CN107898533A - Manually carry the coaxial regeneration vessel stent of medicine and its combination process preparation method - Google Patents
Manually carry the coaxial regeneration vessel stent of medicine and its combination process preparation method Download PDFInfo
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- CN107898533A CN107898533A CN201711174441.3A CN201711174441A CN107898533A CN 107898533 A CN107898533 A CN 107898533A CN 201711174441 A CN201711174441 A CN 201711174441A CN 107898533 A CN107898533 A CN 107898533A
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61F—FILTERS IMPLANTABLE INTO BLOOD VESSELS; PROSTHESES; DEVICES PROVIDING PATENCY TO, OR PREVENTING COLLAPSING OF, TUBULAR STRUCTURES OF THE BODY, e.g. STENTS; ORTHOPAEDIC, NURSING OR CONTRACEPTIVE DEVICES; FOMENTATION; TREATMENT OR PROTECTION OF EYES OR EARS; BANDAGES, DRESSINGS OR ABSORBENT PADS; FIRST-AID KITS
- A61F2/00—Filters implantable into blood vessels; Prostheses, i.e. artificial substitutes or replacements for parts of the body; Appliances for connecting them with the body; Devices providing patency to, or preventing collapsing of, tubular structures of the body, e.g. stents
- A61F2/02—Prostheses implantable into the body
- A61F2/04—Hollow or tubular parts of organs, e.g. bladders, tracheae, bronchi or bile ducts
- A61F2/06—Blood vessels
- A61F2/07—Stent-grafts
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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
- A61L27/16—Macromolecular materials obtained by reactions only involving carbon-to-carbon unsaturated bonds
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- A61L27/00—Materials for grafts or prostheses or for coating grafts or prostheses
- A61L27/14—Macromolecular materials
- A61L27/18—Macromolecular materials obtained otherwise than by reactions only involving carbon-to-carbon unsaturated bonds
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- A—HUMAN NECESSITIES
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- A61L27/00—Materials for grafts or prostheses or for coating grafts or prostheses
- A61L27/14—Macromolecular materials
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- A—HUMAN NECESSITIES
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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
- A61L27/52—Hydrogels or hydrocolloids
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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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- 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
- A61L27/56—Porous materials, e.g. foams or sponges
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- 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
- A61L27/50—Materials characterised by their function or physical properties, e.g. injectable or lubricating compositions, shape-memory materials, surface modified materials
- A61L27/58—Materials at least partially resorbable by the body
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61F—FILTERS IMPLANTABLE INTO BLOOD VESSELS; PROSTHESES; DEVICES PROVIDING PATENCY TO, OR PREVENTING COLLAPSING OF, TUBULAR STRUCTURES OF THE BODY, e.g. STENTS; ORTHOPAEDIC, NURSING OR CONTRACEPTIVE DEVICES; FOMENTATION; TREATMENT OR PROTECTION OF EYES OR EARS; BANDAGES, DRESSINGS OR ABSORBENT PADS; FIRST-AID KITS
- A61F2240/00—Manufacturing or designing of prostheses classified in groups A61F2/00 - A61F2/26 or A61F2/82 or A61F9/00 or A61F11/00 or subgroups thereof
- A61F2240/001—Designing or manufacturing processes
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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
- 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
- A61L2300/204—Biologically active materials used in bandages, wound dressings, absorbent pads or medical devices containing or releasing organic materials with nitrogen-containing functional groups, e.g. aminoxides, nitriles, guanidines
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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
- A61L2300/232—Monosaccharides, disaccharides, polysaccharides, lipopolysaccharides
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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
- A61L2300/404—Biocides, antimicrobial agents, antiseptic agents
- A61L2300/406—Antibiotics
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- A61L2300/00—Biologically active materials used in bandages, wound dressings, absorbent pads or medical devices
- A61L2300/60—Biologically active materials used in bandages, wound dressings, absorbent pads or medical devices characterised by a special physical form
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- A61L2430/00—Materials or treatment for tissue regeneration
- A61L2430/22—Materials or treatment for tissue regeneration for reconstruction of hollow organs, e.g. bladder, esophagus, urether, uterus
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Abstract
The coaxial regeneration vessel stent of medicine and its combination process preparation method are manually carried the invention discloses a kind of, the intravascular stent has three-decker, the preparation of different process method is respectively adopted, wherein inner layer material selects Deferoxamine DFO+PVA sandwich layers solution and PCL shell solution, using coaxial electrically spun forming technology；Intermediate layer material selects PVA+SA mixed solutions, using infusion process forming technology, and is crosslinked after electrospinning outer layer using calcium chloride；Cladding material selects gentamicin GS+PVA sandwich layers solution and PCL shell solution, using coaxial electrically spun forming technology.The present invention prepares three layers of load medicine intravascular stent using two kinds of process combining different materials, and simulates the three-decker of native blood vessels very well, shortens the time needed in vitro culture transplanting and success rate, has broad prospects in clinical practice.
Description
Technical field
The present invention relates to a kind of artificial blood vessel and preparation method thereof, more particularly to a kind of regeneration vessel stent and its compound
Its preparation process, applied to Biotechnology field.
Background technology
Coronary heart disease caused by angiocardiopathy, especially artery sclerosis, the main reason for having become human death at present
One of, one of its main means treated are exactly to carry out vasotransplantation.Due to autologous vein limited source, it is therefore desirable to substantial amounts of
Artificial blood vessel is used for clinic.
With the increase of artificial blood vessel's demand, various techniques prepare artificial blood vessel and produce in succession, wherein biometric print technology
Prepare artificial blood vessel due to its in function with having outstanding performance in efficiency, blood vessel prepare in application gradually increase.
It is prepared by traditional artificial blood vessel, when be made by either simplex skill electrostatic spinning, but biocompatibility or mechanicalness are not
It is very good, second, by electrostatic spinning and the compound preparation artificial blood vessel of biomaterial, but drug delivery technologies are considered in preparation process
Using less.Although the intravascular stent energy simple analog native blood vessels structure prepared at present, often meets during the transplantation process
To infection and histocompatibility issues, thus could be improved in the preparation process of intravascular stent and selection with it is perfect.
The content of the invention
In order to solve prior art problem, it is an object of the present invention to overcome the deficiencies of the prior art, and to provide one kind
The coaxial regeneration vessel stent of medicine and its combination process preparation method are manually carried, is prepared as a result of the compound method of Alternative
Intravascular stent, overcomes the limitation of either simplex skill, shortens the preparation time of stent, and wherein first layer and the second layer utilizes capillary
Phenomenon is combined closely, and the second layer and third layer are combined closely using Chemical Crosslinking Methods, improves the mechanical performance of stent entirety,
As a result of the coaxial technology of medicine is carried, medicine slowly release on time in vivo can be made, reduce the sense of artificial blood vessel after the transfer
Dye rate, since technique employs the biomaterials such as PVA, SA, PCL, improves the histocompatbility of regeneration vessel.
To reach above-mentioned purpose, the present invention adopts the following technical scheme that：
It is a kind of manually to carry the coaxial regeneration vessel stent of medicine, from the coaxial regeneration vessel internal stent of medicine is carried to outside, successively by
Coaxial internal layer, hydrogel intermediate layer and outer layer is combined closely and is formed, and simulates the three-decker of native blood vessels, wherein the internal layer
Combined closely and formed by the compound sandwich layers of Deferoxamine DFO and the first polycaprolactone (PCL) shell successively, form the first polycaprolactone (PCL)
Shell wraps up the lamellar composite endothecium structure of the compound sandwich layers of Deferoxamine DFO, and the compound sandwich layers of Deferoxamine DFO are by Deferoxamine DFO
It is made with the composite material of PVA, the compound sandwich layers of Deferoxamine DFO are directly toward the inner cavity setting for carrying the coaxial regeneration vessel of medicine,
The hydrogel intermediate layer is made of the composite material of sodium alginate SA and PVA, and the outer layer is compound by gentamicin GS successively
Sandwich layer and the second polycaprolactone (PCL) shell are combined closely and are formed, and form the second polycaprolactone (PCL) shell parcel gentamicin GS and answer
Close sandwich layer lamellar composite layer structure, the compound sandwich layers of gentamicin GS by gentamicin GS and PVA composite material system
Into the both sides of the first polycaprolactone (PCL) shell and the compound sandwich layers of gentamicin GS respectively with hydrogel intermediate layer are closely tied
Close, the second polycaprolactone (PCL) shell is directly toward the exterior setting for carrying the coaxial regeneration vessel of medicine.
It is preferred that the constituent mass proportioning of the composite material of the Deferoxamine DFO and PVA of the above-mentioned compound sandwich layers of Deferoxamine DFO is
(15-64):1000；It is preferred that the constituent mass of the composite material of the gentamicin GS and PVA of the above-mentioned compound sandwich layers of gentamicin GS
Match as (3-8):1000.
It is preferred that the constituent mass proportioning of the composite material of the sodium alginate PVA and SA in above-mentioned hydrogel intermediate layer is (2.4-
9.6):(1-5)。
As currently preferred technical solution, the first polycaprolactone (PCL) shell or the second polycaprolactone (PCL) shell
It is made of polycaprolactone (PCL) and basis material blending, described matrix material is by n,N-Dimethylformamide DMF and dichloromethane
DCM is mixed；Wherein N,N-dimethylformamide DMF：The volume ratio of dichloromethane DCM is 1:1, polycaprolactone (PCL) and base
The mass volume ratio of body material is (1-10) g：100ml.
It is preferred that the whole of the coaxial regeneration vessel stent of medicine is manually carried to control by adjusting the thickness in above-mentioned hydrogel intermediate layer
Body wall is thick.
The method for carrying the coaxial regeneration vessel stent of medicine is prepared the present invention provides a kind of combination process, is included the following steps：
A. PVA is dissolved in deionized water, the heating water bath on magnetic stirring apparatus, and stir until PVA be completely dissolved,
The PVA solution that mass percent concentration is 3-8wt.% is made；
B., Deferoxamine DFO is dissolved in the PVA solution prepared in the step a, it is 50- to be configured to mass percent
The DFO internal layer coaxial electrically spun sandwich layer solution of 80wt.%；
C. it is 1 by volume ratio:The mixture of the dichloromethane DCM of 1 n,N-Dimethylformamide DMF, will as solvent
Polycaprolactone (PCL) is dissolved in solvent, and the mass volume ratio for being configured to polycaprolactone (PCL) and solvent is (1-10) g：100ml's
PCL coaxial electrically spun shell solution；
D. sodium alginate SA is dissolved in deionized water, is configured to the SA solution that mass fraction is 1-5wt.%, then
It is in mass ratio by the PVA solution and SA solution that are prepared in the step a (0.8-1.2)：1 ratio mixing, is configured to blood
The intermediate layer solution of pipe holder；
E. gentamicin GS is dissolved in the PVA solution prepared in the step a, it is 0.3- to be configured to mass fraction
The GS outer layer coaxial electrically spun sandwich layer solution of 0.8wt.%；
F. it is respectively adopted and DFO internal layers coaxial electrically spun sandwich layer solution is prepared in the step b and is prepared in the step c
PCL coaxial electrically spun shell solution, and use coaxial electrically spun forming technology, be sequentially prepared on circular shaft PCL coaxial electrically spuns sandwich layer and
Polycaprolactone (PCL) shell, is made and carries medicine regeneration vessel stent internal layer；
G. infusion process forming technology is used, the circle of load medicine regeneration vessel stent internal layer preparation will be completed in the step f
Axis is removed, and is immersed in the intermediate layer solution prepared in the step d, stands 15~60 minutes, will carry medicine green blood again afterwards
The circular shaft of pipe holder internal layer dipping attachment hydrogel takes out, and circular shaft both ends are maked somebody a mere figurehead standing, and until circular shaft, there is no hydrogel
Untill drop drips, then circular shaft is put in -40~-60 DEG C of refrigerator carry out frost 10-15 it is small when after, take out circular shaft, and
When defrosting 4-8 is small at room temperature by circular shaft, places into refrigerator and freezed, so undergo 1-5 freeze-thawing process, from
And the hydrogel intermediate layer shaped in the outside for carrying medicine regeneration vessel stent internal layer；
H. it is respectively adopted and GS outer layers coaxial electrically spun sandwich layer solution is prepared in the step e and is prepared in the step c
PCL coaxial electrically spun shell solution, and coaxial electrically spun forming technology is used, GS outer layer coaxial electrically spun sandwich layers are sequentially prepared on circular shaft
With polycaprolactone (PCL) shell, it is made and carries medicine regeneration vessel stent outer layer, so that the exterior parcel in hydrogel intermediate layer carries medicine again
Green blood pipe holder outer layer, obtains the molding load coaxial regeneration vessel supporting frame prefabrication body of medicine；
I. it is 1- the coaxial regeneration vessel supporting frame prefabrication body of the load medicine prepared in the step h to be immersed in mass fraction
It is chemically crosslinked within 15~60 minutes in the calcium chloride solution of 5wt.%, after completing to be chemically crosslinked, obtains load medicine and coaxially regenerate
Intravascular stent, will take out after then carrying the coaxial regeneration vessel stent freeze-drying of medicine, has just obtained the coaxial regeneration vessel branch of load medicine
Put up product.
As currently preferred technical solution, when preparing intravascular stent ectonexine in the step f and step h, control
The rotary speed of rounding axis is 200-400 revs/min, and it is 0.3-0.6mm/ that X-direction, that is, circular shaft horizontal axis, which moves back and forth speed,
S, the voltage for controlling Electrospun is 10-13KV, electrospinning liquid is fed by micro pump, wherein during electrospinning in the step f
Between be 3-10min, the electrospinning time in the step h is 4-20min, and electrospinning film is adjusted by controlling the Electrospun time
Thickness, so as to adjust the thickness for carrying medicine regeneration vessel stent internal layer or outer layer.The electrospinning time is longer, and electrospinning film is thicker.
As currently preferred technical solution, when preparing intravascular stent intermediate layer in the step g, freeze-thawing
Number determined by required regeneration vessel wall thickness, pass through the water-setting for controlling the number of freeze-thawing to adjust prepared
The thickness in glue intermediate layer, and then regulate and control manually to carry the overall wall thickness of the coaxial regeneration vessel stent of medicine.The number of freeze-thawing is more
Regeneration vessel wall is thicker.
As currently preferred technical solution, when preparing intravascular stent ectonexine in the step f and step h, control
The feeding speed of micro pump processed is 30-60ul/min, and Coaxial nozzle is highly 140-160mm from circular shaft.
As currently preferred technical solution, when preparing intravascular stent ectonexine in the step f and step h, circle
Axis uses conductive stainless steel, a diameter of 2-9mm of circular shaft；The internal diameter of Coaxial nozzle is not more than 0.3mm, and outside diameter is little
In 1mm.
The present invention compared with prior art, has following obvious prominent substantive distinguishing features and remarkable advantage：
1. the intravascular stent of the present invention has biocompatibility, and degradable；
2. the intravascular stent endothecium structure of the present invention is loaded with Deferoxamine DFO, vascular endothelial growth factor can be promoted
(VEGF) generate, because of outer layer covers PCL, DFO can be made slowly to discharge on time, so promote generation of the cell on regeneration vessel with
Seek connections with；
3. the intravascular stent interlayer structure of the present invention freezes-is physical crosslinking generation by PVA, and controllable regeneration vessel
Wall thickness, and the machinery and biological property of regeneration vessel stent can be improved, increase histocompatbility；
4. intravascular stent layer structure of the present invention is loaded with gentamicin GS, by coaxial electrically spun, outer layer covers protective
PCL, can be such that GS slowly discharges on time, reduce the infection risk after intravascular stent moves into vivo；
5. the intravascular stent of the present invention is prepared by electrostatic spinning combination process, there is the characteristic of nanofibrous structures,
And larger porosity and specific surface area, the adhesion to cell create good condition.
Brief description of the drawings
Fig. 1 is the structure diagram that the embodiment of the present invention one manually carries the coaxial regeneration vessel stent of medicine.
Fig. 2 is shaping signal of the embodiment of the present invention one when carrying the coaxial regeneration vessel stent internal layer of medicine and outer layer electrospinning
Figure.
Fig. 3 is that the embodiment of the present invention one is carrying the combined forming process principle signal of the coaxial regeneration vessel stent preparation of medicine
Figure.
Embodiment
Such scheme is described further below in conjunction with specific examples of the implementation, the preferred embodiment of the present invention is described in detail such as
Under：
Embodiment one：
In the present embodiment, referring to Fig. 1~3, a kind of combination process prepares the method for carrying the coaxial regeneration vessel stent of medicine, bag
Include following steps：
A. PVA is dissolved in deionized water, the heating water bath on magnetic stirring apparatus, and stir until PVA be completely dissolved,
The PVA solution 35g that mass percent concentration is 5wt.% is made；
B. 20mg Deferoxamines DFO is dissolved in the PVA solution prepared in the step a of 10g, is configured to DFO internal layers
Coaxial electrically spun sandwich layer solution；
C. it is 1 by volume ratio:The mixture of the dichloromethane DCM of 1 n,N-Dimethylformamide DMF, will as solvent
2g polycaprolactone (PCL)s are dissolved in the above-mentioned solvents of 20ml, are configured to PCL coaxial electrically spun shell solution, then be bisected into two parts；
D. sodium alginate SA is dissolved in deionized water, is configured to the SA solution 10g that mass fraction is 3wt.%, then
It is 1 in mass ratio by the PVA solution and SA solution that are prepared in the step a:1 ratio mixing, is configured to intravascular stent
Intermediate layer solution；
E. gentamicin GS is dissolved in the PVA solution prepared in the step a of 10g, is configured to mass fraction
For the GS outer layer coaxial electrically spun sandwich layer solution of 0.5wt.%；
F. as shown in Figures 1 to 3, it is respectively adopted and DFO internal layers coaxial electrically spun sandwich layer solution is prepared in the step b and in institute
A PCL coaxial electrically spun shell solution prepared in step c is stated, and uses coaxial electrically spun forming technology, wherein DFO internal layers are same
Axis electrospinning sandwich layer solution and PCL coaxial electrically spun shell solution carry out spinning by Coaxial nozzle, and PCL is sequentially prepared on circular shaft U
Coaxial electrically spun sandwich layer and polycaprolactone (PCL) shell, are made and carry medicine regeneration vessel stent internal layer；When preparing intravascular stent internal layer,
The rotary speed for controlling circular shaft U is 300 revs/min, and it is 0.5mm/s that X-direction, that is, circular shaft U horizontal axis, which moves back and forth speed, control
The voltage of Electrospun processed is 10KV, electrospinning liquid is fed by micro pump, and the wherein electrospinning time is 5min, controls the confession of micro pump
Material speed is 35ul/min, and height of the Coaxial nozzle from circular shaft U is 150mm, and circular shaft U uses conductive stainless steel, circle
A diameter of 5mm of axis U；The internal diameter of Coaxial nozzle is 0.3mm, outside diameter 1mm；
G. infusion process forming technology is used, the circle of load medicine regeneration vessel stent internal layer preparation will be completed in the step f
Axis is removed, and is immersed in the intermediate layer solution prepared in the step d, stands 30 minutes, will carry medicine regeneration vessel branch afterwards
The circular shaft of frame internal layer dipping attachment hydrogel takes out, and circular shaft both ends are maked somebody a mere figurehead standing, and until circular shaft, there is no hydrogel drop
Untill dripping, then circular shaft is put in -50 DEG C of refrigerator carry out frost 12 it is small when after, circular shaft is taken out, and by circular shaft in room temperature
It is middle thaw 6 it is small when, place into refrigerator and freezed, so undergo 1 freeze-thawing process so that carry medicine regeneration vessel
The hydrogel intermediate layer that the outside of stent internal layer is shaped；
H. it is respectively adopted and GS outer layers coaxial electrically spun sandwich layer solution is prepared in the step e and is prepared in the step c
Another PCL coaxial electrically spun shell solution, and use coaxial electrically spun forming technology, it is same that GS outer layers are sequentially prepared on circular shaft U
Axis electrospinning sandwich layer and another layer of polycaprolactone (PCL) shell, are made and carry medicine regeneration vessel stent outer layer, so that in hydrogel intermediate layer
Exterior parcel carry medicine regeneration vessel stent outer layer, obtain the molding load coaxial regeneration vessel supporting frame prefabrication body of medicine；Carried preparing
During the coaxial regeneration vessel stent outer layer of medicine, the parameter of circular shaft U, micro pump and nozzle and intravascular stent is prepared in the step f
Process conditions are identical during internal layer, are particular in that it is 15min to prepare the electrospinning time for carrying medicine regeneration vessel stent outer layer；
I. the coaxial regeneration vessel supporting frame prefabrication body of the load medicine prepared in the step h is immersed in mass fraction is
It is chemically crosslinked within 30 minutes in the calcium chloride solution of 3wt.%, after completing to be chemically crosslinked, obtains carrying the coaxial regeneration vessel of medicine
Stent, its structure are taken out as shown in Figure 1, then will carry after the coaxial regeneration vessel stent of medicine is freeze-dried, and it is same just to have obtained load medicine
Axis regeneration vessel stent finished product.
In the present embodiment, referring to Fig. 1-3, prepared using combination process and carry the coaxial regeneration vessel stent of medicine, shown blood vessel
Stent has three-decker, and different manufacturing process is respectively adopted, and wherein internal layer is coaxial electrically spun DFO solution, and PVA is selected in intermediate layer
+ SA solution, is physical crosslinking method, outermost layer is coaxial electrically spun GS solution using freeze-thawing.The present embodiment is compounded with three kinds of differences
Manufacturing process, simulate the three-decker of native blood vessels, the time that intravascular stent is cultivated in vitro shortened, as a result of same
Axis drug delivery technologies, can making intravascular stent, slowly release, the DFO of internal layer can promote the generation of new vessels on time in vivo, middle
Layer can ensure the wall thickness and mechanical strength of blood vessel, and the GS of outer layer can reduce the infection rate of tissue, it is clinically had wide
Application prospect.
Embodiment two：
The present embodiment and embodiment one are essentially identical, are particular in that：
In the present embodiment, a kind of combination process prepares the method for carrying the coaxial regeneration vessel stent of medicine, includes the following steps：
A. PVA is dissolved in deionized water, the heating water bath on magnetic stirring apparatus, and stir until PVA be completely dissolved,
The PVA solution 35g that mass percent concentration is 3wt.% is made；
B. 20mg Deferoxamines DFO is dissolved in the PVA solution prepared in the step a, being configured to mass percent is
The DFO internal layer coaxial electrically spun sandwich layer solution of 50wt.%；
C. it is 1 by volume ratio:The mixture of the dichloromethane DCM of 1 n,N-Dimethylformamide DMF, will as solvent
0.2g polycaprolactone (PCL)s are dissolved in the above-mentioned solvents of 20ml, are configured to PCL coaxial electrically spun shell solution, then be bisected into two parts；
D. sodium alginate SA is dissolved in deionized water, is configured to the SA solution 10g that mass fraction is 1wt.%, then
It is 0.8 in mass ratio by the PVA solution and SA solution that are prepared in the step a:1 ratio mixing, is configured to intravascular stent
Intermediate layer solution；
E. gentamicin GS is dissolved in the PVA solution prepared in the step a of 10g, is configured to mass fraction
For the GS outer layer coaxial electrically spun sandwich layer solution of 0.3wt.%；
F. it is respectively adopted and DFO internal layers coaxial electrically spun sandwich layer solution is prepared in the step b and is prepared in the step c
A PCL coaxial electrically spun shell solution, and use coaxial electrically spun forming technology, wherein DFO internal layers coaxial electrically spun sandwich layer is molten
Liquid and PCL coaxial electrically spun shell solution carry out spinning by Coaxial nozzle, and PCL coaxial electrically spun sandwich layers are sequentially prepared on circular shaft U
With polycaprolactone (PCL) shell, it is made and carries medicine regeneration vessel stent internal layer；When preparing intravascular stent internal layer, the rotation of control circular shaft U
Rotary speed is 400 revs/min, and it is 0.6mm/s that X-direction, that is, circular shaft U horizontal axis, which moves back and forth speed, controls the voltage of Electrospun
For 13KV, electrospinning liquid is set to be fed by micro pump, the wherein electrospinning time is 3min, and the feeding speed for controlling micro pump is 60ul/
Min, height of the Coaxial nozzle from circular shaft U are 160mm, and circular shaft U uses conductive stainless steel, and circular shaft U's is a diameter of
9mm；The internal diameter of Coaxial nozzle is 0.3mm, outside diameter 1mm；
G. infusion process forming technology is used, the circle of load medicine regeneration vessel stent internal layer preparation will be completed in the step f
Axis is removed, and is immersed in the intermediate layer solution prepared in the step d, stands 15 minutes, will carry medicine regeneration vessel branch afterwards
The circular shaft of frame internal layer dipping attachment hydrogel takes out, and circular shaft both ends are maked somebody a mere figurehead standing, and until circular shaft, there is no hydrogel drop
Untill dripping, then circular shaft is put in -40 DEG C of refrigerator carry out frost 10 it is small when after, circular shaft is taken out, and by circular shaft in room temperature
It is middle thaw 4 it is small when, place into refrigerator and freezed, so undergo 3 freeze-thawing processes so that carry medicine regeneration vessel
The hydrogel intermediate layer that the outside of stent internal layer is shaped；
H. it is respectively adopted and GS outer layers coaxial electrically spun sandwich layer solution is prepared in the step e and is prepared in the step c
Another PCL coaxial electrically spun shell solution, and use coaxial electrically spun forming technology, it is same that GS outer layers are sequentially prepared on circular shaft U
Axis electrospinning sandwich layer and another layer of polycaprolactone (PCL) shell, are made and carry medicine regeneration vessel stent outer layer, so that in hydrogel intermediate layer
Exterior parcel carry medicine regeneration vessel stent outer layer, obtain the molding load coaxial regeneration vessel supporting frame prefabrication body of medicine；Carried preparing
During the coaxial regeneration vessel stent outer layer of medicine, the parameter of circular shaft U, micro pump and nozzle and intravascular stent is prepared in the step f
Process conditions are identical during internal layer, are particular in that it is 4min to prepare the electrospinning time for carrying medicine regeneration vessel stent outer layer；
I. the coaxial regeneration vessel supporting frame prefabrication body of the load medicine prepared in the step h is immersed in mass fraction is
It is chemically crosslinked within 60 minutes in the calcium chloride solution of 1wt.%, after completing to be chemically crosslinked, obtains carrying the coaxial regeneration vessel of medicine
Stent, will take out after then carrying the coaxial regeneration vessel stent freeze-drying of medicine, has just obtained the coaxial regeneration vessel branch of load medicine and put up
Product.
In the present embodiment, prepared using combination process and carry the coaxial regeneration vessel stent of medicine, shown intravascular stent there are three layers
Structure, is respectively adopted different manufacturing process, and wherein internal layer is coaxial electrically spun DFO solution, and intermediate layer is selected PVA+SA solution, adopted
Method is physical crosslinking with freeze-thawing, outermost layer is coaxial electrically spun GS solution.The present embodiment is compounded with three kinds of different shaping sides
Method, simulates the three-decker of native blood vessels, shortens the time that intravascular stent is cultivated in vitro, and medicine skill is carried as a result of coaxial
Art, can making intravascular stent, slowly release, the DFO of internal layer can promote the generation of new vessels on time in vivo, and intermediate layer can ensure
The wall thickness and mechanical strength of blood vessel, the GS of outer layer can reduce the infection rate of tissue, before it is clinically had wide application
Scape.
Embodiment three：
The present embodiment is substantially the same as in the previous example, and is particular in that：
In the present embodiment, a kind of combination process prepares the method for carrying the coaxial regeneration vessel stent of medicine, includes the following steps：
A. PVA is dissolved in deionized water, the heating water bath on magnetic stirring apparatus, and stir until PVA be completely dissolved,
The PVA solution 35g that mass percent concentration is 8wt.% is made；
B. 20mg Deferoxamines DFO is dissolved in the PVA solution prepared in the step a, being configured to mass percent is
The DFO internal layer coaxial electrically spun sandwich layer solution of 80wt.%；
C. it is 1 by volume ratio:The mixture of the dichloromethane DCM of 1 n,N-Dimethylformamide DMF, will as solvent
2g polycaprolactone (PCL)s are dissolved in the above-mentioned solvents of 20ml, are configured to PCL coaxial electrically spun shell solution, then be bisected into two parts；
D. sodium alginate SA is dissolved in deionized water, is configured to the SA solution 10g that mass fraction is 5wt.%, then
It is 1.2 in mass ratio by the PVA solution and SA solution that are prepared in the step a:1 ratio mixing, is configured to intravascular stent
Intermediate layer solution；
E. gentamicin GS is dissolved in the PVA solution prepared in the step a of 10g, is configured to mass fraction
For the GS outer layer coaxial electrically spun sandwich layer solution of 0.8wt.%；
F. it is respectively adopted and DFO internal layers coaxial electrically spun sandwich layer solution is prepared in the step b and is prepared in the step c
A PCL coaxial electrically spun shell solution, and use coaxial electrically spun forming technology, wherein DFO internal layers coaxial electrically spun sandwich layer is molten
Liquid and PCL coaxial electrically spun shell solution carry out spinning by Coaxial nozzle, and PCL coaxial electrically spun sandwich layers are sequentially prepared on circular shaft U
With polycaprolactone (PCL) shell, it is made and carries medicine regeneration vessel stent internal layer；When preparing intravascular stent internal layer, the rotation of control circular shaft U
Rotary speed is 200 revs/min, and it is 0.3mm/s that X-direction, that is, circular shaft U horizontal axis, which moves back and forth speed, controls the voltage of Electrospun
For 13KV, electrospinning liquid is set to be fed by micro pump, the wherein electrospinning time is 10min, and the feeding speed for controlling micro pump is 30ul/
Min, height of the Coaxial nozzle from circular shaft U are 140mm, and circular shaft U uses conductive stainless steel, and circular shaft U's is a diameter of
2mm；The internal diameter of Coaxial nozzle is 0.3mm, outside diameter 1mm；
G. infusion process forming technology is used, the circle of load medicine regeneration vessel stent internal layer preparation will be completed in the step f
Axis is removed, and is immersed in the intermediate layer solution prepared in the step d, stands 60 minutes, will carry medicine regeneration vessel branch afterwards
The circular shaft of frame internal layer dipping attachment hydrogel takes out, and circular shaft both ends are maked somebody a mere figurehead standing, and until circular shaft, there is no hydrogel drop
Untill dripping, then circular shaft is put in -60 DEG C of refrigerator carry out frost 15 it is small when after, circular shaft is taken out, and by circular shaft in room temperature
It is middle thaw 8 it is small when, place into refrigerator and freezed, so undergo 5 freeze-thawing processes so that carry medicine regeneration vessel
The hydrogel intermediate layer that the outside of stent internal layer is shaped；
H. it is respectively adopted and GS outer layers coaxial electrically spun sandwich layer solution is prepared in the step e and is prepared in the step c
Another PCL coaxial electrically spun shell solution, and use coaxial electrically spun forming technology, it is same that GS outer layers are sequentially prepared on circular shaft U
Axis electrospinning sandwich layer and another layer of polycaprolactone (PCL) shell, are made and carry medicine regeneration vessel stent outer layer, so that in hydrogel intermediate layer
Exterior parcel carry medicine regeneration vessel stent outer layer, obtain the molding load coaxial regeneration vessel supporting frame prefabrication body of medicine；Carried preparing
During the coaxial regeneration vessel stent outer layer of medicine, the parameter of circular shaft U, micro pump and nozzle and intravascular stent is prepared in the step f
Process conditions are identical during internal layer, are particular in that it is 20min to prepare the electrospinning time for carrying medicine regeneration vessel stent outer layer；
I. the coaxial regeneration vessel supporting frame prefabrication body of the load medicine prepared in the step h is immersed in mass fraction is
It is chemically crosslinked within 15 minutes in the calcium chloride solution of 1wt.%, after completing to be chemically crosslinked, obtains carrying the coaxial regeneration vessel of medicine
Stent, will take out after then carrying the coaxial regeneration vessel stent freeze-drying of medicine, has just obtained the coaxial regeneration vessel branch of load medicine and put up
Product.
In the present embodiment, prepared using combination process and carry the coaxial regeneration vessel stent of medicine, shown intravascular stent there are three layers
Structure, is respectively adopted different manufacturing process, and wherein internal layer is coaxial electrically spun DFO solution, and intermediate layer is selected PVA+SA solution, adopted
Method is physical crosslinking with freeze-thawing, outermost layer is coaxial electrically spun GS solution.The present embodiment is compounded with three kinds of different shaping sides
Method, simulates the three-decker of native blood vessels, shortens the time that intravascular stent is cultivated in vitro, and medicine skill is carried as a result of coaxial
Art, can making intravascular stent, slowly release, the DFO of internal layer can promote the generation of new vessels on time in vivo, and intermediate layer can ensure
The wall thickness and mechanical strength of blood vessel, the GS of outer layer can reduce the infection rate of tissue, before it is clinically had wide application
Scape.
The embodiment of the present invention is illustrated above in conjunction with attached drawing, but the invention is not restricted to above-described embodiment, can be with
The purpose of innovation and creation according to the present invention makes a variety of changes, under all Spirit Essence and principle according to technical solution of the present invention
Change, modification, replacement, combination or the simplification made, should be equivalent substitute mode, as long as meeting the goal of the invention of the present invention,
Without departing from technical principle and the invention of the invention for manually carrying the coaxial regeneration vessel stent of medicine and its combination process preparation method
Design, belongs to protection scope of the present invention.
Claims (10)
1. a kind of manually carry the coaxial regeneration vessel stent of medicine, it is characterised in that：From the coaxial regeneration vessel internal stent of medicine is carried to outer
Portion, is combined closely and is formed by coaxial internal layer, hydrogel intermediate layer and outer layer successively, simulate the three-decker of native blood vessels, its
Described in internal layer combined closely and formed by the compound sandwich layers of Deferoxamine DFO and the first polycaprolactone (PCL) shell successively, it is poly- to form first
Caprolactone PCL shells wrap up the compound sandwich layers of Deferoxamine DFO lamellar composite endothecium structure, the compound sandwich layers of Deferoxamine DFO by
The composite material of Deferoxamine DFO and PVA are made, and the compound sandwich layers of Deferoxamine DFO, which are directly toward, carries the coaxial regeneration vessel of medicine
Inner cavity is set, and the hydrogel intermediate layer is made of the composite material of sodium alginate SA and PVA, and the outer layer is big mould by celebrating successively
The compound sandwich layer of plain GS and the second polycaprolactone (PCL) shell are combined closely and are formed, and it is big to form the parcel celebrating of the second polycaprolactone (PCL) shell
The lamellar composite layer structure of the compound sandwich layers of mycin GS, the compound sandwich layers of gentamicin GS the answering by gentamicin GS and PVA
Condensation material is made, the first polycaprolactone (PCL) shell and the compound sandwich layers of gentamicin GS respectively with hydrogel intermediate layer two
Side is combined closely, and the second polycaprolactone (PCL) shell is directly toward the exterior setting for carrying the coaxial regeneration vessel of medicine.
2. the coaxial regeneration vessel stent of medicine is manually carried according to claim 1, it is characterised in that：The Deferoxamine DFO is compound
The constituent mass proportioning of the composite material of the Deferoxamine DFO and PVA of sandwich layer is (15-64):1000；The gentamicin GS is compound
The constituent mass proportioning of the composite material of the gentamicin GS and PVA of sandwich layer is (3-8):1000.
3. according to claim 1 or claim 2 manually carry the coaxial regeneration vessel stent of medicine, it is characterised in that：Among the hydrogel
The constituent mass proportioning of the composite material of the sodium alginate PVA and SA of layer is (2.4-9.6):(1-5).
4. according to claim 1 or claim 2 manually carry the coaxial regeneration vessel stent of medicine, it is characterised in that：Described first gathers in oneself
Ester PCL shells or the second polycaprolactone (PCL) shell are made of polycaprolactone (PCL) and basis material blending, described matrix material by
N,N-dimethylformamide DMF and dichloromethane DCM are mixed；Wherein N,N-dimethylformamide DMF：Dichloromethane DCM
Volume ratio be 1:1, the mass volume ratio of polycaprolactone (PCL) and basis material is (1-10) g：100ml.
5. according to claim 1 or claim 2 manually carry the coaxial regeneration vessel stent of medicine, it is characterised in that：By adjusting hydrogel
The thickness in intermediate layer manually carries the overall wall thickness of the coaxial regeneration vessel stent of medicine to control.
6. a kind of combination process prepares the method for carrying the coaxial regeneration vessel stent of medicine, it is characterised in that includes the following steps：
A. PVA is dissolved in deionized water, the heating water bath on magnetic stirring apparatus, and stir until PVA be completely dissolved, be made
Mass percent concentration is the PVA solution of 3-8wt.%；
B., Deferoxamine DFO is dissolved in the PVA solution prepared in the step a, it is 50-80wt.% to be configured to mass percent
DFO internal layer coaxial electrically spun sandwich layer solution；
C. it is 1 by volume ratio:The mixture of the dichloromethane DCM of 1 n,N-Dimethylformamide DMF will gather oneself as solvent
Lactone PCL is dissolved in solvent, and the mass volume ratio for being configured to polycaprolactone (PCL) and solvent is (1-10) g：The PCL of 100ml is same
Axis electrospinning shell solution；
D. sodium alginate SA is dissolved in deionized water, is configured to the SA solution that mass fraction is 1-5wt.%, then will be
The PVA solution and SA solution prepared in the step a is (0.8-1.2) in mass ratio：1 ratio mixing, is configured to blood vessel branch
The intermediate layer solution of frame；
E. gentamicin GS is dissolved in the PVA solution prepared in the step a, it is 0.3- to be configured to mass fraction
The GS outer layer coaxial electrically spun sandwich layer solution of 0.8wt.%；
F. it is respectively adopted and DFO internal layers coaxial electrically spun sandwich layer solution is prepared in the step b and prepares PCL in the step c
Coaxial electrically spun shell solution, and use coaxial electrically spun forming technology, be sequentially prepared on circular shaft (U) PCL coaxial electrically spuns sandwich layer and
Polycaprolactone (PCL) shell, is made and carries medicine regeneration vessel stent internal layer；
G. infusion process forming technology is used, the circular shaft for completing to carry the preparation of medicine regeneration vessel stent internal layer in the step f is taken
Under, and be immersed in the intermediate layer solution prepared in the step d, 15~60 minutes are stood, medicine regeneration vessel branch will be carried afterwards
The circular shaft of frame internal layer dipping attachment hydrogel takes out, and circular shaft both ends are maked somebody a mere figurehead standing, and until circular shaft, there is no hydrogel drop
Untill dripping, then circular shaft is put in -40~-60 DEG C of refrigerator carry out frost 10-15 it is small when after, take out circular shaft, and will circle
Axis is placed into refrigerator and freezed when defrosting 4-8 is small at room temperature, so undergoes 1-5 freeze-thawing process, so that
Carry the hydrogel intermediate layer that the outside of medicine regeneration vessel stent internal layer is shaped；
H. the preparation GS outer layers coaxial electrically spun sandwich layer solution in the step e is respectively adopted and preparation PCL is same in the step c
Axis electrospinning shell solution, and use coaxial electrically spun forming technology, be sequentially prepared on circular shaft (U) GS outer layer coaxial electrically spun sandwich layers and
Polycaprolactone (PCL) shell, is made and carries medicine regeneration vessel stent outer layer, so that the exterior parcel in hydrogel intermediate layer carries medicine regeneration
Intravascular stent outer layer, obtains the molding load coaxial regeneration vessel supporting frame prefabrication body of medicine；
I. it is 1- the coaxial regeneration vessel supporting frame prefabrication body of the load medicine prepared in the step h to be immersed in mass fraction
It is chemically crosslinked within 15~60 minutes in the calcium chloride solution of 5wt.%, after completing to be chemically crosslinked, obtains load medicine and coaxially regenerate
Intravascular stent, will take out after then carrying the coaxial regeneration vessel stent freeze-drying of medicine, has just obtained the coaxial regeneration vessel branch of load medicine
Put up product.
7. combination process prepares the method for carrying the coaxial regeneration vessel stent of medicine according to claim 6, it is characterised in that：Institute
State when preparing intravascular stent ectonexine in step f and step h, the rotary speed for controlling circular shaft (U) is 200-400 revs/min, X
It is 0.3-0.6mm/s that direction, that is, circular shaft (U) horizontal axis, which moves back and forth speed, and the voltage for controlling Electrospun is 10-13KV, makes electricity
Liquid is spun to be fed by micro pump；The electrospinning time wherein in the step f is 3-10min, during electrospinning in the step h
Between be 4-20min, adjust the thickness of electrospinning film by controlling the Electrospun time, thus adjust carry medicine regeneration vessel stent internal layer
Or the thickness of outer layer.
8. combination process prepares the method for carrying the coaxial regeneration vessel stent of medicine according to claim 6, it is characterised in that：Institute
State when intravascular stent intermediate layer is prepared in step g, the number of freeze-thawing is determined by required regeneration vessel wall thickness, is led to
Cross the number of control freeze-thawing and lead to the thickness in prepared hydrogel intermediate layer to adjust, and then regulate and control manually to carry medicine coaxially again
The overall wall thickness of green blood pipe holder.
9. combination process prepares the method for carrying the coaxial regeneration vessel stent of medicine according to any one in claim 6~8, its
It is characterized in that：When preparing intravascular stent ectonexine in the step f and step h, the feeding speed for controlling micro pump is 30-
60ul/min, Coaxial nozzle are highly 140-160mm from circular shaft (U).
10. combination process prepares the method for carrying the coaxial regeneration vessel stent of medicine according to any one in claim 6~8,
It is characterized in that：When preparing intravascular stent ectonexine in the step f and step h, circular shaft (U) uses conductive stainless steel
Material, a diameter of 2-9mm of circular shaft (U)；The internal diameter of Coaxial nozzle is not more than 0.3mm, and outside diameter is not more than 1mm.
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| CN113842504A (en) * | 2021-09-22 | 2021-12-28 | 苏州大学附属第二医院 | Preparation method of multifunctional electrospinning bracket for bone regeneration |
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