
With the group of acetic acid aqueous solution composition, preferably acetic acid aqueous solution.
In some preferred embodiments, the concentration of the acid solution be 0.45 to 0.55mM (such as 0.45,0.50 or
0.55mM)。
In some preferred embodiments, the pH in step (4) is adjusted using sodium hydrate aqueous solution, preferably
It is that the concentration of the sodium hydrate aqueous solution is 0.9M to 1.1M (such as 0.9,1.0 or 1.1M).
In some preferred embodiments, the calcium ions aqueous solution and/or phosphorus-containing acid ion aqueous solution plus
Enter and is carried out by the way of being stirred when being slowly added dropwise.
In some preferred embodiments, pH is adjusted to 8 and then stirring 8 to 24 hours, shape in step (4)
At white suspension, stand 12 to 24 hours (such as 12,18 or 24 hours), remove supernatant, then plus purified water to original volume after
It stirs 5 to 10 times (such as 5,6,7,8,9 or 10 times), removes supernatant after standing 1 to 3 hour, continuous wash 3 to 7 times, Zhi Daoshang
The pH value of clear liquid is between 7.0~7.5 (7.0,7.1,7.2,7.3,7.4 or 7.5).
In some preferred embodiments, the concentration of the calcium ions aqueous solution be 0.45M to 0.55M (such as
0.45,0.50 or 0.55M).
In some preferred embodiments, the concentration of the phosphorus-containing acid ion aqueous solution be 0.4M to 0.6M (such as
0.40,0.50 or 0.60M).
In some preferred embodiments, the type i collagen fiber is the I type bovine collagen fibers of ungelled state.
In some preferred embodiments, it is used to prepare the calcium salt of the calcium ions aqueous solution and is used to prepare phosphorous
The microcosmic salt of acid ion aqueous solution is the other calcium salt of medicinal or pharmaceutic adjuvant grade and microcosmic salt.
The present invention provides mineralized collagen dry powder obtained made from first aspect present invention the method in second aspect.
The present invention provides made from first aspect present invention the method or second aspect of the present invention in the third aspect
Application of the mineralized collagen dry powder in the preparation compound porous bone material of mineralized collagen-macromolecule.
The present invention provides a kind of method for preparing the compound porous bone material of mineralized collagen-macromolecule, institute in fourth aspect
The method of stating includes the following steps:
(a) macromolecule polymer solution that mass-volume concentration is 0.05 to 0.10g/ml is prepared, the solvent used is choosing
Any one of free 1,4- dioxane, chloroform or group of dimethyl sulfoxide composition；
(b) mineralized collagen dry powder, obtained polymer/calcium-phosphorus salt/collagen/mixed are added in Xiang Suoshu macromolecule polymer solution
Close solution, the mass ratio of the mineralized collagen dry powder and the high molecular polymer be 1:2~3:2 (such as 1:2,2:2 or 3:
2)；
(c) under normal pressure by polymer/calcium-phosphorus salt/collagen/mixed solution in -30~-20 DEG C (such as -30, -25 or -20
DEG C) pre-freeze is carried out, then distil under vacuum conditions in -10~0 DEG C (such as -10, -5 or 0 DEG C)；
(d) remaining organic solvent and drying after being lyophilized are removed by way of supersonic extraction；
(e) it sterilizes to the material for eliminating organic solvent, so that it is compound porous that the mineralized collagen-macromolecule is made
Bone material.
In some preferred embodiments, in step (d) and (e) between further include whether Determination of Residual Organic Solvents is in
Detecting step within the scope of qualified residue.
In some preferred embodiments, the qualified residue range is less than or equal to 50ppm.
In some preferred embodiments, the detecting step carries out in the following way: weighing the secondary jelly of 1.0g
Dry collagen-based composite, detection Determination of Residual Organic Solvents whether be less than or equal to 50ppm (such as no more than 50,40,30,20 or
10ppm), if it is greater than 50ppm, then repeatedly step (d), until Determination of Residual Organic Solvents is less than or equal to 50ppm.
In some preferred embodiments, the high molecular polymer is PLA and/or PCL, in some preferred realities
It applies in mode, the viscosity average molecular weigh (hereinafter sometimes simply referred to as molecular weight) of the PLA is 100,000~300,000 (for example,
100,000,200,000 or 300,000).In another preferred embodiment, the inherent viscosity 1.0 of the PCL to
2.5dl/g (for example, 1.0,1.5,2.0 or 2.5).
In some preferred embodiments, the supersonic extraction carries out in the following way: sample is placed into beaker
In, after being totally submerged at least 2 hours with dehydrated alcohol；Ultrasonic cleaning is outwelled behind 2 to 10min (such as 2,5,7 or 10 minutes)
Cleaning solution, after repeating ultrasound 2 to 5 times；Be packed into centrifuge loading filter bag, be centrifuged primary (3 to 20 seconds) (such as 3,5,7,
10,15 or 20 seconds), centrifuge speed is 1000 to 4000r/min (such as 1000,2000,3000 or 4000r/min), then
It is dried in vacuo.
In some preferred embodiments, it is described sterilizing by using 15kgy to 40kgy (such as 15,20,25,30,
35 or 40kgy) -60 irradiation dose of cobalt carry out.
In some preferred embodiments, the mineralized collagen dry powder is method system described in first aspect present invention
?.
The present invention additionally provides a kind of method for preparing the compound porous bone material of mineralized collagen-macromolecule at the 5th aspect,
Described method includes following steps:
(1) type i collagen fiber is configured to the collagen solution that concentration is 0.0001 to 0.001g/ml using acid solution；
(2) that calcium ions are added into the collagen solution with 0.08 to 0.12 mole of calcium ion/gram collagen amount is water-soluble
Liquid, so that calcium/collagen composite solution be made；
(3) phosphorus-containing acid ion aqueous solution is added into the calcium/collagen composite solution with the amount of Ca/P=1.2~1.8,
To which calcium/phosphorus/collagen composite solution be made；
(4) calcium/phosphorus/collagen composite solution is adjusted to pH value is 8, and stirring, precipitation and separation, water washing and precipitating are until supernatant
PH value 7.0~7.5, to obtain sediment；
(5) using purifying water washing precipitate and filtering, filter cake is obtained；
(6) it under normal pressure by filter cake in -30~-20 DEG C of progress pre-freezes, then is risen under vacuum conditions in -10~0 DEG C
China, to be freeze-dried to filter cake；
(7) the mineralized collagen dry powder that partial size is not more than 80 mesh will be ground by the filter cake of freeze-drying；
(8) macromolecule polymer solution that mass-volume concentration is 0.05 to 0.10g/ml is prepared, the solvent used is choosing
Any one of free 1,4- dioxane, chloroform or group of dimethyl sulfoxide composition；
(9) mineralized collagen dry powder, obtained polymer/calcium-phosphorus salt/collagen/mixed are added in Xiang Suoshu macromolecule polymer solution
Close solution, the mass ratio of the mineralized collagen dry powder and the high molecular polymer be 1:2~3:2 (such as 1:2,2:2 or 3:
2)；
(10) under normal pressure by polymer/calcium-phosphorus salt/collagen/mixed solution in -30~-20 DEG C (such as -30, -25 or -
20 DEG C) pre-freeze is carried out, then distil under vacuum conditions in -10~0 DEG C (such as -10, -5 or 0 DEG C)；
(11) remaining organic solvent and drying after being lyophilized are removed by way of supersonic extraction；
(12) it sterilizes to the material for eliminating organic solvent, so that it is compound more that the mineralized collagen-macromolecule is made
Hole bone material.
In some preferred embodiments, the acid solution in step (1) is selected from by aqueous hydrochloric acid solution, aqueous solution of nitric acid
With the group of acetic acid aqueous solution composition, preferably acetic acid aqueous solution.
In some preferred embodiments, the concentration of the acid solution is 0.45 to 0.55mM.
In some preferred embodiments, the pH in step (4) is adjusted using sodium hydrate aqueous solution, preferably
It is that the concentration of the sodium hydrate aqueous solution is 0.9M to 1.1M.
In some preferred embodiments, the calcium ions aqueous solution and/or phosphorus-containing acid ion aqueous solution plus
Enter and is carried out by the way of being stirred when being slowly added dropwise.
In some preferred embodiments, pH is adjusted to 8 and then stirring 8 to 24 hours, shape in step (4)
At white suspension, stand 12 to 24 hours, remove supernatant, then plus purified water to original volume after stir 5 to 10 times, standing 1 to 3
Supernatant is removed after hour, continuous wash 3 to 7 times, until the pH value of supernatant is between 7.0~7.5.
In some preferred embodiments, the concentration of the calcium ions aqueous solution is 0.45M to 0.55M.
In some preferred embodiments, the concentration of the phosphorus-containing acid ion aqueous solution is 0.4M to 0.6M.
In some preferred embodiments, the type i collagen fiber is the I type bovine collagen fibers of ungelled state.
In some preferred embodiments, the calcium salt and phosphorus-containing acid ion of the calcium ions aqueous solution are used to prepare
The calcium salt of aqueous solution and the microcosmic salt for being used to prepare phosphorus-containing acid ion aqueous solution are medicinal or pharmaceutic adjuvant auxiliary material rank calcium salt
And microcosmic salt.
In some preferred embodiments, further include whether Determination of Residual Organic Solvents is located between step (11) and (12)
Detecting step within the scope of qualified residue.
In some preferred embodiments, the qualified residue range is less than or equal to 50ppm.
In some preferred embodiments, the detecting step carries out in the following way: weighing the secondary jelly of 1.0g
Dry collagen-based composite, detection Determination of Residual Organic Solvents whether be less than or equal to 50ppm (such as no more than 50,40,30,20 or
10ppm), if it is greater than 50ppm, then repeatedly step (d), until Determination of Residual Organic Solvents is less than or equal to 50ppm.
In some preferred embodiments, the high molecular polymer is PLA and/or PCL.In some preferred realities
It applies in mode, the molecular weight of the PLA is 100,000~300,000 (for example, 100,000,200,000 or 300,000).?
In another preferred embodiment, the inherent viscosity 1.0 of the PCL to 2.5dl/g (for example, 1.0,1.5,2.0 or
2.5)。
In some preferred embodiments, the supersonic extraction carries out in the following way: sample is placed into beaker
In, after being totally submerged at least 2 hours with dehydrated alcohol；Ultrasonic cleaning is outwelled behind 2 to 10min (such as 2,5,7 or 10 minutes)
Cleaning solution, after repeating ultrasound 2 to 5 times；Be packed into centrifuge loading filter bag, be centrifuged primary (3 to 20 seconds) (such as 3,5,7,
10,15 or 20 seconds), centrifuge speed is 1000 to 4000r/min (such as 1000,2000,3000 or 4000r/min), then
It is dried in vacuo.
In some preferred embodiments, it is described sterilizing by using 15kgy to 40kgy -60 irradiation dose of cobalt into
Row.
In some more specific embodiments, of the invention is used to prepare the compound porous aggregate of mineralized collagen-macromolecule
The method of material includes the following steps:
(1) it is configured to the acid solution of type i collagen fiber, acid therein is in the group being made of hydrochloric acid, nitric acid or acetic acid
Any one, the concentration of the collagen in acid solution is 0.0001 to 0.001g/ml；
(2) solution containing calcium ion is slowly added dropwise in acid solution, dripping quantity is that calcium ion 0.08 is added dropwise in every gram of collagen
To 0.12mol, dropwise addition while, is stirred；
(3) aqueous solution containing phosphate anion, the phosphate radical of addition is slowly added dropwise while stirring in above-mentioned solution
The molar ratio of the amount of the calcium ion of the amount and addition of ion is Ca:P=1.2 to 1.8；
(4) be added dropwise while stirring in the solution that phosphate anion has been added dropwise NaOH solution to pH value be 8, be stirred for 8 to
24 hours, form white suspension, stand 12 to 24 hours, remove supernatant, then plus purified water to original volume stir, stand 1 to 3
Supernatant is removed after hour, continuous wash 5 to 10 times, until the pH value of supernatant is between 7.0~7.5；
(5) pumping and filtering device is opened, sediment is poured into batches, is filtered, is washed repeatedly filter cake 2 to 4 times with purified water；
(6) it is put into freeze dryer in -30~-20 DEG C of progresss pre-freezes (not vacuumizing), -10~0 DEG C is distilled and (takes out very
It is empty) it is dry；
(7) it then grinds and mineralized collagen dry powder is made, the screening of 80 mesh stainless steel mesh is spare；
(8) PCL for the PLA or inherent viscosity 1.0 to 2.5dl/g for being 100,000~300,000 by molecular weight is placed in reaction
In kettle, solvent is added, prepares the solution that mass-volume concentration is 0.05 to 0.10g/ml at room temperature, wherein solvent is Isosorbide-5-Nitrae-
Any one of dioxane, chloroform or dimethyl sulfoxide；
(9) mineralized collagen dry powder is added into polymer solution, is uniformly mixed, wherein mineralized collagen dry powder and polymer
Mass ratio be 1:2~3:2, be made polymer/calcium-phosphorus salt/collagen/mixed solution；
(10) solution of acquisition is lyophilized, parameter used condition when being prepared with mineralized collagen dry powder；
(11) by way of supersonic extraction, remaining organic solvent after freeze-drying is removed, wherein sample is placed into beaker
In, after being totally submerged at least 2 hours with dehydrated alcohol；Ultrasonic cleaning outwells cleaning solution after 2 to 10min, repeats ultrasound 2
To after 5 times；It is packed into centrifuge loading filter bag, is centrifuged primary (3 to 20 seconds), centrifuge speed is 1000 to 4000r/min, after
It is dried in vacuo；
(12) sterilizing processing is carried out using 40kgy -60 irradiation dose of cobalt below, can be obtained mineralized collagen/macromolecule
Compound porous bone material.
The present invention provides mineralized collagen-made from fourth aspect present invention or the 5th aspect the method at the 6th aspect
The compound porous bone material of macromolecule.
Seventh aspect present invention provides the compound porous bone material of mineralized collagen-macromolecule described in sixth aspect present invention
Application in manufacture bone implant.
In the present invention, the present inventor at least improve at following aspect and has achieved corresponding technical effect:
(1) calcium ion 0.08 is added dropwise to 0.12mol using every gram of collagen；Very few calcium can sometimes be added dropwise in the prior art
Ion, sometimes even as low as 0.01mol；Excessive calcium ion sometimes is added dropwise again, sometimes up to 0.16mol/g；Excessive calcium
Ion, which will lead to, remains extra free calcium ions in the waste or material of calcium salt, so that subsequent cleaning becomes complexity even
Difficulty may cause mineralized collagen deficiency if calcium ion is very few, and intensity is lower, and material and bone tissue difference are big, influence material
The bone guided and inductivity of material.
(2) using collagen solution concentration, under the collagen concentration, whole solution viscosity is more for 0.0001 to 0.001g/ml
Add it is moderate, caused by avoiding because of viscosity localized ion concentration it is excessive or too small brought by influence.
(3) use calcium phosphorus molar feed ratio for Ca:P=1.2 to 1.8, so that the inorganic salts composition of material is more in line with hydroxyl
The theoretical calcium-phosphorus ratio 1.667 of base apatite improves the utilization rate of calcium salt and microcosmic salt, reduces calcium ion and/or phosphate anion
Residual in the material reduces the operation difficulties such as subsequent wash, improves time efficiency, avoids free calcium ions or phosphate anion
The problem of the deficiency of the strength of materials caused by residual.
(4) pH value is used to substantially reduce for 8 precipitating terminal so that mineralized collagen precipitates more quickly more abundant
Stirring and time of repose；In the prior art even using up to 9 or down to 7 pH so that only time of repose is even up to 5 days
Time.In addition, since NaOH solution is often added dropwise to mixed system pH=6~8 in the past, and observe as pH=5~6, it mixes
Zoarium system starts to precipitate, and as pH=7, white suspension occurs in mixed system, therefore NaOH to pH is often added dropwise on 7 left sides
It is right.The present inventor has found through overtesting, and NaOH solution is added dropwise to mixed system pH=8, is washed again after precipitating to 7.0 to 7.5, instead
And the time required to sedimentation effect can be improved and reduce.
(5) it is mixed using 80 mesh mineralized collagen dry powder below with high molecular polymer, the dry powder under the mesh number is easier
Dispersed in polymer solution system.
(6) use the mass volume ratio of high molecular polymer and organic solvent for 0.05 to 0.10g/ml, according to as group
The porous support of weaver's journey, different histocytes have different requirements to aperture, and bone and cartilage tissue engineered, need
100 to 250 μm of aperture is preferred 200 to 400 μm of degradable stephanoporate stent material, therefore organic solvent pore-creating under the concentration
Aperture is close to theoretical value, and porosity is 70 to 95%；
(7) it uses room temperature and the swelling under good solvent is carried out to high molecular polymer, because using the good molten of polymer
Agent, therefore swelling can be completed at room temperature, reduce temperature compared with risks such as the volatilization of relative superiority or inferiority solvent and toxicity；
(8) it uses 40kgy irradiation dose below and carries out sterilizing processing, because material is absorbable polymer and collagen
Deng the material for radiation sensitive, in order to significantly reduce the mechanical property and degradation time of material, using low irradiation dose
Carrying out sterilizing is very important.
In addition, the present inventor also in quality system establishment process, has carried out following improvement to part raw material and has obtained
Corresponding technical effect:
(1) I-type collagen is used, the fiber of ungelled state is avoided as collage raw material because containing admittedly in gel
That measures is inconsistent, causes inventory inconsistent；Type I collagen is a kind of structural proteins found in animal body simultaneously, is self
The main organic component of bone.I-type collagen is the major structural protein of spinal animals, is osteoblast in osteogenetic process
The extracellular matrix of secretion is the bracket of calcium deposition and the template of the promotor of bone matrix mineralising, mineralising；Cell can be promoted to move
It moves, absorption, differentiation, and is adjustable cell growth, approved by U.S. FDA as biomaterial, and there are a series of collagens to be implanted into
Product, including bone implant product.
(2) the other calcium salt of medicinal or pharmaceutic adjuvant grade, microcosmic salt are used, is avoided because the rank of raw material causes impurity, again
Metal, ash content it is exceeded, also influence material biocompatibility.
(3) replace PLGA as macromolecular scaffold material using l-lactic acid (PLA) and polycaprolactone (PCL), because
The degradation time of PLGA is copolymerized in PLA section with PGA sections, and degradation time is very fast, is unfavorable for osteanagenesis, the present invention individually makes
With PLA and PCL degradation time is matched with bone remoulding.
In addition, the present inventor also uses suction filtration mode to carry out the separation of supernatant and sediment, and clean to neutrality.This hair
Bright people's discovery, compared with centrifugation in the prior art, can so make porous material obtain contacting with purified water again
Chance, after repeatedly washing, the pH value of leaching liquor can level off to neutrality for inside and outside.
In addition, the present inventor is removed solvent and pore-creating, i.e., (is not vacuumized) under normal pressure by way of segmentation freeze-drying
Pre-freeze is first carried out at -30~-20 DEG C in freeze dryer, is then just distilled at -10~0 DEG C in vacuum condition, enables material
It is enough to solidify at a lower temperature, it distils at slightly higher temperature, shortens freeze-drying time significantly.
In addition, organic solvent residual is always technical problem in obtained repair materials.The present inventor is taken out by ultrasound
The mode mentioned removes the residual (Isosorbide-5-Nitrae-dioxane, dimethyl sulfoxide, chloroform) of organic solvent after freeze-drying, relatively uses Vacuum solutions
The mass transfer balance that is also easy to produce when analysing porous material and the disadvantages of the time is long, efficiency lower (residual quantity is sometime up to 150ppm),
Ultrasound make extracting solution well into and inside contact material, substantially reduce the time, after supersonic extraction, improve efficiency,
And residual quantity is less than 50ppm.
It is important to note that the numberical range of this specification indicate the upper limit value of the numberical range, lower limit value and
Any numerical value or the subrange being within the numberical range.Therefore, if not otherwise specified, it is related in the present specification
Just no longer itemizing when numberical range includes specific value in the numberical range.
Embodiment
The present invention will be hereafter illustrated by way of embodiment, but protection scope of the present invention should not be by
It is considered limited to these embodiments.
Embodiment 1
(1) using the acid solution of the type i collagen fiber of the acetic acid solutions 0.0005g/ml of 0.5M；
(2) the calcium ions aqueous solution of 0.5M is slowly added dropwise in Xiang Suoshu acid solution while stirring, additional amount is every gram of glue
Calcium ion 0.10mol is added dropwise in original, to form calcium/collagen composite solution；
(3) the phosphorus-containing acid ion aqueous solution of 0.5M, the phosphoric acid of addition is added dropwise while stirring in calcium/collagen composite solution
The molar ratio of the amount of the calcium ion of the amount and addition of radical ion is Ca/P=1.5；
(4) NaOH aqueous solution to the pH value that 1M is added dropwise while stirring in the solution that phosphate anion has been added dropwise is 8, then is stirred
Mix 16 hours, form white suspension, stand 18 hours, remove supernatant, then plus purified water to original volume after stir, it is small to stand 2
When after remove supernatant, continuous wash 5 times, through detecting, the pH value of supernatant at this time is 7.2 (being between 7.0~7.5), is obtained
To sediment；
(5) pumping and filtering device is opened, sediment is poured into, is filtered, is washed repeatedly filter cake 3 times with purified water, obtains filter cake；
(6) filter cake is put into freeze drier and carries out pre-freeze under normal pressure at -25 DEG C, under vacuum conditions -5 DEG C of progress
Distillation is to realize freeze-drying；
(7) at the quartering of filter cake periphery and filter cake central location carry out five point samplings, its homogenieity of observation analysis；So
The filter cake by freeze-drying is ground afterwards, mineralized collagen dry powder is made, the screening of 80 mesh stainless steel mesh is spare.
Embodiment 2 to 4
Other than the content of the following table 1, carried out in mode substantially the same manner as Example 1.
Embodiment 5
It other than the content of the following table 1, is carried out in mode substantially the same manner as Example 1, wherein sedimentation time 36 hours
After measure pH.
Embodiment 6
It other than the content of the following table 1, is carried out in mode substantially the same manner as Example 1, wherein when detection pH reaches 7.2
It is required to staticly settle the time, if pH does not reach 7.2, repeats addition water and staticly settle again, record stands required
Time.
Embodiment 7
(7') mineralized collagen dry powder is prepared in the same manner as example 1；
(8) it is that 100,000PLA is placed in reaction kettle by molecular weight, solvent is added, prepares mass-volume concentration at room temperature
For the solution of 0.05g/ml, wherein solvent is Isosorbide-5-Nitrae-dioxane；
(9) the mineralized collagen dry powder is added into PLA solution, is uniformly mixed, the matter of mineralized collagen dry powder and polymer
Polymer/calcium-phosphorus salt/collagen/mixed solution is made than being 1:2 in amount；
(10) under normal pressure by PLA/ calcium-phosphorus salt/collagen/mixed solution in -20 DEG C of progress pre-freezes, then under vacuum conditions
It distils in -5 DEG C；
(11) by way of supersonic extraction, remaining organic solvent after freeze-drying is removed, wherein sample is placed into beaker
In, after being totally submerged 2 hours with dehydrated alcohol；Ultrasonic cleaning outwells cleaning solution after 5min, after repeating ultrasound 3 times；It is packed into
Centrifuge loading filter bag, be centrifuged one time 10 seconds, centrifuge speed 25000r/min, after be dried in vacuo；It takes out about
0.1g does organic solvent residual test, as a result, it has been found that Determination of Residual Organic Solvents is less than 50ppm；
(12) sterilizing processing is carried out using 20kgy -60 irradiation dose of cobalt below, can be obtained mineralized collagen/macromolecule
Compound porous bone material.
Then the average pore size and porosity of the obtained compound porous bone material of mineralized collagen-macromolecule are measured.
Embodiment 8-12
Other than the content shown in the following table 2, carried out in mode same as Example 7.
Embodiment 13
Other than the content shown in the following table 2, carried out in mode substantially the same manner as Example 7, but in step (12),
Without supersonic extraction, but solvent is removed by vacuumizing.
Embodiment 14
It is carried out in mode substantially the same manner as Example 7, difference is to go out using -60 irradiation dose of cobalt of 60kgy
Bacterium processing, then measures the mechanical property of embodiment 7 and the resulting compound porous bone material of mineralized collagen-macromolecule of embodiment 14
And degradation time.As a result, it has been found that the former compression strength is 2 times of the latter, the latter is often below 0.5MPa, degradation time the former be
1.5 times of the latter, the latter is often below 3 months.
Claims (11)
1. a kind of method for preparing the compound porous bone material of mineralized collagen-macromolecule, which is characterized in that the method includes as follows
Step:
(1) type i collagen fiber is configured to the collagen solution that concentration is 0.0001 to 0.001g/ml using acid solution；
(2) calcium ions aqueous solution is added into the collagen solution with 0.08 to 0.12 mole of calcium ion/gram collagen amount, from
And calcium/collagen composite solution is made；
(3) phosphorus-containing acid ion aqueous solution is added into the calcium/collagen composite solution with the amount of Ca/P=1.2~1.8, thus
Calcium/phosphorus/collagen composite solution is made；
(4) by calcium/phosphorus/collagen composite solution be adjusted to pH value be 8, stirring, precipitation and separation, water washing and precipitating until supernatant pH
Value is 7.0~7.5, to obtain sediment, wherein pH is adjusted to 8 and then stirring 8 to 24 hours, it is suspended to form white
Liquid, stand 12 to 24 hours, remove supernatant, then plus purified water to original volume after stir 5 to 10 times, standing 1 to 3 hour after remove
Supernatant, continuous wash 3 to 7 times, until the pH value of supernatant is between 7.0~7.5；
(5) using purifying water washing precipitate and filtering, filter cake is obtained；
(6) it under normal pressure by filter cake in -30~-20 DEG C of progress pre-freezes, then distils under vacuum conditions in -10~0 DEG C, from
And filter cake is freeze-dried；
(7) the mineralized collagen dry powder that partial size is not more than 80 mesh will be ground by the filter cake of freeze-drying；
(8) prepare mass-volume concentration be 0.05 to 0.10g/ml macromolecule polymer solution, the solvent used be selected from by
Any one of 1,4- dioxane, chloroform or group of dimethyl sulfoxide composition；
(9) mineralized collagen dry powder is added in Xiang Suoshu macromolecule polymer solution, it is molten that polymer/calcium-phosphorus salt/collagen/mixing is made
The mass ratio of liquid, the mineralized collagen dry powder and the high molecular polymer is 1:2 to 3:2；
(10) under normal pressure by polymer/calcium-phosphorus salt/collagen/mixed solution in -30 to -20 DEG C of progress pre-freezes, then in vacuum item
It distils under part in -10 to 0 DEG C；
(11) remaining organic solvent and drying after being lyophilized are removed by way of supersonic extraction；
(12) it sterilizes to the material for eliminating organic solvent, so that the compound porous bone of mineralized collagen-macromolecule be made
Material.
2. the method according to claim 1, wherein the acid solution in step (1) be selected from by aqueous hydrochloric acid solution,
The group of aqueous solution of nitric acid and acetic acid aqueous solution composition, preferably acetic acid aqueous solution.
3. according to the method described in claim 1, it is characterized by:
The concentration of the acid solution is 0.45 to 0.55mM；
PH in step (4) is adjusted using the sodium hydrate aqueous solution that concentration is 0.9M to 1.1M；
The concentration of the calcium ions aqueous solution is 0.45M to 0.55M；And/or
The concentration of the phosphorus-containing acid ion aqueous solution is 0.4M to 0.6M.
4. according to the method in any one of claims 1 to 3, which is characterized in that the type i collagen fiber is ungelled
The I type bovine collagen fibers of state.
5. according to the method in any one of claims 1 to 3, which is characterized in that also wrapped between step (11) and (12)
Include the detecting step whether Determination of Residual Organic Solvents is within the scope of qualified residue.
6. according to the method described in claim 5, it is characterized in that, it is described qualification residue range be less than or equal to
50ppm。
7. according to the method in any one of claims 1 to 3, which is characterized in that described to sterilize by using 15kgy extremely
- 60 irradiation dose of cobalt of 40kgy carries out.
8. according to the method in any one of claims 1 to 3, which is characterized in that the high molecular polymer is polylactic acid
And/or pla-pcl；
Preferably, the weight average molecular weight of the polylactic acid is 100,000~300,000；And/or the pla-pcl characteristic is viscous
Degree 1.0 to 2.5dl/g.
9. according to the method in any one of claims 1 to 3, which is characterized in that step (11) carries out in the following way:
Sample is placed into beaker, after being totally submerged at least 2 hours with dehydrated alcohol；Ultrasonic cleaning, 2 is clear to outwelling after ten minutes
Washing lotion, after repeating ultrasound 2 to 5 times；Be packed into centrifuge loading filter bag, be centrifuged 3 to 20 seconds, centrifuge speed be 1000 to
It 4000 revs/min, is then dried in vacuo.
10. the compound porous bone material of mineralized collagen-macromolecule made from the method as described in any one of claims 1 to 9.
11. the compound porous bone material of mineralized collagen-macromolecule made from the method as described in any one of claims 1 to 9 exists
Manufacture the application in bone implant.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201910736390.1A CN110292661A (en) | 2016-10-31 | 2016-10-31 | Compound porous bone material of mineralized collagen/macromolecule and preparation method thereof |
Applications Claiming Priority (2)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201910736390.1A CN110292661A (en) | 2016-10-31 | 2016-10-31 | Compound porous bone material of mineralized collagen/macromolecule and preparation method thereof |
| CN201610928746.8A CN106421927A (en) | 2016-10-31 | 2016-10-31 | Mineralized collagen/polymer composite porous bone material and preparation method thereof |
Related Parent Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201610928746.8A Division CN106421927A (en) | 2016-10-31 | 2016-10-31 | Mineralized collagen/polymer composite porous bone material and preparation method thereof |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN110292661A true CN110292661A (en) | 2019-10-01 |
Family
ID=58178084
Family Applications (2)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201910736390.1A Pending CN110292661A (en) | 2016-10-31 | 2016-10-31 | Compound porous bone material of mineralized collagen/macromolecule and preparation method thereof |
| CN201610928746.8A Pending CN106421927A (en) | 2016-10-31 | 2016-10-31 | Mineralized collagen/polymer composite porous bone material and preparation method thereof |
Family Applications After (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201610928746.8A Pending CN106421927A (en) | 2016-10-31 | 2016-10-31 | Mineralized collagen/polymer composite porous bone material and preparation method thereof |
Country Status (1)
| Country | Link |
|---|---|
| CN (2) | CN110292661A (en) |
Cited By (10)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN112158817A (en) * | 2020-09-14 | 2021-01-01 | 湖北赛罗生物材料有限公司 | Bone tissue repair material and preparation method and application thereof |
| CN113913961A (en) * | 2021-11-16 | 2022-01-11 | 清华大学 | Mineralized collagen nanofiber doped with active elements and preparation method thereof |
| CN114366857A (en) * | 2022-02-08 | 2022-04-19 | 博纳格科技(天津)有限公司 | Preparation method of degradable artificial bone composite material |
| CN114732958A (en) * | 2022-05-09 | 2022-07-12 | 洛阳理工学院 | Bionic collagen mineralized bone powder and preparation method thereof |
| CN114886623A (en) * | 2022-06-02 | 2022-08-12 | 奥精医疗科技股份有限公司 | Preparation method of interbody fusion cage |
| CN115006589A (en) * | 2022-06-28 | 2022-09-06 | 奥精医疗科技股份有限公司 | Carbon nanotube modified mineralized collagen material and preparation method and application thereof |
| CN115120774A (en) * | 2022-06-28 | 2022-09-30 | 奥精医疗科技股份有限公司 | Carbon nano tube strengthened mineralized collagen material and preparation method and application thereof |
| CN115137883A (en) * | 2022-08-03 | 2022-10-04 | 尧舜泽生物医药(南京)有限公司 | Bionic composite mineralized scaffold and preparation method thereof |
| CN115414530A (en) * | 2022-10-24 | 2022-12-02 | 深圳市致美生物科技有限公司 | Injectable filler for accelerating collagen regeneration induction and preparation and application thereof |
| CN115887776A (en) * | 2021-09-24 | 2023-04-04 | 中国科学院理化技术研究所 | Composite porous scaffold formed on basis of gelatin-induced magnesium phosphate deposition and preparation method and application thereof |
Families Citing this family (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN108159500A (en) * | 2017-12-27 | 2018-06-15 | 天津宝坻紫荆科技有限公司 | A kind of artificial bone renovating material of 3D printing and preparation method thereof |
| CN108421088B (en) * | 2018-04-19 | 2021-06-11 | 奥精医疗科技股份有限公司 | Mineralized collagen-based medium-strength artificial bone repair material and preparation method thereof |
| CN114601975B (en) * | 2022-04-02 | 2022-12-06 | 奥精医疗科技股份有限公司 | Polyether-ether-ketone composite mineralized collagen material as well as preparation method and application thereof |
Citations (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US20010037014A1 (en) * | 1999-08-28 | 2001-11-01 | Sung-Tsuen Liu | Thin mineralized collagen membrane and method of making same |
| CN1325734A (en) * | 2001-06-29 | 2001-12-12 | 清华大学 | Process for preparing nm-phase calcium-phosphorus salt/collagen/high-molecular bone compounded porous material |
| CN102205150A (en) * | 2011-05-19 | 2011-10-05 | 清华大学 | Preparation method for anti-infectious nano collagen/ calcium phosphate bone repair material |
| CN103830775A (en) * | 2012-11-27 | 2014-06-04 | 北京奥精医药科技有限公司 | High-strength collagen base artificial bone repair material |
| US20140194363A1 (en) * | 2013-01-08 | 2014-07-10 | Beijing Allgens Medical and Technology Co, Ltd | Compression-resistant collagen-based artificial bone repair material |
| CN104258465A (en) * | 2014-10-13 | 2015-01-07 | 北京奥精医药科技有限公司 | Mineralized collagen based skull repairing device and manufacturing method thereof |
Family Cites Families (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101417145B (en) * | 2008-12-04 | 2012-11-07 | 山东大学 | Bracket material for bone tissue engineer and preparation method thereof |
| US20100268227A1 (en) * | 2009-04-15 | 2010-10-21 | Depuy Products, Inc. | Methods and Devices for Bone Attachment |
| EP2524707A1 (en) * | 2011-05-16 | 2012-11-21 | Beijing Allgens Medical Science & Technology Co., Ltd. | Injectible, biocompatible synthetic bone growth composition |
-
2016
- 2016-10-31 CN CN201910736390.1A patent/CN110292661A/en active Pending
- 2016-10-31 CN CN201610928746.8A patent/CN106421927A/en active Pending
Patent Citations (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US20010037014A1 (en) * | 1999-08-28 | 2001-11-01 | Sung-Tsuen Liu | Thin mineralized collagen membrane and method of making same |
| CN1325734A (en) * | 2001-06-29 | 2001-12-12 | 清华大学 | Process for preparing nm-phase calcium-phosphorus salt/collagen/high-molecular bone compounded porous material |
| CN102205150A (en) * | 2011-05-19 | 2011-10-05 | 清华大学 | Preparation method for anti-infectious nano collagen/ calcium phosphate bone repair material |
| CN103830775A (en) * | 2012-11-27 | 2014-06-04 | 北京奥精医药科技有限公司 | High-strength collagen base artificial bone repair material |
| US20140194363A1 (en) * | 2013-01-08 | 2014-07-10 | Beijing Allgens Medical and Technology Co, Ltd | Compression-resistant collagen-based artificial bone repair material |
| CN104258465A (en) * | 2014-10-13 | 2015-01-07 | 北京奥精医药科技有限公司 | Mineralized collagen based skull repairing device and manufacturing method thereof |
Non-Patent Citations (1)
| Title |
|---|
| 顾林等主编: "《"十三五"普通高等教育本科部委级规划教材 食品机械与设备》", 30 June 2016, 中国纺织出版社 * |
Cited By (13)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN112158817A (en) * | 2020-09-14 | 2021-01-01 | 湖北赛罗生物材料有限公司 | Bone tissue repair material and preparation method and application thereof |
| CN115887776A (en) * | 2021-09-24 | 2023-04-04 | 中国科学院理化技术研究所 | Composite porous scaffold formed on basis of gelatin-induced magnesium phosphate deposition and preparation method and application thereof |
| CN113913961A (en) * | 2021-11-16 | 2022-01-11 | 清华大学 | Mineralized collagen nanofiber doped with active elements and preparation method thereof |
| CN114366857A (en) * | 2022-02-08 | 2022-04-19 | 博纳格科技(天津)有限公司 | Preparation method of degradable artificial bone composite material |
| CN114732958A (en) * | 2022-05-09 | 2022-07-12 | 洛阳理工学院 | Bionic collagen mineralized bone powder and preparation method thereof |
| CN114886623A (en) * | 2022-06-02 | 2022-08-12 | 奥精医疗科技股份有限公司 | Preparation method of interbody fusion cage |
| CN115006589A (en) * | 2022-06-28 | 2022-09-06 | 奥精医疗科技股份有限公司 | Carbon nanotube modified mineralized collagen material and preparation method and application thereof |
| CN115120774A (en) * | 2022-06-28 | 2022-09-30 | 奥精医疗科技股份有限公司 | Carbon nano tube strengthened mineralized collagen material and preparation method and application thereof |
| CN115120774B (en) * | 2022-06-28 | 2024-05-28 | 奥精医疗科技股份有限公司 | Carbon nano tube reinforced mineralized collagen material and preparation method and application thereof |
| CN115137883A (en) * | 2022-08-03 | 2022-10-04 | 尧舜泽生物医药(南京)有限公司 | Bionic composite mineralized scaffold and preparation method thereof |
| CN115137883B (en) * | 2022-08-03 | 2023-12-29 | 尧舜泽生物医药(南京)有限公司 | Bionic composite mineralization bracket and preparation method thereof |
| CN115414530A (en) * | 2022-10-24 | 2022-12-02 | 深圳市致美生物科技有限公司 | Injectable filler for accelerating collagen regeneration induction and preparation and application thereof |
| CN115414530B (en) * | 2022-10-24 | 2024-01-12 | 广西小分子医疗科技有限公司 | Injectable filler for accelerating induced collagen regeneration and preparation and application thereof |
Also Published As
| Publication number | Publication date |
|---|---|
| CN106421927A (en) | 2017-02-22 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN110292661A (en) | Compound porous bone material of mineralized collagen/macromolecule and preparation method thereof | |
| CN110368531A (en) | Mineralized collagen base alveolar bone repairing material and preparation method thereof | |
| Ibekwe et al. | Synthesis and characterization of chitosan/gum arabic nanoparticles for bone regeneration | |
| EP1799277B1 (en) | Porous biomaterial-filler composite and a method for making the same | |
| CN107899086B (en) | Hyaluronic acid oligosaccharide modified collagen nanofiber vascular repair material and preparation method thereof | |
| CN104788559B (en) | A kind of extracting method of bio-medical rat tail collagen protein | |
| CN115536866A (en) | Injectable hydrogel for cartilage defect repair and preparation method thereof | |
| Jolly et al. | Bioactive Phoenix dactylifera seeds incorporated chitosan/hydroxyapatite nanoconjugate for prospective bone tissue engineering applications: A bio-synergistic approach | |
| Poddar et al. | Influence of varying concentrations of chitosan coating on the pore wall of polycaprolactone based porous scaffolds for tissue engineering application | |
| CA2339978C (en) | Process for obtaining highly-purified alginates | |
| Vu et al. | Fabrication of in situ crosslinking hydrogels based on oxidized alginate/N, O-carboxymethyl chitosan/β-tricalcium phosphate for bone regeneration | |
| Smetana Jr et al. | Implantation of p (HEMA)-collagen composite into bone | |
| CN105327394A (en) | Mineralized collagen-based biological tooth root scaffold material and preparation method thereof | |
| RU2472516C1 (en) | Biomaterial for bone defect replacement | |
| EP1171175B8 (en) | Active agent coated endoprosthesis with long-term stability | |
| CN108888806B (en) | Osteoinductive material based on fish skin collagen aggregate and preparation method and application thereof | |
| EP3426827B1 (en) | Jellyfish extract nanofibers | |
| EP1766038A1 (en) | Method for purifying hyaluronic acid using calcium salt and phosphate salt, or calcium | |
| CN108619552B (en) | Absorbable wound repair material for adsorbing GGTAl gene knockout pig collagen and preparation method thereof | |
| CN113521387A (en) | Preparation method and application of strontium-doped modified natural hydroxyapatite scaffold material | |
| Xu et al. | Application of new bone cement biomaterials in osteoporotic compression fractures | |
| CN112812576A (en) | Novel zinc-containing biogenic collagen film and preparation method and application thereof | |
| KR102074038B1 (en) | Preparation Method of Customized Bone Graft for 3D Printing | |
| CN115920135B (en) | Bone repair material and preparation method thereof | |
| CN113521386A (en) | Injectable rhBMP-2-containing bone repair hydrogel and preparation method thereof |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| PB01 | Publication | ||
| PB01 | Publication | ||
| SE01 | Entry into force of request for substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| RJ01 | Rejection of invention patent application after publication | ||
| RJ01 | Rejection of invention patent application after publication |
Application publication date: 20191001 |