CN110339403A - Spherical nano hydroxyapatite/natural polymer biomimetic scaffolds and preparation method - Google Patents
Spherical nano hydroxyapatite/natural polymer biomimetic scaffolds and preparation method Download PDFInfo
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- A61L27/00—Materials for grafts or prostheses or for coating grafts or prostheses
- A61L27/40—Composite materials, i.e. containing one material dispersed in a matrix of the same or different material
- A61L27/44—Composite materials, i.e. containing one material dispersed in a matrix of the same or different material having a macromolecular matrix
- A61L27/46—Composite materials, i.e. containing one material dispersed in a matrix of the same or different material having a macromolecular matrix with phosphorus-containing inorganic fillers
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- A61L—METHODS OR APPARATUS FOR STERILISING MATERIALS OR OBJECTS IN GENERAL; DISINFECTION, STERILISATION OR DEODORISATION OF AIR; CHEMICAL ASPECTS OF BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES; MATERIALS FOR BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES
- A61L27/00—Materials for grafts or prostheses or for coating grafts or prostheses
- A61L27/50—Materials characterised by their function or physical properties, e.g. injectable or lubricating compositions, shape-memory materials, surface modified materials
- A61L27/52—Hydrogels or hydrocolloids
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- A61L27/00—Materials for grafts or prostheses or for coating grafts or prostheses
- A61L27/50—Materials characterised by their function or physical properties, e.g. injectable or lubricating compositions, shape-memory materials, surface modified materials
- A61L27/56—Porous materials, e.g. foams or sponges
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61L—METHODS OR APPARATUS FOR STERILISING MATERIALS OR OBJECTS IN GENERAL; DISINFECTION, STERILISATION OR DEODORISATION OF AIR; CHEMICAL ASPECTS OF BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES; MATERIALS FOR BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES
- A61L27/00—Materials for grafts or prostheses or for coating grafts or prostheses
- A61L27/50—Materials characterised by their function or physical properties, e.g. injectable or lubricating compositions, shape-memory materials, surface modified materials
- A61L27/58—Materials at least partially resorbable by the body
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61L—METHODS OR APPARATUS FOR STERILISING MATERIALS OR OBJECTS IN GENERAL; DISINFECTION, STERILISATION OR DEODORISATION OF AIR; CHEMICAL ASPECTS OF BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES; MATERIALS FOR BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES
- A61L2300/00—Biologically active materials used in bandages, wound dressings, absorbent pads or medical devices
- A61L2300/60—Biologically active materials used in bandages, wound dressings, absorbent pads or medical devices characterised by a special physical form
- A61L2300/602—Type of release, e.g. controlled, sustained, slow
- A61L2300/604—Biodegradation
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61L—METHODS OR APPARATUS FOR STERILISING MATERIALS OR OBJECTS IN GENERAL; DISINFECTION, STERILISATION OR DEODORISATION OF AIR; CHEMICAL ASPECTS OF BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES; MATERIALS FOR BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES
- A61L2430/00—Materials or treatment for tissue regeneration
- A61L2430/02—Materials or treatment for tissue regeneration for reconstruction of bones; weight-bearing implants
Abstract
The invention discloses spherical nano hydroxyapatite/natural polymer biomimetic scaffolds and preparation method, solve the problem of that hydroxyapatite particle morphology, size and content are uncontrollable and be unevenly distributed in the prior art.The preparation method of a kind of spherical nano hydroxyapatite/natural polymer biomimetic scaffolds of the present invention, comprising the following steps: natural polymer is dissolved in deionized water, Polymer Solution is made；Polymer Solution is cross-linked to form high-molecular gel；Solubility calcium salting liquid is added drop-wise to gel surface, is spread；Soluble solution containing phosphate is added drop-wise to gel surface again, continues to spread；The solution of gel surface, by soak in alkaline solution, then with phosphate buffer solution (PBS), deionized water successively washs gel after the completion of removal diffusion, is freeze-dried and obtains three-dimensional porous biomimetic scaffolds.Design science of the present invention, method is simple, easy to operate.
Description
Technical field
The invention belongs to biomedical materials fields, and in particular to a kind of spherical nano hydroxyapatite/natural polymer
Biomimetic scaffolds and preparation method.
Background technique
Due to wound, tumour, population aging etc., bone defect patient is more and more, and bone defect disease has become prestige
Coerce the major disease of human health.Bone matrix mainly has organic matter and inanimate matter to constitute, organic matter mainly by collagenous fibres structure
At uniform sequentially the main component of inanimate matter is hydroxyl lime stone, and nano hydroxyl phosphorite crystal is arranged in the length of collagenous fibres
Axis direction.For the Nomenclature Composition and Structure of Complexes of natural imitation bone, by natural organic high-molecular (such as collagen, gelatin etc.) and inorganic hydroxyl phosphorus
The compound next bionical building bone defect damage repair materials of lime stone particle have great importance.
Currently, the method for preparing biomimetic scaffolds mainly has physical blending process, biomineralization method, in situ deposition method.Physics is total
Mixed method is directly to adopt natural organic high-molecular and inorganic nano hydroxyapatite physical mixed, such as 105521524 A of patent CN
A kind of gelatin Hydroxyapatite Nanocomposites are prepared for physical blending process, but there are nanoparticles to reunite sternly for this method
The problem of weight, inorganic phase are unevenly distributed, this will affect the mechanical property of bracket and Bone Defect Repari performance.Biomineralization method is to pass through
Material is immersed in the simulated solutions such as simulated body fluid (SBF), macromolecule surface formed one layer of mineralized layer, but the method preparation
The ratio of inorganic constituents is very low in bracket, bioactivity and poor (the CN 105688288A of bone formation performance；
Adv.Funct.Mater.2018,28,1804730).In-situ deposition rule is by the way that calcium salt (chlorine is added in Polymer Solution
Change calcium, calcium nitrate etc.) and microcosmic salt (disodium hydrogen phosphate, ammonium dihydrogen phosphate etc.), hydroxyapatite is grown in high molecular material,
Although being uniformly distributed for inorganic phase may be implemented in this method, and content is adjustable, but there are hydroxyapatite pattern and partial sizes not
Controllably, there are still the problem of growing of reuniting for hydroxyapatite particle.(CN 107929812A；CN 1106861C；ACS
Appl.Mater.Interfaces 2015,7,10386) therefore, how to realize hydroxyapatite particle morphology, size and content
Controllably, the technical issues of and being evenly distributed in organic matter, be current urgent need to resolve.
Summary of the invention
Technical problem solved by the present invention is providing a kind of spherical nano hydroxyapatite/natural polymer biomimetic scaffolds
Preparation method, solve in the prior art hydroxyapatite particle morphology, size and content it is uncontrollable, it is unevenly distributed to ask
Topic.
The present invention also provides using the bionical branch of spherical nano hydroxyapatite/natural polymer made from the preparation method
Frame.
The technical solution adopted by the invention is as follows:
A kind of preparation method of spherical nano hydroxyapatite/natural polymer biomimetic scaffolds of the present invention, including
Following steps:
Natural polymer is dissolved in deionized water by step 1., is prepared into Polymer Solution；
Polymer Solution obtained in step 1 is cross-linked to form high-molecular gel by step 2.；
Solubility calcium salting liquid is added drop-wise to gel surface made from step 2 by step 3., diffusion；It again will be soluble phosphorous molten
Drop is added to gel surface, continues to spread；
The solution of gel surface after the completion of step 4. removal diffusion, by soak in alkaline solution, then it is slow with phosphate
Rush solution (PBS) solution, deionized water successively washs gel, freeze-drying obtain three-dimensional porous biomimetic scaffolds.
It is diffused as spreading under cryogenic conditions in the step 3, be spread under the conditions of preferably 2-6 DEG C.
Further, the natural polymer selects collagen, gelatin, hyaluronic acid, cellulose, chitin, chitosan, fibroin
Any one or a few mixing in albumen and its modified product.
Further, the Polymer Solution mass concentration in the step 1 is 0.5%~10%.
Further, knot of the crosslinking method in one or both of chemical crosslinking, physical crosslinking in the step 2
It closes.
Further, the crosslinking agent used that is chemically crosslinked is glutaraldehyde, Geniposide, one of sodium glycero-phosphate.
Further, the soluble calcium salt is one of calcium chloride, calcium nitrate, and the concentration of soluble calcium salt is
0.05~1.5mol/L.
Further, the soluble phosphorus-containing compound is one of sodium dihydrogen phosphate, phosphoric acid, and solubility is phosphorous molten
The concentration of liquid is 0.04~1mol/L.
Further, in the step 3, solubility calcium salting liquid is added drop-wise to gel surface made from step 2, diffusion 2
~12 hours；Soluble solution containing phosphate is added drop-wise to gel surface again, continues diffusion 2~12 hours.
Spherical nano hydroxyapatite made of above-mentioned preparation method of the present invention/natural polymer biomimetic scaffolds.
Preferably, which has the porous structure of three-dimensional perforation, and aperture size is 100~800 μm, porosity 40%
~90%；Hydroxyapatite average particle size is 20~200nm in the bracket.
Preferably, the mass content of hydroxyapatite particle is 5%~80% in the bracket.
Compared with prior art, the invention has the following advantages:
Design science of the present invention, method is simple, easy to operate.The present invention creatively uses " first plastic-deposits afterwards " side
Method successfully realizes the regulation to hydroxyapatite pattern, size and content.Firstly, solubility calcium salting liquid is added drop-wise to gel table
Soluble solution containing phosphate after low temperature diffusion, then is added drop-wise to gel surface by face, and low temperature diffusion, passes through the pH of regulation system again
Value makes to generate nanoscale hydroapatite particles inside gel, and granule-morphology is spherical shape, and is evenly distributed in gel.
In addition, the soaking time by control gel in aqueous slkali, realizes the adjusting of hydroapatite particles particle size, thus full
The a variety of different clinical demands of foot.
Using spherical nano hydroxyapatite made of the method for the present invention/natural polymer biomimetic scaffolds, passed through with three-dimensional
Logical porous structure is conducive to the migration of cell and blood vessel and grows into；With good biocompatibility and biological degradability,
Degradation rate is adjustable, and nanometer hydroxyapatite particle can with it is high molecular degradation and slow release, be conducive to new bone again
It is raw.
Detailed description of the invention
Attached drawing 1 is spherical nano hydroxyapatite of the invention/natural polymer biomimetic scaffolds scanning electron microscope (SEM) photograph.Its
In, a, b, c are the stereoscan photograph of manufactured bracket in embodiment 2: figure a is 100 times of amplification, and figure b is amplification 20000
Times, figure c is 50000 times of amplification.Scheme the stereoscan photograph that d is the bracket that embodiment 3 uses, amplification factor is 20000 times.
Attached drawing 2 is that spherical nano hydroxyapatite/natural polymer biomimetic scaffolds and medulla mesenchyma are dry in test example one
1,3,7 day MTT result figure after cell (B MSCs) co-cultivation.
Attached drawing 3 is that spherical nano hydroxyapatite in test example two/natural polymer biomimetic scaffolds implantation Rat calvarial lacks
The slice that materials carry out Masson trichrome stain after 8 weeks at damage amplifies 10 times of result figure.
Specific embodiment
Below by specific embodiment to spherical nano hydroxyapatite of the present invention/natural polymer biomimetic scaffolds
It is further described.
Embodiment 1
Present embodiment discloses spherical nano hydroxyapatite of the invention/natural polymer biomimetic scaffolds preparation sides
Method selects gelatin as natural polymer raw material, and soluble calcium salt is calcium chloride, the glutaraldehyde solution that mass concentration is 0.25%
For crosslinking agent, implementation steps are as follows:
(1) 1g gelatin and 18mL deionized water are mixed and is placed in 60 DEG C of water-baths heating stirring 30 minutes, obtained
Bright uniform gelatin solution, mass fraction 5%.
(2) (glutaraldehyde is final concentration of in the glutaraldehyde solution addition gelatin solution for being 2.5% by 2mL mass fraction
0.25%), solution is poured into plastic culture dish after mixing, refrigerator 12 hours for being placed in 4 DEG C become gel.It will
It after the dropwise addition of 3.6mL calcium chloride solution is paved with gel surface, places it in 4 DEG C of refrigerators and spreads 12 hours, take out, then again will
The dropwise addition of 3.6mL phosphoric acid solution is paved with gel surface, continues diffusion 12 hours in 4 DEG C of refrigerators.Wherein, calcium chloride solution concentration is
0.267mol/L, phosphoric acid concentration 0.16mol/L, Ca:P=1.67:1.
(3) liquid for removing gel surface, by soak in 15% ammonia spirit 3h.It takes out bracket and is soaked in 0.1M's
Arginine solution to remove remaining glutaraldehyde in bracket, then with phosphate buffer (PBS), deionized water successively to gel into
Row washing, freeze-drying obtain three-dimensional porous biomimetic scaffolds, and aperture is 100-300 μm.Hydroxy-apatite stone grain in bracket
Son is the spheric granules being evenly distributed, partial size 50-70nm.Nanometer hydroxyapatite quality/gelatin high molecule mass in bracket
Ratio is 5.26%.
Embodiment 2
Select gelatin as natural polymer raw material, soluble calcium salt is calcium nitrate, 0.1% glutaraldehyde as cross linker,
Implementation steps are as follows:
(1) it is placed in 60 DEG C of water-baths heating stirring 30 minutes, obtains after mixing 1g gelatin and 18mL deionized water
The transparent uniform gelatin solution that mass fraction is 5%.
(2 by 2mL mass fraction be 1% glutaraldehyde solution be added gelatin solution in (final concentration of 0.1%) of glutaraldehyde,
Solution is poured into plastic culture dish after mixing, refrigerator 12 hours for being placed in 4 DEG C become gel.By 6mL nitric acid
After calcium solution dropwise addition is paved with gel surface, places it in 4 DEG C of refrigerators and spread 12 hours, then 6mL phosphoric acid solution is added dropwise again
It is paved with gel surface, continues diffusion 12 hours in 4 DEG C of refrigerators.Wherein, nitrification calcium solution concentration is 1.326mol/L, phosphoric acid
Concentration is 0.794mol/L, Ca:P=1.67:1.
(3) liquid for removing gel surface, by soak in 15% ammonia spirit 3h.Bracket is taken out to be soaked in
The arginine solution of 0.1mol/L successively washes gel to remove remaining glutaraldehyde in bracket, then with PBS, deionized water
It washs, freeze-drying obtains three-dimensional porous biomimetic scaffolds, and aperture is 100-300 μm (see Fig. 1 a).Hydroxyapatite in bracket
Particle is the spheric granules being evenly distributed, and partial size is 50-70nm (see Fig. 1 b, c).Nanometer hydroxyapatite quality in bracket/bright
Glue high molecule mass ratio is 27%.
Embodiment 3
Select gelatin as natural polymer raw material, soluble calcium salt is calcium chloride, 0.25% glutaraldehyde as cross linker
Implementation steps are as follows:
(1) it is placed in 60 DEG C of water-baths and stirs 30 minutes after mixing 1g gelatin and 18mL deionized water, obtain transparent
Uniform gelatin solution, mass fraction 5%.
(2) solution is poured into plastic culture dish, being placed in 4 DEG C of refrigerator 2h makes solution be solidified into colloid, takes out culture dish
In colloid to be soaked in 20mLL concentration be glutaraldehyde solution 12 hours of 0.25%.Then the dropwise addition of 6mL calcium chloride solution is paved with
After gel surface, places it in and spread in 4 DEG C of refrigerators 6 hours, the dropwise addition of 6mL phosphoric acid solution is then paved with gel surface again, 4
Continue diffusion 6 hours in DEG C refrigerator.Wherein, calcium chloride solution concentration 1.32mol/L, phosphoric acid concentration 0.794mol/L, Ca:
P=1.67:1.
(3) liquid for removing gel surface, by soak in 15% ammonia spirit 6h.Bracket is taken out to be soaked in
The arginine solution of 0.05mol/L successively carries out gel to remove remaining glutaraldehyde in bracket, then with PBS, deionized water
Washing, freeze-drying obtain three-dimensional porous biomimetic scaffolds.Hydroxyapatite particle in bracket is spherical to be evenly distributed
Grain, partial size are 200nm (see Fig. 1 d).
Embodiment 4
Select gelatin as natural polymer raw material, soluble calcium salt is calcium chloride, does not add crosslinking agent and passes through gelatin
Low-temperature setting plastic, implementation steps are as follows:
(1) it is placed in 60 DEG C of water-baths heating stirring 30 minutes, obtains after mixing 0.6g gelatin and 18mL deionized water
The transparent uniform gelatin solution for being 3% to mass fraction.
(2) gelatin solution is poured into plastic culture dish, being placed in 4 DEG C makes its low-temperature setting become solidifying for refrigerator 12 hours
Glue.Then it after the dropwise addition of 6mL calcium chloride solution being paved with gel surface, places it in and is spread in 4 DEG C of refrigerators 2 hours, then again will
The dropwise addition of 6mL phosphoric acid solution is paved with gel surface, continues diffusion 2 hours in 4 DEG C of refrigerators.Wherein, calcium chloride solution concentration is
0.792mol/L, phosphoric acid concentration 0.4764mol/L, Ca:P=1.67:1.
(3) liquid for removing gel surface, by soak in 15% ammonia spirit 3h.It is successively right with PBS, deionized water
Gel is washed, and freeze-drying obtains three-dimensional porous biomimetic scaffolds.
Embodiment 5
Select gelatin as natural polymer raw material, soluble calcium salt is calcium chloride, and Geniposide is crosslinking agent, implementation steps
It is as follows:
(1) it is placed in 60 DEG C of water-baths and stirs 30 minutes after mixing 2g gelatin and 18mL deionized water, obtain transparent
Uniform gelatin solution, mass fraction 10%.
(2) (Geniposide is final concentration of in the genipin solution addition gelatin solution for being 2.5% by 2mL mass fraction
0.25%), solution is poured into plastic culture dish after mixing, refrigerator 12 hours for being placed in 4 DEG C become gel.So
After the dropwise addition of 12mL calcium chloride solution is paved with gel surface afterwards, places it in 4 DEG C of refrigerators and spread 2 hours, then again by 12mL
Sodium dihydrogen phosphate dropwise addition is paved with gel surface, continues diffusion 12 hours in 4 DEG C of refrigerators.Wherein, calcium chloride solution concentration
For 1.32mol/L, phosphate dihydrogen sodium concentration 0.794mol/L, Ca:P=1.67:1.
(3) liquid of gel surface, the sodium hydroxide solution 3h for being 8 in pH by soak are removed.Bracket is taken out to impregnate
In the arginine solution of 0.05mol/L to remove remaining glutaraldehyde in bracket, then with PBS, deionized water successively to gel into
Row washing, freeze-drying obtain three-dimensional porous biomimetic scaffolds.
Embodiment 6
Select collagen as natural polymer raw material, soluble calcium salt is calcium chloride, and Geniposide is crosslinking agent, implementation steps
It is as follows:
(1) under condition of ice bath, the capital that 2mL concentration is 2.5% is added in the collagen solution that 18mL mass fraction is 10%
The flat solution of Buddhist nun places 3 hours plastics.
(2) it after the dropwise addition of 1.5mL calcium chloride solution being paved with gel surface, places it in 4 DEG C of refrigerators and spreads 12 hours, and
The dropwise addition of 1.5mL phosphoric acid solution is paved with gel surface again afterwards, continues diffusion 2 hours in 4 DEG C of refrigerators.Wherein, calcium chloride solution
Concentration is 1.32mol/L, phosphoric acid concentration 0.794mol/L, Ca:P=1.67:1.
(3) liquid for removing gel surface, by soak in 20% ammonia spirit 3h, then successively with PBS, deionized water
Gel is washed, freeze-drying obtains three-dimensional porous biomimetic scaffolds.
Embodiment 7
Selecting sulfhydryl modified hyaluronic acid (HA-SH) is natural polymer raw material, and soluble calcium salt is calcium chloride implementation
Steps are as follows:
(1) 150mgHA-SH is weighed in deionized water, and oscillation dissolution forms uniform solution.It is 0.3mol/L's with concentration
NaOH aqueous solution adjusts pH to 7, and placing 6h at room temperature makes solution plastic.
(2) it after the dropwise addition of 1.5mL calcium chloride solution being paved with gel surface, places it in 4 DEG C of refrigerators and spreads 6 hours, and
The dropwise addition of 1.5mL phosphoric acid solution is paved with gel surface again afterwards, continues diffusion 2 hours in 4 DEG C of refrigerators.Wherein, calcium chloride solution
Concentration is 0.66mol/L, phosphoric acid concentration 0.397mol/L, Ca:P=1.67:1.
(3) liquid for removing gel surface, by soak in 20% ammonia spirit 3h, then successively with PBS, deionized water
Gel is washed, freeze-drying obtains three-dimensional porous biomimetic scaffolds.
Embodiment 8
Chitosan is selected, soluble calcium salt is calcium chloride, and implementation steps are as follows:
(1) 150mg chitosan is weighed in 9mL deionized water, and oscillation dissolves it sufficiently, is then added into solution
The glutaraldehyde that 1mL concentration is 2.5%.Solution is poured into plastic culture dish after mixing, is placed in 4 DEG C of refrigerator 12 hours
Become gel.
(2) it after the dropwise addition of 1.5mL calcium chloride solution being paved with gel surface, places it in 4 DEG C of refrigerators and spreads 2 hours, and
The dropwise addition of 1.5mL sodium dihydrogen phosphate is paved with gel surface again afterwards, continues diffusion 6 hours in 4 DEG C of refrigerators.Wherein, chlorination
Calcium solution concentration is 1.32mol/L, phosphate dihydrogen sodium concentration 0.794mol/L, Ca:P=1.67:1.
(3) liquid for removing gel surface, by soak in 20% ammonia spirit 3h, then successively with PBS, deionized water
Gel is washed, freeze-drying obtains three-dimensional porous biomimetic scaffolds.
Test example one, cell compatibility test
1, subjects: choosing mesenchymal stem cell (BMSCs), cultivates the preservation committee by Chinese Academy of Sciences typical case
Cell bank (Chinese Shanghai) provides.
2, test material: spherical nano hydroxyapatite/natural polymer biomimetic scaffolds prepared by embodiment 1.
3, test method:
Mesenchymal stem cell (BMSCs) is recovered, is passed on, proliferation.The BMSCs to grow fine is taken to be seeded in embodiment
(sterilizing of 75% ethyl alcohol) observes the growth of cell after material co-cultures 1,3,5 day with cell on the bionical compound rest of 1 preparation
Situation.
4, test result is as shown in Figure 2
The cell proliferative conditions on biomimetic scaffolds are detected using tetrazolium salts colorimetric test (MTT), materials in vitro in
After cell co-cultures 1,3,5 day, 0.5mg/mL MTT (200 hole μ L/) is added into orifice plate, is discarded supernatant after being incubated for 4h at 37 DEG C
Liquid after softly being rinsed with PBS, transfers material into 24 new orifice plates, and dimethyl sulfoxide (DMSO) oscillation, which is added, makes blue knot
Brilliant product dissolution, then measures the absorbance of solution in microplate reader under 490nm wavelength.MTT testing result, embodiment 3 are made
Standby spherical nano hydroxyapatite/natural polymer biomimetic scaffolds can promote the proliferation of BMSCs, no obvious inhibiting effect.
Test result shows using spherical nano hydroxyapatite provided by the invention/natural polymer biomimetic scaffolds to normal cell
It has no toxic side effect.
The osteogenic activity of test example two, animal et al. Ke evaluation material
1, test material: spherical nano hydroxyapatite/natural polymer biomimetic scaffolds prepared by embodiment 3.
2, it experimental subjects: SD rat 3, is provided by Sichuan University's West China Experimental Animal Center.
3, test method: choosing 3 healthy SD rats, manufactures two diameters in every Rat calvarial and is 5mm, is highly
The cylindrical defect of 2mm.The sample of size identical with defect, layer-by-layer suture muscle and skin are implanted into each defect point.Postoperative 8
Week materials, sample through fixation, dehydration, transparent, paraffin embedding and etc. the paraffin sections of 5 μ m-thicks is made, using tri- color of Masson
The bone formation performance of dyeing observation material.
4, test result is as shown in Figure 3.
It can be seen from Masson stained slice after implantation muscle 8 weeks, there is newborn bone tissue to generate in material hole,
Illustrate that this material has preferable bone regeneration capability, there is good application value.
The above-described embodiments merely illustrate the principles and effects of the present invention, and is not intended to limit the present invention.It is any ripe
The personage for knowing this technology all without departing from the spirit and scope of the present invention, carries out modifications and changes to above-described embodiment.Cause
This, institute is complete without departing from the spirit and technical ideas disclosed in the present invention by those of ordinary skill in the art such as
At all equivalent modifications or change, should be covered by the claims of the present invention.
Claims (10)
1. a kind of spherical nano hydroxyapatite/natural polymer biomimetic scaffolds preparation method, which is characterized in that including following
Step:
Natural polymer is dissolved in deionized water by step 1., is prepared into Polymer Solution；
Polymer Solution obtained in step 1 is cross-linked to form high-molecular gel by step 2.；
Solubility calcium salting liquid is added drop-wise to gel surface made from step 2 by step 3., diffusion；Soluble solution containing phosphate is dripped again
It is added to gel surface, continues to spread；
The solution of gel surface after the completion of step 4. removal diffusion, by soak in alkaline solution, then it is molten with phosphate-buffered
Liquid, deionized water successively wash gel, and freeze-drying obtains three-dimensional porous biomimetic scaffolds.
2. a kind of preparation method of spherical nano hydroxyapatite/natural polymer biomimetic scaffolds according to claim 1,
It is characterized in that, the natural polymer select collagen, gelatin, hyaluronic acid, cellulose, chitin, chitosan, fibroin albumen and
Any one or a few mixing in its modified product.
3. a kind of preparation method of spherical nano hydroxyapatite/natural polymer biomimetic scaffolds according to claim 2,
It is characterized in that, the Polymer Solution mass concentration in the step 1 is 0.5%~10%.
4. a kind of preparation method of spherical nano hydroxyapatite/natural polymer biomimetic scaffolds according to right 1, special
Sign is that crosslinking method is selected from the combination of one or both of chemical crosslinking, physical crosslinking in the step 2.
5. a kind of preparation method of spherical nano hydroxyapatite/natural polymer biomimetic scaffolds according to claim 4,
It is characterized in that, the crosslinking agent that uses of being chemically crosslinked is glutaraldehyde, and Geniposide, one of sodium glycero-phosphate.
6. a kind of preparation method of spherical nano hydroxyapatite/natural polymer biomimetic scaffolds according to claim 1,
It is characterized in that, the soluble calcium salt is one of calcium chloride, calcium nitrate, the concentration of soluble calcium salt is 0.05~
1.5mol/L。
7. a kind of preparation method of spherical nano hydroxyapatite/natural polymer biomimetic scaffolds according to claim 1,
It is characterized in that, the soluble phosphorus-containing compound be one of sodium dihydrogen phosphate, phosphoric acid, soluble solution containing phosphate it is dense
Degree is 0.04~1mol/L.
8. a kind of preparation method of spherical nano hydroxyapatite/natural polymer biomimetic scaffolds according to claim 1,
It is characterized in that, solubility calcium salting liquid is added drop-wise to gel surface made from step 2, diffusion 2~12 is small in the step 3
When；Soluble solution containing phosphate is added drop-wise to gel surface again, continues diffusion 2~12 hours.
9. spherical nano hydroxyapatite/natural polymer made of preparation method described in claim 1-8 any one is imitative
Raw bracket.
10. spherical nano hydroxyapatite as claimed in claim 9/natural polymer biomimetic scaffolds, which is characterized in that have three
The porous structure of perforation is tieed up, aperture size is 100~800 μm, and porosity is 40%~90%；Hydroxyapatite in the bracket
Average particle size is 20~200nm.
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| CN111603607A (en) * | 2020-05-21 | 2020-09-01 | 武汉理工大学 | Fibroin-calcium hydrogen phosphate compound obtained by induction with fibroin as template and preparation method thereof |
| CN117887134A (en) * | 2024-03-14 | 2024-04-16 | 四川大学 | Hydrogel biomimetic mineralization stent and preparation method and application thereof |
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| CN111603607A (en) * | 2020-05-21 | 2020-09-01 | 武汉理工大学 | Fibroin-calcium hydrogen phosphate compound obtained by induction with fibroin as template and preparation method thereof |
| CN111603607B (en) * | 2020-05-21 | 2021-01-05 | 武汉理工大学 | Fibroin-calcium hydrogen phosphate compound obtained by induction with fibroin as template and preparation method thereof |
| CN117887134A (en) * | 2024-03-14 | 2024-04-16 | 四川大学 | Hydrogel biomimetic mineralization stent and preparation method and application thereof |
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| CN110339403B (en) | 2020-06-12 |
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