CN104027844A - Novel method for coating artificial blood vessel with Astragalus polysaccharide - Google Patents
Novel method for coating artificial blood vessel with Astragalus polysaccharide Download PDFInfo
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- CN104027844A CN104027844A CN201410138204.1A CN201410138204A CN104027844A CN 104027844 A CN104027844 A CN 104027844A CN 201410138204 A CN201410138204 A CN 201410138204A CN 104027844 A CN104027844 A CN 104027844A
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Abstract
The invention provides a novel method for coating an artificial blood vessel with Astragalus polysaccharide. The artificial blood vessel is prepared by coating a terylene (with a chemical name of polyethylene terephthalate) woven or knitted elastic porous tube-shaped or Y-shaped synthesized blood vessel prosthesis with an Astragalus polysaccharide mixed solution 3 to 10 times; the mixed solution mainly comprises Astragalus polysaccharide and a plasticizer and is uniformly distributed on the whole layer of the blood vessel prosthesis through infiltration; and the coated blood vessel prosthesis is placed in a drying box containing carbodiimide steam for cross-linking and fixation.
Description
Affiliated technical field
The present invention relates to a kind of pre-coagulation coating method that can be used for replacing artificial blood vessel's prosthese of pathological changes human vas, especially use the astragalus polyose solution coating people blood vessel prosthesis that is mixed with plasticiser, make it not leak blood and regenerate with promotion vascular tissue, thereby exempt the method that in operation process, pre-blood coagulation operates and promotion vascular tissue regenerates.
Background technology
At present, using artificial vascular replacement human lesion blood vessel is an important surgical procedures technology.Artificial blood vessel has polytype and has various material.The artificial blood vessel of terylene (formal name used at school polyethylene terephthalate) material uses the most general in modern vascular replacement operation, artificial blood vessel's tube wall by terylene woven has hole in various degree conventionally, is convenient to receptor tissue's intyrusive, parcel artificial blood vessel's prosthese short its healing.But meanwhile, artificial blood vessel's hole also has its disadvantageous one side, while implanting, can cause a large amount of blood by leaktightness of wall of tubes in body cavity, obviously increase operation risk.
Pre-coagulation coating refers to when the terylene material artificial blood vessel who uses at present replaces body lesion vessels, because the artificial blood vessel of terylene material itself has many seepages, the guided bone of inorganizable growth simultaneously, if coat applicable biological tissue's compatibility chemical substance as terylene material artificial blood vessel's coating (this process is called pre-solidifying), get final product the seepage of anti-Hemostatic Oral Liquid, also can promote the material of vascular smooth muscle cell and migration of fibroblast cells regeneration, i.e. pre-coagulation coating simultaneously.At present clinically normally used pre-coagulation coating be mainly degradable biological material, comprise autologous plasma, albumin, collagen, fibroin albumen and gelatin etc. (Shen Zonglin. introduce a kind of terylene blood vessel solidifying method in advance. Chinese circulation magazine .1990; Feng Muxian. all syndrome cardiovascular diseasess' of horse Operation cooperation. south of the Five Ridges cardiovascular diseases's magazine .1998; Yellow Fu Hua. fimbrin artificial blood vessel's development. Chinese Academy of Medical Sciences's thesis for the doctorate; Kuijpers et al, Journal of Biomaterials Science:Polymer Edition, 11,225,2000; Folk JE.Transglutaminases.Biochemistry1980,49:517～531; Falcone P, Serafini D, Del Duca S.Comparative studies of transglutaminase activity and substrates in different organs of helianthus tuverosus.Plant Physiol1993,142:263～273; Yasueda lI, Kumazawa Y, Motoki M.Purification and characterization of a tissue-type transglutaminase from red sea bream.Bioscience Biotechnology Biochemistry1994,58:2041～2045).But the above coating material is unstable, extract and purification difficult, expensive (J.H.Lee of while, W.G.Kim, and S.S.Kim, " Development and characterization ok an alginate impregnated polyester vascular graft, " J Biomed Mater Res36,200-208 (1997)).
Summary of the invention
Unstable for overcoming coating material, extract and purification difficult, expensive difficulty of while, the present invention adopts degradable biological material astragalus polysaccharides mixed solution coating artificial blood vessel prosthese, reach equally and exempt pre-blood coagulation operation in operation process, reduce and leak blood, promote the object that vascular tissue reinvents simultaneously.This artificial blood vessel forms with the astragalus polyose solution that is mixed with plasticiser is crosslinked by having concrete dynamic modulus tubulose or the Y shape synthetic vessel prosthese that biocompatible material forms, and makes artificial blood vessel's prosthese not leak blood, thereby exempts the pre-solidifying operation in operation process.This concrete dynamic modulus artificial blood vessel prosthese is woven or knitting forming by terylene material.
From Chinese medicine astragalus, refine and obtain, the former powder of highly purified astragalus polysaccharides, by proportioning, making suspension by methods such as microwave-assisted and precipitations.Highly purified astragalus polyose solution is evenly applied to the interior outside of synthetic vessel prosthese, makes to be full of on artificial blood vessel structure elasticity and possess handling and extending longitudinally ability preferably.Use astragalus polysaccharides protein mixed solution coating 3 to 10 times, be exposed to after dry in the drying baker of carbonization diimine steam crosslinked fixing.
For keeping artificial blood vessel's prosthese to keep softness not subside, there is elasticity under drying regime, before using astragalus polyose solution coating artificial blood vessel prosthese, need the plasticiser of arabic gum or the other biological compatibility.In mixed coating material, astragalus polysaccharides concentration is 20 μ g/ml, and plasticiser concentration is 20-25% (percentage by weight).
Synthetic artificial blood vessel knitting or woven type uses according to method of the present invention and obtains most important feature after the mixed liquor coating of astragalus polysaccharides and plasticiser is to make the seepage rate of the vascular stroma of original multi-pore structure approximately be down to zero, and is conducive to the growth of vascular tissue.Experiment in contrast, gets the not terylene blood vessel of 10 synthetic woven types of coating at random, and average seepage rate is: 357.9 ± 20.4ml/min*cm^2, is down to zero according to seepage rate after coating of the present invention.The terylene blood vessel of simultaneously getting 5 synthetic woven types of coating was not implanted in experimental animals after 6 hours, and pathological examination shows and vesselless tissue's growth, according to the growth of coating Hou You of the present invention vascular tissue.
Thus, the object of the invention is to provide a kind of method of not leaking blood and promoting artificial blood vessel's prosthese of vascular tissue's regeneration of preparing.
Further aim of the present invention is to provide a kind of method that astragalus polyose solution coating synthetic vessel prosthese makes it not leak blood and can promote vascular tissue's regeneration of preparing.
Other object and benefit of the present invention will be shown in the example explanation following clearly.
Relation between the feature that the present invention comprises, character and relative theory and operating procedure and each step is set forth the details in following instantiation one by one, and scope of the present invention will illustrate in the claims.
Accompanying drawing explanation
For a better understanding of the present invention, must be with reference to Figure of description and explanation:
Fig. 1 is the cross section of the linear type artificial blood vessel prosthese part of the astragalus polyose solution coating prepared according to method of the present invention;
Fig. 2 is the cross section of the Y type artificial blood vessel prosthese part of the astragalus polyose solution coating prepared according to method of the present invention;
Fig. 3 represents that the seepage rate of artificial blood vessel's prosthese is along with use astragalus polyose solution coating increased frequency progressively to reduce according to method of the present invention;
Fig. 4 represents to use the growth performance of the upper vascular tissue of method astragalus polysaccharides coating artificial blood vessel of the present invention.
Fig. 5 represents the impact of variable concentrations astragalus polysaccharides on cell proliferation.
The specific embodiment
Under the synthetic people who prepares according to the present invention, blood vessel prosthesis as shown in Figure 1: comprise cylinder structure main body 01, it is the multi-pore structure body that the knitting or woven substrate forming 02 of the Fiber Composite Material by tool biocompatibility, especially polyethylene terephthalate (being terylene) forms. substrate 02 has the two-layer matte 04 in interior outside.Terylene forms cylinder structure main body 01, and makes it possess vascular tissue's intyrusive that mushy structure allows receptor, keeps open luminal structure to allow that blood is smooth and easy to be passed through simultaneously.At matte 04 medial surface, be coated with astragalus polyose solution coating 06, astragalus polyose solution coating 06 need to be used astragalus polysaccharides and plasticiser mixing material material coating 3-10 time and be cross-linked and form by being exposed in carbonization diimine steam.
Fig. 2 demonstrates artificial blood vessel's prosthese 12 of a Y-shaped astragalus polyose solution coating, and blood vessel prosthesis 12 comprises main tubular structure 14 and two branch tubular bodies 16.Main tubular structure 14 and branch tubular body 16 consist of polyester knitting or the woven substrate forming 18, and substrate 18 inner side mattes are coated with astragalus polyose solution coating 20, need to use astragalus polyose solution coating 3-10 time.
According to artificial blood vessel's prosthese substrate of multi-pore structure of the present invention by terylene by the knitting or woven optimum that forms.Conventionally the artificial blood vessel's prosthese seepage rate scope forming by the woven braiding of terylene is at 357.9 ± 20.4ml/min*cm^2.The method of crosslinked astragalus polyose solution is that the mixed liquor of astragalus polysaccharides and plasticiser is evenly coated in artificial blood vessel's prosthese substrate, remove unnecessary coating material and place be exposed in carbonization diimine steam after dry crosslinked, after at drying baker inner drying, dewater.The artificial blood vessel's of the astragalus polyose solution coating of preparing according to the present invention seepage rate (unit: ml/min*cm^2) close to zero.Conventionally the artificial blood vessel's prosthese forming by the braiding of terylene substrate is implanted in experimental animals, and in 6 hours, on artificial blood vessel's prosthese, vascular tissue does not grow substantially.The artificial blood vessel's prosthese that contains astragalus polysaccharides coating of making by the present invention is implanted in experimental animals, and after 6 hours, on artificial blood vessel prosthesis, there is obvious growth in vascular tissue.
Following instance is by explanation astragalus polysaccharides extraction and purification and process of preparing astragalus polyose solution coating artificial blood vessel prosthese according to invention from the plant Radix Astragali.The object of illustrative example is to set forth patent rather than limit its scope.
Example 1: astragalus polysaccharides extracting method
The Radix Astragali is worn in flour mill to the Powdered of even thickness, then getting 100g Radix Astragali powder adds appropriate distilled water to dissolve stirring formation solution, after by solution reflux, extract, 3 times in 70 ℃ of constant temperature, each 3h that extracts, add purification water yield 40ml and 95% ethanol to final concentration 90% and carry out the standing 4h of precipitate with ethanol, collecting precipitation thing.The precipitate of collecting forms Powdered astragalus polysaccharides after dry 24h in drying baker, and now the former powder concentration of gained astragalus polysaccharides is 70-80% (percentage by weight), finally adds suitable distilled water to be deployed into the required concentration of experiment.
Example 2: astragalus polyose solution coating artificial blood vessel
By contain astragalus polysaccharides (extracting according to example 1) concentration from the Radix Astragali, be that the astragalus polysaccharides mixed liquor of 20 μ g/ml is filled with in the syringe of 50ml, this astragalus polysaccharides mixed liquor contains 20% arabic gum and distilled water, and viscosity is 30000-37000 centipoise.Syringe is connected with artificial blood vessel's prosthese one end, and astragalus polysaccharides mixed liquor is injected into artificial blood vessel chamber and made coating solution be uniformly coated on lumen wall, and unnecessary astragalus polysaccharides mixed liquor flows out in the other one end of artificial blood vessel's prosthese.After coating, artificial blood vessel is dried 1 hour in the drying baker of 50 degrees Celsius, and coating is dried needs repeatable operation 3-10 time.
After coating 3-10 time, make artificial blood vessel be exposed to crosslinked being placed in 50 degrees Celsius of drying baker in carbonization diimine steam and be dried 24 hours.
Example 3: artificial blood vessel's prosthese blood penetration rate comparison before and after coating
The artificial blood vessel of the astragalus polyose solution coating of preparing according to the method for example 2 detects as follows, getting at random 10 diameters is 8mm, length is the synthetic woven artificial blood vessel after 10cm coating and is connected with blood reservior, under 120mmHg pressure, the blood of heparinization is passed through in artificial blood vessel, collect blood the metering (unit: ml/min*cm^2) of seepage.10 artificial blood vessels' of testing result blood leaching rate is respectively: 0.10,0.05,0.01,0.04,0.05,0.01,0.0,0.01,0.03,0.02, and average blood leaching rate is 0.032ml/min*cm^2, close to zero.
For with not woven artificial blood vessel's comparison of coating, choose the not artificial blood vessel of coating and repeat above-mentioned experimental implementation, recording average blood leaching rate is 30.1ml/min*cm^2.
Example 4: be coated with the artificial blood vessel's prosthese seepage rate comparison of hierachy number front and back
Artificial blood vessel is down to below 6% through seepage rate after astragalus polysaccharides mixed liquor coating 3 times, and after coating 10 times, seepage rate is down to below 1%.The seepage rate measuring of standard is under the water column pressure of 120mmHg, to pass through the circular hole of 2.44cm^2 area, and the sample of artificial blood vessel's prosthese is fixed on to lmin on circular hole, collects and measure water percolating capacity.Each sample is measured respectively for several times, and measurement result represents as follows: seepage rate=ml/ (min*cm^2).Woven artificial blood vessel's seepage rate is the seepage rate of 600ml/min*cm^2 after different number of times coatings.
Change as follows:
The astragalus polyose solution using in the coating each time of above-mentioned experiment is all described and is extracted according to example 1 from the Radix Astragali, and coating process is all described operation according to example 2.Result as shown in Figure 3.Artificial blood vessel's seepage rate after result astragalus polysaccharides coating declines to a great extent, close to zero.
Compare with the artificial blood vessel of coating not, the artificial blood vessel of astragalus polyose solution coating has not only reduced seepage rate, and promotes vascular tissue's growth.Following examples prove is compared astragalus polyose solution coating artificial blood vessel prosthese and is promoted significantly vascular tissue's growth with matched group.
Example 5: the upper vascular tissue of artificial blood vessel growth comparative experiments (experiment in body) before and after coating
In astragalus polysaccharides coating artificial blood vessel prosthese body, (experiment new zealand white rabbit) experiment is as follows.Animal is preoperative weigh stable, the rear urethane 3ml/kg intravenous anesthesia that is 20% by concentration.Before each animal implantable artificial blood vessel, through vein, giving 100ug/kg heparin avoids Intravascular Thrombus to form.Abdominal part median incision, the ventral aorta 3-6cm of the free following 1cm of about renal artery opening, with aortic occlusion clamp, block up and down and excise 1-2 ventral aorta, with 6-0 Prolene line, adopt end to end anastomosis to implant astragalus polysaccharides coating blood vessel prosthese, art finish with in protamine and heparin (1: 1) prevent postoperative hemorrhage.In art, give antibiotic head and embrace quiet the pushing away of azoles quinoline 100mg, postoperative same dose intramuscular injection 3 days, routine feeding before putting to death.
According to take aforesaid operations, repeat to perform the operation as 5 laboratory animals.And the astragalus polysaccharides coating artificial blood vessel that totally 5 time points took out in bodies at 6 hours, 1 day, 1 week, 1 month, 3 months respectively carries out Histological section, and the mode of clicking is assessed the growing state of the upper vascular tissue of artificial blood vessel.
Take out on schedule graft, longitudinal incision blood vessel prosthesis, rinses so that PBS liquid is careful, takes pictures and observes blood vessel profile, the long specimen of the about 1cm of microscopic analysis blood vessel prosthesis proximal anastomosis mouth, blood vessel stage casing and far-end anastomotic stoma.The fixing 2h of 2% formaldehyde for specimen, paraffin mass embedding after lyophilization, the section of 100pm thickness, with dyeing with the single-minded Yihong of Lignum Sappan after fixing on slide, the healing feature of stained preparation is by observation by light microscope inflammation, FBGCR, the degree of intimal fibrosis.Simple saying is divided into Three Estate by the extent of reaction, and with the corresponding grade of parameter from 0 to 2 expression, the grade of inflammation is according to grafting vessel plasma cell around, lymphocyte and polymorphonuclear cell number estimate, 0=is without to slightly; 1=is light to moderate; 2=moderate is to severe.FBGCR is estimated in blood vessel fabric situation around according to multinucleated giant cell and histiocyte (macrophage): the extremely slight reaction of 0=extends to graft less than 1/3 thickness from center line, and 1=foreign body reaction arrives graft from center line and reaches 2/3 thickness; 2 two foreign body reactions reach the thickness of whole graft wall, and " the Fibrotic analysis of blood vessel prosthesis is according to the Fibrotic degree of intimal surface: the fibrin of the 0=overwhelming majority and erythrocyte; Some fibroblasts of 1=and fibrin; The 2=overwhelming majority is fibroblast and collagen.The neointima of blood vessel prosthesis inwall forms and evaluates by optical microscope and sem observation.
Concrete outcome is as follows:
For with not woven artificial blood vessel's comparison of coating, choose the not artificial blood vessel of coating and repeat above-mentioned experimental implementation.
Concrete outcome is as follows:
Based on above-mentioned experimental result, the artificial terylene blood vessel of astragalus polysaccharides coating significantly promotes vascular tissue's growth, and significantly more than not coagulating in advance group, this may can promote that vascular tissue's growth is relevant with astragalus polysaccharides.
Example 6: variable concentrations astragalus polysaccharides promotes human umbilical vein endothelial cell experiment (experiment in vitro)
Human umbilical vein endothelial cell proliferation experiment is as follows.The trophophase cell of taking the logarithm, cell suspension is made in conventional digestion.Re-suspended cell, is inoculated in 96 orifice plates with about 1 * 10^4 every 200 μ g of cell.After 24h, after cell attachment, give respectively biomembrane 4 and process, and blank when blank culture fluid group is set as colorimetric, each concentration arranges 3 multiple holes.Continue to cultivate 24h.Every hole adds CCK820LL, puts 5%CO2 incubator and continues to cultivate 24h.By microplate reader, detect the optical density value (OD value) at 450nm place.
The impact of astragalus polysaccharides variable concentrations on cell proliferation: astragalus polysaccharides concentration is respectively 0,10,20,40,80,120Lg/ml processes human umbilical vein endothelial cell and adds CCK820LL after 24 hours, 37e, after 1h, 490nm detects its OD value result.Repeat 3 times (n=3) and get average and standard deviation.The Promote cell's growth effect when 20ug/ml of astragalus polysaccharides concentration is the most obvious, and after this along with the rising of concentration, somatotrophic effect reduces gradually, but is all significantly higher than control group (P<0.01); Astragalus polysaccharides endothelial cell proliferation rate when 20ug/ml almost reaches 2 times of control groups, and is significantly higher than other each groups (P<0.01).
Concrete outcome as shown in Figure 5.
All examples to sum up, artificial blood vessel's prosthese of the synthetic woven tool multi-pore structure of mixed solution coating of astragalus polysaccharides and plasticiser has been provided by the pre-blood coagulation operation in operation process, the intrusion and the parcel growth that provide desirable substrate to be convenient to vascular tissue simultaneously.Synthetic knitting or weaving polyester artificial blood vessel makes its soft flexible operability having possessed after using astragalus polyose solution coating according to the present invention.
According to above method operation of setting forth, the object of the invention can objective effective realization thus.But do not deviating from spirit and scope of the invention situation, above-mentioned operation concrete steps and some clause can have certain change, the Cucumber material that therefore above-mentioned example description and accompanying drawing are described related method and use is to set forth invention for demonstration, and should not be construed restriction, invents the scope containing.
Claims (6)
1. a method of preparing artificial blood vessel's prosthese of pre-coagulation coating, the cated artificial blood vessel's prosthese of described tool comprises artificial blood vessel's prosthese and is coated in the coating on artificial blood vessel's prosthese, it is characterized in that described coating contains astragalus polysaccharides and plasticiser.
2. the method for preparing artificial blood vessel according to claim 1, be characterised in that the mixed solution that contains astragalus polysaccharides and plasticiser is evenly coated in to artificial blood vessel's holostrome, then in the drying baker that contains carbonization diimine steam, be cross-linked and fix, the described artificial blood vessel's prosthese of final acquisition.
3. the method for preparing artificial blood vessel's prosthese according to claim 1 and 2, is characterized in that plasticiser is selected from arabic gum.
4. the method for preparing artificial blood vessel's prosthese according to claim 1 and 2, it is characterized in that artificial blood vessel's prosthese made by biocompatible materials, it is terylene material (formal name used at school polyethylene terephthalate) and has elasticity concrete dynamic modulus cast or the synthetic fabric of Y-shaped structure, can be woven type or knit type.
5. the method for preparing artificial blood vessel's prosthese according to claim 1 and 2, the concentration that it is characterized in that astragalus polysaccharides is 20 μ g/mL, plasticiser concentration is 20-25% (percentage by weight).
6. the method for preparing artificial blood vessel's prosthese according to claim 1 and 2, wherein applying coating solution is generally wanted 3-10 layer.
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Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
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| CN106264779A (en) * | 2016-08-05 | 2017-01-04 | 江苏百优达生命科技有限公司 | A kind of artificial blood vessel with pre-coagulation coating |
| FR3086089A1 (en) * | 2018-09-17 | 2020-03-20 | Arnaud Collignon | ARTIFICIAL THROMBUS POSITIONED IN AN ARTIFICIAL VEIN OR ARTERY. |
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| Publication number | Priority date | Publication date | Assignee | Title |
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| CN106264779A (en) * | 2016-08-05 | 2017-01-04 | 江苏百优达生命科技有限公司 | A kind of artificial blood vessel with pre-coagulation coating |
| FR3086089A1 (en) * | 2018-09-17 | 2020-03-20 | Arnaud Collignon | ARTIFICIAL THROMBUS POSITIONED IN AN ARTIFICIAL VEIN OR ARTERY. |
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Application publication date: 20140910 |