CN106730021A - A kind of bioactivity glass modified gelatin composite aquogel and preparation method thereof - Google Patents
A kind of bioactivity glass modified gelatin composite aquogel and preparation method thereof Download PDFInfo
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Abstract
The invention discloses a kind of bioactivity glass modified gelatin composite aquogel and preparation method thereof, belong to biomedical materials field.The bioactivity glass modified gelatin composite aquogel is obtained by following methods：Bioactive glass powder is added into the modified gelatin solution containing light trigger, mould is poured into after ultrasound, dispersed with stirring are uniform, bioactivity glass modified gelatin composite aquogel is obtained after ultraviolet light certain hour.The bioactivity glass modified gelatin composite aquogel has good bioactivity, osteoinductive and high-moisture percentage（More than 80%）, can be used for the fields such as non-bearing bone defect healing, medicament slow release and bone tissue engineer.
Description
Technical field
The invention belongs to the preparation field of bio-medical material, and in particular to a kind of bioactivity glass-modified gelatin is multiple
Heshui gel and preparation method thereof.
Background technology
The reasons such as wound, tumor resection, malformation correction and infection easily cause Cranial defect after making bone loss sclerotin.At present,
The treatment topmost method of Cranial defect is bone collection, including autologous bone and allograph bone.Autologous bone transplanting need not consider tissue compatible
Property and immunological rejection, but supply bone limited source；Allograph bone wide material sources, but there is immunological rejection and infection haematogenous virus
Equivalent risk.Accordingly, it would be desirable to research and develop novel artificial bone renovating material to meet clinical demand.
The main component of bone tissue is collagen and hydroxyapatite.Based on bionical angle, using similar to natural bone component
Organic and inorganic matter prepare bone renovating material and turned into one of current study hotspot.Bioactivity glass is used as typical silicon
Silicate material, with good biocompatibility, bone guided and osteoinductive, has a wide range of applications in bone defect healing.
Gelatin is the collagenous portion product that obtains of hydrolysis, and biocompatibility is good, at the same on chain electronegative carboxylic group can as into
Core site, control and induction hydroxyapatite are formed.Bioactivity glass/gelatin composite material in composition with people's bone tissue class
Seemingly, and with the biological characteristics such as good biocompatibility and induced osteogenesis effect, Bone Defect Repari material is met to a certain extent
The requirement of material.
At present, the common method for preparing bioactivity glass/gelatin composite material is by a certain amount of bioactivity glass
Powder is added into gelatin solution, is poured into mould after stirring, freeze-drying shaping, then carries out chemical place with crosslinking agent
Reason, then freeze-drying is carried out, finally obtain compound support frame material.However, the composite bone repairing material prepared by the above method
There is problems with：（1）Preparation technology is complicated, need to be through the multiple step such as overmolding, freeze-drying and chemical crosslinking；（2）Support
Needed after shaping by being chemically crosslinked raising stability, but crosslinking agent（Such as glutaraldehyde）Easily residual in the bracket, causes cell
Toxicity；（3）Compound rest moisture content is relatively low, is unfavorable for the transport of nutriment and metabolite.
Modified gelatin is, by active amino and methacrylic acid the anhydride reactant gained of gelatin side chain, light-initiated to be issued ultraviolet
Raw Raolical polymerizable forms hydrogel, the shortcomings of improve gelatin poor mechanical property and fast degradation rate.Gelatin hydrogel
With good biocompatibility and excellent processability, while there is similar structure with extracellular matrix, can be glutinous for cell
Attached, propagation and differentiation provide good microenvironment.There is researcher by adding inorganic nano-particle in gelatin hydrogel（Nanometer
Gold grain, silica and hydroxyapatite etc.）, to improve the bioactivity and osteogenic action of composite aquogel.
The present invention prepares bioactivity glass-modified gelatin composite aquogel timbering material using photopolymerization reaction principle.
The composite aquogel has good biocompatibility and self-bone grafting ability；Bioactivity glass is dispersed in three-dimensional network knot
In structure, combined closely with modified gelatin, silicon, calcium and phosphorus plasma dissolution product can slowly discharge；High-moisture percentage can ensure nutrition
The normal transport of material and metabolite；Preparation process is simple, without using chemical cross-linking agent, has in bone tissue engineer field
Wide application prospect.
The content of the invention
Present invention aim at the deficiency for prior art and material, there is provided a kind of bioactivity glass-modified gelatin
Composite aquogel and preparation method thereof, obtains the composite aquogel with good biological activity and self-bone grafting ability, can be used for non-
The treatment of weight bearing area Cranial defect, realizes the reparation and regeneration of bone tissue.
The present invention is achieved through the following technical solutions.
A kind of preparation method of bioactivity glass-modified gelatin composite aquogel, comprises the following steps：
1）By modified gelatin and light trigger 2- hydroxyls -4 ' -（2- hydroxy ethoxies）- 2 methyl phenyl ketones（IRGACURE 2959）Fill
Divide and be dissolved in PBS, obtain modified gelatin pre-polymer solution；
2）Bioactive glass powder, ultrasonic disperse and magnetic agitation are added in modified gelatin pre-polymer solution, is disperseed
Uniform mixed slurry；
3）Mixed slurry is poured into Teflon mould, triggers Raolical polymerizable in ultraviolet light, after reaction
Product it is swelling removing unreacted monomer and light trigger in PBS, obtain bioactivity glass-modified
Gelatin-compounded hydrogel.
Further, step 1）The methacryl amido substitution value of the modified gelatin is 40%-80%.
Further, step 1）Concentration of the middle modified gelatin prepolymer in modified gelatin pre-polymer solution is 7-15 w/
V%, preferably 10 w/v%.
Further, step 1）Concentration of the described light trigger in PBS is 0.1-1 w/v%.
Further, step 2）Described bioactive glass powder is using organic formwork combination sol-gel legal system
Standby micro-/ nano spherical powder.
Further, step 2）The ultrasonic disperse 30min and h of magnetic agitation 4.
Further, step 2）Concentration of the described bioactive glass powder in mixed slurry is 1-10 w/v%.
Further, step 3）The wavelength of the ultraviolet light is 254-400 nm, and intensity is 5-20 mW/cm2, irradiation
Time is 1-20 min.Optimal wavelength is 365 nm, and intensity is 5m W/cm2.
A kind of bioactivity glass-modified gelatin the composite aquogel obtained by above-described preparation method.The biology
Activity glass-modified gelatin composite aquogel can be applied to the treatment at non-bearing Cranial defect position, promote the reparation of bone tissue with
Regeneration.
Compared with prior art, the beneficial effects of the invention are as follows：
（1）The present invention chooses two kinds of materials of bioactivity glass and modified gelatin and is combined, and is issued in ultraviolet light and is conigenous
Composite aquogel is formed by base polymerisation.The composite aquogel has good biocompatibility and self-bone grafting ability；Synthesis
Material source is extensively, cheap；Preparation method is simple and convenient, and without using chemical cross-linking agent.
（2）Bioactivity glass is uniformly embedded into the three dimensional network of modified gelatin hydrogel formation in composite aquogel of the invention
In network structure, physics mode is passed through with modified gelatin（Hydrogen bond）Combine closely, silicon, calcium and phosphorus plasma is slowly discharged.And it is multiple
Heshui gel moisture content is higher（More than 80%）, it is ensured that the transport of nutriment and metabolic waste.
（3）Modified gelatin as good cell carrier, room temperature, in the short time can plastic, filled between marrow can be combined
The bone tissue engineer of the constructing function such as matter stem cell and growth factor.
（4）Bioactivity glass of the invention-modified gelatin composite aquogel can be applied to bone defect healing, medicament slow release
The field such as carrier and bone tissue engineer.
Brief description of the drawings
Fig. 1 a, Fig. 1 b are the gained bioactivity glass of the embodiment of the present invention 1-modified gelatin composite aquogels in different amplifications
The scanning electron microscope (SEM) photograph of multiple；
Fig. 1 c, Fig. 1 d are the gained bioactivity glass of the embodiment of the present invention 2-modified gelatin composite aquogels in different amplification
Scanning electron microscope (SEM) photograph；
Fig. 1 e, Fig. 1 f are the gained bioactivity glass of the embodiment of the present invention 3-modified gelatin composite aquogels in different times magnifications
Several scanning electron microscope (SEM) photographs；
Fig. 2 is the infared spectrum of the gained bioactivity glass of the embodiment of the present invention 2-modified gelatin composite aquogel；
Fig. 3 is the moisture content figure of the gained bioactivity glass of the embodiment of the present invention 1,2,3-modified gelatin composite aquogel；
Fig. 4 is dry with the mouse bone marrow cells mesenchyma thin gained bioactivity glass of the embodiment of the present invention 2-modified gelatin composite aquogel
Born of the same parents co-culture the dead colored graph of work after 24 h.
Specific embodiment
With reference to embodiment and accompanying drawing, the present invention is described in further detail, but embodiments of the present invention are not limited
In this.
Embodiment 1
（1）The preparation process of bioactive glass powder：
Bioactive glass powder is obtained by organic formwork combination sol-gel process, and specific building-up process is as follows：First by 40 g ten
Diamines is added into the mixed solution of 250 ml deionized waters and 800 ml absolute ethyl alcohols, is stirred in 40 DEG C of water-baths to complete
Dissolving；202 ml tetraethyl orthosilicates are subsequently added, 30 min are stirred；24 ml triethyl phosphates are added, 30 min are stirred；Add
137 g four water-calcium nitrates；The milky white solution that will be obtained continues to stir 3 h；The centrifugation of glass colloidal sol is finally obtained into white precipitate,
It is placed in 60 DEG C of drying boxes and dries 24 h, then be placed in 650 DEG C of 3 h of sintering of Muffle furnace, obtains final product bioactive glass powder.
（2）Modified gelatin（Substitution value is 40%）Preparation process：
Modified gelatin is by the active amino of gelatin side chain with obtained by methacrylic acid anhydride reactant, and specific building-up process is as follows：First by 2
G gelatin is added into 20 ml PBSs, is stirred in 60 DEG C of thermostat water baths, is made 10% gelatin solution；Treat bright
Glue is completely dissolved, and adds 0.25 ml methacrylic anhydrides, continues to react 4 h；In going to 40 DEG C of thermostat water baths after having reacted,
Add 40 ml PBSs to dilute terminating reaction, continue to stir 30 min；It is subsequently transferred in the bag filter of 12k thoroughly
7 d are analysed, to remove unreacted methacrylic anhydride and inorganic salts in solution；Solution is finally placed in pre-freeze in -80 DEG C of refrigerators,
Modified gelatin is obtained after freeze-drying（Substitution value is 40%）.
（3）Bioactivity glass-specific building-up process of modified gelatin composite aquogel is as follows：First by 1 g modified gelatins（Take
Dai Du is 40%）Added into 10 ml PBSs with 0.01 g light triggers IRGACURE 2959, in 60 DEG C of perseverances
Stirred in warm water bath to being completely dissolved；0.1 g bioactive glass powders are subsequently added, the min of ultrasonic disperse 30, magnetic force is stirred
4 h are mixed, makes bioactivity glass dispersed in the solution；Finally mixed slurry is added into Teflon mould,
365 nm、5 mW/cm2The min of ultraviolet light 20, obtain bioactivity glass-modified gelatin composite aquogel.Gained is answered
Heshui gel in PBS swelling 24 h removing unreacted monomer and light trigger.
Embodiment 2
（1）The preparation process of bioactive glass powder：
Bioactive glass powder is obtained by organic formwork combination sol-gel process, and specific building-up process is as follows：First by 4 g ten
Diamines is added into the mixed solution of 25 ml deionized waters and 80 ml absolute ethyl alcohols, stirred in 40 DEG C of water-baths to completely it is molten
Solution；20.2 ml tetraethyl orthosilicates are subsequently added, 30 min are stirred；2.4 ml triethyl phosphates are added, 30 min are stirred；Add
13.7 g four water-calcium nitrates；The milky white solution that will be obtained continues to stir 3 h；The centrifugation of glass colloidal sol is finally obtained into white heavy
Form sediment, be placed in 60 DEG C of drying boxes and dry 24 h, then be placed in 650 DEG C of 5 h of sintering of Muffle furnace, obtain final product bioactive glass powder.
（2）Modified gelatin（Substitution value is 60%）Preparation process：
Modified gelatin is by the active amino of gelatin side chain with obtained by methacrylic acid anhydride reactant, and specific building-up process is as follows：First will
20 g gelatin are added into 200 ml PBSs, are stirred in 60 DEG C of thermostat water baths, are made 10% gelatin solution；
Treat that gelatin is completely dissolved, add 10 ml methacrylic anhydrides, continue to react 4 h；40 DEG C of thermostat water baths are gone to after having reacted
In, 500 ml PBSs dilution terminating reaction is added, continue to stir 30 min；It is subsequently transferred to the bag filter of 12k
7 d of middle dialysis, to remove unreacted methacrylic anhydride and inorganic salts in solution；Finally solution is placed in -80 DEG C of refrigerators
Pre-freeze, obtains modified gelatin after freeze-drying（Substitution value is 60%）.
（3）Bioactivity glass-specific building-up process of modified gelatin composite aquogel is as follows：First by 1 g modified gelatins（Take
Dai Du is 60%）Added into 10 ml PBSs with 0.05 g light triggers IRGACURE 2959, in 60 DEG C of perseverances
Stirred in warm water bath to being completely dissolved；0.5 g bioactive glass powders are subsequently added, the min of ultrasonic disperse 30, magnetic force is stirred
4 h are mixed, makes bioactivity glass dispersed in the solution；Finally mixed slurry is added into Teflon mould,
365 nm、5 mW/cm2The min of ultraviolet light 10, obtain bioactivity glass-modified gelatin composite aquogel.Gained is answered
Heshui gel in PBS swelling 24 h removing unreacted monomer and light trigger.
Embodiment 3
（1）The preparation process of bioactive glass powder：
Bioactive glass powder is obtained by organic formwork combination sol-gel process, and specific building-up process is as follows：First by 8 g ten
Diamines is added into the mixed solution of 50 ml deionized waters and 160 ml absolute ethyl alcohols, is stirred in 40 DEG C of water-baths to complete
Dissolving；40.4 ml tetraethyl orthosilicates are subsequently added, 30 min are stirred；4.8 ml triethyl phosphates are added, 30 min are stirred；Plus
Enter 27.4 g four water-calcium nitrates；The milky white solution that will be obtained continues to stir 3 h；The centrifugation of glass colloidal sol is finally obtained into white heavy
Form sediment, be placed in 60 DEG C of drying boxes and dry 24 h, then be placed in 650 DEG C of 3 h of sintering of Muffle furnace, obtain final product bioactive glass powder.
（2）Modified gelatin（Substitution value is 80%）Preparation process：
Modified gelatin is by the active amino of gelatin side chain with obtained by methacrylic acid anhydride reactant, and specific building-up process is as follows：First by 5
G gelatin is added into 50 ml PBSs, is stirred in 60 DEG C of thermostat water baths, is made 10% gelatin solution；Treat bright
Glue is completely dissolved, and adds 5 ml methacrylic anhydrides, continues to react 4 h；Gone to after having reacted in 40 DEG C of thermostat water baths, added
100 ml PBSs dilute terminating reaction, continue to stir 30 min；It is subsequently transferred to dialyse in the bag filter of 12k 7
D, to remove unreacted methacrylic anhydride and inorganic salts in solution；Solution is finally placed in pre-freeze in -80 DEG C of refrigerators, is freezed
Modified gelatin is obtained after drying（Substitution value is 80%）.
（3）Bioactivity glass-specific building-up process of modified gelatin composite aquogel is as follows：First by 1 g modified gelatins（Take
Dai Du is 80%）Added into 10 ml PBSs with 0.1 g light triggers IRGACURE 2959, in 60 DEG C of constant temperature
Stirred in water-bath to being completely dissolved；It is subsequently added 1 g bioactive glass powders, the min of ultrasonic disperse 30, magnetic agitation 4
H, makes bioactivity glass dispersed in the solution；Finally mixed slurry is added into Teflon mould, 365
nm、5 mW/cm2The min of ultraviolet light 1, obtain bioactivity glass-modified gelatin composite aquogel.Gained Compound Water
Gel in PBS swelling 24 h removing unreacted monomer and light trigger.
Bioactivity glass-modified gelatin composite aquogel to the gained of embodiment 1,2,3 carries out performance characterization, and gained is swept
Electron microscope is retouched as shown in Fig. 1 a, Fig. 1 b, Fig. 1 c, Fig. 1 d, Fig. 1 e, Fig. 1 f, three kinds of bioactivity glass-modified gelatin compound water congealings
Glue is presented the connected porous structure of nest honeybee shape, and pore diameter range is 200-500 μm, and bioactive glass particle is uniformly embedded into
In the three-dimensional network of composite aquogel.Fig. 2 is red for the gained bioactivity glass of embodiment 2-modified gelatin composite aquogel
Outer collection of illustrative plates, wherein a curves are the infrared spectrum of 10% modified gelatin, show acid amides peak specific to typical glutin, its
In be included in 1702 cm-1The flexible peaks of the C=O of amide I band, in 1540 cm-1The N-H stretching vibration peaks of acid amides II bands.B curves
It is the infared spectrum of bioactivity glass, 1091,802 and 474 cm-1Make a living typical Si-O-Si in thing activity glass respectively at place
Asymmetric stretching vibration peak, the symmetrical stretching vibration peak of Si-O and the symmetric curvature vibration peak of Si-O-Si.C curves are compound
The infared spectrum of hydrogel, while there is the characteristic peak of bioactivity glass and modified gelatin, it is same in provable composite aquogel
When there is bioactivity glass and modified gelatin.The bioactivity glass of other embodiments-modified gelatin composite aquogel it is red
Outer collection of illustrative plates is similar to Fig. 2, can prove that in composite aquogel while there is bioactivity glass and modified gelatin.Fig. 3 displays are real
Bioactivity glass-modified gelatin the composite aquogel for applying example 1,2,3 is respectively provided with moisture content higher（More than 80%）, it is ensured that
The transport of nutriment and metabolic waste.Fig. 4 is bioactivity glass-change of the Marrow Mesenchymal Stem Cells in embodiment 2
The dead colored graph of work of the gelatin-compounded h of hydrogel culture 24 of property, it is seen that cell can stick in hydrogel surface, in fusiformis shape, have
Preferable cell viability.The dead staining conditions of work and reality of the bioactivity glass of other embodiments-modified gelatin composite aquogel
Example 2 is applied similar, equal visible cell can stick in hydrogel surface, in fusiformis shape, with preferable cell viability.
Claims (9)
1. the preparation method of a kind of bioactivity glass-modified gelatin composite aquogel, it is characterised in that comprise the following steps：
1）By modified gelatin and light trigger 2- hydroxyls -4 ' -（2- hydroxy ethoxies）- 2 methyl phenyl ketones are completely dissolved in phosphate
In cushioning liquid, modified gelatin pre-polymer solution is obtained；
2）Bioactive glass powder, ultrasonic disperse and magnetic agitation are added in modified gelatin pre-polymer solution, is disperseed
Uniform mixed slurry；
3）Mixed slurry is poured into Teflon mould, Raolical polymerizable is triggered under ultraviolet light, will reacted
Product afterwards is swelling removing unreacted monomer and light trigger in PBS, obtains bioactivity glass-change
The gelatin-compounded hydrogel of property.
2. a kind of preparation method of bioactivity glass-modified gelatin composite aquogel according to claim 1, its feature
It is：Step 1）Methacryl amido substitution value is 40%-80% in the modified gelatin.
3. a kind of preparation method of bioactivity glass-modified gelatin composite aquogel according to claim 1, its feature
It is：Step 1）Concentration of the middle modified gelatin prepolymer in modified gelatin pre-polymer solution is 7-15 w/v%.
4. a kind of preparation method of bioactivity glass-modified gelatin composite aquogel according to claim 1, its feature
It is：Step 1）Concentration of the described light trigger in PBS is 0.1-1 w/v%.
5. a kind of preparation method of bioactivity glass-modified gelatin composite aquogel according to claim 1, its feature
It is：Step 2）Described bioactive glass powder is the micro-/ nano ball prepared using organic formwork combination sol-gel process
Shape powder.
6. a kind of preparation method of bioactivity glass-modified gelatin composite aquogel according to claim 1, its feature
It is：Step 2）Concentration of the described bioactive glass powder in mixed slurry is 1-10 w/v%.
7. a kind of preparation method of bioactivity glass-modified gelatin composite aquogel according to claim 1, its feature
It is：Step 3）The wavelength of the ultraviolet light is 254-400 nm, and intensity is 5-20 mW/cm2, irradiation time is 1-20
min。
8. a kind of preparation method of bioactivity glass-modified gelatin composite aquogel according to claim 7, its feature
It is：The wavelength of the ultraviolet light is 365 nm, and intensity is 5m W/cm2。
9. a kind of bioactivity glass-modified gelatin Compound Water that the preparation method as described in claim any one of 1-8 is obtained
Gel.
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| CN107663377A (en) * | 2017-08-24 | 2018-02-06 | 浙江大学 | It is a kind of that there is temperature sensitive and the mixing hydrogel and its 3D printing method of light sensitive characteristic |
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