CN108478863A - The preparation method and products thereof of compound small-caliber artificial blood vessel - Google Patents
The preparation method and products thereof of compound small-caliber artificial blood vessel Download PDFInfo
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- CN108478863A CN108478863A CN201810374391.1A CN201810374391A CN108478863A CN 108478863 A CN108478863 A CN 108478863A CN 201810374391 A CN201810374391 A CN 201810374391A CN 108478863 A CN108478863 A CN 108478863A
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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
- A61L27/50—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
-
- 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
- A61F2/062—Apparatus for the production of blood vessels made from natural tissue or with layers of living cells
-
- 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
- A61L27/18—Macromolecular materials obtained otherwise than by reactions only involving carbon-to-carbon unsaturated bonds
-
- 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
- A61L27/22—Polypeptides or derivatives thereof, e.g. degradation products
- A61L27/222—Gelatin
-
- 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
- A61L27/26—Mixtures of macromolecular compounds
-
- 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
- A61L27/36—Materials for grafts or prostheses or for coating grafts or prostheses containing ingredients of undetermined constitution or reaction products thereof, e.g. transplant tissue, natural bone, extracellular matrix
- A61L27/3604—Materials for grafts or prostheses or for coating grafts or prostheses containing ingredients of undetermined constitution or reaction products thereof, e.g. transplant tissue, natural bone, extracellular matrix characterised by the human or animal origin of the biological material, e.g. hair, fascia, fish scales, silk, shellac, pericardium, pleura, renal tissue, amniotic membrane, parenchymal tissue, fetal tissue, muscle tissue, fat tissue, enamel
- A61L27/3625—Vascular tissue, e.g. heart valves
-
- 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
- A61L27/36—Materials for grafts or prostheses or for coating grafts or prostheses containing ingredients of undetermined constitution or reaction products thereof, e.g. transplant tissue, natural bone, extracellular matrix
- A61L27/3683—Materials for grafts or prostheses or for coating grafts or prostheses containing ingredients of undetermined constitution or reaction products thereof, e.g. transplant tissue, natural bone, extracellular matrix subjected to a specific treatment prior to implantation, e.g. decellularising, demineralising, grinding, cellular disruption/non-collagenous protein removal, anti-calcification, crosslinking, supercritical fluid extraction, enzyme treatment
- A61L27/3687—Materials for grafts or prostheses or for coating grafts or prostheses containing ingredients of undetermined constitution or reaction products thereof, e.g. transplant tissue, natural bone, extracellular matrix subjected to a specific treatment prior to implantation, e.g. decellularising, demineralising, grinding, cellular disruption/non-collagenous protein removal, anti-calcification, crosslinking, supercritical fluid extraction, enzyme treatment characterised by the use of chemical agents in the treatment, e.g. specific enzymes, detergents, capping agents, crosslinkers, anticalcification agents
-
- 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
- A61L33/00—Antithrombogenic treatment of surgical articles, e.g. sutures, catheters, prostheses, or of articles for the manipulation or conditioning of blood; Materials for such treatment
- A61L33/0005—Use of materials characterised by their function or physical properties
- A61L33/0011—Anticoagulant, e.g. heparin, platelet aggregation inhibitor, fibrinolytic agent, other than enzymes, attached to the substrate
-
- 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
- A61L33/00—Antithrombogenic treatment of surgical articles, e.g. sutures, catheters, prostheses, or of articles for the manipulation or conditioning of blood; Materials for such treatment
- A61L33/06—Use of macromolecular materials
- A61L33/08—Polysaccharides
-
- 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
- A61F2210/00—Particular material properties of prostheses classified in groups A61F2/00 - A61F2/26 or A61F2/82 or A61F9/00 or A61F11/00 or subgroups thereof
- A61F2210/0076—Particular material properties of prostheses classified in groups A61F2/00 - A61F2/26 or A61F2/82 or A61F9/00 or A61F11/00 or subgroups thereof multilayered, e.g. laminated structures
-
- 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
- A61F2210/00—Particular material properties of prostheses classified in groups A61F2/00 - A61F2/26 or A61F2/82 or A61F9/00 or A61F11/00 or subgroups thereof
- A61F2210/0085—Particular material properties of prostheses classified in groups A61F2/00 - A61F2/26 or A61F2/82 or A61F9/00 or A61F11/00 or subgroups thereof hardenable in situ, e.g. epoxy resins
Abstract
The present invention relates to a kind of preparation methods and products thereof of compound small-caliber artificial blood vessel, and preparation method includes the following steps：Coronary artery is subjected to de- cell processing, obtains and takes off cellular vascular；Then by de- cellular vascular test tube of hepari, then with the mixing electrospun solution of gelatin and poly- L lactides caprolactone (PLCL) carries out spinning, removal organic solvent obtains compound small-caliber artificial blood vessel；Artificial blood vessel obtained is double-layer tissue engineering blood vessel, using de- cell porcine coronary as artificial blood vessel's internal layer, has good compliance and cell compatibility, can promote sticking, be proliferated and break up, sprawling for cell, complete the endothelialization again of blood vessel；Outer layer is the blended layer of PLCL and gelatin, increases the mechanical property of blood vessel, is conducive to the isocellular intrusion of smooth muscle, forms the middle level of blood vessel；The inner surface of internal layer can use different drug modifications, such as heparin, improve artificial blood vessel's antithrombotic and endangium is promoted to be formed, be conducive to the reconstruction of blood vessel.
Description
Technical field
The invention belongs to medical instrument preparation fields, are related to the preparation method of compound small-caliber artificial blood vessel, further relate to by
Product made from this method.
Background technology
As the number for suffering from angiocardiopathy (Coronary artery disease, CAD) constantly rises, artificial blood vessel
Transplanting demand increases year by year.Clinically, heavy caliber blood vessel has been obtained for applying well, but uses identical material system
Small-caliber vascular (the diameter obtained<It 6mm) is but easy to happen thrombosis problem, reason may be with graft material performance itself
It is related.Autologous vein is considered as the goldstandard clinically used, but autologous vein take to patient there are huge wound and
Desirable limited amount, therefore find, the blood of good biocompatibility and its abundance similar with human body native blood vessel performance
Tube material is particularly important.
De- cellular vascular, which eliminates heterogenous cell, can efficiently reduce rejection, while remain extracellular matrix, mainly
For collagen and elastomer, these substances have very high homology in mammals, and species variation is small, and this blood vessel is due to source
In native blood vessels, so having good compliance and cell compatibility, sticking, be proliferated and break up, spreading for cell can be promoted
Exhibition, completes the endothelialization again of blood vessel.But vascular grafts, after de- cell processing, the endothelial cell of blood vessel is gone
It removing, collagen exposes under inner membrance, can activate blood coagulation system, thus if de- cellular vascular is directly used as alternative materials implant
It is interior, easily cause thrombus, patency rate is not high, and through take off cell processing after vascular mechanics reduced performance, elastin degradation
Accelerate etc., so as to cause aneurysmal formation.
Therefore, it is badly in need of a kind of antithrombotic artificial blood vessel, and there is preferable mechanical property.
Invention content
In view of this, the purpose of the present invention is to provide a kind of preparation method of compound small-caliber artificial blood vessel, pass through use
Drug such as heparin (heparin) modification takes off cell porcine coronary blood vessel (Decellularized porcine coronary
Artery, DPCA), achieve the purpose that antithrombotic.Then electrostatic spinning technique is recycled, by Poly L-lactide-caprolactone (Poly
(L-Lactide-co-caprolactone), abridge PLCL) and the mixed solution of gelatin (gelatin) spin on the surfaces DPCA, make
For at compound small-caliber artificial blood vessel, i.e. double-layer tissue engineering blood vessel (Bilayer tissue-engineered vascular
Graft, BTEV), increase the mechanical property of blood vessel.The second object of the present invention is to provide made from the preparation method
Compound small-caliber artificial blood vessel.
In order to achieve the above objectives, the present invention provides the following technical solutions：
1. the preparation method of compound small-caliber artificial blood vessel, includes the following steps：Coronary artery is subjected to de- cell processing,
It obtains and takes off cellular vascular；Then in the inner wall modified medicaments of de- cellular vascular, then with the mixing electrospun solution of gelatin and PLCL into
Row spinning removes organic solvent, obtains compound small-caliber artificial blood vessel.
Preferably, the de- cell processing is to take out and be placed in -80 DEG C of ice coronary artery water Hypotonic treatment in 4h
Case 30min, then 18~25 DEG C of placement 30min, are repeated 2 times；Hypotonic treatment uses the pancreatin 37 of mass fraction 0.125% afterwards for 24 hours
DEG C shaking table 1.5h, removes remaining pancreatin；Last 4 DEG C are stored in containing in the dual anti-sterile PBS of Pen .- Strep or passing through
It is preserved after vacuum freeze drier drying, wherein penicillin concn is 100U/ml, a concentration of 0.1mg/ml of streptomysin.
Preferably, the drug is heparin.
It is furthermore preferred that the method for de- cellular vascular modification test tube of hepari is as follows：2- (the N- of pH5.6,0.05mol/L will be taken
Coffee quinoline) ethanesulfonic acid buffer, heparin, 1- (3- dimethylamino-propyls) -3- ethyl-carbodiimide hydrochlorides, N- hydroxysuccinimidyls is added
Acid imide to final concentration is respectively 0.2g/mL, 0.3834g/mL, 0.23g/mL, and pre-reaction 10min makes the carboxylic of heparin at 37 DEG C
Base activates；De- cellular vascular is added, 37 DEG C of response light microseisms are swung for 24 hours, are then used PBS neutralization reactions and are removed unbonded liver
Element.Preferably, the mixing electrospun solution is by volume ratio 2：8 gelatin solution and PLCL solution compositions.
It is furthermore preferred that the mass fraction of institute's gelatine solution is 10%, the mass fraction of the PLCL solution is 10%.
It is formulated it is furthermore preferred that 20ml hexafluoroisopropanols are added by every 2g PLCL in the PLCL solution；The gelatin is molten
Liquid is added 20ml hexafluoroisopropanols by every 2g gelatin and is formulated.
It is furthermore preferred that electrospun solution flow velocity is 1ml/h to the spinning condition in order to control, rotating speed 1000rpm, voltage are
Electrospinning under the conditions of 12kV.
2. the compound small-caliber artificial blood vessel made from the preparation method.
Preferably, the compound small-caliber artificial blood vessel includes de- cell porcine coronary and PLCL and gelatin from the inside to the outside
Blended layer, wherein de- cell porcine coronary inner wall is modified with heparin.It is furthermore preferred that the drug is heparin.
The beneficial effects of the present invention are：The invention discloses the preparation method of compound small-caliber artificial blood vessel, using de-
Cell porcine coronary and electrostatic spinning prepare compound small-caliber artificial blood vessel (diameter<6mm)；Compound small-caliber artificial obtained
Blood vessel is double-layer tissue engineering blood vessel, including one section of de- cell porcine coronary has suitable well as artificial blood vessel's internal layer
Answering property and cell compatibility can promote sticking, be proliferated and break up, sprawling for cell, complete the endothelialization again of blood vessel；Outer layer is
The blended layer of PLCL and gelatin increase the mechanical property of blood vessel, are conducive to the isocellular intrusion of smooth muscle, are formed in blood vessel
Layer；The inner surface of internal layer can use different drug modifications, improve artificial blood vessel's antithrombotic and endangium is promoted to be formed, be conducive to
The reconstruction of blood vessel.
Description of the drawings
In order to keep the purpose of the present invention, technical solution and advantageous effect clearer, the present invention provides following attached drawing and carries out
Explanation：
Fig. 1 is the de- cell coronary artery after vacuum freeze drying.
Fig. 2 is compound small-caliber artificial blood vessel (BTEV) and de- cell coronary artery (DPCA) figure.
Fig. 3 is the scanning electron microscope (SEM) photograph of compound small-caliber artificial blood vessel cross section (B is the partial enlarged view of A).
Fig. 4 is compound small-caliber artificial blood vessel cross-sectional view.
Fig. 5 is the histotomy figure of blood vessel graft and native blood vessels.
Specific implementation mode
Below in conjunction with attached drawing, the preferred embodiment of the present invention is described in detail.
The material Pigs Hearts used in the embodiment of the present invention is from the double upright stone tablet slaughterhouses of Chongqing Lian Xin Food Co., Ltd.
The preparation method of embodiment 1, compound small-caliber artificial blood vessel
The preparation method of compound small-caliber artificial blood vessel, is as follows：
(1) collection of vascular grafts
It is cleaned immediately with sterile saline after taking out heart, is subsequently placed in frost physiological saline and is transported to laboratory, with
Afterwards using the careful separation Pigs Hearts blood vessel of tweezers and scissors, after removing fat and adherent tissue, with sterile cold saline pair
Coronary artery carries out cleaning down；
(2) preparation of cellular vascular is taken off
The coronary artery of separation is placed in shaking table Hypotonic treatment in sterile distilled water for 24 hours, to take out in 4h and be placed in -80 DEG C
Refrigerator 30min, is then placed at room temperature for 30min, and the above operation is repeated 2 times；Hypotonic treatment for 24 hours afterwards with 0.125% 37 DEG C of pancreatin
Shaking table 1.5h, then 3h is handled with distilled water shaking table, thoroughly remove remaining pancreatin；Last 4 DEG C are stored in containing dual anti-(penicillin
A concentration of 0.1mg/ml of a concentration of 100U/ml, streptomysin) sterile PBS in it is spare, or after vacuum freeze drier is dried
It is stored in spare in drying box；Blood vessel after freeze-dried is as shown in Figure 1.
(3) test tube of hepari of cellular vascular is taken off
Heparin is made using 1- (3- dimethylamino-propyls) -3- ethyl-carbodiimide hydrochlorides and n-hydroxysuccinimide
Amino forms stable amido bond with remaining carboxyl on acellular matrix.Concrete operations are：First take 2- (N- morpholines) second sulphur
Acid buffer (0.05mol/L, pH5.6) 50mL, is added 0.1g heparin, 0.1917g EDC and 0.115g NHS, pre- anti-at 37 DEG C
10min is answered, by the activated carboxylic of heparin；De- cellular vascular is added, 37 DEG C of response light microseisms are swung for 24 hours, are then used in PBS and anti-
Should and it remove unbonded heparin.
(4) preparation of electrospun solution
2g PLCL are weighed every time to be added in 25ml reagent bottles, and 20ml hexafluoroisopropanol organic solvents are added with liquid-transfering gun, match
Mass fraction processed be 10% (the PLCL solution of polymer quality (g)/solvent volume (m1), at room temperature magnetite be stirred overnight or extremely
Polymer is completely dissolved；Same method prepares the gelatin solution that mass fraction is 10%.Then use mass fraction be 10% it is bright
The PLCL solution that sol solution and mass fraction are 10% by volume 2：8 configuration gelatin-PLCL mix electrospun solution.
(5) electrospinning prepares the blended layer of PLCL and gelatin
The de- cellular vascular of test tube of hepari is through dimension artificial blood vessel to receive on stick, is put into vacuum freeze drier mistake
At night, it is 1ml/h, electrospinning under the conditions of rotating speed 1000rpm, voltage are 12kV, after electrospinning then to control electrospun solution flow velocity
72h or more is dried in vacuo to remove organic solvent as possible, obtains compound small-caliber artificial blood vessel.
Compound small-caliber artificial blood vessel obtained is as shown in Figures 2 and 3, after the sterilizing of 1% Peracetic acid, is placed in sterile physiological
It is spare in brine.
The cross-sectional view of its artificial blood vessel is as shown in Figure 4.As seen from Figure 4, compound small-caliber artificial blood vessel by
Interior to outer includes the blended layer of de- cell porcine coronary and PLCL and gelatin, wherein de- cell porcine coronary inner wall is modified with
Heparin.Therefore, compound small-caliber artificial blood vessel inner wall produced by the present invention is modified with heparin, the purpose with antithrombotic；Outer layer
PLCL and gelatin blended layer have increase blood vessel mechanical property.
Embodiment 2, artificial blood vessel are implanted into rat body
Using weight 300g or so SD rats as experimental subjects, 48h is fasted before artificial vessel replacement's experiment, uses grape instead
Syrup is raised.0.5ml/100g dosage intraperitoneal injection of anesthesia is pressed with the chloraldurate of mass fraction 7%, waits for SD rat holonarcosis
Afterwards, the hair that abdomen is then removed with shaver, is in dorsal position by rat, and four limbs stretching, extension is fixed on operating table.By Surgery
75% medicinal alcohol of position volume fraction sterilizes, and opens shadowless lamp and is directed at abdomen, cuts off skin, and abdomen is opened further along ventrimeson
Stomach is pushed into both sides by chamber with wet cotton ball, then exposure abdominal aorta goes out abdominal aorta blood vessel with tweezers careful separation.With
100U/kg dosage injecting heparin sodium is pressed from tail vein afterwards, whole body test tube of hepari processing is carried out to rat.Thin vessels ligature in time, prevent
Only accidental haemorrhage.After isolating abdominal aorta, inferior caval vein lower end and common iliac artery upper end are clamped with hopkins' vascular clamp, scissors for vessels is therefrom
Between cut blood vessel.End to end anastomosis artificial blood vessel and rat aorta, uniformly take the needle, one week about 8 needle, first loose after coincideing
The blood vessel clip of distal end is opened, situation of coincideing is observed, sees if there is leakage blood, if occurring at apparent leakage blood, is mended at leakage blood
Needle；Then the blood vessel clip of proximal part is unclamped again, repeats to mend needle operation, until without leakage blood phenomenon.Wait for that both ends blood vessel clip unclamps
Afterwards, without apparent leakage blood, and artificial blood vessel is well then considered as the success of artificial vessel replacement's autologous vein with autologous vein beating, then
It takes out cotton balls, clear up abdominal cavity with physiological saline, musculature of coincideing step by step is finally closed skin.Entire vascular anastomosis is excessively program-controlled
System is within 50min.
The middle film of blood vessel graft remolds process as shown in figure 5, with the extension of time, cell is gradually total to blood vessel pipe to people
Infiltration in wall.After the transfer first week, blood vessel was wrapped up by cambium, is had a large amount of cells in cambium and is migrated by outer membrane
To middle film, the degradation of outer layer electrostatic spinning shows part cavity；In second week, tube wall inner cell increases, and middle film and inner membrance are equal
Have cellular infiltration, and form new intima, and the cell of middle film present it is certain put in order, the gap after electrostatic spinning degradation
It is filled by cambium；When to 4th week, middle film inner cell aligns, and inner membrance is covered by cambium, realizes endothelialization, bullet
Power plate shows waveform, entire blood vessel structure and is intended to normal vascular tissues.Illustrate composite double layer small-caliber artificial blood vessel
The reconstruction being implanted with conducive to blood vessel.
Finally illustrate, preferred embodiment above is merely illustrative of the technical solution of the present invention and unrestricted, although logical
It crosses above preferred embodiment the present invention is described in detail, however, those skilled in the art should understand that, can be
Various changes are made to it in form and in details, without departing from claims of the present invention limited range.
Claims (10)
1. the preparation method of compound small-caliber artificial blood vessel, which is characterized in that include the following steps：Coronary artery is carried out de- thin
Born of the same parents are handled, and are obtained and are taken off cellular vascular；Then in the inner wall modified medicaments of de- cellular vascular, then with gelatin and Poly L-lactide-oneself
The mixing electrospun solution of lactone carries out spinning, removes organic solvent, obtains compound small-caliber artificial blood vessel.
2. the preparation method of compound small-caliber artificial blood vessel according to claim 1, it is characterised in that：The de- cell processing
It is that coronary artery water Hypotonic treatment is taken out in 4h and is placed in -80 DEG C of refrigerator 30min, then 18~25 DEG C of placements
30min is repeated 2 times；Hypotonic treatment uses 37 DEG C of shaking table 1.5h of pancreatin of mass fraction 0.125% afterwards for 24 hours, removes remaining pancreas
Enzyme；Last 4 DEG C are stored in containing being preserved in the dual anti-sterile PBS of Pen .- Strep or after vacuum freeze drying, wherein
Penicillin concn is 100U/ml, a concentration of 0.1mg/ml of streptomysin.
3. the preparation method of compound small-caliber artificial blood vessel according to claim 1, it is characterised in that：The drug is liver
Element.
4. the preparation method of compound small-caliber artificial blood vessel according to claim 3, it is characterised in that：De- cellular vascular modification
The method of test tube of hepari is as follows：2- (N- morpholines) ethanesulfonic acid buffer of pH5.6,0.05mol/L will be taken, heparin, 1- (3- is added
Dimethylamino-propyl) -3- ethyl-carbodiimide hydrochlorides, n-hydroxysuccinimide to final concentration be respectively 0.2g/mL,
0.3834g/mL, 0.23g/mL, pre-reaction 10min makes the activated carboxylic of heparin at 37 DEG C；Add de- cellular vascular, 37 DEG C
Response light microseism is swung for 24 hours, is then used PBS neutralization reactions and is removed unbonded heparin.
5. the preparation method of compound small-caliber artificial blood vessel according to claim 1, it is characterised in that：The mixing electrospinning is molten
Liquid is by volume ratio 2：8 gelatin solution and Poly L-lactide-caprolactone solution composition.
6. the preparation method of compound small-caliber artificial blood vessel according to claim 4, it is characterised in that：Institute's gelatine solution
Mass fraction is 10%, and the mass fraction of the Poly L-lactide-caprolactone solution is 10%.
7. the preparation method of compound small-caliber artificial blood vessel according to claim 1, it is characterised in that：The poly- L- third is handed over
Ester-caprolactone solution is added 20ml hexafluoroisopropanols by every 2g Poly L-lactide-caprolactone and is formulated；Institute's gelatine solution is pressed
20ml hexafluoroisopropanols are added per 2g gelatin to be formulated.
8. the preparation method of compound small-caliber artificial blood vessel according to claim 1, it is characterised in that：The spinning condition is
Control electrospun solution flow velocity is 1ml/h, electrospinning under the conditions of rotating speed 1000rpm, voltage are 12kV.
9. the compound small-caliber artificial blood vessel made from claim 1~8 any one of them preparation method.
10. compound small-caliber artificial blood vessel according to claim 9, it is characterised in that：The compound small-caliber artificial blood
Pipe includes the blended layer of de- cell porcine coronary, Poly L-lactide-caprolactone and gelatin from the inside to the outside, wherein de- cell pig hat
Shape Wall of Artery is modified with drug.
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| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201810374391.1A CN108478863A (en) | 2018-04-24 | 2018-04-24 | The preparation method and products thereof of compound small-caliber artificial blood vessel |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201810374391.1A CN108478863A (en) | 2018-04-24 | 2018-04-24 | The preparation method and products thereof of compound small-caliber artificial blood vessel |
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| Publication Number | Publication Date |
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| CN108478863A true CN108478863A (en) | 2018-09-04 |
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Cited By (6)
| Publication number | Priority date | Publication date | Assignee | Title |
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| CN109498839A (en) * | 2018-11-13 | 2019-03-22 | 南开大学 | A kind of biology composite artificial blood vessel and application |
| CN110755174A (en) * | 2019-10-31 | 2020-02-07 | 重庆大学 | Biological mixed type artificial blood vessel and preparation method thereof |
| CN111850818A (en) * | 2019-04-30 | 2020-10-30 | 深圳市罗湖区人民医院 | Preparation method and product of conjugate electrospun nanofiber artificial small-caliber intravascular stent |
| CN112274699A (en) * | 2019-07-24 | 2021-01-29 | 西安交通大学医学院第一附属医院 | Small-caliber tissue improved composite artificial blood vessel |
| CN113855859A (en) * | 2021-05-28 | 2021-12-31 | 首都医科大学宣武医院 | Small-caliber tissue engineering blood vessel constructed by acellular vascular matrix and capable of promoting rapid endothelialization |
| CN115137881A (en) * | 2022-07-27 | 2022-10-04 | 天津大学温州安全(应急)研究院 | Three-layer bionic artificial blood vessel with antithrombotic and tissue regeneration promoting functions and preparation method thereof |
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Cited By (9)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN109498839A (en) * | 2018-11-13 | 2019-03-22 | 南开大学 | A kind of biology composite artificial blood vessel and application |
| CN111850818A (en) * | 2019-04-30 | 2020-10-30 | 深圳市罗湖区人民医院 | Preparation method and product of conjugate electrospun nanofiber artificial small-caliber intravascular stent |
| CN111850818B (en) * | 2019-04-30 | 2022-07-15 | 深圳市罗湖区人民医院 | Preparation method and product of conjugate electrospun nanofiber artificial small-caliber intravascular stent |
| CN112274699A (en) * | 2019-07-24 | 2021-01-29 | 西安交通大学医学院第一附属医院 | Small-caliber tissue improved composite artificial blood vessel |
| CN110755174A (en) * | 2019-10-31 | 2020-02-07 | 重庆大学 | Biological mixed type artificial blood vessel and preparation method thereof |
| CN110755174B (en) * | 2019-10-31 | 2021-10-15 | 重庆大学 | Biological mixed type artificial blood vessel and preparation method thereof |
| CN113855859A (en) * | 2021-05-28 | 2021-12-31 | 首都医科大学宣武医院 | Small-caliber tissue engineering blood vessel constructed by acellular vascular matrix and capable of promoting rapid endothelialization |
| CN115137881A (en) * | 2022-07-27 | 2022-10-04 | 天津大学温州安全(应急)研究院 | Three-layer bionic artificial blood vessel with antithrombotic and tissue regeneration promoting functions and preparation method thereof |
| CN115137881B (en) * | 2022-07-27 | 2023-08-25 | 天津大学温州安全(应急)研究院 | Three-layer bionic artificial blood vessel for resisting thrombus and promoting tissue regeneration and preparation method thereof |
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