CN109224134A - A kind of novel inducting osseous tissue regeneration duplicature and preparation method thereof - Google Patents
A kind of novel inducting osseous tissue regeneration duplicature and preparation method thereof Download PDFInfo
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- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
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- A61L27/00—Materials for grafts or prostheses or for coating grafts or prostheses
- A61L27/14—Macromolecular materials
- A61L27/22—Polypeptides or derivatives thereof, e.g. degradation products
- A61L27/24—Collagen
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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
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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
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Abstract
The invention discloses a kind of novel inducting osseous tissue regeneration duplicatures and preparation method thereof, the duplicature for preparing including I-type collagen and polycaprolactone and coated in -8 coating of nano-sized zeolites imidazate framework material on duplicature, preparation method includes the following steps: S1) prepare duplicature；S2) dry disinfection；S3) -8 coating of coated with nano grade zeolitic imidazolate framework material.The present invention solves traditional collagem membrane bad mechanical property and easily collapses, too fast technological deficiency of degrading in vivo, have good biocompatibility, be conducive to cell grow into and soft tissue healing.
Description
Technical field
Oral medical Material Field of the present invention, in particular to a kind of novel inducting osseous tissue regeneration duplicature and
Preparation method.
Background technique
As the improvement of people's living standards, more patients start to consider that plantation is repaired to restore defect of dentition or denture
Missing, and then restore chewing, function of pronunciation and beauty.In plantation is repaired, sufficient sclerotin bone amount is that implant operation is successful
Guarantee, and Guided Bone Regeneration (guided bone regeneration GBR) technology is to be implanted into bone grafting in the region of Bone mineral change
Material, and prevent from growing faster epithelial cell and fibroblast as physical barriers in surface covering Guided Bone Regeneration film
It grows into, the bone tissue and blood vessel to grow slower provide space.Whether inducting osseous tissue regeneration film foundation can be absorbed and be divided into
Two classes, nonabsorable film and absorbability film.Nonabsorable film mainly includes polytetrafluoroethylene film, titanium film and miillpore filter
Deng because can not be absorbed by organisms, second operation being needed to take out, made troubles to patient and financial burden.Absorbability degradation
Film is not necessarily to take out after implanting, and reduces the pain and operating difficulty of patient, and current degradable biological film reported in the literature has
Two kinds, one kind is collagem membrane (such as commercialized product Bio-Gide collagem membrane) made of animal collagen, and another kind is with poly-
Lactic acid, the artificial membrane of the synthesis of polymer materials such as polycaprolactone, such as patent CN201410812046.3 and
The guide tissue regeneration film of CN201620047619.2 report.Collagem membrane is because it is with preferable biocompatibility, and in clinic
In be widely used, but still come with some shortcomings, such as expensive, there is enough mechanical screens at rigid implant surgery position
Barrier effect, but be likely to occur mechanical strength within a short period of time and decline rapidly, it supports barrier action can gradually weaken to disappear, occurs
Film is collapsed and is shifted, degradable, and without antibacterial activity, is influenced the effect of osteanagenesis, limited its clinical application.
Metal-organic framework (Metal-organic frameworks, MOFs) is a kind of using metal ion as matching
Centrical novel crystal class material has high specific surface area, and high porosity, structure diversity and unsaturated metal are matched
Position key, is conducive to modify or be grafted other molecular substances, gets in recent years in the application that field of biomedicine especially carries medicine field
Come more extensive, ZIF-8 is a kind of MOFs material with self-bone grafting ability, has structure function adjustability, osteoinductive
With load medicine potential, gradually it is taken seriously in organizational project.
Polycaprolactone (polycaprolactone, PCL) is a kind of synthesis polyphosphazene polymer ester material developed in recent years, tool
There are the features such as good Physical Mechanical, chemical stability, histocompatbility, drug passability and easy processing shape, and warp
A kind of biodegradable polymer for crossing Food and Drug Adminstration of the US (FDA) certification, in its natural state then easily by microorganism
Enzyme decomposition is carried out, final complete decomposition product is water and carbon dioxide nontoxic to the human body.
I-type collagen (COL) is the most abundant one kind in various collagens contained in human body, be present in muscle,
Skin, arterial wall in fibrocartilage, can be extracted from animal tissue, have excellent biological property, as low antigenicity,
The behaviors such as adherency, proliferation, the differentiation of hemoglutination and adjustable ganglion cell are best substitution material in terms of biocompatibility
Material, and be conducive to cell grow into and soft tissue healing.
Zinc is one of the essential trace elements of the human body, and the Zn-ef ficiency of human body about 50% is distributed among tooth and bone, health
The daily zinc intake of adult is 15mg.Zn has important adjustment effect to normal development, the metabolism of bone.The study found that
Alkaline phosphatase (Alkaline phosphatase, ALP) is adjusting enzyme important in skeletonization mineralization process, and zinc is as ALP's
Prothetic group plays an important role in bone mineralization process.In addition, Zn-ef ficiency is mixed the bone renovating material of calcium phosphate by a large amount of scholar
In, assign its Zn2+Slow release effect, result of study shows Zn2+Addition improve this kind of bone renovating material skeletonization effect
Fruit.In addition, zinc can also promote bone marrow mesenchymal stem cells secrete cytokines stimulating endothelial cell at blood vessel function, have indirectly
At the effect of blood vessel.
Summary of the invention
The purpose of the present invention is to provide a kind of novel inducting osseous tissue regeneration duplicatures, and it is mechanical to solve traditional collagem membrane
Performance difference is easily collapsed, and too fast technological deficiency of degrading in vivo has good biocompatibility, is conducive to cell and is grown into and soft group
Knit healing.
The present invention also provides a kind of preparation methods of novel inducting osseous tissue regeneration duplicature, using high-voltage electrostatic spinning
Method prepares duplicature in two times, then uses hydro-thermal method -8 coating of dip-coating nano-sized zeolites imidazate framework material on duplicature,
Obtain the finely dispersed regeneration duplicature of -8 coating of nano-sized zeolites imidazate framework material.
The present invention is achieved through the following technical solutions: a kind of novel inducting osseous tissue regeneration duplicature, including type i collagen egg
The duplicature of white and polycaprolactone preparation and coated in -8 coating of nano-sized zeolites imidazate framework material on duplicature.
Further, -8 coating of nano-sized zeolites imidazate framework material is prepared by hydro-thermal method.
Further, the duplicature is prepared using high-voltage electrostatic spinning method.The duplicature uses high-pressure electrostatic
Spin processes are prepared twice: high-voltage electrostatic spinning method preparation the first tunic (I-type collagen-polycaprolactone film) is first passed through,
The second tunic is prepared using high-voltage electrostatic spinning method on the first tunic prepared again, it is preferably double-deck to obtain mechanical performance
Film.
The novel inducting osseous tissue regeneration duplicature is prepared by the following method:
S1 it) prepares duplicature: I-type collagen and polycaprolactone is dissolved in formation blend spinning liquid, magnetic force in hexafluoroisopropanol
Stirring to blend spinning liquid is clarified, and is then extracted blend spinning liquid with syringe, is prepared first layer by high-voltage electrostatic spinning method
Film；Then blend spinning liquid is extracted with syringe again, the second tunic is prepared by high-voltage electrostatic spinning method on the first tunic, is obtained
To duplicature；
S2) dry disinfection: the duplicature of preparation is dried in 35 ~ 45 DEG C of baking ovens again after ventilation is dried, after disinfection to reserve；
S3) -8 coating of coated with nano grade zeolitic imidazolate framework material: take 2-methylimidazole and zinc nitrate hexahydrate be dissolved in from
In sub- water, crystal growth liquid is stirred to get, then immerses the duplicature after disinfection in step S2) in crystal growth liquid, is stood
30min takes out, and obtains product after elution is dry.
Further, in the blend spinning liquid the sum of I-type collagen and the mass concentration of polycaprolactone be 7.5 ~
8.5g/ml。
Further, the step S1) in I-type collagen and gather oneself in the blend spinning liquid that uses of the first tunic of preparation
The mass ratio of lactone is 1:1；Prepare the mass ratio of I-type collagen and polycaprolactone in the blend spinning liquid of the second tunic use
For 1.5:1.
Further, it is used during the first layer film preparation and injects speed as 0.25mL/h, reception speed is 120r/
min；What is used during the second layer film preparation injects speed as 0.20mL/h, and reception speed is 120r/min.
Further, the molar ratio of 2-methylimidazole and zinc nitrate hexahydrate is 60 ~ 80:1, institute in the crystal growth liquid
The concentration for stating 2-methylimidazole in crystal growth liquid is 6 ~ 8g/L, and the concentration of the zinc nitrate hexahydrate is 0.2 ~ 0.5g/L.
Further, the step S2) in, the sterilization method of duplicature uses the Co of 20kGy metering60Irradiation 5 ~
8h。
Compared with prior art, the present invention have the following advantages that and the utility model has the advantages that
(1) the present invention provides a kind of novel inducting osseous tissue regeneration duplicature, metal-organic framework materials coating is answered for the first time
For the modification of polycaprolactone-I-type collagen duplicature surface and osteanagenesis field, solves traditional collagem membrane mechanical performance
Poor easily to collapse, too fast technological deficiency of degrading in vivo has good biocompatibility, is conducive to that cell is grown into and soft tissue is cured
It closes.
(2) a kind of novel inducting osseous tissue regeneration duplicature provided by the invention and preparation method thereof, raw material are easy to obtain
, competitively priced, machinability is good, and operating method is simple, is suitable for large range of osteanagenesis application.
Detailed description of the invention
Fig. 1 is the electron microscope of novel inducting osseous tissue regeneration duplicature prepared by the present invention；
Fig. 2 is the electron microscope of collagem membrane；
Fig. 3 is that the stretching of novel inducting osseous tissue regeneration duplicature prepared by the present invention and collagem membrane detects comparison diagram；
Fig. 4 is the elasticity modulus comparison diagram of novel inducting osseous tissue regeneration duplicature and collagem membrane prepared by the present invention；
Fig. 5 is the degradation property comparison diagram of novel inducting osseous tissue regeneration duplicature and collagem membrane prepared by the present invention；
Fig. 6 is the cell compatibility comparison diagram of novel inducting osseous tissue regeneration duplicature and collagem membrane prepared by the present invention；
Fig. 7 is novel inducting osseous tissue regeneration duplicature prepared by the present invention and collagem membrane and mesenchymal stem cell adhesiveness
It can comparison diagram；
Fig. 8 is novel inducting osseous tissue regeneration duplicature prepared by the present invention and collagem membrane and Human umbilical vein endothelial cells adhesiveness
It can comparison diagram；
Fig. 9 is novel inducting osseous tissue regeneration duplicature prepared by the present invention and collagem membrane skeletonization comparison diagram；
Figure 10 is novel inducting osseous tissue regeneration duplicature prepared by the present invention and collagem membrane into blood vessel comparison diagram.
Specific embodiment
The embodiment of the present invention is described below in detail.The embodiments described below with reference to the accompanying drawings are exemplary, purport
It is being used to explain the present invention, and is being not considered as limiting the invention.
Embodiment 1:
A kind of novel inducting osseous tissue regeneration duplicature, the duplicature prepared including I-type collagen and polycaprolactone and coating
- 8 coating of nano-sized zeolites imidazate framework material on duplicature.
The novel inducting osseous tissue regeneration duplicature the preparation method is as follows:
S1 it) prepares duplicature: weighing I-type collagen and polycaprolactone, be according to I-type collagen and polycaprolactone mass ratio
The ratio of 1:1 is dissolved in the first blend spinning liquid for being prepared into that the mass fraction of solute is 7.5% in hexafluoroisopropanol, according to I type glue
The ratio that former albumen and polycaprolactone mass ratio are 1:1.5 is dissolved in that the mass fraction of solute is prepared into hexafluoroisopropanol is 7.5%
The second blend spinning liquid, after magnetic agitation 8h, first extract the first blend spinning liquid of 20ml 1:1 respectively with two syringes,
It is fixed on constant flow pump, the anode of HV generator is connected on syringe needle, cathode is connected on receiver board, receiver board upper berth
There is one layer of aluminium-foil paper, for receiving electrostatic spinning fiber, sets and inject flow velocity as 0.25ml/h, reception speed is 120r/min, is spun
Silk is completed to obtain the first tunic, then the second blend spinning liquid of the 1:1.5 with other two syringe extraction 20ml, is fixed on perseverance
It on stream pump, sets and injects flow velocity as 0.20mL/h, reception speed is that 120r/min removes aluminium-foil paper after the completion of spinning, is obtained
Duplicature.
S2) dry disinfection: the duplicature of preparation is placed on 3 ~ 5h of placement in ventilating kitchen, sets in 37 DEG C of baking ovens and dries for 24 hours, solely
The Co measured after vertical packaging by 20kGy60It is spare after 5 h of illumination-based disinfection；
S3 2-methylimidazole and six) -8 coating of coated with nano grade zeolitic imidazolate framework material: are weighed according to the molar ratio of 60:1
Nitric hydrate zinc is dissolved in deionized water, stir to get 2-methylimidazole concentration be 6g/L, the zinc nitrate hexahydrate it is dense
Degree is the crystal growth liquid of 0.2g/L, continues stirring to crystal growth liquid and is creamy white, then will be double after disinfection in step S2)
Tunic immerses in crystal growth liquid, stands 30min, takes out, and after being eluted three times with deionized water, is dried at room temperature for obtaining product
1。
Embodiment 2:
The novel inducting osseous tissue regeneration duplicature the preparation method is as follows:
S1 it) prepares duplicature: weighing I-type collagen and polycaprolactone, be according to I-type collagen and polycaprolactone mass ratio
The ratio of 1:1 is dissolved in the first blend spinning liquid for being prepared into that the mass fraction of solute is 8.5% in hexafluoroisopropanol, according to I type glue
The ratio that former albumen and polycaprolactone mass ratio are 1:1.5 is dissolved in that the mass fraction of solute is prepared into hexafluoroisopropanol is 8.5%
The second blend spinning liquid, after magnetic agitation 12h, first extract the first blend spinning of 20ml 1:1 respectively with two syringes
Liquid is fixed on constant flow pump, and the anode of HV generator is connected on syringe needle, and cathode is connected on receiver board, receiver board
On be covered with one layer of aluminium-foil paper, for receiving electrostatic spinning fiber, set and inject flow velocity as 0.25ml/h, reception speed is 120r/
Min, spinning are completed to obtain the first tunic, then the second blend spinning liquid of the 1:1.5 with other two syringe extraction 20ml, Gu
It is scheduled on constant flow pump, sets and inject flow velocity as 0.20mL/h, reception speed is that 120r/min removes aluminium foil after the completion of spinning
Paper obtains duplicature.
S2) dry disinfection: the duplicature of preparation is placed on 3 ~ 5h of placement in ventilating kitchen, sets in 45 DEG C of baking ovens and dries for 24 hours, solely
The Co measured after vertical packaging by 20kGy60It is spare after 8 h of illumination-based disinfection；
S3 2-methylimidazole and six) -8 coating of coated with nano grade zeolitic imidazolate framework material: are weighed according to the molar ratio of 80:1
Nitric hydrate zinc is dissolved in deionized water, stir to get 2-methylimidazole concentration be 8g/L, the zinc nitrate hexahydrate it is dense
Degree is the crystal growth liquid of 0.5g/L, continues stirring to crystal growth liquid and is creamy white, then will be double after disinfection in step S2)
Tunic immerses in crystal growth liquid, stands 20min, takes out, and after being eluted three times with deionized water, is dried at room temperature for obtaining product
2。
Embodiment 3:
S1 it) prepares duplicature: weighing I-type collagen and polycaprolactone, be according to I-type collagen and polycaprolactone mass ratio
The ratio of 1:1 is dissolved in the first blend spinning liquid for being prepared into that the mass fraction of solute is 8% in hexafluoroisopropanol, according to type i collagen
The ratio that albumen and polycaprolactone mass ratio are 1:1.5 be dissolved in be prepared into hexafluoroisopropanol solute mass fraction be 8%
Two blend spinning liquid after magnetic agitation 10h, first extract the first blend spinning liquid of 20ml 1:1 respectively with two syringes, Gu
It is scheduled on constant flow pump, the anode of HV generator is connected on syringe needle, cathode is connected on receiver board, is covered on receiver board
One layer of aluminium-foil paper sets for receiving electrostatic spinning fiber and injects flow velocity as 0.25ml/h, reception speed is 120r/min, spinning
Completion obtains the first tunic, then the second blend spinning liquid of the 1:1.5 with other two syringe extraction 20ml, is fixed on constant current
It on pump, sets and injects flow velocity as 0.20mL/h, reception speed is that 120r/min removes aluminium-foil paper after the completion of spinning, is obtained
Duplicature.
S2) dry disinfection: the duplicature of preparation is placed on 3 ~ 5h of placement in ventilating kitchen, sets in 37 DEG C of baking ovens and dries for 24 hours, solely
The Co measured after vertical packaging by 20kGy60It is spare after 6 h of illumination-based disinfection；
S3 2-methylimidazole and six) -8 coating of coated with nano grade zeolitic imidazolate framework material: are weighed according to the molar ratio of 74:1
Nitric hydrate zinc is dissolved in deionized water, stir to get 2-methylimidazole concentration be 7g/L, the zinc nitrate hexahydrate it is dense
Degree is the crystal growth liquid of 0.35g/L, continues stirring to crystal growth liquid and is creamy white, then will be after disinfection in step S2)
Duplicature immerses in crystal growth liquid, stands 10min, takes out, and after being eluted three times with deionized water, is dried at room temperature for being produced
Product 3.
Contrast sample 1
Collagem membrane as a comparison sample 1 is used in the embodiment of the present invention, the collagen is membrane derived for the positive marine growth sea Austria in Yantai
Commercially available oral restoration film, main component are collagens.
Sample detection
Wherein, POL-COL-ZIF8 is novel inducting osseous tissue regeneration duplicature prepared by the present invention, and it is not that material, which is not added,
The cell tissue of novel inducting osseous tissue regeneration duplicature prepared by the present invention is added.
Sample 3 obtained in embodiment 4 and contrast sample 1 are obtained into electron microscope, institute as shown in Figure 1, Figure 2 by electron-microscope scanning
Show, the duplicature prepared by high-voltage electrostatic spinning method forms criss-cross fibrous layer, mechanical performance by electrostatic spinning fiber
Preferably, it is not easy to collapse, while nano-sized zeolites imidazate framework material -8 is evenly distributed on the electrostatic spinning fibre to form duplicature
In dimension, the specific surface area of duplicature is increased, structure diversity is made it have, is conducive to and cell adherence.
By in embodiment 4 sample 3 and contrast sample 1 carry out extension test and elasticity modulus test, test method is such as
Under: at room temperature, sample 3 and contrast sample 1 are cut into the cuboid of 10*30*0.35mm, using the omnipotent test of Instron
Instrument tests its tensile strength and elasticity modulus, and test results are shown in figure 3.In fig. 3 a as can be seen that sample 3 is in dry shape
Maximum tension load under state and moisture state is higher than collagem membrane, shows that sample 3 has compared to collagem membrane and preferably draws
Stretch performance；Simultaneously it can be seen in figure 3 b that elasticity modulus of the sample 3 under drying regime and moisture state more than collagem membrane
Height shows that sample 3 has better elastic property compared to collagem membrane, and therefore, novel inducting osseous tissue disclosed by the invention is again
Raw duplicature has better mechanical performance compared to collagem membrane.
By in embodiment 4 sample 3 and contrast sample 1 carry out degradation test, test method are as follows: by film immerse PBS in,
37 DEG C of constant temperature are placed, and are taken the film out after 1 week, 2 weeks, 3 weeks, 4 weeks, 5 weeks, 6 weeks, 7 weeks, are weighed after dry.It is dropped in 42d
Mass excess rate is as shown in Figure 5 after solution.In figure 4, it can be seen that be degraded after 42d, the mass excess rate of sample 3 less than 20%,
And the mass excess rate of collagem membrane is more than 60%, the mass excess rate of sample 3 shows well below the mass excess rate of collagem membrane
Sample 3 is easier to be degraded and absorbed.
By in embodiment 4 sample 3 and contrast sample 1, contrast sample 1 by carry out cell compatibility test, test
Method are as follows: take two groups of samples 3 and contrast sample 1 respectively, be inoculated with mesenchymal stem cell, people's navel respectively on two groups of samples 3
Venous endothelial cell carries out CCK-8 detection increment detection in 1d, 4d, 7d, between being inoculated with marrow respectively in two groups of contrast samples 1
Mesenchymal stem cells, Human umbilical vein endothelial cells carry out CCK-8 detection increment detection in 1d, 4d, 7d, test sample 3, comparison
The cell compatibility of sample 1 and contrast sample 1 and mesenchymal stem cell, Human umbilical vein endothelial cells, test result is such as
Shown in Fig. 5, Fig. 6.From Fig. 5, Fig. 6 as can be seen that sample 3 and mesenchymal stem cell, Human umbilical vein endothelial cells it is thin
Cell phase capacitive is slightly above the cell compatibility of collagem membrane and mesenchymal stem cell, Human umbilical vein endothelial cells, also above
The cell compatibility of contrast sample 1 and mesenchymal stem cell, Human umbilical vein endothelial cells is therefore, disclosed by the invention new
Type inducting osseous tissue regeneration duplicature has good biocompatibility, and the addition of nano-sized zeolites imidazate framework material -8
Can increase regeneration duplicature cell compatibility, be conducive to cell grow into and soft tissue healing.
By in embodiment 4 sample 3 and contrast sample 1 carry out mesenchymal stem cell respectively with sample 3 and comparative sample
The adhesiveness of product 1 is tested, test method are as follows: be inoculated in mesenchymal stem cell respectively in sample 3 and contrast sample 1 and train
It supports 1 day, uses mass fraction to fix for 4% paraformaldehyde, rinsed using PBS, distinguish using DAPI and FITC- phalloidine
Nucleus and cell myofilament F-actin skeleton to mesenchymal stem cell carry out Fluorescent Staining Observation, experimental result such as Fig. 7
It is shown.As shown in fig. 7, adherency density of the mesenchymal stem cell on sample 3 is higher than mesenchymal stem cell in collagen
Density is adhered on film, is shown compared to collagem membrane, and mesenchymal stem cell is easier to be adhered to novel guidance disclosed by the invention
On bone tissue regeneration duplicature.
By in embodiment 4 sample 3 and contrast sample 1 by carry out Human umbilical vein endothelial cells respectively with sample 3 and right
Adhesiveness than sample 1 is tested, test method are as follows: Human umbilical vein endothelial cells are inoculated in sample 3 and contrast sample respectively
On, it cultivates 1 day, uses mass fraction to fix for 4% paraformaldehyde, rinsed using PBS, using DAPI and FITC- phalloidine
Fluorescent Staining Observation, experiment knot are carried out to the nucleus of Human umbilical vein endothelial cells and cell myofilament F-actin skeleton respectively
Fruit is as shown in Figure 8.As shown in figure 8, adherency density of the Human umbilical vein endothelial cells on sample 3 is higher than Human umbilical vein endothelial cells
Density is adhered on collagem membrane, is shown compared to collagem membrane, and Human umbilical vein endothelial cells are easier to be adhered to disclosed by the invention new
On type inducting osseous tissue regeneration duplicature.
By in embodiment 4 sample 3 and contrast sample 1 carry out the promotion to mesenchymal stem cell early stage skeletonization effect
Fruit, test method are as follows: mesenchymal stem cell (BMSCs) is inoculated on film, osteogenic induction, cultivates 7d, detect early period
Skeletonization marker alkaline phosphatase (ALP), the image after alkaline phosphatase staining 7d are as shown in Figure 9.As shown in figure 9, sample 3
Collagem membrane is substantially better than to the osteogenic induction effect of mesenchymal stem cell to imitate the osteogenic induction of mesenchymal stem cell
Fruit shows that, compared to collagem membrane, novel inducting osseous tissue regeneration duplicature prepared by the embodiment of the present invention 4 has better skeletonization
Activity.
Test sample 3 and contrast sample 1 are to Human umbilical vein endothelial cells at pipe ability, and matrigel is at the figure after pipe 6h
As shown in Figure 10.As shown in Figure 10, sample 3 is substantially better than collagem membrane at pipe effect to the promotion of Human umbilical vein endothelial cells
Promote into pipe effect, show compared to collagem membrane, novel inducting osseous tissue regeneration duplicature disclosed by the invention has better
Angiogenesis.
Therefore, novel inducting osseous tissue regeneration duplicature disclosed by the invention has skeletonization at blood vessel economic benefits and social benefits activity, significantly
Improve the predictability of film inducting osseous tissue regeneration.
The above is only presently preferred embodiments of the present invention, not does limitation in any form to the present invention, it is all according to
According to technical spirit any simple modification to the above embodiments of the invention, equivalent variations, protection of the invention is each fallen within
Within the scope of.
Claims (9)
1. a kind of novel inducting osseous tissue regeneration duplicature, it is characterised in that: prepared including I-type collagen and polycaprolactone
Duplicature and coated in -8 coating of nano-sized zeolites imidazate framework material on duplicature.
2. a kind of novel inducting osseous tissue regeneration duplicature according to claim 1, it is characterised in that: the nanoscale boiling
- 8 coating of stone imidazate framework material is prepared by hydro-thermal method.
3. a kind of novel inducting osseous tissue regeneration duplicature according to claim 1, it is characterised in that: the duplicature is adopted
It is prepared with high-voltage electrostatic spinning method.
4. the preparation method of described in any item a kind of novel inducting osseous tissue regeneration duplicatures according to claim 1 ~ 3, special
Sign is: the following steps are included:
S1 it) prepares duplicature: I-type collagen and polycaprolactone is dissolved in formation blend spinning liquid, magnetic force in hexafluoroisopropanol
Stirring to blend spinning liquid is clarified, and is then extracted blend spinning liquid with syringe, is prepared first layer by high-voltage electrostatic spinning method
Film；Then blend spinning liquid is extracted with syringe again, the second tunic is prepared by high-voltage electrostatic spinning method on the first tunic, is obtained
To duplicature；
S2) dry disinfection: the duplicature of preparation is dried in 35 ~ 45 DEG C of baking ovens again after ventilation is dried, after disinfection to reserve；
S3) -8 coating of coated with nano grade zeolitic imidazolate framework material: take 2-methylimidazole and zinc nitrate hexahydrate be dissolved in from
In sub- water, crystal growth liquid is stirred to get, then immerses the duplicature after disinfection in step S2) in crystal growth liquid, is stood
30min takes out, and obtains product after elution is dry.
5. a kind of preparation method of novel inducting osseous tissue regeneration duplicature according to claim 4, it is characterised in that: institute
Stating the sum of I-type collagen and the mass concentration of polycaprolactone in blend spinning liquid is 7.5 ~ 8.5g/ml.
6. a kind of preparation method of novel inducting osseous tissue regeneration duplicature according to claim 5, it is characterised in that: institute
State step S1) in the blend spinning liquid that uses of the first tunic of preparation the mass ratio of I-type collagen and polycaprolactone for 1:1；
The mass ratio of I-type collagen and polycaprolactone is prepared in the blend spinning liquid that the second tunic uses as 1.5:1.
7. a kind of preparation method of novel inducting osseous tissue regeneration duplicature according to claim 4, it is characterised in that: institute
It states use during first layer film preparation and injects speed as 0.25mL/h, reception speed is 120r/min；The second tunic system
What is used during standby injects speed as 0.20mL/h, and reception speed is 120r/min.
8. a kind of preparation method of novel inducting osseous tissue regeneration duplicature according to claim 4, it is characterised in that: institute
Stating the molar ratio of 2-methylimidazole and zinc nitrate hexahydrate in crystal growth liquid is 60 ~ 80:1,2- first in the crystal growth liquid
The concentration of base imidazoles is 6 ~ 8g/L, and the concentration of the zinc nitrate hexahydrate is 0.2 ~ 0.5g/L.
9. a kind of preparation method of novel inducting osseous tissue regeneration duplicature according to claim 4, it is characterised in that: institute
State step S2) in, the sterilization method of duplicature uses the Co of 20kGy metering60Irradiate 5 ~ 8h.
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