CN102895702B - Composite manual bile duct and preparation method thereof - Google Patents
Composite manual bile duct and preparation method thereof Download PDFInfo
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- CN102895702B CN102895702B CN201210447616.4A CN201210447616A CN102895702B CN 102895702 B CN102895702 B CN 102895702B CN 201210447616 A CN201210447616 A CN 201210447616A CN 102895702 B CN102895702 B CN 102895702B
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Abstract
The invention discloses a composite manual bile duct. A tubular acellular matrix made of a degradable high-molecular polymer material is coated on the inner surface and outer surface of the composite manual bile duct, wherein 2-20 mg of the degradable high-molecular polymer material is uniformly distributed on the inner surface of the acellular matrix for each square centimeter, and 0-20 mg of the degradable high-molecular polymer material is uniformly distributed on the outer surface of the acellular matrix for each square centimeter. The manual bile duct provided by the invention has the advantages of low immunogenicity, good in-vivo repairing effect and good clinical application prospect.
Description
Technical field
The present invention relates to a kind of combined artificial bile duct and preparation method thereof.
Background technology
Extrahepatic bile ducts is the descending unique passage of bile, clinically, in the diseases such as tumor of bile duct, bile duct injury, bile duct congenital malformation and liver transplantation treatment, conventionally adopts gallbladder intestinal anastomosis operation.This operation can be dredged biliary tract, but operation has changed the normal physiological routes of biliary tract, can make Oddi sphincter function lose, the complication such as postoperative easy generation retrograde infection, anastomotic stricture.Therefore the R and D that, can be used for the artificial choledochus of clinical treatment have very important realistic meaning.
At present, a lot of for the material category of manufacturing artificial bile duct, according to the degradability of material, can be divided into Nondegradable material and the large class of degradable materials two.
Comparatively common non-degradable material is the material of the inanimate object activity of synthetic, has silicone rubber, polyurethane, fluorubber, politef etc.Application number: 200810033513.7, denomination of invention: the Patent Application Publication of " plug-in type inside and outside wall dual structure politef artificial choledochus and preparation thereof " a kind of preparation method of plug-in type outer wall dual structure politef artificial choledochus, this artificial choledochus adopts two-layer macromolecular material to be composited, internal layer is teflon hose, and outer is that chemical fibre forms with knitting braiding.Application number: 200710170515.6, denomination of invention: the Patent Application Publication of " a kind of politef/fluorubber combined artificial bile duct and preparation method thereof " a kind of artificial choledochus, this artificial choledochus is a kind of composite and flexible tube, be followed successively by from inside to outside polytetrafluoroethylene film inner bag basic unit, naphthalene-sodium-tetrahydrofuran solution processing layer and fluorubber cast layer.Non-degradable material non-degradable and mechanical property and normal group are woven with larger difference, cannot grow and tissue reconstruction by sustenticular cell, can only use as the bile duct substitute of short-term, can not be for a long time in body.
Biodegradable material can be degraded in tissue repair, generates the small-molecule substance of inanimate object toxicity, and excretes with cellular metabolism, for the research and development of artificial choledochus open up a new way.Common degradable material has the macromolecule polymer material of synthetic, as, polycaprolactone, also has natural material, as chitin, chitosan and natural acellular matrix.The degradation material of synthetic has good degradability, but cannot grow and tissue reconstruction by sustenticular cell, and in body, repairing effect is good not.Natural acellular matrix material has good degradability and biocompatibility, and contain can Promote cell's growth and the cytokine of tissue repair, and its research of applying in organizational project has been subject to paying close attention to widely.
The people such as Rosen, " Rosen M; Ponsky J; Petras R; et al.Novel bile duct repairf0rbleed ing biliary anastomatic varices:case report and literature review ", JGastrointest Surg, 2005,9 (2): 832] applying the enough biliary tracts of conventional acellular matrix material-submucous layer of small intestine (Small intestinal submucosa, SIS) reparation succeeds.SIS implants after 5 months degradable, and by the bile duct tissue of regenerating is replaced, the reaction of proliferation of fibrous tissue around of the bile duct tissue of regeneration is extremely slight.But Gomez etc., " Comez NA; Zapafier JA, Vargas PE.Re: " Smallintestinal submucosa as a bioscafold for biliary tract regeneration ", Surgery; 2004; 135 (4): 460] its result of study is queried, think that SIS, for repairing bile duct wall defects, can produce seepage of bile and cause tissue adhesion; the inflammation that strong immunoreation causes, and other complicated problems.According to applicant's research, use separately bile duct acellular matrix to repair bile duct defect, also can produce seepage of bile and cause tissue adhesion, there is biliary tract stenosis, and the inflammation that has strong immunoreation to cause, finally cause that laboratory animal is dead.
Summary of the invention
In order to address the above problem, the invention provides a kind of new combined artificial bile duct being formed by synthetic material and natural material and preparation method thereof.
Combined artificial bile duct of the present invention, it is the tubulose acellular matrix that surfaces externally and internally is coated with degradable high polymer material, wherein, on acellular matrix inner surface, every square centimeter is uniformly distributed 2 ~ 20mg degradable high polymer material, and on outer surface, every square centimeter is uniformly distributed 0 ~ 20mg degradable high polymer material.
Described tubulose acellular matrix, refers to the acellular matrix that is shaped as tubulose, as pig, and the acellular matrix of the mammiferous bile duct such as monkey, urethra or blood vessel.
Preferably, on acellular matrix outer surface every square centimeter be uniformly distributed 2 ~ 20mg degradable high polymer material.
Wherein, described artificial choledochus diameter is everywhere identical, is 4 ~ 13mm.
Preferably, described artificial choledochus is cylindrical, and diameter is 4 ~ 13mm; Or be truncated cone-shaped, handle face diameter is 4 ~ 13mm, large bottom surface diameter is at most than the large 2mm of handle face diameter.
Wherein, the long 10 ~ 150mm of described artificial choledochus, pipe thickness is 0.5 ~ 2mm.
Preferably, described degradable high polymer material is polyurethane or polyglycolic acid.
Preferably, the molecular weight of described polyurethane is 2 ~ 50,000 g/mol, and the molecular weight of described polyglycolic acid is 3 ~ 50,000 g/mol.
Polyurethane of the present invention is degradability polyurethane, can be according to the patent No.: ZL 200610022715.2, and denomination of invention: the method preparation that the patent of " method for preparing aqueous nontoxic degradable polyurethane elastomer " is recorded.
The preparation method of the artificial bile duct of the present invention, it comprises the steps:
(1) get tubulose acellular matrix and the degradable high polymer material of aforementioned proportioning;
(2), by tubulose acellular matrix, be inserted in and be not stained with on mould, lyophilization;
The described grinding tool of not being stained with is cylindrical, and diameter is identical with acellular matrix diameter; Or be truncated cone-shaped, handle face diameter is identical with acellular matrix diameter, large bottom surface diameter is at most than the large 2mm of handle face diameter;
(3) get degradable high polymer material, make the emulsion that concentration is 5 ~ 20wt%, the tubulose acellular matrix after coating step (2) is processed, 30 ~ 37 ℃ are dry, sterilizing.
Be not stained with mould, refer to and the NA mould of acellular matrix.
Wherein, the described degradable high polymer material of step (1) is polyurethane or polyglycolic acid.
Wherein, the molecular weight of described polyurethane is 2-5 ten thousand g/mol, and the molecular weight of described polyglycolic acid is 3-5 ten thousand g/mol.
Wherein, described in step (2), not being stained with mould adopts tetrafluoroethene material to be prepared from.
Wherein, in described step (3), the concentration of polyurethane solutions is 15wt%.
Wherein, in described step (3), temperature when dry is 37 ℃.
The immunogenicity of combined artificial bile duct of the present invention is low, and in body, repairing effect is good, can overcome the defect of Traditional Man bile duct, has good potential applicability in clinical practice.
Obviously, according to foregoing of the present invention, according to ordinary skill knowledge and the customary means of this area, not departing under the above-mentioned basic fundamental thought of the present invention prerequisite, can also make modification, replacement or the change of other various ways.
The specific embodiment of form, is described in further detail foregoing of the present invention again by the following examples.But this should be interpreted as to the scope of the above-mentioned theme of the present invention only limits to following example.All technology realizing based on foregoing of the present invention all belong to scope of the present invention.
Accompanying drawing explanation
Fig. 1 combined artificial biliary tract prosthesis cross sectional representation;
Fig. 2 combined artificial biliary tract prosthesis longitudinal section schematic diagram;
Fig. 3 is in the structure of 45 ° of C baking oven bakings combined artificial bile duct after 6 hours;
Fig. 4 is in the structure of 20 ° of C baking oven bakings combined artificial bile duct after 6 hours;
Fig. 5 is in the structure of 37 ° of C baking oven bakings combined artificial bile duct of the present invention after 6 hours;
Immunogenicity in the embedding test of Fig. 6 combined artificial biliary tract prosthesis muscle detects.A is acellular matrix support; B is polyurethane; C is the artificial gallbladder pipe of the present invention;
Fig. 7 matched group is repaired the internal anatomy of bile duct defect;
Fig. 8 combined artificial bile duct of the present invention is implanted into Adult Pig in vivo test result.(A) the normal HE of bile duct tissue dyeing; (B) combined artificial bile duct is repaired HE dyeing after 90 days.
The specific embodiment
Experiment material:
Polyurethane, according to the patent No.: ZL 200610022715.2, denomination of invention: the method preparation that the patent of " method for preparing aqueous nontoxic degradable polyurethane elastomer " is recorded.
Polyglycolic acid, purchased from Aldrich.
Embodiment 1 preparation combined artificial bile duct of the present invention
1, the preparation of tubulose acellular matrix:
Get an Adult Pig bile duct, reject bile duct lumen organization fat and connective tissue around, cutting into length is the long pipeline of 30mm, diameter 4mm, thick 1mm;
Pipeline stirring and washing 2 hours in PBS liquid, in 1%SDS solution, soaking and stirring is 24 hours;
Take out PBS and clean after 2 hours in 1%Triton-100 solution soaking and stirring 24 hours;
Support after de-cell cleans 2 hours in PBS, on the teflon rod that to be fixed on outside diameter and to be 6mm, end diameter be 4mm, obtains dry acellular matrix support after lyophilization.
2, at dry de-cytoskeleton inner surface, apply the aqueous emulsion of polyurethane that concentration is 5wt%, control valve wall thickness is 0.5mm, 30 ℃ of dry 72h, pack, γ ray sterilization, obtains combined artificial bile duct of the present invention, can be used for repairing the pathological changes bile duct that diameter is less.
Embodiment 2 preparation combined artificial bile duct of the present invention
1, the preparation of tubulose acellular matrix:
Get an Adult Pig bile duct, reject bile duct lumen organization fat and connective tissue around, cutting into length is the long pipeline of 10mm, diameter 6mm, thick 2mm;
Pipeline stirring and washing 2 hours in PBS liquid, in 1%SDS solution, soaking and stirring is 24 hours;
Take out PBS and clean after 2 hours in 1%Triton-100 solution soaking and stirring 24 hours;
Support after de-cell cleans 2 hours in PBS, on the teflon rod that to be fixed on outside diameter and to be 8mm, end diameter be 6mm, obtains dry acellular matrix support after lyophilization.
2, at dry de-cytoskeleton inner surface, apply the aqueous emulsion of polyurethane that concentration is 15wt%, control valve wall thickness is 1mm, 37 ℃ of dry 24h, pack, γ ray sterilization, obtains combined artificial bile duct of the present invention, can be used for substituting the pathological changes bile duct of intermediate diameters.
Embodiment 3 preparation combined artificial bile duct of the present invention
1, the preparation of tubulose acellular matrix:
Get an Adult Pig bile duct, reject bile duct lumen organization fat and connective tissue around, cutting into length is the long pipeline of 10mm, diameter 6mm, thick 2mm;
Pipeline stirring and washing 2 hours in PBS liquid, in 1%SDS solution, soaking and stirring is 24 hours;
Take out PBS and clean after 2 hours in 1%Triton-100 solution soaking and stirring 24 hours;
Support after de-cell cleans 2 hours in PBS, and being fixed on diameter is on the cylindrical teflon rod of 6mm, obtains dry acellular matrix support after lyophilization.
2, at dry de-cytoskeleton inner surface, apply the aqueous emulsion of polyurethane that concentration is 15wt%, control valve wall thickness is 1mm, 37 ℃ of dry 24h, pack, γ ray sterilization, obtains combined artificial bile duct of the present invention, can be used for substituting the pathological changes bile duct of intermediate diameters.
Embodiment 4 preparation combined artificial bile duct of the present invention
1, the preparation of tubulose acellular matrix:
Get an Adult Pig bile duct, reject bile duct lumen organization fat and connective tissue around, cut into the pipeline that length is 150 millimeters long, diameter 13mm, thick 2mm;
Pipeline stirring and washing 2 hours in PBS liquid, in 1%SDS solution, soaking and stirring is 24 hours;
Take out PBS and clean after 2 hours in 1%Triton-100 solution soaking and stirring 24 hours;
Support after de-cell cleans 2 hours in PBS, on the teflon rod that to be fixed on outside diameter and to be 15mm, end diameter be 13mm, obtains dry acellular matrix support after lyophilization.
2, at dry de-cytoskeleton inner surface, apply the polyglycolic acid aqueous emulsion that concentration is 20wt%, control valve wall thickness is 2mm, 35 ℃ of dry 48h, pack, γ ray sterilization, obtains combined artificial bile duct of the present invention, can be used for substituting larger-diameter pathological changes bile duct.
Embodiment 5 preparation combined artificial bile duct of the present invention
1, the preparation of tubulose acellular matrix:
Get an Adult Pig bile duct, reject bile duct lumen organization fat and connective tissue around, cut into the pipeline that length is 100 millimeters long, diameter 8mm, thick 2mm;
Pipeline stirring and washing 2 hours in PBS liquid, in 1%SDS solution, soaking and stirring is 24 hours;
Take out PBS and clean after 2 hours in 1%Triton-100 solution soaking and stirring 24 hours;
Support after de-cell cleans 2 hours in PBS, on the teflon rod that to be fixed on outside diameter and to be 10mm, end diameter be 8mm, obtains dry acellular matrix support after lyophilization.
2, at dry de-cytoskeleton inner surface and outer surface, apply the aqueous emulsion of polyurethane that concentration is 12wt%, control valve wall thickness is 2mm, 37 ℃ of dry 24h, pack, γ ray sterilization, obtains combined artificial bile duct of the present invention, can be used for substituting the pathological changes bile duct of intermediate diameters.
Embodiment 6 temperature screening experiment
1, experimental technique
(1) preparation of tubulose acellular matrix:
Get an Adult Pig bile duct, reject bile duct lumen organization fat and connective tissue around, cut into the pipeline that length is 100 millimeters long, diameter 6mm, thick 1.5mm;
Pipeline stirring and washing 2 hours in PBS liquid, in 1%SDS solution, soaking and stirring is 24 hours;
Take out PBS and clean after 2 hours in 1%Triton-100 solution soaking and stirring 24 hours;
Support after de-cell cleans 2 hours in PBS, on the teflon rod that to be fixed on outside diameter and to be 8mm, end diameter be 6mm, obtains dry acellular matrix support after lyophilization.
(2) at dry de-cytoskeleton inner surface, apply the aqueous emulsion of polyurethane that concentration is 12wt%, control valve wall thickness is 1mm, at 20 ℃, 37 ℃ and 45 ℃ of dry 48h, packs respectively, and γ ray sterilization, obtains combined artificial bile duct.
2, experimental result
Result as shown in Fig. 3 ~ Fig. 5, when temperature is 45 ℃, excess Temperature, supporting structure is destroyed; When temperature is 20 ℃, temperature is too low, and part is thicker, and polyurethane is inhomogeneous at porous support face coat; When temperature is 37 ℃, temperature is suitable, and polyurethane forms uniform coating at lumen wall, and supporting structure is complete.
Description of test, excess Temperature or too lowly all can not obtain the acellular matrix that has applied even degradable high polymer material, many experiments is found, only, in the scope of 30 ~ 37 ℃ of temperature, just can obtain the artificial bile duct of the present invention---applied the acellular matrix of even degradable high polymer material.
Mode with experimental example illustrates beneficial effect of the present invention below:
The immunogenicity of experimental example 1 combined artificial bile duct of the present invention detects
1, experimental technique
By the diameter 8mm of the embodiment of the present invention 2 preparations, thick 2mm acellular matrix support and artificial choledochus, and polyurethane timbering material is embedded in spinal column both sides, the adult rabbits back latissimus dorsi m. of the about 2.5kg of body weight.Before operation starts, every laboratory animal injection 40Wan unit penicillin, the chloral hydrate of back part of animal operative site preserved skin pneumoretroperitoneum injection 3ml/kg is anaesthetized.At back part of animal, cut 8-10cm longitudinal incision, after spinal column two lateral incision muscle peplos, expose muscle of back, in embedded material position, cut off 1cm osculum, by smooth the inserting wherein of timbering material.Then, 5-0 absorbable suture interrupted suture is inserted in rearing-box and is raised after animal revives, and close observation animal state.Material is implanted rear first, second and third day to every rabbit injection 40Wan unit penicillin.In postoperative the 7th day, laboratory animal lumbar injection pentobarbital sodium inj (100mg/kg) is put to death.Dissect to take out and transplant specimen and perienchyma, specimens paraffin embedding slices is carried out HE dyeing, observes transplanting specimen and rabbit normal structure contact site inflammatory cell and assembles situation.
2, experimental result
As shown in Figure 6, acellular matrix material and muscular tissue contact site have a large amount of inflammatory cells to assemble to experimental result; On polyurethane timbering material, can only observe the inflammatory cell of minute quantity; On combined artificial biliary tract prosthesis of the present invention, also have the gathering of inflammatory cell, but cell quantity is less.
The immunogenicity of the artificial bile duct of experimental result explanation the present invention is low, suitable with polyurethane porous support, significantly lower than acellular matrix.
Reparative experiment in experimental example 2 combined artificial bile duct of the present invention body
1, experimental technique
Select the Neijiang Pig of local closed colony, preoperative 2.5% pentobarbital sodium (1ml/kg) intravenous anesthesia.Free common bile duct, apart from duodenum upper limb 1cm left and right, place cuts about 1cm common bile duct, inserts the combined artificial bile duct of the embodiment of the present invention 2 preparations, 5-0 absorbable suture interrupted suture, needle gage 1.5mm, back gauge 1.5mm.Check that anastomotic stoma has or not seepage of bile, in art, infusion 0.9% normal saline 500ml adds gentamycin 160,000 U.Postoperative animal fasting on the same day, vein supplements 10% liquid of glucose and 5% sugar-salt-water, and second day after operation starts feed stream juice or semifluid diet, and is transitioned into gradually normal diet.
After transplanting, in 90 days, laboratory animal is implemented to the anesthesia of intraperitoneal pentobarbital sodium, dissect to take out and transplant specimen and perienchyma, with 10% formaldehyde, fix, carry out HE dyeing.
Matched group: free common bile duct, place cuts about 1cm common bile duct apart from duodenum upper limb 1cm left and right, inserts the acellular matrix support of embodiment 2 preparations of having sterilized, 5-0 absorbable suture interrupted suture, needle gage 1.5mm, back gauge 1.5mm.Check that anastomotic stoma has or not seepage of bile, in art, infusion 0.9% normal saline 500ml adds gentamycin 160,000 U.Postoperative animal fasting on the same day, vein supplements 10% liquid of glucose and 5% sugar-salt-water, and second day after operation starts feed stream juice or semifluid diet, and is transitioned into gradually normal diet.
2, experimental result
As shown in Figure 7, there is seepage of bile in matched group, causes surrounding tissue adhesion, occurs biliary tract stenosis, dead after 7 days.
Experimental group animal progressively recovers normally, gallbladder does not occur after experiment and leak, and does not occur biliary tract stenosis, and injury region bile duct is progressively repaired.After transplanting, in the experimental result of 90 days HE dyeing as shown in Figure 8, contrast knownly with normal bile duct tissue, after the artificial bile duct of the present invention is repaired bile duct defect, artificial choledochus support is degraded, and bile duct cell epimatrix forms, and repairs still in continuation.
Experimental result explanation, the effect that the artificial bile duct of the present invention is repaired bile duct wall defects is good, there will not be the side effect such as tissue adhesion that seepage of bile causes and biliary tract stenosis.
Combined artificial bile duct immunogenicity of the present invention is low, there will not be the side effect such as seepage of bile and biliary tract stenosis for bile duct defect, and the effect of repairing bile duct defect in body is good, and potential applicability in clinical practice is good.
Claims (10)
1. a combined artificial bile duct, it is characterized in that: it is the tubulose acellular matrix that surfaces externally and internally is coated with degradable high polymer material, wherein, on acellular matrix inner surface, every square centimeter is uniformly distributed 2 ~ 20mg degradable high polymer material, and on outer surface, every square centimeter is uniformly distributed 0 ~ 20mg degradable high polymer material;
Described degradable high polymer material is polyurethane or polyglycolic acid.
2. artificial choledochus according to claim 1, is characterized in that: on acellular matrix outer surface, every square centimeter is uniformly distributed 2 ~ 20mg degradable high polymer material.
3. according to the artificial choledochus described in claim 1 or 2, it is characterized in that: described artificial choledochus is cylindrical, diameter is 4 ~ 13mm; Or be truncated cone-shaped, handle face diameter is 4 ~ 13mm, large bottom surface diameter is at most than the large 2mm of handle face diameter.
4. artificial choledochus according to claim 1, is characterized in that: the long 10 ~ 150mm of described artificial choledochus, pipe thickness is 0.5 ~ 2mm.
5. artificial choledochus according to claim 4, is characterized in that: the molecular weight of described polyurethane is 2 ~ 50,000 g/mol, and the molecular weight of described polyglycolic acid is 3 ~ 50,000 g/mol.
6. a method of preparing artificial choledochus described in claim 1 ~ 5 any one, is characterized in that: it comprises the steps:
(1) according to proportioning described in claim 1 or 2, get tubulose acellular matrix and degradable high polymer material;
(2), by tubulose acellular matrix, be inserted in and be not stained with on mould, lyophilization;
The described grinding tool of not being stained with is cylindrical, and diameter is identical with acellular matrix diameter; Or be truncated cone-shaped, handle face diameter is identical with acellular matrix diameter, large bottom surface diameter is at most than the large 2mm of handle face diameter;
(3) get degradable high polymer material, make the emulsion that concentration is 5 ~ 20 wt%, the tubulose acellular matrix after coating step (2) is processed, 30 ~ 37 ℃ are dry, sterilizing;
The described degradable high polymer material of step (1) is polyurethane or polyglycolic acid.
7. method according to claim 6, is characterized in that: the molecular weight of described polyurethane is 2-5 ten thousand g/mol, and the molecular weight of described polyglycolic acid is 3-5 ten thousand g/mol.
8. method according to claim 6, is characterized in that: described in step (2), be not stained with mould and adopt tetrafluoroethene material to be prepared from.
9. method according to claim 6, is characterized in that: in described step (3), the concentration of emulsion is 15wt%.
10. method according to claim 6, is characterized in that: in described step (3), temperature when dry is 37 ℃.
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| CN104473706B (en) * | 2014-12-09 | 2017-02-22 | 金华市人民医院 | Biodegradable composite type tubular urethral stent and preparation method |
| CN105012050A (en) * | 2015-07-16 | 2015-11-04 | 清华大学 | Method and special mould for preparing tissue and organ precursor with multi-branch channels |
| CN105999405A (en) * | 2016-05-19 | 2016-10-12 | 李世荣 | Absorbable fundal mesh composite and preparation method thereof |
| CN106075582A (en) * | 2016-06-27 | 2016-11-09 | 暨南大学 | A kind of engineering blood vessel support and construction method thereof |
| CN108721697B (en) * | 2018-07-24 | 2020-10-13 | 中国人民解放军总医院 | Modified acellular vascular stent with surface compounded with 3D printed chitosan and preparation method thereof |
| CN113144286B (en) * | 2021-04-21 | 2022-12-09 | 四川大学华西医院 | Degradable self-supporting artificial bile duct and preparation method thereof |
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| CN101244293A (en) * | 2008-02-04 | 2008-08-20 | 皋岚雅 | Jack-in inside and outside wall dual structure polytetrafluoroethylene artificial bile duct and preparation |
| CN101618233B (en) * | 2009-08-14 | 2013-04-03 | 东华大学 | Polytetrafluorethylene/fluoroelastomer composite artificial bile duct and preparation thereof |
| CN101732117B (en) * | 2009-12-17 | 2012-01-04 | 武汉科技学院 | Bile duct support and preparation method thereof |
| CN202477899U (en) * | 2011-12-21 | 2012-10-10 | 许军 | Biological material tube |
| CN102641160B (en) * | 2012-04-24 | 2014-12-24 | 浙江大学 | Artificial bile duct bionic stent with double-layered compound structure and preparation method of artificial bile duct bionic stent |
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