CN108543115A - A kind of osteoinductive collagen-based composite hydrogel and preparation method thereof - Google Patents
A kind of osteoinductive collagen-based composite hydrogel and preparation method thereof Download PDFInfo
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Abstract
The invention discloses a kind of osteoinductive collagen-based composite hydrogels and preparation method thereof, hide glue original solution is prepared using acidic buffer, it is sufficiently stirred and is disperseed with hydrotropy solution, then the solution ph is slowly adjusted to 6.0~8.0 using lye, it is subsequently placed into the hatching of Constant temperature hatch case, obtains collagen hydrogel substrate；Collagen hydrogel substrate is immersed into chemical modification agent solution, dopamine is added, and oxidant is added and fully reacts, obtains the collagen hydrogel substrate of dopamine surface modification；Nanometer fish-bone is added, adjusts temperature and pH, is fully washed using ultra-pure water after reaction, subsequent irradiation sterilization, obtain osteoinductive collagen-based composite hydrogel.The advantage of the invention is that：The interface compatibility in prepared hydrogel between particulate cancellous bone and organic phase hydrogel is set to be greatly improved；So that prepared hydrogel has both excellent osteoinductive；New field is opened for the high level conversion of collagen and fish-bone.
Description
Technical field
The present invention relates to technical field of biological materials, more particularly to a kind of osteoinductive collagen-based composite hydrogel and its system
Preparation Method.
Background technology
Bone defect healing caused by the reasons such as wound, tumour, inflammation and surgical operation is always orthopedist and material
The a great problem that scholar is faced.In clinical treatment now, the structure for being limited to bulk bone in bone graft has delayed Bone Defect Repari
The mass exchange and signal transduction of middle material and cell, the design and development of particle bone renovating material come into being, nature bone
Main component is collagenous fibres and hydroxide radical phosphorite nanocrystalline body, since autologous bone tissue sampling is limited, particulate cancellous bone main source
In animal bone tissue (ox bone, pig bone, coral bone, fish-bone etc.) or hydroxyapatite, calcium silicates, phosphoric acid with similar component
Calcium, calcium carbonate etc..But in particulate cancellous bone application process there are the insufficient inherent shortcoming of spatial stability (it is loosely organized, easily by blood
Liquid or flushing liquor are broken up) (Kim J.A., Yun H.S., Choi Y.A., Kim J.E., Choi S.Y., Kwon T.G., Kim
Y.K.,Kwon T.Y.,Bae M.A.,Kim N.J.,Bae Y.C.,Shin H.I.,Park E.K.Magnesium
phosphate ceramics incorpo rating a novel indene compound promote osteoblast
differentiation in vitro and bone regeneration in vivo[J].Biomaterials,2018,
157:51-61；Sun Jing, Xiao Yumei, model Korean pine, Zhang Xingdong hydroxyapatites are to the preparation of calcium alginate hydrogel and performance
Influence [J] Journal of Inorganic Materials, 2010,25 (10):1087-1091).
Research direction is then focused on the research of osteoinductive hydrogel by people, is expected that by the mode of hydrogel load
The preferable spatial stability of imparting particulate cancellous bone (Chang C.W., Wu Y.R., Chang K.C., Ko C.L., Lin D.J.,
Chen W.C.In vitro characte rization of porous calcium phosphate scaffolds
capped with crosslinke d hydrogels to avoid inherent brittleness[J].Ceramics
International,2018,44(2):1575-1582；Hosseinzadeh H.,Ramin S.Fabrication of
starc h-graft-poly(acrylamide)/graphene oxide/hydroxyapatite nanocomposite
hydrogel adsorbent for removal of malachite green dye from aqueous solution
[J].International Journal of Biological Macromolecules,2018,106:101-115).For example
Chitosan is subjected to carboxymethyl modification, then compound with tricalcium silicate, the spatial stability of particle bone material can be improved to a certain degree
Property；The hyaluronic acid of different molecular weight is either prepared into hydrogel for loading calcium phosphate or calcium carbonate etc.；For another example pass through
The molecular weight for changing the main chain of Poly(D,L-lactide-co-glycolide and the chemical constitution of collateral group, polymer is used for calcium carbonate
Material load, prepare based on synthesize high molecular bone repair hydrogel (Zhao Ningbo hyaluronic acid composite bone substitution materials promote at
Bone effect and Mechanism Study [D] .2016, The Fourth Military Medical University, Ph.D. Dissertation).In view of nature bone itself be by inorganic and
The height complex of organic matter composition, basic component units are the hydroxide radical phosphorite nanocrystalline bodies containing collagenous fibres, from group
At the angle of bionic structure, the composite hydrogel based on collagen and hydroxyapatite has received widespread attention, including by bionical
The mode of mineralising organically combines collagen hydrogel and hydroxyapatite, or directly loads nanometer or μ-hydroxyapatite
In collagen hydrogel, be used to prepare osteoinductive hydrogel (Gong T., Xie J., Liao J., Zhang T., Lin S.,
Lin Y.Nanomaterials and bone regeneration[J].Bone Research,2015,3:1-7；
Meskinfam M.,Bertoldi S.,Albanese N.,Cerri A.,Tanzi M.C.,Imani R.,Baheiraei
N.,Farokhi M.,Fare S.Polyurethane foam/nano hydroxyapatite composite as a
bone tissue regeneration[J].Mater ials Science&Engineering C-Materials for
Biological Applications,2018,82:130-140).In general, using hydrogel as excipient, with particulate cancellous bone
Material (micron or nanoscale) carry out organic composite, or be expected solve particulate cancellous bone repair process in spatial stability it is insufficient solid
It is defective.However, organic phase (macromolecule hydrogel) and particulate cancellous bone interface compatibility official post obtain the effect of synergistic effect between the two
Fruit is not satisfactory.Currently, with the bionical method of macromolecule, by imitative mussel adhesion protein Functional Unit (3,4- dihydroxy benzenes propylamine,
Dopamine) it is organically combined with high molecular material, the excellent interface adhesion properties of mussel adhesion protein are replicated accordingly, because its is very strong
Structure designability receives much attention that (Li Hui China is repaiied based on poly-dopamine adhesion layer biology with ease-to-operate
Adorn polylactic acid and its cytocompatibility Journal of Sex Research [D] .2016, Ji'nan University, master's thesis).But current research is limited to more
In synthesis macromolecule and dopamine interaction, final product biology inducing action is limited, based on bioactive polymer into
It is still inadequate in terms of row mussel Bionic Design osteoinductive hydrogel.
In conclusion currently existing technology is confined to the simple load of hydrogel and particulate cancellous bone, organic phase and particulate cancellous bone more
Interface compatibility is poor, in spite of utilizing relevant report of the dopamine for interfacial adhesion, but be confined to synthesis macromolecule with it is more
The interaction of bar amine, osteoinductive is obviously insufficient, especially based on " collagen chemistry modification " and " assembling of dopamine autohemagglutination "
The osteoinductive hydrogel of bionical structure load nanometer fish-bone and preparation method thereof aspect is not yet seen in report.
Invention content
The present invention in view of the drawbacks of the prior art, provides a kind of based on " collagen chemistry modification " and " assembling of dopamine autohemagglutination "
The osteoinductive hydrogel and preparation method thereof of bionical structure load nanometer fish-bone.This method not only makes in prepared hydrogel
Interface compatibility between particulate cancellous bone and organic phase hydrogel is greatly improved, and it is excellent so that prepared hydrogel has both
Different osteoinductive.This not only develops new bone repair hydrogel material, but also the high level conversion for collagen and fish-bone is opened
New field is warded off.
In order to realize the above goal of the invention, the technical solution adopted by the present invention is as follows：
A kind of osteoinductive collagen-based composite hydrogel and preparation method thereof, preparation process is as follows：
(1) in the case where solution temperature is 0~10 DEG C, hide glue original solution, the concentration model of buffer solution are prepared using acidic buffer
It is 2.0~5.0 to enclose for 0.1~0.5mol/L, pH；Collagen solution concentration is 1~15mg/mL；Collagen be pigskin, ox-hide,
Sheepskin, donkey hide and one kind in fish-skin type i collagen or in which several mixtures；
It is sufficiently stirred and is disperseed with hydrotropy solution, the hide glue original solution is slowly then adjusted using the lye of a concentration of 0.1mol/L
PH value is subsequently placed into the Constant temperature hatch case that temperature is 30~40 DEG C and hatches 4~12h, obtain collagen hydrogel to 6.0~8.0
Base material；
(2) by collagen hydrogel substrate be 20~37 DEG C in temperature at immerse chemical modification agent solution 4~for 24 hours, it is described
Chemical modifier be oxidized hyaluronic acid, any one of condensed tannin, the dosage of chemical modifier is collagen water-setting
1~8wt% of gum base materials dry weight；
Dopamine is then added, dopamine dosage is collagen hydrogel substrate 5~10wt% of dry weight, and dopamine is added
The oxidant of 0.1~3wt% of weight fully reacts 1~4h, and dopamine is allowed to carry out molecule autohemagglutination in collagen hydrogel substrate surface
It closes and assembles, obtain the collagen hydrogel substrate of dopamine surface modification；
(3) nanometer fish-bone well prepared in advance is added after dopamine reaction, the dosage of nanometer fish-bone is collagen water-setting
1~10wt% of gum base materials dry weight, it is 20~37 DEG C to adjust load temperature, and pH is 5.0~8.0, using super after 1~12h of reaction
Pure water fully washs, and then packs and is sterilized to get to osteoinductive collagen-based composite hydrogel using -60 x ray irradiation x of cobalt.
Preferably, the collagen described in step 1 is ox-hide type i collagen, it is different from its catabolite gelatin or collagen
Small peptide, the type i collagen have complete triple helix structure；The acidic buffer is acetic acid-sodium acetate acidic buffer,
Its a concentration of 0.25mol/L, pH 2.5, it is 4 DEG C that acidic buffer, which prepares hide glue original solution solution temperature, hide glue original solution
A concentration of 10mg/mL.
Preferably, the lye described in step 1 is the NaOH solution of a concentration of 0.1mol/L, Constant temperature hatch box temperature degree is excellent
37 DEG C are selected, brooding time 8h.
Preferably, the chemical modifier described in step 2 is oxidized hyaluronic acid, the dosage of chemical modifier is hide glue
The 5wt% of raw water gel base dry weight.
Preferably, when collagen hydrogel substrate being immersed chemical modification agent solution in step 2, reaction temperature 30
DEG C, reaction time 8h.
Preferably, the dopamine dosage is the 8wt% of collagen hydrogel substrate dry weight,
Preferably, the oxidant described in step 2 is H2O2, dosage is the 0.5wt% of dopamine weight, and oxidation is added
The agent reaction time is 3h.
Preferably, the nanometer fish-bone described in step 3 is Tilapia bone, the salmon prepared using high energy wet ball grinding
One kind in the fish-bones such as bone, grass carp bone, silver carp bone, black carp bone or mixture, grain size are 20~100nm；The use of nanometer fish-bone
Amount is the 5wt% of collagen hydrogel substrate dry weight, and it is 30 DEG C, pH 7.0 to adjust load temperature, and ultra-pure water is used after reacting 4h
Fully washing 3 times.
Preferably, the addition opportunity of the nanometer fish-bone described in step 3 replace with it is synchronous with dopamine addition.
Preferably, the osteoinductive collagen-based composite hydrogel, nanometer fish-bone is by more with collagen hydrogel
Bar amine is loaded and is fixed, and avoids the phase separation of organic phase and inorganic phase, the payload rate of nanometer fish-bone 80~
95% (on the basis of nanometer fish-bone adds weight), appearance are light grey or black gelinite, and the impurity being visible by naked eyes should
Hydrogel mean porosities are that 50%~90%, pH is 7.0~7.5, and cytotoxicity is no more than 0 grade, specific retention 80%~95%,
Swelling ratio 50%~300%, elasticity modulus 100Pa~3000Pa.
Compared with prior art the advantage of the invention is that：
1, osteoinductive collagen-based composite hydrogel prepared by the present invention has fine compared with simple particle bone material
Spatial stability, be not easy to be broken up by blood or flushing liquor.
2, the addition of crosslinking agent of the present invention improves the mechanical property of hydrogel, meanwhile, it is used with traditional collagen hydrogel
Glutaraldehyde cross-linking agent compare, the present invention used in crosslinking agent belong to bion crosslinking agent, avoid traditional chemical crosslinking agent (penta
Dialdehyde) the potential source biomolecule toxicity that is introduced from source.
3, compared with directly extracting mussel adhesion protein from natural mussel, the raw material sources used in the present invention are extensive, institute
The cost for obtaining product is relatively lower.
4, compared with using traditional synthesis macromolecule hydrogel load particulate cancellous bone, osteoinductive collagen prepared by the present invention
Base composite hydrogel has excellent tissue inducing action, and organic phase and inorganic phase interface compatibility are more preferable.
Description of the drawings
Fig. 1 is that one group of elastic modulus detection result of osteoinductive collagen-based composite hydrogel of the embodiment of the present invention is linear
Figure；
Fig. 2 is that one group of loss moduli measurement result of osteoinductive collagen-based composite hydrogel of the embodiment of the present invention is linear
Figure.
Specific implementation mode
To make the objectives, technical solutions, and advantages of the present invention more comprehensible, develop simultaneously embodiment referring to the drawings, right
The present invention is described in further details.
Embodiment 1
Using 0.1mol/L, the acetic acid that pH is 2.0-sodium acetate solution dissolves pigskin collagen, is sufficiently stirred at 4 DEG C to help
Dissolving dispersion, obtain the collagen solution of a concentration of 15mg/mL, then use 0.1mol/L NaOH solution adjust solution ph to
7.0, then the Constant temperature hatch 4h at 30 DEG C, obtains collagen hydrogel substrate；Prepared collagen hydrogel substrate is existed
It is immersed at 20 DEG C anti-in the chemical modifier containing oxidized hyaluronic acid (dosage is the 5wt% of collagen hydrogel substrate dry weight)
Answer 8h.Dopamine and H is then added2O2, dopamine dosage is collagen hydrogel substrate dry weight 5wt%, H2O2Dosage is DOPA
Amine weight 0.5wt% reacts 3h.It is subsequently added into a nanometer Tilapia bone (dosage is the 5wt% of collagen hydrogel substrate dry weight),
It is 30 DEG C, pH 7.0 to adjust load temperature, after reacting 4h, is fully washed 3 times using ultra-pure water, then packs and uses cobalt -60
X ray irradiation x sterilizing is to get to osteoinductive collagen-based composite hydrogel；The osteoinductive collagen-based that the present invention is prepared is multiple
The results are shown in Table 1 for one group of performance characterization of Heshui gel.
Table 1
Embodiment 2
Using 0.1mol/L, acetic acid-sodium acetate solution that pH is 2.0 dissolves sheepskin collagen, be sufficiently stirred at 10 DEG C with
Hydrotropy solution is disperseed, and the collagen solution of a concentration of 10mg/mL is obtained, and the NaOH solution of 0.1mol/L is then used to adjust solution ph
To 7.5, then the Constant temperature hatch 8h at 37 DEG C, obtains collagen hydrogel substrate；By prepared collagen hydrogel substrate
It is immersed at 30 DEG C in the chemical modifier containing oxidized hyaluronic acid (dosage is the 6wt% of collagen hydrogel substrate dry weight)
React 8h.Dopamine and H is then added2O2, dopamine dosage is collagen hydrogel substrate dry weight 10wt%, H2O2Dosage is
Dopamine weight 1.0wt% reacts 3h.Being subsequently added into a nanometer salmon fish-bone, (dosage is collagen hydrogel substrate dry weight
10wt%), it is 30 DEG C, pH 7.5 to adjust load temperature, after reacting 8h, is fully washed 3 times using ultra-pure water, is then packed simultaneously
It is sterilized to get to osteoinductive collagen-based composite hydrogel, osteoinductive collagen of the embodiment of the present invention using -60 x ray irradiation x of cobalt
One group of elastic modulus detection result Line Chart of base composite hydrogel is as shown in Figure 1.
Embodiment 3
Using 0.15mol/L, acetic acid-sodium acetate solution that pH is 2.5 dissolves ox-hide collagen, be sufficiently stirred at 4 DEG C with
Hydrotropy solution is disperseed, and the collagen solution of a concentration of 5mg/mL is obtained, and the NaOH solution of 0.1mol/L is then used to adjust solution ph
To 8.0, then the Constant temperature hatch 6h at 37 DEG C, obtains collagen hydrogel substrate；By prepared collagen hydrogel substrate
It is immersed at 30 DEG C anti-in the chemical modifier containing condensed tannin (dosage is the 5wt% of collagen hydrogel substrate dry weight)
Answer 8h.Dopamine and H is then added2O2, dopamine dosage is collagen hydrogel substrate dry weight 6wt%, H2O2Dosage is DOPA
Amine weight 2.0wt% reacts 2h.It is subsequently added into a nanometer grass carp fish-bone (dosage is the 8wt% of collagen hydrogel substrate dry weight),
It is 30 DEG C, pH 6.5 to adjust load temperature, after reacting 12h, is fully washed 3 times using ultra-pure water, then packs and uses cobalt-
For the sterilizing of 60 x ray irradiation xs to get to osteoinductive collagen-based composite hydrogel, osteoinductive collagen-based of the embodiment of the present invention is compound
One group of loss moduli measurement result Line Chart of hydrogel is as shown in Figure 2.
Embodiment 4
Using 0.2mol/L, the acetic acid that pH is 3.0-sodium acetate solution dissolves donkey skin collagens, is sufficiently stirred at 4 DEG C to help
Dissolving dispersion, obtain the collagen solution of a concentration of 12mg/mL, then use 0.1mol/L NaOH solution adjust solution ph to
7.0, then the Constant temperature hatch 8h at 37 DEG C, obtains collagen hydrogel substrate；Prepared collagen hydrogel substrate is existed
It immerses in the chemical modifier containing condensed tannin (dosage is the 4wt% of collagen hydrogel substrate dry weight) and reacts at 30 DEG C
8h.Dopamine and H is then added2O2, dopamine dosage is collagen hydrogel substrate dry weight 5wt%, H2O2Dosage is dopamine
Weight 3.0wt% reacts 3h.It is subsequently added into a nanometer silver carp fish-bone (dosage is the 5wt% of collagen hydrogel substrate dry weight), is adjusted
It is 30 DEG C, pH 7.0 to save load temperature, after reacting 4h, is fully washed 3 times using ultra-pure water, then packs and is penetrated using cobalt -60
Line irradiation sterilization is to get to osteoinductive collagen-based composite hydrogel.
Embodiment 5
Using 0.1mol/L, acetic acid-sodium acetate solution that pH is 2.0 dissolves fish skin collagen, be sufficiently stirred at 10 DEG C with
Hydrotropy solution is disperseed, and the collagen solution of a concentration of 10mg/mL is obtained, and the NaOH solution of 0.1mol/L is then used to adjust solution ph
To 7.0, then the Constant temperature hatch 8h at 37 DEG C, obtains collagen hydrogel substrate；By prepared collagen hydrogel substrate
It is immersed at 30 DEG C anti-in the chemical modifier containing condensed tannin (dosage is the 5wt% of collagen hydrogel substrate dry weight)
Answer 8h.Dopamine, nanometer black carp fish-bone and H then is added simultaneously2O2, dopamine dosage is collagen hydrogel substrate dry weight
5wt%, H2O2Dosage is dopamine weight 3.0wt%, and (dosage is collagen hydrogel substrate dry weight to nanometer black carp fish-bone
5wt%), it is 30 DEG C, pH 7.0 to adjust load temperature, after reacting 4h, is fully washed 3 times using ultra-pure water, then packs and adopt
It is sterilized to get to osteoinductive collagen-based composite hydrogel with -60 x ray irradiation x of cobalt.
Embodiment 6
Using 0.1mol/L, the acetic acid that pH is 2.0-sodium acetate solution dissolves fish skin collagen, is sufficiently stirred at 4 DEG C to help
Dissolving dispersion, obtain the collagen solution of a concentration of 8mg/mL, then use 0.1mol/L NaOH solution adjust solution ph to
7.5, then the Constant temperature hatch 8h at 30 DEG C, obtains collagen hydrogel substrate；Prepared collagen hydrogel substrate is existed
It is immersed at 30 DEG C anti-in the chemical modifier containing oxidized hyaluronic acid (dosage is the 5wt% of collagen hydrogel substrate dry weight)
Answer 8h.Dopamine, nanometer grass carp fish-bone and H then is added simultaneously2O2, dopamine dosage is collagen hydrogel substrate dry weight
6wt%, H2O2Dosage is dopamine weight 1.0wt%, and (dosage is collagen hydrogel substrate dry weight to nanometer grass carp fish-bone
8wt%), it is 37 DEG C, pH 6.5 to adjust load temperature, after reacting 3h, is fully washed 3 times using ultra-pure water, then packs and adopt
It is sterilized to get to osteoinductive collagen-based composite hydrogel with -60 x ray irradiation x of cobalt.
Those of ordinary skill in the art will understand that in each method embodiment of the present invention, the sequence of each step
The sequencing that number can not be used to limit each step is not paying creative labor for those of ordinary skill in the art
Under the premise of dynamic, the priority of each step is changed also within protection scope of the present invention.The embodiments described herein be in order to
Reader is helped to understand the implementation of the present invention, it should be understood that protection scope of the present invention is not limited to such especially old
It states and embodiment.Those skilled in the art can make various do not depart from according to the technical disclosures disclosed by the invention
Various other specific variations and combinations of present invention essence, these variations and combinations are still within the scope of the present invention.
Claims (10)
1. a kind of osteoinductive collagen-based composite hydrogel and preparation method thereof, it is characterised in that：The preparation process is as follows：
(1) in the case where solution temperature is 0~10 DEG C, hide glue original solution is prepared using acidic buffer, the concentration range of buffer solution is
0.1~0.5mol/L, pH are 2.0~5.0；Collagen solution concentration is 1~15mg/mL；Collagen be pigskin, ox-hide, sheepskin,
Donkey hide and one kind in fish-skin type i collagen or in which several mixtures；
It is sufficiently stirred and is disperseed with hydrotropy solution, the collagen solution ph is slowly then adjusted using the lye of a concentration of 0.1mol/L
To 6.0~8.0, it is subsequently placed into the Constant temperature hatch case that temperature is 30~40 DEG C and hatches 4~12h, obtain collagen hydrogel substrate；
(2) by collagen hydrogel substrate be 20~37 DEG C in temperature at immerse chemical modification agent solution 4~for 24 hours, the change
Modifying agent is any one of oxidized hyaluronic acid, condensed tannin, and the dosage of chemical modifier is collagen water-setting matrix
1~8wt% of material dry weight；
Dopamine is then added, dopamine dosage is collagen hydrogel substrate 5~10wt% of dry weight, and dopamine weight is added
The oxidant of 0.1~3wt% fully reacts 1~4h, allow dopamine collagen hydrogel substrate surface carry out molecule auto polymerization with
Assembling, obtains the collagen hydrogel substrate of dopamine surface modification；
(3) nanometer fish-bone well prepared in advance is added after dopamine reaction, the dosage of nanometer fish-bone is collagen water-setting matrix
1~10wt% of material dry weight, it is 20~37 DEG C to adjust load temperature, and pH is 5.0~8.0, and ultra-pure water is used after reacting 1~12h
Fully washing is then packed and is sterilized to get to osteoinductive collagen-based composite hydrogel using -60 x ray irradiation x of cobalt.
2. a kind of osteoinductive collagen-based composite hydrogel according to claim 1 and preparation method thereof, it is characterised in that：
Collagen described in step 1 is ox-hide type i collagen；The acidic buffer is acetic acid-sodium acetate acidic buffer, dense
Degree is 0.25mol/L, and pH 2.5, it is 4 DEG C that acidic buffer, which prepares hide glue original solution solution temperature, the concentration of hide glue original solution
For 10mg/mL.
3. a kind of osteoinductive collagen-based composite hydrogel according to claim 1 and preparation method thereof, it is characterised in that：
Lye described in step 1 is the NaOH solution of a concentration of 0.1mol/L, preferably 37 DEG C of Constant temperature hatch box temperature degree, and brooding time is
8h。
4. a kind of osteoinductive collagen-based composite hydrogel according to claim 1 and preparation method thereof, it is characterised in that：
Chemical modifier described in step 2 is oxidized hyaluronic acid, and the dosage of chemical modifier is collagen hydrogel substrate dry weight
5wt%.
5. a kind of osteoinductive collagen-based composite hydrogel according to claim 1 and preparation method thereof, it is characterised in that：
When collagen hydrogel substrate being immersed chemical modification agent solution in step 2, reaction temperature is 30 DEG C, reaction time 8h.
6. a kind of osteoinductive collagen-based composite hydrogel according to claim 1 and preparation method thereof, it is characterised in that：
Dopamine dosage described in step 2 is the 8wt% of collagen hydrogel substrate dry weight.
7. a kind of osteoinductive collagen-based composite hydrogel according to claim 1 and preparation method thereof, it is characterised in that：
Oxidant described in step 2 is H2O2, dosage is the 0.5wt% of dopamine weight, and the addition oxidant reaction time is 3h.
8. a kind of osteoinductive collagen-based composite hydrogel according to claim 1 and preparation method thereof, it is characterised in that：
Nanometer fish-bone described in step 3 be the Tilapia bone prepared using high energy wet ball grinding, salmon bone, grass carp bone, silver carp bone,
One kind in the fish-bones such as black carp bone or mixture, grain size are 20~100nm；The dosage of nanometer fish-bone is collagen water-setting matrix
The 5wt% of material dry weight, it is 30 DEG C, pH 7.0 to adjust load temperature, is fully washed 3 times using ultra-pure water after reacting 4h.
9. a kind of osteoinductive collagen-based composite hydrogel according to claim 1 and preparation method thereof, it is characterised in that：
The addition opportunity of nanometer fish-bone described in step 3 replaces with synchronous with dopamine addition.
10. a kind of osteoinductive collagen-based composite hydrogel according to claim 1 and preparation method thereof, feature exists
In：The osteoinductive collagen-based composite hydrogel, nanometer fish-bone and collagen hydrogel be by dopamine carry out load and
It is fixed, the phase separation of organic phase and inorganic phase is avoided, for the payload rate of nanometer fish-bone 80~95%, appearance is light gray
Color or black gelinite, the impurity being visible by naked eyes, the hydrogel mean porosities are that 50%~90%, pH is 7.0~7.5,
Cytotoxicity is no more than 0 grade, specific retention 80%~95%, swelling ratio 50%~300%, elasticity modulus 100Pa~3000Pa.
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