CN108404215A - Small-caliber artificial blood vessel preparation method based on electrostatic spinning - Google Patents
Small-caliber artificial blood vessel preparation method based on electrostatic spinning Download PDFInfo
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- CN108404215A CN108404215A CN201810559364.1A CN201810559364A CN108404215A CN 108404215 A CN108404215 A CN 108404215A CN 201810559364 A CN201810559364 A CN 201810559364A CN 108404215 A CN108404215 A CN 108404215A
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- A—HUMAN NECESSITIES
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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
- A61L27/14—Macromolecular materials
- A61L27/22—Polypeptides or derivatives thereof, e.g. degradation products
- A61L27/227—Other specific proteins or polypeptides not covered by A61L27/222, A61L27/225 or A61L27/24
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- A61L27/00—Materials for grafts or prostheses or for coating grafts or prostheses
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61L—METHODS OR APPARATUS FOR STERILISING MATERIALS OR OBJECTS IN GENERAL; DISINFECTION, STERILISATION OR DEODORISATION OF AIR; CHEMICAL ASPECTS OF BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES; MATERIALS FOR BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES
- A61L27/00—Materials for grafts or prostheses or for coating grafts or prostheses
- A61L27/14—Macromolecular materials
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- A61L27/24—Collagen
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- A61L27/00—Materials for grafts or prostheses or for coating grafts or prostheses
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- A61L27/00—Materials for grafts or prostheses or for coating grafts or prostheses
- A61L27/50—Materials characterised by their function or physical properties, e.g. injectable or lubricating compositions, shape-memory materials, surface modified materials
- A61L27/507—Materials characterised by their function or physical properties, e.g. injectable or lubricating compositions, shape-memory materials, surface modified materials for artificial blood vessels
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- A61L27/00—Materials for grafts or prostheses or for coating grafts or prostheses
- A61L27/50—Materials characterised by their function or physical properties, e.g. injectable or lubricating compositions, shape-memory materials, surface modified materials
- A61L27/54—Biologically active materials, e.g. therapeutic substances
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- D—TEXTILES; PAPER
- D01—NATURAL OR MAN-MADE THREADS OR FIBRES; SPINNING
- D01D—MECHANICAL METHODS OR APPARATUS IN THE MANUFACTURE OF ARTIFICIAL FILAMENTS, THREADS, FIBRES, BRISTLES OR RIBBONS
- D01D5/00—Formation of filaments, threads, or the like
- D01D5/0007—Electro-spinning
- D01D5/0015—Electro-spinning characterised by the initial state of the material
- D01D5/003—Electro-spinning characterised by the initial state of the material the material being a polymer solution or dispersion
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- D—TEXTILES; PAPER
- D01—NATURAL OR MAN-MADE THREADS OR FIBRES; SPINNING
- D01D—MECHANICAL METHODS OR APPARATUS IN THE MANUFACTURE OF ARTIFICIAL FILAMENTS, THREADS, FIBRES, BRISTLES OR RIBBONS
- D01D5/00—Formation of filaments, threads, or the like
- D01D5/0007—Electro-spinning
- D01D5/0061—Electro-spinning characterised by the electro-spinning apparatus
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- D—TEXTILES; PAPER
- D01—NATURAL OR MAN-MADE THREADS OR FIBRES; SPINNING
- D01D—MECHANICAL METHODS OR APPARATUS IN THE MANUFACTURE OF ARTIFICIAL FILAMENTS, THREADS, FIBRES, BRISTLES OR RIBBONS
- D01D5/00—Formation of filaments, threads, or the like
- D01D5/0007—Electro-spinning
- D01D5/0061—Electro-spinning characterised by the electro-spinning apparatus
- D01D5/0069—Electro-spinning characterised by the electro-spinning apparatus characterised by the spinning section, e.g. capillary tube, protrusion or pin
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- D—TEXTILES; PAPER
- D01—NATURAL OR MAN-MADE THREADS OR FIBRES; SPINNING
- D01D—MECHANICAL METHODS OR APPARATUS IN THE MANUFACTURE OF ARTIFICIAL FILAMENTS, THREADS, FIBRES, BRISTLES OR RIBBONS
- D01D5/00—Formation of filaments, threads, or the like
- D01D5/0007—Electro-spinning
- D01D5/0061—Electro-spinning characterised by the electro-spinning apparatus
- D01D5/0076—Electro-spinning characterised by the electro-spinning apparatus characterised by the collecting device, e.g. drum, wheel, endless belt, plate or grid
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- A61L2300/00—Biologically active materials used in bandages, wound dressings, absorbent pads or medical devices
- A61L2300/20—Biologically active materials used in bandages, wound dressings, absorbent pads or medical devices containing or releasing organic materials
- A61L2300/23—Carbohydrates
- A61L2300/236—Glycosaminoglycans, e.g. heparin, hyaluronic acid, chondroitin
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- A61L2300/00—Biologically active materials used in bandages, wound dressings, absorbent pads or medical devices
- A61L2300/40—Biologically active materials used in bandages, wound dressings, absorbent pads or medical devices characterised by a specific therapeutic activity or mode of action
- A61L2300/42—Anti-thrombotic agents, anticoagulants, anti-platelet agents
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Abstract
The small-caliber artificial blood vessel preparation method based on electrostatic spinning that the invention discloses a kind of, includes the following steps：(1) sandwich layer solution is conveyed in electrospinning device and carry out coaxial electrostatic spinning, build artificial blood vessel's internal layer；The sandwich layer solution is heparin sodium aqua, VEGF solution and BSA solution according to 20:20:60 mass ratio mixes；(2) shell solution is conveyed in electrospinning device and carry out coaxial electrostatic spinning, build artificial blood vessel's outer layer；The shell solution is P (LLA CL), collagen and elastin according to 80:10:10 mass ratio is made after being completely dissolved in solvent.The experimental results showed that using artificial blood vessel prepared by the method for the present invention there is good histocompatbility, the postoperative anti-freezing for being not necessarily to anti-freezing or only needing low degree of patient to reduce the risk of drug complication to mitigate minimal invasive treatment's burden.
Description
Technical field
The present invention relates to a kind of small-caliber artificial blood vessel preparation methods more particularly to a kind of based on the small-bore of electrostatic spinning
Artificial blood vessel's preparation method.
Background technology
Small-bore (bore is less than 6mm) artificial blood vessel is the surgical material that current department of cardiac surgery is badly in need of, its technology requirement
It is not only enough intensity and toughness, it is often more important that the influence to blood flow, if blood injury ingredient, and whether form blood
Bolt.Current market sales of mainstream artificial blood vessel has terylene (Dacron) and expanded polytetrafluoroethylsealing (ePTFE), but because interior
Wall easily forms lateral thrombus and loses work(rapidly.Therefore, patient is postoperative must take anticoagulant for a long time or even all the life, this is not only
Increase living burden, also increases the risk of drug complication, the especially use of resistance drug, be the main of hemorrhage of digestive tract
Reason or even threat to life.
Electrostatic spinning technique is a prior art, it is so that polymer solution is charged using plus high-voltage field, works as electricity
When field intensity is powerful enough, the drop of capillary tip overcomes surface tension to form jet stream, and polymer jet stream is from capillary nozzle
Tip is stretched during projecting to receiving terminal, and nano-scale fiber is formed after solvent volatilization.When receiving terminal be rotation roller bearing,
It can be obtained tubular structure.
Invention content
In view of this, the purpose of the present invention is to provide a kind of small-caliber artificial blood vessel preparation side based on electrostatic spinning
Method has good histocompatbility using artificial blood vessel prepared by this method, and patient is postoperative to be not necessarily to anti-freezing or only need low journey
The anti-freezing of degree reduces the risk of drug complication to mitigate minimal invasive treatment's burden.
The small-caliber artificial blood vessel preparation method based on electrostatic spinning of the present invention, includes the following steps：
(1) sandwich layer solution is conveyed in electrospinning device and carry out coaxial electrostatic spinning, build artificial blood vessel's internal layer；It is described
Sandwich layer solution is heparin sodium aqua, VEGF solution and BSA solution according to 20:20:60 mass ratio mixes；
(2) shell solution is conveyed in electrospinning device and carry out coaxial electrostatic spinning, build artificial blood vessel's outer layer；It is described
Shell solution is P (LLA-CL), collagen and elastin according to 80:10:10 mass ratio is made after being completely dissolved in solvent.
Preferably, in step (1), the mass percentage concentration of the heparin sodium aqua is 30%, the heparin sodium aqua
The molecular weight of middle heparin sodium is 20000.
Preferably, in step (1), the VEGF solution is the VEGF of 100ug/ml165Solution.
Preferably, in step (1), the mass percentage concentration of the BSA solution is 0.1%.
Preferably, in step (2), the collagen is ox source type i collagen, molecular weight 100000.
Preferably, in step (2), the solvent is poly- hexafluoroisopropanol, is formed by the quality percentage of shell solution
A concentration of 10%.
Preferably, in step (1) and step (2), the operating voltage of electrospinning device is 16KV, and room temperature is 26 DEG C,
Air humidity is 30%.
Preferably, in step (1) and step (2), the receiving terminal of electrospinning device is the stainless steel shaft of 4mm, described
The rotating speed of stainless steel shaft is 800r/min.
Preferably, in step (1), the fltting speed of sandwich layer solution is 0.1ml/h.
Preferably, in step (2), the fltting speed of shell solution is 1.0ml/h.
By above disclosure, the present invention has following advantageous effects：
The small-caliber artificial blood vessel preparation method based on electrostatic spinning of the present invention, the artificial blood vessel prepared using this method
With good histocompatbility, the postoperative anti-freezing for being not necessarily to anti-freezing or only needing low degree of patient, to alleviate minimal invasive treatment
Burden, reduces the risk of drug complication.
Description of the drawings
Fig. 1 is the structural schematic diagram of electrospinning device used in the present invention；
Fig. 2 is artificial blood vessel's pictorial diagram using bore 4mm prepared by preparation method of the present invention；
Fig. 3 is using artificial blood vessel prepared by preparation method of the present invention and Human saphenous vein comparison diagram；
Fig. 4 is ultra microstructure figure under the mirror for using artificial blood vessel prepared by preparation method of the present invention to amplify 1000 times；
Fig. 5 is the platelet attachment experiment result of ePTEE blood vessels；
Fig. 6 is the platelet attachment experiment result for carrying heparin Static Spinning diaphragm；
Fig. 7 is that (sequential of expanded PTFE is without VEGF for cell proliferation experimental result picture165The condition of culture solution
Under)；
Fig. 8 is that (negative control does not carry the electrostatic film of drug+be free of VEGF to cell proliferation experimental result picture165Culture solution)；
Fig. 9 is cell proliferation experimental result picture (positive control：The Static Spinning diaphragm of drug+contain 10ng/ml is not carried
VEGF165Solution)；
Figure 10 is that cell proliferation experimental result picture (carries VEGF165Static Spinning diaphragm)；
Figure 11 is cell proliferation experimental result picture (endothelial cell is in ePTFE membrane on piece)；
Figure 12 is cell proliferation experimental result picture (endothelial cell is on Static Spinning film)；
Figure 13 is vein transplantation model and evaluation figure under the artificial blood vessel's row rabbit kidney prepared using the present invention；
Figure 14 is the artificial blood vessel's row sheep carotid artery transplantation result figure prepared using the present invention；
Figure 15 is the artificial blood vessel's row sheep carotid artery transplantation vascular ultrasound figure prepared using the present invention；
Figure 16 is performance figure under mirror after being implanted into 3 months using artificial blood vessel's row sheep carotid artery transplantation for preparing of the present invention.
Specific implementation mode
To make those skilled in the art more fully understand technical scheme of the present invention, in the following with reference to the drawings and specific embodiments
The present invention is described in further detail.
Embodiment
Electrospinning device
As shown in Figure 1, electrospinning device used by the present embodiment includes micro pump 1, the first syringe in structure
2, the second syringe 3, woven hose 4, coaxial spinneret 5, high voltage power supply 6 and receiving terminal 8；The electrospinning device is in principle and makes
With consistent with the prior art in method；First syringe 2, the second syringe 3 push piston motion, capacity by micro pump 1
It can be 10ml, fltting speed is controlled by micro pump 1；The outlet end of first syringe 2 and the first feed liquor of coaxial spinneret 5
End connection, the outlet end of the second syringe 3 are connect with the second liquid feeding end of coaxial spinneret 5；It can be in the first syringe 2 when use
Middle injection shell solution injects sandwich layer solution in the second syringe 3；Plus high-voltage field is formed using high voltage power supply 6, is made same
Polymer solution electrification in axis spinneret 5, when electric field strength is powerful enough, the drop of 5 capillary tip of coaxial spinneret
Surface tension is overcome to form jet stream, polymer jet stream is stretched during being projected from capillary nozzle tip to receiving terminal, molten
Nano-scale fiber (forming spinning 7) is formed after agent volatilization.
Small-caliber artificial blood vessel preparation method
The small-caliber artificial blood vessel preparation method based on electrostatic spinning of the present embodiment, includes the following steps：
(1) sandwich layer solution is conveyed in electrospinning device and carry out coaxial electrostatic spinning, build artificial blood vessel's internal layer；It is described
Sandwich layer solution is heparin sodium aqua, VEGF (vascular endothelial growth factor) solution and BSA (bovine serum albumin(BSA)) solution according to 20:
20:60 mass ratio mixes.
In sandwich layer solution, the mass percentage concentration of the heparin sodium aqua is 30%, heparin in the heparin sodium aqua
The molecular weight of sodium is 20000, and supplier is Hebei Changshan pharmaceutical factory.The VEGF solution is the VEGF of 100ug/ml165Solution,
Supplier is Tepro Tech companies of the U.S..The mass percentage concentration of the BSA solution is 0.1%, and supplier is U.S. R＆D public
Department.
In the preparation process of artificial blood vessel's internal layer, the operating voltage of electrospinning device is 16KV, and room temperature is 26 DEG C, empty
Air humidity degree is 30%, and the fltting speed of sandwich layer solution is 0.1ml/h；The receiving terminal of electrospinning device is the stainless steel shaft of 4mm,
The rotating speed of the stainless steel shaft is 800r/min when preparation.The secondary spinning time (the specific time can be depending on required thickness) be
3 hours.
(2) shell solution is conveyed in electrospinning device and carry out coaxial electrostatic spinning, build artificial blood vessel's outer layer；It is described
Shell solution is P (LLA-CL), collagen and elastin according to 80:10:After 10 mass ratio is completely dissolved in solvent at room temperature
It is made.
P (LLA-CL) i.e. Poly L-lactic acid-caprolactones are a kind of degradable macromolecule polyalcohols, are ratified to use by FDA
In manufacture body implant, selects the material to no longer need to do toxicological experiment, be widely used in artificial tracheae, tendon, nerve, skin
Equal tissue engineering brackets, good mechanical property.However, P (LLA-CL) is unsatisfactory as the biocompatibility of artificial blood vessel, and lead to
Crossing collagen and elastin then can effectively promote its biocompatibility, and in quality, P (LLA-CL)：Collagen：Elastin=
80:10:10, it is best mass ratio.
Collagen is ox source type i collagen, and molecular weight 100000, supplier is the happy biological products company of Chengdu section.It is described
Solvent is poly- hexafluoroisopropanol, and supplier is that Shanghai reaches auspicious fine chemistry industry, and the mass percentage concentration for being formed by shell solution is
10%.
In the preparation process of artificial blood vessel's shell, the operating voltage of electrospinning device is 16KV, and room temperature is 26 DEG C, empty
Air humidity degree is 30%, and the fltting speed of shell solution is 1.0ml/h.；The receiving terminal of electrospinning device is the stainless steel of 4mm
Axis, when preparation the rotating speed of the stainless steel shaft be 800r/min.Secondary spinning time (the specific time can according to required thickness and
It is fixed) it is 3 hours.
The artificial blood vessel's average thickness obtained after spinning is 0.44 ± 0.55mm, will be in tube body at room temperature vacuum desiccator
For 24 hours, 4 DEG C preserve for volatilization.Fig. 2 is artificial blood vessel's pictorial diagram of the bore 4mm prepared using this method.Fig. 3 is using this method
The artificial blood vessel of preparation and Human saphenous vein comparison diagram, as can be seen from the figure the two is identical.Fig. 4 is using our legal system
Ultra microstructure figure (1000 times of amplification) under standby artificial blood vessel's mirror.
Verification to artificial blood vessel's indices of above-mentioned preparation
1) maximum pressure is 1158 ± 197mmH before connection highly pressurised liquid measures rupture；It is measured using 6-0Prolene sutures
1cm hanging wire pulling force is 1.3 ± 0.5N.
2) measurement of heparin sustained release：The load heparin Static Spinning diaphragm of three groups of 200mg is weighed, after ultraviolet irradiation sterilizing respectively
It is fitted into 10ml centrifuge tubes, 4mlPBS liquid is added in every group of sample, and (phosphate buffer particle, Beijing Suo Laibao biotechnologys are limited
Company), it is leached in 37 DEG C of shaking tables；According to time point 1h, 3h, 6h, the 12h of setting, for 24 hours, the 2-7 days, the 9th, 11,13,15
It takes out 2ml leachates for testing, and the fresh PBS liquid of 2ml is supplemented after taking out 2ml leachates every time.1. heparin sodium titer
Configuration：Heparin sodium 500mg+PBS liquid 50ml obtains 10mg/ml heparin mother liquid of sodium, takes 1000ul that 100mlPBS liquid is added and obtains
100ug/ml heparin sodium titers；2. the configuration of toluidine blue reagent, 500mg toluidine blues powder (AMERSCO companies of the U.S.)
It is dissolved in 500ml deionized waters and is configured to 1mg/ml mother liquors.It takes 25ml mother liquors to be settled to 500ml with deionized water, obtains a concentration of
0.005% toluidine blue reagent.1g sodium chloride is added and obtains 0.2% 0.005% toluidine blue liquid；3. 0.25ml is taken,
0.5ml, 0.75ml, 1.0ml, 1.25ml heparin sodium titer are placed in 1-5 test tubes, are adjusted dissolving to 2ml using PBS liquid,
3ml toluidine blue reagents are added, 37 DEG C of water-baths 2 hours after concussion；So that toluidine blue is fully reacted with heparin, generates complexing
Object.1 hour extraction complex compound is stood after the concussion of 5ml n-hexanes is added；Lower layer's water phase is left and taken using separatory funnel.Measure lower water
The mutually absorbance at 629nm.Using absorbance as the longitudinal axis, heparin concentration is horizontal axis, draws standard curve (see Fig. 5).4. sample
The measurement of heparin sodium：3ml toluidine blue reagents, 37 DEG C of water-baths after concussion is added in the 2mlPBS leachates that each sample point is acquired
Reaction 2 hours.Toluidine blue and heparin is set fully to be complexed.1 hour extraction complex compound is stood after the vibrations of 5ml n-hexanes are added.Make
Lower layer's water phase is left and taken with separatory funnel, area's 100ul subnatants are added to 96 orifice plates, and each sample does 3 multiple holes；Use reading version
Instrument measures absorbance of lower layer's water phase at 629nm, and heparin amount is calculated according to standard curve；As a result the nanometer of shell-and-core structure is shown
Fiber can sustained release heparin nearly 4 weeks, total 6.93 ± 1.03mg of burst size, the 4th weekend amount to release 77% loading
Amount.Whether the heparin for verification release is active, we determine the hemolysis rate for carrying heparin Static Spinning in vitro and blood platelet is viscous
Attached situation is repeatedly measured three times, and according to hemolysis rate calculation formula, the hemolysis rate for carrying the Static Spinning film of heparin is 0.92%, no
The Static Spinning film hemolysis rate for carrying heparin is 1.54%, and varicosity tetrafluoroethene is 2.97%.The Static Spinning film for carrying heparin often regards
Wild platelet count 3.2, varicosity tetrafluoroethene are per 17, the visual field.
3) hemolysis rate of heparin Static Spinning film is carried：1. preparing human whole blood's dilution：9ml fresh whole bloods, 1ml 3.8%
Sodium citrate anti-freezing, mixing, normal saline dilution to 22.5ml；2. blank control：EPTFE blood vessels, negative control：-.2ml is dilute
It releases blood and 10ml physiological saline, positive control is added：10ml distilled water is added in 0.2ml diluted bloods.Three groups of a diameter of 1cm circles are carried
Heparin diaphragm does not carry heparin diaphragm, varicosity tetrafluoroethene blood vessel piece with after 75% ethyl alcohol immersion 10min, and PBS solution rinses three
It is secondary, it is placed in 50ml physiological saline and impregnates 12h, be added in 10ml physiological saline test tubes after further taking out, 37 C water bath 30min,
Hemodilution 2ml is added in micropipettor, keeps 37 C water bath 1 hour.1000r/min centrifuges 5min, takes supernatant.Make
With OD values at ultraviolet light spectrophotometer measuring and calculating A540nm, hemolysis rate is calculated.
4) activated clotting time measures：1. preparing blood vessel piece：It is straight that heparin sodium Static Spinning diaphragm, which will be carried, and cut out using punching pin
Diameter 5mm disks, control group are diameter 4mm varicosity tetrafluoroethene blood vessel pieces, are flattened after longitudinal axis incision, diameter 5mm disks are made,
Two groups are rinsed three times with PBS, and 4ml physiological saline impregnates 24 hours；2. ACT is measured, it is arranged three groups, it is quiet to carry heparin for blank group
Electrospinning film group, varicosity tetrafluoroethene diaphragm group, test specimen is taken out, and is placed in ACT test bottom of the tube, is rapidly added fresh whole blood, on
Machine tests activated coagulation time.As a result show that the Static Spinning diaphragm ACT times exceed machine range 360s, varicosity tetrafluoroethene
For 149 ± 72s, blank control group is 118 ± 46s.
5) VEGF165 sustained releases measure：1. the load VEGF nano electrostatics for weighing three groups of 200mg spin diaphragm, ultraviolet irradiation sterilizing
After be respectively charged into 10ml centrifuge tubes, 4mlPBS liquid is added in every group of sample, is leached in 37 DEG C of shaking tables, by preset time point,
1h, 3h, 6h, 12h the 2-7 days, take out 2ml soaks on the 9th, 11,13,15 day for testing, take out 2ml every time and leach for 24 hours
The fresh PBS liquid of 2ml is supplemented after liquid.It is frozen for -80 DEG C after leachate addition protease inhibitors.2. leachate VEGF contents ELASA
Method measures：The configuration of titer：0.5ml distilled waters are added in 20ngVEGF liquid, are configured to 40ng/ml solution, established standards pipe 8
Pipe.First pipe plus titer are diluted to 900ul, the second to eight pipe plus dilution 500ul, and titer is added into the first pipe
100ul shakes up after dilution.500ul is extracted after mixing, the second pipe is added, half-and-half dilute repeatedly, until the 7th pipe discards.8th
Pipe is blank control.Cleaning solution is 1:20 distilled waters dilute.It examines：Sample-adding：Sample to be tested or standard items 100ul is added in each hole,
Reaction plate is mixed well and is placed on 37 DEG C of 120min → board-washings：Reaction plate is washed 6 times with cleaning solution, printed on filter paper it is dry →
First antibody 100ul is added per hole, reacts 60min → board-washings for 37 DEG C after mixing：With the 2nd step → enzyme labeling antibody working solution in every hole
100ul, 37 DEG C of reaction 30min → board-washings → substrate working solution 100ul, 37 DEG C of dark place reaction 15min → add per hole are added per hole
Enter 100ul terminate liquids → absorbance at microplate reader measurement 450nm → and utilizes standard curve determination concentration of specimens.The results show that core
Layer structure nanofiber can sustained release VEGF, first 24 hours, release 30%, rear sustained release 28 days.
Surrounding releases 60% loading, total 22.17 ± 5.5ug of burst size.
6) platelet attachment experiment：1. preparing platelet rich plasma：It is prepared by PRP：18ml fresh and healthy adult whole bloods are added
3.8% sodium citrate anti-freezings (9 of 2ml:1).100r/min, 25 degrees Celsius of centrifugation 5min, obtains Platelet-rich plasm；2. growing
It spends 1cm ePTFE blood vessels pieces and carries heparin Static Spinning vessel segment ethyl alcohol soaking disinfection 10min, after cleaning three times, physiology salt water logging
Bubble 24 hours, is placed in siliconized tubes, and 2mlPRP, 37 degrees Celsius of static reactions 1 hour is added.3. PBS liquid is softly washed after taking out
It washs, 4 degrees Celsius of 2% glutaraldehydes of precooling fix 24 hours；4. being dehydrated：Ethyl alcohol is dehydrated step by step：50%, 70%, 90%, absolute ethyl alcohol
It is dehydrated step by step, each 10min；5. being dried in vacuo ware to stay overnight；6. metal spraying：Observe the metal spraying that faces upward；7. scanning electron microscope (× 1000)
Platelet adhesion reaction situation is observed, takes pictures, as a result sees Fig. 5 and Fig. 6.
7) endothelial cell adherency test：1. preparing cell suspension：Using culture medium dilution, (0.4% is expected trypan blue dye liquor
Blue dye liquor：Cell suspension=1:1), automated cell calculating instrument prepares the cell suspension that cell density is 1 × 105/ml, calculates thin
Born of the same parents' activity.2. using punching pin by P (LLA-CL) Static Spinning blood vessel piece, the blended Static Spinning blood of P (LLA-CL)-collagen-elastin
Diameter 14mm circular films are made in section of jurisdiction, varicosity tetrafluoroethene blood vessel piece, can be smooth seamless applying with 24 orifice plate bottoms, sterile
Two groups of blood vessel pieces are laid in 24 orifice plate bottoms by interior disinfection by ultraviolet light, and to be pushed down with stainless steel steel ring after high pressure sterilization, (outer ring is straight
Diameter 14mm, inner ring diameter 10mm, inner ring volume 500ul).3. being inoculated with 1 × 104 endothelial cell inside each steel ring, longest is carried out
For culture in 12 hours blood vessel piece was rinsed using 2ml culture mediums at the time point of each setting.It is calculated using cell counter
Into the endothelial cell of adherency loosely in rinsing liquid, setting time point is to be inoculated with half an hour after, 1 hour, 2 hours, 4 hours, 6
Hour, 10 hours, 12 hours.The results show that, P (LLA-CL)-collagen-bullet the most notable in the 2nd hour cell adherence rate difference
Power protein blended spinning electrostatic film, pure P (LLA-CL) Static Spinnings film, ePTFE membrane cell adhesion rate be respectively 83.26 ± 8.02%,
67.07 ± 4.16%, 46.87 ± 8.85% (P<0.001).
8) endothelial cell proliferation is tested：Endothelial cell is assessed in Static Spinning using CCK-8 methods, fluorescence Electronic Speculum, scanning electron microscope
Moral proliferative conditions on blood vessel.1. preparing cell suspension：Prepare cell suspension：Use culture medium dilution, trypan blue dye liquor (0.4%
Trypan blue dye liquor：Cell suspension=1:1), automated cell calculating instrument prepares the cell suspension that cell density is 1 × 105/ml, meter
Calculate cell viability.2. using punching pin by P (LLA-CL) Static Spinning blood vessel piece, P (the LLA-CL)-blended electrostatic of collagen-elastin
Diameter 14mm circular films are made in spinning blood vessel piece, varicosity tetrafluoroethene blood vessel piece, can be smooth seamless applying with 24 orifice plate bottoms, nothing
Two groups of blood vessel pieces are laid in 24 orifice plate bottoms by disinfection by ultraviolet light in bacterium platform, (outer to be pushed down with stainless steel steel ring after high pressure sterilization
Loop diameter 14mm, inner ring diameter 10mm, inner ring volume 500ul).3. cell Proliferation designs four groups of condition of culture：Positive control：No
The Static Spinning diaphragm of load drug+be free of VEGF165 culture solutions；Positive control：The Static Spinning diaphragm of drug+contain 10ng/ml is not carried
VEGF165 culture solutions；Carry VEGF Static Spinning diaphragms：Without VEGF culture solutions；Expanded PTFE：Without VEGF culture solutions.
Above-mentioned four groups of condition of culture are inoculated with 1 × 104 human aorta endothelial cell.37 DEG C, in the saturated humidity incubator containing 5%CO2
Continuous culture, every group is divided equally two parts, and portion is detected for CCK-8 methods, and portion is used for fluorescence microscope.4. in culture the 1st,
CCK-8 methods on probation assess cell Proliferation within 3,7 days：50ulCCK-8 solution is added to every hole, 24 well culture plates are put into incubator relaying
Continuous culture 3 hours.The liquid medium 100ul that micropipettor on probation collects the discoloration per hole injects 96 orifice plates, careful to inject, must not
There is bubble, OD values at its 450nm are measured with microplate reader.5. fluorescence microscope：DAPI is dyed, film-making flow：From 24 orifice plates
Middle taking-up steel ring, diaphragm, softly methanol impregnates 2 minutes → 70% ethyl alcohol immersion, 2 minutes → PBS drifts to PBS three times → 100% for rinsing
It washes 3 minutes → DAPI and dyes 2 minutes → PBS rinsings 3 times → dropwise addition glycerine, resin mounting；6. scanning electron microscopic observation.As a result see figure
7- Figure 12.
9) the unobstructed situation under the conditions of non-anti-freezing：1. actively by the synthetic vascular grafts of the invention prepared to rabbit kidney lower abdomen
Arteries and veins, clinical follow 4 weeks do not give any antiplatelet drug, general anesthesia materials after 4 weeks；2. the grafting vessel of taking-up is indulged along long axis
Row is rived, soft to rinse, and portion is preserved with the special glutaraldehyde fixer of precooling Electronic Speculum, a to be protected with 4% paraformaldehyde fixer
It deposits；3. HE is dyed：The slice that thickness is 4um, dewaxing, dyeing is made in wax embedding fixing organization；4. prepared by electron microscope specimen：New osmium
The ethanol solution of acid soak 120 minutes → 3 times → 50% ethanol solution of distillation water washing → 70% → 100% ethanol solution → epoxy
Propane (26 DEG C, twice, 10 minutes every time) → propylene oxide：Resin=1:1 (28 DEG C, 40 minutes) → propylene oxide：Resin=
1:3 (28 DEG C, 40 minutes) → pure resins (28 DEG C, 40 minutes) → embedding：36 DEG C 5 hours → 60 DEG C 5 hours → 80 DEG C 5 hours →
Patch platform → metal spraying → observation is taken pictures.
The result is shown in Figure 13：In figure, A：Static Spinning blood vessel appearance；B：Artery transplantation model under rabbit kidney；C：After transplanting one month
Static Spinning blood vessel stage casing is without thrombosis (× 200)；D：Neoendothelial cells (× 400) near previous anastomotic；E：Varicosity tetrafluoro second
The a large amount of thrombus of alkene Endovascular (× 200)；F：Varicosity tetrafluoroethene vascular anastomosis has no endothelial cell growth (× 400)；G：
Static Spinning blood vessel neointima has a common boundary (× 1000) with not yet endothelialization region；H：Varicosity tetrafluoroethene blood vessel surface is covered without endothelium
Lid, adheres to a large amount of thrombus (× 1000).
10) small-caliber artificial blood vessel prepared by the present invention grows section transplantation effect：1. with artificial blood vessel's row proposed by the present invention
The carotid artery transplantation of sheep；2. postoperative do not give anticoagulant；3. the postoperative 1st, 2, March, footline vascular ultrasound, was assessed unobstructed
Narrow situation at rate and lateral thrombus and previous anastomotic；4. postoperative 90 days rows DSA is checked；5. after postoperative 90 days rows DSA is checked, taking out
Grafting vessel, row general pathology, light microscopic, Electronic Speculum inspection.
As a result it shows：
Ultrasonic aspect：All transplanting objects are unobstructed in post-transplantation early stage blood flow, the 1st, 2, footline ultrasonic examination in March
See, the no blood signal of varicosity tetrafluoroethene 1 month after surgery；P (LLA-CL)-collagen-elastin mixing Static Spinning blood vessel
3 months after surgery still unobstructed, and DSA checks that display is unobstructed；In terms of gross specimen：Varicosity tetrafluoroethene blood vessel has entirely shut, P
(LLA-CL) there are still and have no thrombosis to-collagen-elastin mixing Static Spinning vessel lumen；Light microscopy checking is shown in varicosity
It is all thrombus covering in tetrafluoroethene tube chamber, and mixes the intravascular stage casing skin layer cellulose covering of Static Spinning, previous anastomotic is attached
Nearly visible new intima；Electronic Speculum is shown in：Varicosity tetrafluoroethene whole process thrombus and wadding fibrous element have no endothelial cell, mix electrostatic
It is thin layer cellulose to spin sequential stage casing, and the fragmentary island endothelium in part covers, and previous anastomotic has formed the covering of " paving stone " sample endothelium.Knot
Fruit sees Figure 14-Figure 16.
In fig. 14, A：Animal；B：Arteria carotis is implanted into 6cm Static Spinning blood vessels；C：Outer surface is by newborn group after implantation 3 months
Knit package；D：It is implanted into 6cm varicosity tetrafluoroethene blood vessels；E：Static Spinning is intravascular unobstructed；F：Varicosity tetrafluoroethene internal blood vessel is big
Measure thrombus.
In fig.15, A：Self artery；B：Month after operation；C：3 months after operation.
In figure 16, A：Varicosity tetrafluoroethene blood vessel, thrombus and cellulose filling (× 100) in tube chamber；B：Static Spinning blood
The visible fragmentary endothelial cell C in pipe stage casing：Static Spinning vessel endothelialisation region has a common boundary with stage casing；D：Varicosity tetrafluoroethene blood under Electronic Speculum
Blood platelet, cellulose covering in pipe；F：Static Spinning vascular anastomosis proximal end forms intact endothelium.
To sum up, using the small-caliber artificial blood vessel prepared by the method for the present invention, anti-freezing is can avoid, there is tissue phase well
Capacitive.
Finally illustrate, applying specific case herein, principle and implementation of the present invention are described, with
The explanation of upper embodiment is merely used to help understand the core idea of the present invention, without departing from the principles of the present invention, also
Can several improvements and modifications are made to the present invention, these improvement and modification are also fallen within the scope of the present invention.
Claims (10)
1. a kind of small-caliber artificial blood vessel preparation method based on electrostatic spinning, which is characterized in that include the following steps：
(1) sandwich layer solution is conveyed in electrospinning device and carry out coaxial electrostatic spinning, build artificial blood vessel's internal layer；The sandwich layer
Solution is heparin sodium aqua, VEGF solution and BSA solution according to 20:20:60 mass ratio mixes；
(2) shell solution is conveyed in electrospinning device and carry out coaxial electrostatic spinning, build artificial blood vessel's outer layer；The shell
Solution is P (LLA-CL), collagen and elastin according to 80:10:10 mass ratio is made after being completely dissolved in solvent.
2. the small-caliber artificial blood vessel preparation method according to claim 1 based on electrostatic spinning, it is characterised in that：In step
Suddenly in (1), the mass percentage concentration of the heparin sodium aqua is 30%, and the molecular weight of heparin sodium is in the heparin sodium aqua
20000。
3. the small-caliber artificial blood vessel preparation method according to claim 1 based on electrostatic spinning, it is characterised in that：In step
Suddenly in (1), the VEGF solution is the VEGF of 100ug/ml165Solution.
4. the small-caliber artificial blood vessel preparation method according to claim 1 based on electrostatic spinning, it is characterised in that：In step
Suddenly in (1), the mass percentage concentration of the BSA solution is 0.1%.
5. the small-caliber artificial blood vessel preparation method according to claim 1 based on electrostatic spinning, it is characterised in that：In step
Suddenly in (2), the collagen is ox source type i collagen, molecular weight 100000.
6. the small-caliber artificial blood vessel preparation method according to claim 1 based on electrostatic spinning, it is characterised in that：In step
Suddenly in (2), the solvent is poly- hexafluoroisopropanol, and the mass percentage concentration for being formed by shell solution is 10%.
7. the small-caliber artificial blood vessel preparation method according to any one of claims 1 to 6 based on electrostatic spinning, feature
It is：In step (1) and step (2), the operating voltage of electrospinning device is 16KV, and room temperature is 26 DEG C, and air humidity is
30%.
8. the small-caliber artificial blood vessel preparation method according to claim 7 based on electrostatic spinning, it is characterised in that：In step
Suddenly in (1) and step (2), the receiving terminal of electrospinning device is the stainless steel shaft of 4mm, and the rotating speed of the stainless steel shaft is
800r/min。
9. the small-caliber artificial blood vessel preparation method according to claim 8 based on electrostatic spinning, it is characterised in that：In step
Suddenly in (1), the fltting speed of sandwich layer solution is 0.1ml/h.
10. the small-caliber artificial blood vessel preparation method according to claim 9 based on electrostatic spinning, it is characterised in that：
In step (2), the fltting speed of shell solution is 1.0ml/h.
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Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN111603267A (en) * | 2020-06-12 | 2020-09-01 | 西安交通大学医学院第一附属医院 | Method for manufacturing coaxial electrostatic spinning magnetic anastomosis artificial blood vessel |
| CN112657051A (en) * | 2020-12-25 | 2021-04-16 | 上海交通大学医学院附属上海儿童医学中心 | Biological type ventricular assist pump based on conjugate coaxial electrostatic spinning technology and preparation method thereof |
Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2012006072A2 (en) * | 2010-06-28 | 2012-01-12 | Virginia Commonwealth University | Air impedance electrospinning for controlled porosity |
| CN103893817A (en) * | 2014-03-20 | 2014-07-02 | 北京大学第三医院 | Coaxial electrostatic spinning fibrous scaffold and preparation method thereof |
| CN105233339A (en) * | 2015-11-06 | 2016-01-13 | 东华大学 | Preparation method of heparin and twin factor synergistically regulated P(LLA-CL)/collagen bilayer intravascular stent |
| CN105363076A (en) * | 2015-09-30 | 2016-03-02 | 东华大学 | Method for preparing polylactic acid caprolactone-collagen two-layer bionic vascular scaffold |
| CN106421921A (en) * | 2015-11-10 | 2017-02-22 | 江苏工程职业技术学院 | Preparation method of double-layer intravascular stent |
-
2018
- 2018-06-01 CN CN201810559364.1A patent/CN108404215A/en active Pending
Patent Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2012006072A2 (en) * | 2010-06-28 | 2012-01-12 | Virginia Commonwealth University | Air impedance electrospinning for controlled porosity |
| CN103893817A (en) * | 2014-03-20 | 2014-07-02 | 北京大学第三医院 | Coaxial electrostatic spinning fibrous scaffold and preparation method thereof |
| CN105363076A (en) * | 2015-09-30 | 2016-03-02 | 东华大学 | Method for preparing polylactic acid caprolactone-collagen two-layer bionic vascular scaffold |
| CN105233339A (en) * | 2015-11-06 | 2016-01-13 | 东华大学 | Preparation method of heparin and twin factor synergistically regulated P(LLA-CL)/collagen bilayer intravascular stent |
| CN106421921A (en) * | 2015-11-10 | 2017-02-22 | 江苏工程职业技术学院 | Preparation method of double-layer intravascular stent |
Non-Patent Citations (1)
| Title |
|---|
| 杨力: "《组织修复生物力学》", 31 December 2017 * |
Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN111603267A (en) * | 2020-06-12 | 2020-09-01 | 西安交通大学医学院第一附属医院 | Method for manufacturing coaxial electrostatic spinning magnetic anastomosis artificial blood vessel |
| CN112657051A (en) * | 2020-12-25 | 2021-04-16 | 上海交通大学医学院附属上海儿童医学中心 | Biological type ventricular assist pump based on conjugate coaxial electrostatic spinning technology and preparation method thereof |
| CN112657051B (en) * | 2020-12-25 | 2023-08-04 | 上海交通大学医学院附属上海儿童医学中心 | Biological type ventricular assist pump based on conjugate coaxial electrostatic spinning technology and preparation method thereof |
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