CN104740682B - Hydroxylapatite/gelatin-carboxymethyl chitosan composite scaffold - Google Patents
Hydroxylapatite/gelatin-carboxymethyl chitosan composite scaffold Download PDFInfo
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Abstract
The invention discloses a hydroxylapatite/gelatin-carboxymethyl chitosan composite scaffold. The hydroxylapatite/gelati-carboxymethyl chitosan composite scaffold is prepared from hydroxylapatite, gelatin and carboxymethyl chitosan, wherein the mass ratio of the hydroxylapatite to the gelatin to the carboxymethyl chitosan is (60-80):(10-20):(10-20). The invention also provides a preparation method of the composite scaffold. According to the preparation method, the hydroxylapatite is synthesized in situ by use of the gelatin and the carboxymethyl chitosan as templates, and then the composite scaffold material is prepared by virtue of particle percolation in combination with a freeze-drying method. In the obtained composite scaffold, the weak crystalline nano-hydroxylapatite is evenly dispersed in an organic high polymer network to form a communicated porous structure having relatively high strength, and the communicated porous structure is similar to the extracellular matrix in morphology and advantageous for cells to attach and grow; meanwhile, the degradation speed and porosity of the communicated porous structure are adjustable so that the material can be used as a bone repair material.
Description
Technical field
The present invention relates to technical field of biological materials, particularly a kind of hydroxyapatite/gelatin-carboxymethyl chitosan is combined
Support and preparation method thereof.
Background technology
It is one of common difficult disease of Orthopedic Clinical by the Cranial defect that the reasons such as wound, tumour or birth defect cause, answering
Have become as the tissue transplanting being only second to transfuse blood with bone collection in quantity.With medical science applied development, people are permissible
Treat Cranial defect using autologous bone, allograph bone and artificial bone substitute materials, but these materials have not enough, the immune row of donor
Scold, the mechanical performance of transmission and host bone and formation rate mismatch and cannot degrade etc. very important problem.Group
Knit engineering science and provide brand-new Research Thinking and important selectivity strategy for bone defect healing.
As one of organizational project fundamental, timbering material plays vital effect, be connect seed cell and
The bridge of regenerating tissues.Preferably support should simulate the reset condition being repaired position as far as possible.Because nature bone is in microcosmic
On be uniform on a molecular scale by hydroxyapatite nanoparticle (nano-hydroxyapatite, n-HA) and collagenous fibres
A kind of active very vascular tissue of ordered arrangement, hydroxyapatite/collagen composite is in bone tissue work
Cause extremely wide research in journey.Wang etc. will be containing phosphatic collagen solution and hydroxide using microwave radiation technology titration
Calcium solution is mixed with out collagen/hydroxyapatite biomimetic scaffolds, finds the hydroxyapatite whisker of low-crystallinity in composite
With collagen for template in-situ preparation, gained support has higher porosity and big aperture to grain.The research such as Chen finds former
Add fibroin albumen that the elastic modelling quantity of collagen/hydroxyapatite compound rest can be greatly improved in the building-up process of position, and make it
There is good biocompatibility.Although collagen can provide requisite three-dimensional structure for calcified tissue, to mineral deposition
Play inducing action, and with the sticking, grow of cell, express and all have substantial connection；But, there is bad mechanical property, price in it
The problems such as expensive, degradation time is difficult to control to and antigenicity eliminates uncertain, these limit collagen/hydroxyapatite
The development further of composite.
Gelatin (gelatin, Gel) be collagenous portion hydrolysis after denatured products, compared with collagen, possess antigenicity low,
The features such as abundance, specific surface area height, low cost, Stability Analysis of Structures.Based on its surface and medium characteristic, gelatin can pass through electrostatic
Realize ionic adsorption with hydrophobic effect, additionally, different gelatin has different isoelectric points, thus can select quiet by ion
Electro ultrafiltration binds albumen composition.In recent years, for the research of hydroxyapatite/gelatin composite material have been achieved with larger enter
Exhibition, Mohammad etc. prepares hydroxyapatite/gelatin powder by coprecipitation, then utilizes Freeze Drying Technique to obtain power
Learn the compound rest of function admirable.But hydroxyapatite/gelatin composite material plasticity is low, elastic modelling quantity is mated with human body bone
Property is poor.
In order to improve its performance, many scholars start to add second organic in hydroxyapatite/gelatin composite material
Polymer is as enhancing phase.Chitosan derivative, in chitin, is nature only one cationic polysaccharide, because it has biology
Compatibility, biodegradability, antibiotic property, no cytotoxicity and excellent albumen compatibility, in biological medicine especially medicine
Thing carrier aspect has broad application prospects.Zahra etc. prepares hydroxyapatite/gelatin-chitosan by situ Precipitation
Sugared composite, finds that hydroxyapatite forms crystalline substance by the interaction between the function key on calcium ion and macromolecular chain
Core, then grows up in situ；They find that the addition of shitosan can reduce the grain size of hydroxyapatite crystal, thus carrying simultaneously
The mechanical property of high composite.Mohamed etc. have studied hydroxyapatite/chitosan-gelatin composite material in simulated body fluid
In degradability it was demonstrated that this material has good degradability and bone inductive effect, can be used as osseous tissue renovating material.China is specially
Sharp CN1326574C " nanometer hydroxyapatite/chitosan/gelatin porous scaffold and preparation method thereof " is by the skill that is separated
Art is prepared for shitosan/gelatin support, and then its alternate immersion obtains in calcium salt and phosphate solution compound support frame material.
Chinese patent CN101401961B " preparation method of gelatine-chitosan-hydroxyapatite spherical honeycombed grain material " provides
A kind of prepare spherical honeycombed grain material with homogeneous distributed granule diameter and preparation method thereof and device, obtain according to this patent
Be conducive to the flowing of tissue fluid and moving of cell after the gelatine-chitosan obtaining-hydroxyapatite spherical honeycombed grain material filling
Colonization is long.However, in addition to the aqueous solution such as watery hydrochloric acid, spirit of vinegar, shitosan is water insoluble and organic solvent；Early-stage Study table simultaneously
When bright shitosan is used for timbering material, its degradation rate is relatively low with the growth rate matching degree of regenerating tissues and catabolite is in human body
Inside it is difficult to absorb.Therefore, it is necessary to transform to the chemical constitution of shitosan, thus improving its solubility property, widening it should
Use scope.Carboxymethyl chitosan (carboxymethyl chitosan, CMCS) is in shitosan chain using carboxymethylation
On introduce the product after carboxymethyl, be a kind of water soluble chitosan, it had both remained the advantage of shitosan, changed simultaneously and greatly
It has been apt to it water-soluble.In addition, carboxymethyl chitosan is a kind of polyampholyte, compared with shitosan its do not contain only amino,
The groups such as hydroxyl, also add carboxyl on its strand in addition, are thus advantageous to increase and are drawn by hydrogen bond, electrostatic with other molecules
Power etc. forms the molecular recognition site of multiple interaction.Early-stage Study shows that carboxymethyl chitosan antibiotic property is higher, is a kind of
Good organizational project and Thermosensitive Material Used for Controlled Releasing of Medicine, but carboxymethyl chitosan is organized with hydroxyapatite and gelatin-compounded preparation
The research of engineering scaffold material has not yet to see report.
Content of the invention
For above-mentioned prior art, the invention provides a kind of hydroxyapatite/gelatin-carboxymethyl chitosan composite support
Frame, in this timbering material, the nanometer hydroxyapatite of weak crystallization is dispersed in organic polymer network, and formation has higher
The loose structure of the UNICOM of intensity, its pattern is similar to extracellular matrix, sticking and grow beneficial to cell；Meanwhile, its degraded
Speed and adjustable porosity are so that this material is suitable as bone renovating material.
Present invention also offers a kind of preparation method of hydroxyapatite/gelatin-carboxymethyl chitosan compound rest.
The present invention is achieved by the following technical solutions：
A kind of hydroxyapatite/gelatin-carboxymethyl chitosan compound rest, by hydroxyapatite, gelatin, carboxymethyl chitosan
Sugar is constituted, and wherein, hydroxyapatite, gelatin, the mass ratio of carboxymethyl chitosan are (60～80): (10～20)：(10～20).
The preparation method of above-mentioned compound rest, comprises the following steps：
1) four water-calcium nitrate is dissolved in water, four water-calcium nitrate is dissolved in water, obtaining concentration is the water-soluble of 0.2～3mol/L
Liquid, is designated as solution a；
2) weigh gelatin and carboxymethyl chitosan, add it to solution a, under 500～1000rmp, stir 20～40min,
The mass ratio obtaining solution b, wherein gelatin and carboxymethyl chitosan is (1～5)：1；
3) prepare ammonium dibasic phosphate solution；
4) solution b is placed in heating water bath agitating device, is stirred continuously at 20～90 DEG C, by step 3) phosphoric acid prepared
Hydrogen two ammonium salt solution is added dropwise over, and wherein, in solution b, Ca ion and the mol ratio of P ion in added ammonium dibasic phosphate solution are
1.67, after completion of dropping, continue stirring 0.5～2h, adjusting reacting liquid pH value in course of reaction is 8～12；
5) by step 4) mixed solution that obtains is in 110～180 DEG C of hydro-thermal reaction 0.5～6h；Reaction terminate after, spend from
Sub- water washing uses absolute ethanol washing, drying again to neutrality, obtains hydroxyapatite/gelatin-carboxymethyl chitosan composite powder
Body；
6) above-mentioned hydroxyapatite/gelatin-carboxymethyl chitosan composite granule is added in absolute ethyl alcohol, 500～
1000rmp stirs 0.5～2h, obtains hydroxyapatite/gelatin-carboxymethyl chitosan composite colloid, wherein hydroxyapatite/bright
Glue-carboxymethyl chitosan composite granule is 1 with the mass ratio of absolute ethyl alcohol：(1～3)；
7) sodium chloride particle and Geniposide are added in above-mentioned complex sol, stirring, injection molding, room temperature stands 2～4h,
Wherein hydroxyapatite/gelatin-carboxymethyl chitosan composite granule and the mass ratio of sodium chloride are 1：(0.1～2), hydroxy-apatite
Stone/gelatin-carboxymethyl chitosan composite granule is 1 with the mass ratio of Geniposide：(0.01～0.1)；Then -80 DEG C of pre-freezes 10～
24h, -53 DEG C of freeze-drying 15～24h, obtain hydroxyapatite/gelatin-carboxymethyl chitosan composite；
8) by after gained hydroxyapatite/gelatin-carboxymethyl chitosan composite demoulding, be placed under room temperature from
2～6h, then freeze-drying is soaked in sub- water；Repeat this step 3～5 time, will be clean for sodium chloride particle leaching, you can obtain hydroxyl
Base apatite/gelatin-carboxymethyl chitosan compound support frame material.
Step 3) in, the molar concentration of described diammonium hydrogen phosphate is 0.1～1.5mol/L.
In step 4, the regulation of pH value is adjusted by ammoniacal liquor, and wherein the mass concentration of ammoniacal liquor is 2%～15%.
Step 7) in sodium chloride particle particle diameter be 150～500 μm.
Beneficial effects of the present invention：From bionical angle, the main component according to nature bone is hydroxy-apatite to the present invention
Stone, collagen and carbohydrate, using hydroxyapatite, gelatin and the respective advantage of carboxymethyl chitosan, prepare a kind of new
Osseous tissue renovating material.The present invention imitates original position crystallization mistake under the regulation and control of organic substrate macromolecular for the inorganic phase during bon e formation
Journey, prepares hydroxyapatite/gelatin-carboxymethyl chitosan composite granule using Situ Hydrothermal method.Carboxymethyl shell in preparation process
Glycan can be acted on by Electrostatic complexation with gelatin, pre-assembled organic formwork, then utilize the polar functional group of the two pass through from
Son coordination and electrostatic attraction effect are be combined with each other with calcium ion and phosphate anion, and the located growth for follow-up mineral crystal provides
Effectively nucleation site is so that inorganic phase realizes nanoscale dispersion in timbering material, thus improving inorganic phase and organic phase
Interface compatibility, improve the mechanical property of support.On this basis, freeze drying process system is combined using particle leaching
Standby compound support frame material, can control porosity and the degree of cross linking by adjusting pore creating material and dosage of crosslinking agent in the process, obtain
To the compound rest with different hydrophilic and mechanical property.Compared with prior art, respectively become in gained compound rest of the present invention
Divide increasingly similar with the structure of nature bone, constituent, scaffold degradation rate and porosity are easier to control.
Brief description
Fig. 1 is the HA/Gel-CMCS composite XRD analysis of the embodiment of the present invention 1 preparation；
Fig. 2 is the HA/Gel-CMCS composite FT-IR analysis of the embodiment of the present invention 1 preparation；
Fig. 3 is the HA/Gel-CMCS compound rest sem analysis of the embodiment of the present invention 1 preparation.
Specific embodiment
With reference to embodiment, the present invention is further illustrated.
Embodiment 1
A kind of hydroxyapatite/gelatin-carboxymethyl chitosan compound rest, by hydroxyapatite, gelatin, carboxymethyl chitosan
Sugar is constituted, and wherein, hydroxyapatite, gelatin, the mass ratio of carboxymethyl chitosan are 7：2：1.
The preparation method of above-mentioned compound rest, comprises the following steps：
1) accurately weigh four water-calcium nitrate 23.6g, be dissolved in 500ml deionized water, obtain solution a；
2) weigh gelatin and carboxymethyl chitosan, be added into solution a, stir 30min under 500rmp, obtain solution b, its
The mass ratio of middle gelatin and carboxymethyl chitosan is 2：1；
3) preparation molar concentration is the ammonium dibasic phosphate solution of 1.0mol/L；
4) solution b is placed in heating water bath agitating device, under the conditions of being stirred continuously at 70 DEG C, by above-mentioned diammonium hydrogen phosphate
Solution is added dropwise over, and wherein, in solution b, Ca ion and the mol ratio of P ion in added ammonium dibasic phosphate solution are 1.67, drip
Add after finishing, continue stirring 2h, adjusting reacting liquid pH value by the ammoniacal liquor that mass concentration is 5% in course of reaction is 9；
5) above-mentioned mixed solution is added in hydrothermal reaction kettle, 120 DEG C of hydro-thermal reactions 4h；Reaction terminate after, spend from
Sub- water washing uses absolute ethanol washing, drying again to neutrality, obtains hydroxyapatite/gelatin-carboxymethyl chitosan composite powder
Body；
6) compare composite granule according to quality：Absolute ethyl alcohol=1：1 ratio, by above-mentioned hydroxyapatite/gelatin-carboxymethyl
Shitosan composite granule is added in absolute ethyl alcohol, and 500rmp stirs 0.5h, obtains hydroxyapatite/gelatin-carboxymethyl chitosan
Sugared composite colloid；
7) compare composite granule according to quality：Sodium chloride=1：0.5, composite granule is 1 with the mass ratio of Geniposide：0.05,
By particle diameter be 150～200 μm sodium chloride particle and Geniposide be added in above-mentioned complex sol, stirring, injection molding, room temperature is quiet
Put 4h；Then -80 DEG C of pre-freeze 10h, -53 DEG C of freeze-drying 24h, obtain composite；
8) immersion 4h, then freeze-drying in deionized water will after the gained composite demoulding, be placed under room temperature；Weight
Multiple this step 3 time, sodium chloride particle leaching is clean, obtain hydroxyapatite/gelatin-carboxymethyl chitosan compound rest material
Material.
Embodiment 1 gained hydroxyapatite/gelatin-carboxymethyl chitosan compound rest is carried out XRD (Shimadzu,
XRD6100) test, test condition is：Cu target, 30Kv, test result is as shown in Figure 1.To XRD analysis of spectrum, find 26.1 °,
31.4 °, 32.3 °, 32.8 ° is the characteristic peak of hydroxyapatite；22.1 °～23.5 ° is organic phase diffusing scattering peak.To embodiment 1
Gained hydroxyapatite/gelatin-carboxymethyl chitosan compound rest carries out FT-IR analysis (Brooker, EQUINOX55) test,
Test scope 400～4000cm-1, result is as shown in Figure 2.Analysis understands 597cm-1、613cm-1Locate the phosphoric acid for hydroxyapatite
Base；1085cm-1、1160cm-1、1456cm-1Typical functional groups for carboxymethyl chitosan；1550cm-1、1630cm-1With gelatin
Typical functional groups match.It can thus be appreciated that compound rest is made up of hydroxyapatite, gelatin, carboxymethyl chitosan three-phase；?
1354cm-1With 1456cm-1The peak value at place show hydroxyapatite pass through carboxymethyl and amino respectively with carboxymethyl chitosan and bright
Glue is connected, and between each phase, adhesion is stronger, beneficial to the overall performance improving composite.Shown by the sem analysis of Fig. 3, this Shen
Compound rest please has the loose structure of connection, and, more than 75%, average pore size is at 120～200 μm for the porosity；Compression experiment table
Bright gained support mean compressive strength, more than 170MPa, meets the requirement of bone holder material.
Embodiment 2
A kind of hydroxyapatite/gelatin-carboxymethyl chitosan compound rest, by hydroxyapatite, gelatin, carboxymethyl chitosan
Sugar is constituted, and wherein, hydroxyapatite, gelatin, the mass ratio of carboxymethyl chitosan are 6：2：2.
The preparation method of above-mentioned compound rest, comprises the following steps：
1) accurately weigh four water-calcium nitrate 59.0g, be dissolved in 500ml deionized water, obtain solution a；
2) weigh gelatin and carboxymethyl chitosan, be added into solution a, stir 30min under 500rmp, obtain solution b, its
The mass ratio of middle gelatin and carboxymethyl chitosan is 2：1；
3) preparation molar concentration is the ammonium dibasic phosphate solution of 1.0mol/L；
4) solution b is placed in heating water bath agitating device, under the conditions of being stirred continuously at 20 DEG C, by above-mentioned diammonium hydrogen phosphate
Solution is added dropwise over, and wherein, in solution b, Ca ion and the mol ratio of P ion in added ammonium dibasic phosphate solution are 1.67, drip
Add after finishing, continue stirring 0.5h, adjusting reacting liquid pH value by the ammoniacal liquor that mass concentration is 2% in course of reaction is 8；
5) above-mentioned mixed solution is added in hydrothermal reaction kettle, 110 DEG C of hydro-thermal reactions 6h；Reaction terminate after, spend from
Sub- water washing uses absolute ethanol washing, drying again to neutrality, obtains hydroxyapatite/gelatin-carboxymethyl chitosan composite powder
Body；
6) compare composite granule according to quality：Absolute ethyl alcohol=1：1.5 ratio, by above-mentioned hydroxyapatite/gelatin-carboxylic first
Base enclosure glycan composite granule is added in absolute ethyl alcohol, and 700rmp stirs 0.75h, obtains hydroxyapatite/gelatin-carboxymethyl shell
Glycan composite colloid；
7) compare composite granule according to quality：Sodium chloride=1：0.1, composite granule is 1 with the mass ratio of Geniposide：0.02,
By particle diameter be 200～250 μm sodium chloride particle and Geniposide be added in above-mentioned complex sol, stirring, injection molding, room temperature is quiet
Put 4h；Then -80 DEG C of pre-freeze 14h, -53 DEG C of freeze-drying 15h, obtain composite；
8) immersion 2h, then freeze-drying in deionized water will after the gained composite demoulding, be placed under room temperature；Weight
Multiple this step 5 time, sodium chloride particle leaching is clean, obtain hydroxyapatite/gelatin-carboxymethyl chitosan compound rest material
Material.
Embodiment 3
A kind of hydroxyapatite/gelatin-carboxymethyl chitosan compound rest, by hydroxyapatite, gelatin, carboxymethyl chitosan
Sugar is constituted, and wherein, hydroxyapatite, gelatin, the mass ratio of carboxymethyl chitosan are 7：2：1.
The preparation method of above-mentioned compound rest, comprises the following steps：
1) accurately weigh four water-calcium nitrate 118g, be dissolved in 500ml deionized water, obtain solution a；
2) weigh gelatin and carboxymethyl chitosan, be added into solution a, stir 30min under 500rmp, obtain solution b, its
The mass ratio of middle gelatin and carboxymethyl chitosan is 3：1；
3) preparation molar concentration is the ammonium dibasic phosphate solution of 1.0mol/L；
4) solution b is placed in heating water bath agitating device, under the conditions of being stirred continuously at 40 DEG C, by above-mentioned diammonium hydrogen phosphate
Solution is added dropwise over, and wherein, in solution b, Ca ion and the mol ratio of P ion in added ammonium dibasic phosphate solution are 1.67, drip
Add after finishing, continue stirring 1h, adjusting reacting liquid pH value by the ammoniacal liquor that mass concentration is 5% in course of reaction is 9；
5) above-mentioned mixed solution is added in hydrothermal reaction kettle, 140 DEG C of hydro-thermal reactions 3h；Reaction terminate after, spend from
Sub- water washing uses absolute ethanol washing, drying again to neutrality, obtains hydroxyapatite/gelatin-carboxymethyl chitosan composite powder
Body；
6) compare composite granule according to quality：Absolute ethyl alcohol=1：2 ratio, by above-mentioned hydroxyapatite/gelatin-carboxymethyl
Shitosan composite granule is added in absolute ethyl alcohol, and 700rmp stirs 0.7h, obtains hydroxyapatite/gelatin-carboxymethyl chitosan
Sugared composite colloid；
7) compare composite granule according to quality：Sodium chloride=1：1, composite granule is 1 with the mass ratio of Geniposide：0.04, bright
Glue is 1 with the mass ratio of Geniposide：0.2, by particle diameter be 250～300 μm sodium chloride particle and Geniposide be added to above-mentioned
In complex sol, stirring, injection molding, room temperature stands 4h；Then -80 DEG C of pre-freeze 17h, -53 DEG C of freeze-drying 18h, obtain composite wood
Material；
8) immersion 4h, then freeze-drying in deionized water will after the gained composite demoulding, be placed under room temperature；Weight
Multiple this step 3 time, sodium chloride particle leaching is clean, obtain hydroxyapatite/gelatin-carboxymethyl chitosan compound rest material
Material.
Embodiment 4：
A kind of hydroxyapatite/gelatin-carboxymethyl chitosan compound rest, by hydroxyapatite, gelatin, carboxymethyl chitosan
Sugar is constituted, and wherein, hydroxyapatite, gelatin, the mass ratio of carboxymethyl chitosan are 8：2：2.
The preparation method of above-mentioned compound rest, comprises the following steps：
1) accurately weigh four water-calcium nitrate 236g, be dissolved in 500ml deionized water, obtain solution a；
2) weigh gelatin and carboxymethyl chitosan, be added into solution a, stir 40min under 1000rmp, obtain solution b,
The mass ratio of wherein gelatin and carboxymethyl chitosan is 4：1；
3) preparation molar concentration is the ammonium dibasic phosphate solution of 1.5mol/L；
4) solution b is placed in heating water bath agitating device, under the conditions of being stirred continuously at 70 DEG C, by above-mentioned diammonium hydrogen phosphate
Solution is added dropwise over, and wherein, in solution b, Ca ion and the mol ratio of P ion in added ammonium dibasic phosphate solution are 1.67, drip
Add after finishing, continue stirring 1.5h, adjusting reacting liquid pH value by the ammoniacal liquor that mass concentration is 10% in course of reaction is 10；
5) above-mentioned mixed solution is added in hydrothermal reaction kettle, 160 DEG C of hydro-thermal reactions 2h；Reaction terminate after, spend from
Sub- water washing uses absolute ethanol washing, drying again to neutrality, obtains hydroxyapatite/gelatin-carboxymethyl chitosan composite powder
Body；
6) compare composite granule according to quality：Absolute ethyl alcohol=1：2 ratio, by above-mentioned hydroxyapatite/gelatin-carboxymethyl
Shitosan composite granule is added in absolute ethyl alcohol, and 1000rmp stirs 0.5h, obtains hydroxyapatite/gelatin-carboxymethyl chitosan
Sugared composite colloid；
7) compare composite granule according to quality：Sodium chloride=2：3, composite granule is 1 with the mass ratio of Geniposide：0.07, will
Particle diameter is 350～400 μm of sodium chloride particle and Geniposide is added in above-mentioned complex sol, stirring, injection molding, and room temperature stands
4h；Then -80 DEG C of pre-freeze 20h, -53 DEG C of freeze-drying 20h, obtain composite；
8) immersion 4h, then freeze-drying in deionized water will after the gained composite demoulding, be placed under room temperature；Weight
Multiple this step 4 time, sodium chloride particle leaching is clean, obtain hydroxyapatite/gelatin-carboxymethyl chitosan compound rest material
Material.
Embodiment 5
A kind of hydroxyapatite/gelatin-carboxymethyl chitosan compound rest, by hydroxyapatite, gelatin, carboxymethyl chitosan
Sugar is constituted, and wherein, hydroxyapatite, gelatin, the mass ratio of carboxymethyl chitosan are 6：1：1.
The preparation method of above-mentioned compound rest, comprises the following steps：
1) accurately weigh four water-calcium nitrate 354g, be dissolved in 500ml deionized water, obtain solution a；
2) weigh gelatin and carboxymethyl chitosan, be added into solution a, stir 20min under 500rmp, obtain solution b, its
The mass ratio of middle gelatin and carboxymethyl chitosan is 5：1；
3) preparation molar concentration is the ammonium dibasic phosphate solution of 0.1mol/L；
4) solution b is placed in heating water bath agitating device, under the conditions of being stirred continuously at 90 DEG C, by above-mentioned diammonium hydrogen phosphate
Solution is added dropwise over, and wherein, in solution b, Ca ion and the mol ratio of P ion in added ammonium dibasic phosphate solution are 1.67, drip
Add after finishing, continue stirring 2h, adjusting reacting liquid pH value by the ammoniacal liquor that mass concentration is 15% in course of reaction is 8；
5) above-mentioned mixed solution is added in hydrothermal reaction kettle, 180 DEG C of hydro-thermal reactions 0.5h；After reaction terminates, spend
Ion water washing uses absolute ethanol washing, drying again to neutrality, obtains hydroxyapatite/gelatin-carboxymethyl chitosan and is combined
Powder；
6) compare composite granule according to quality：Absolute ethyl alcohol=1：3 ratio, by above-mentioned hydroxyapatite/gelatin-carboxymethyl
Shitosan composite granule is added in absolute ethyl alcohol, and 1000rmp stirs 0.5h, obtains hydroxyapatite/gelatin-carboxymethyl chitosan
Sugared composite colloid；
7) compare composite granule according to quality：Sodium chloride=1：2, composite granule is 1 with the mass ratio of Geniposide：0.1, by grain
Footpath is 450～500 μm of sodium chloride particle and Geniposide is added in above-mentioned complex sol, stirring, injection molding, and room temperature stands
4h；Then -80 DEG C of pre-freeze 24h, -53 DEG C of freeze-drying 24h, obtain composite；
8) immersion 6h, then freeze-drying in deionized water will after the gained composite demoulding, be placed under room temperature；Weight
Multiple this step 3 time, sodium chloride particle leaching is clean, obtain hydroxyapatite/gelatin-carboxymethyl chitosan compound rest material
Material.
Although the above-mentioned accompanying drawing that combines is described to the specific embodiment of the present invention, not model is protected to the present invention
The restriction enclosed, one of ordinary skill in the art should be understood that on the basis of technical scheme, and those skilled in the art are not
Need to pay the various modifications that creative work can make or deformation still within protection scope of the present invention.
Claims (1)
1. hydroxyapatite/gelatin-carboxymethyl chitosan compound rest is it is characterised in that by hydroxyapatite, gelatin, carboxylic first
Base enclosure glycan is constituted, and wherein, hydroxyapatite, gelatin, the mass ratio of carboxymethyl chitosan are (60～80)：(10～20)：(10
～20)；
The preparation method of described hydroxyapatite/gelatin-carboxymethyl chitosan compound rest, comprises the following steps：
1) four water-calcium nitrate is dissolved in water, obtains the aqueous solution that concentration is 0.2～3mol/L, be designated as solution a；
2) weigh gelatin and carboxymethyl chitosan, add it to solution a, stir 20～40min under 500～1000rmp, obtain
The mass ratio of solution b, wherein gelatin and carboxymethyl chitosan is (1～5)：1；
3) prepare ammonium dibasic phosphate solution；
4) solution b is placed in heating water bath agitating device, is stirred continuously at 20～90 DEG C, by step 3) the phosphoric acid hydrogen two prepared
Ammonium salt solution is added dropwise over, and wherein, in solution b, Ca ion and the mol ratio of P ion in added ammonium dibasic phosphate solution are 1.67,
After completion of dropping, continue stirring 0.5～2h, adjusting reacting liquid pH value in course of reaction is 8～12；
5) by step 4) mixed solution that obtains is in 110～180 DEG C of hydro-thermal reaction 0.5～6h；After reaction terminates, deionized water
Wash to neutrality and use absolute ethanol washing, drying again, obtain hydroxyapatite/gelatin-carboxymethyl chitosan composite granule；
6) above-mentioned hydroxyapatite/gelatin-carboxymethyl chitosan composite granule is added in absolute ethyl alcohol, 500～1000rmp
Stirring 0.5～2h, obtains hydroxyapatite/gelatin-carboxymethyl chitosan composite colloid, wherein hydroxyapatite/gelatin-carboxylic first
Base enclosure glycan composite granule is 1 with the mass ratio of absolute ethyl alcohol：(1～3)；
7) sodium chloride particle and Geniposide are added in above-mentioned complex sol, stirring, injection molding, room temperature stands 2～4h, wherein
Hydroxyapatite/gelatin-carboxymethyl chitosan composite granule is 1 with the mass ratio of sodium chloride：(0.1～2), hydroxyapatite/
Gelatin-carboxymethyl chitosan composite granule is 1 with the mass ratio of Geniposide：(0.01～0.1)；Then -80 DEG C of pre-freezes 10～
24h, -53 DEG C of freeze-drying 15～24h, obtain hydroxyapatite/gelatin-carboxymethyl chitosan composite；
8) by after gained hydroxyapatite/gelatin-carboxymethyl chitosan composite demoulding, under room temperature, it is placed on deionized water
Middle immersion 2～6h, then freeze-drying；Repeat this step 3～5 time, sodium chloride particle leaching is clean, that is, obtain hydroxy-apatite
Stone/gelatin-carboxymethyl chitosan compound support frame material；
Wherein, step 3) in, the molar concentration of described diammonium hydrogen phosphate is 0.1～1.5mol/L；
Step 4) in pH value adjusted by ammoniacal liquor, wherein the mass concentration of ammoniacal liquor be 2%～15%；
Step 7) in sodium chloride particle particle diameter be 150～500 μm.
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| CN107137766B (en) * | 2017-05-03 | 2019-06-11 | 中国矿业大学 | The preparation method at antimicrobial artificial arthrodesis interface |
| CN107236145A (en) * | 2017-06-04 | 2017-10-10 | 西北师范大学 | The preparation and application of soybean protein composite hydroxylapatite net high-polymer material |
| CN107281547A (en) * | 2017-07-03 | 2017-10-24 | 扬州大学 | A kind of preparation method of injection bone renovating material |
| CN107376014A (en) * | 2017-07-20 | 2017-11-24 | 莆田学院 | A kind of method of the dicarboxylic acids modification of chitosan bio-mimetic syntheses hydroxyapatite of imidazoles 4,5 |
| CN108892124B (en) * | 2018-06-25 | 2021-11-05 | 广东药科大学附属第一医院 | Method for preparing graphene/nano hydroxyapatite composite material and application |
| CN113209376B (en) * | 2021-05-05 | 2022-07-26 | 西北工业大学 | Normal-temperature neutral preparation method of functional HA/CMCS composite biological ceramic bone scaffold with toughness |
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