CN104473706B - Biodegradable composite type tubular urethral stent and preparation method - Google Patents
Biodegradable composite type tubular urethral stent and preparation method Download PDFInfo
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- CN104473706B CN104473706B CN201410750552.4A CN201410750552A CN104473706B CN 104473706 B CN104473706 B CN 104473706B CN 201410750552 A CN201410750552 A CN 201410750552A CN 104473706 B CN104473706 B CN 104473706B
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Abstract
The invention provides a biodegradable composite type tubular urethral stent and a preparation method. By the biodegradable composite type tubular urethral stent and the preparation method, the shortcomings that the using quantity of the existing patch materials is limited and complications are easily caused on sampled portions are overcome. The biodegradable tissue engineering stent comprises an inner layer and an outer layer, wherein the inner layer is tubular; the outer layer is arranged on the outer surface of the inner layer; the inner layer and the outer layer are made of degradable high polymer materials and acellular stromata; the degradable high polymer materials are polylactic acid- polylactic-co-glycolic acid (PLGA) films; the acellular stromata are small intestinal submucosa (SIS) films; and the polylactic acid- polylactic-co-glycolic acid (PLGA) films and the small intestinal submucosa (SIS) films are connected to one another. By the biodegradable composite type tubular urethral stent, autologous sampling is not required during urethral canal compounding and reestablishment, and the complications which are possibly caused on the sampled portions are avoided.
Description
Technical field
The present invention relates to medicine technology field, particularly a kind of degradable composite tubular urethra rack and preparation side
Method.
Background technology
Traumatic or iatrogenic urethral injury, gonococcal or NUS and congenital urethra pathology etc. various because
The ankylurethria that element causes, is one of difficult problem that urological surgeon faces.Conventional treatments include urethral dilatation, in urethra
Incision, end-to-end anastomosis of urethra etc. are only applicable to the shorter ankylurethria patient of distance, for the patient master of long section ankylurethria
Will be using substituting operative treatment, genital skin, bladder mucosa, albuginea penis, mucous membrane of mouth and lingual mucous membrane are all used for making
Patching material for urethra prosthesis；But draw materials and would generally cause the complication at position of drawing materials, and the amount of drawing materials is restricted.
Content of the invention
The purpose of the present invention is to cause wound, material to take for when existing and draw materials in existing long section ankylurethria treatment method
Measure weak point limited, that position of drawing materials easily causes complication, a kind of degradable composite tubular urethra rack and system are provided
Preparation Method.
The present invention completes in the following way：A kind of degradable composite tubular urethra rack, this is degradable
Composite tubular urethra rack is a kind of degradable tissue engineering bracket, and including internal layer and outer layer, internal layer is tubulose, and outer layer sets
On the outer surface of internal layer；Described internal layer and outer layer are all made using degradable macromolecular material and acellular matrix.
In described a kind of degradable composite tubular urethra rack, described make the degradable of internal layer and outer layer
Macromolecular material adopts Poly(D,L-lactide-co-glycolide PLGA film, and described acellular matrix adopts small intestine to take off cell base
Matter SIS film；It is interconnected between Poly(D,L-lactide-co-glycolide PLGA film and small intestine acellular matrix SIS film
Together.
In described a kind of degradable composite tubular urethra rack, described Poly(D,L-lactide-co-glycolide
It is combined by linear slit between PLGA film and small intestine acellular matrix SIS film；Described poly lactic-co-glycolic acid copolymerization
Thing PLGA film and small intestine acellular matrix SIS film are all in strip structure；Described line is using 9/0 nylon wire.
In described a kind of degradable composite tubular urethra rack, described outer layer is wrapped in by spiral way
On the outer surface of internal layer.
Polylactic acid-glycolic base second in described a kind of degradable composite tubular urethra rack, on described internal layer
Acid copolymer PLGA film and small intestine acellular matrix SIS film horizontal positioned, longitudinally spaced arrangement is stitched together in the form of sheets,
Synthesize tubulose around catheter external latasuture.
Polylactic acid-glycolic base second in described a kind of degradable composite tubular urethra rack, on described outer layer
Acid copolymer PLGA film and small intestine acellular matrix SIS film level are intervally arranged and are stitched together in strip.
In described a kind of degradable composite tubular urethra rack, described internal layer is by the poly- breast of two rectangles
The small intestine acellular matrix SIS film longitudinally spaced arrangement composition of acid-co-glycolic acid PLGA film and two rectangles；Institute
The outer layer stated takes off cell base by two rectangular-shaped Poly(D,L-lactide-co-glycolide PLGA films and two rectangular-shaped small intestines
Matter SIS film level is intervally arranged and is stitched together.
The preparation method of a kind of degradable composite tubular urethra rack of the present invention comprises the following steps：
(1) macromolecular material polylactic acid PLA and polyglycolic acid PGA are pressed 50:50～85:15 mixed in molar ratio, adopts
Poly(D,L-lactide-co-glycolide PLGA nanoscale electrospun fiber membrane is obtained with electrostatic spinning technique, in prepared electrospinning fibre
Film surface grafting I-type collagen is modified, and obtains modified Poly(D,L-lactide-co-glycolide PLGA film；
(2) small intestinal submucosa cell tissue is soaked in de-cell liquid, de-cell liquid is 0.2% by volumetric concentration
Triton X-100-100 (Triton X-100) and the ammoniacal liquor that volumetric concentration is 0.03% are mixed, and vibration is de- thin
Carry out cryogenic freezing after born of the same parents and be vacuum dried and obtain small intestine acellular matrix SIS film at a temperature of -40 DEG C～-50 DEG C；
(3) according to urethra caliber require, the Poly(D,L-lactide-co-glycolide PLGA film that will be obtained by step (1) and
The small intestine acellular matrix SIS film that step (2) obtains makes the rectangle of rule, is closed with linear slit and obtains internal layer and outer layer.
In the preparation method of described a kind of degradable composite tubular urethra rack, described line adopts nylon
Line.
In the preparation method of described a kind of degradable composite tubular urethra rack, described polylactic acid PLA and
The mol ratio of polyglycolic acid PGA is 75:25.
In the preparation method of described a kind of degradable composite tubular urethra rack, described step (1) is obtained
Poly(D,L-lactide-co-glycolide PLGA nanoscale electrospinning fibre film thickness be 0.35～0.45mm, porosity be 85%
～95%, a diameter of 300～450nm of Electrospun, aperture is 60～100 μm.
In the preparation method of described a kind of degradable composite tubular urethra rack, described is a kind of degradable
The preparation method of composite tubular urethra rack inoculates VEGF using liquid phase cultural method on internal layer
(VEGF) fat mesenchymal stem cell of 165 gene transfections, inoculates by smoothing that fat mesenchymal stem cell converts on outer layer
Myocyte.
A kind of support microstructure of degradable composite tubular urethra rack of the present invention is three-dimensional porous shape, has good
Good biocompatibility and mechanical property, have both elasticity and pliability, can keep stable tube-like condition in vivo；The present invention adopts
It is polymeric biomaterial and acellular matrix, is conducive to seed cell to stick, breeds and break up, promote surrounding tissue
Growth；Bi-material progressively can be decomposed into small-molecule substance by respective mode in vivo and naturally absorb, and need not take out.
Urethra reparation and reconstruction are carried out using the present invention, autologous drawing materials need not be carried out, it is to avoid position of drawing materials may cause
Complication；Mucous membrane of urethra epithelial cell, stem cell, smooth muscle cell etc. can adhere on support and breed, and can be directly used for
Urethra is repaired and is rebuild, suitable clinical practice.
Brief description
Fig. 1 is a kind of structural representation of degradable composite tubular urethra rack internal layer of the present invention.
Fig. 2 is a kind of structural representation of degradable composite tubular urethra rack outer layer of the present invention.
Fig. 3 is the expanded schematic diagram of Fig. 1.
Fig. 4 is the expanded schematic diagram of Fig. 2.
Fig. 5 is the electron microscope of Poly(D,L-lactide-co-glycolide PLGA film.
Fig. 6 is the electron microscope of small intestine acellular matrix SIS film.
In accompanying drawing 1～6,1 expression internal layer；2 expression outer layers；3 expression Poly(D,L-lactide-co-glycolide PLGA films；4
Represent small intestine acellular matrix SIS film.
Specific embodiment
Below against accompanying drawing 1～6, by embodiment, the invention will be further described.
Embodiment 1
A kind of preparation method of degradable composite tubular urethra rack, this preparation method comprises the following steps：
(1) by macromolecular material polylactic acid PLA and polyglycolic acid PGA in molar ratio 50:50 ratio mixing, using quiet
Electrospinning is obtained Poly(D,L-lactide-co-glycolide PLGA nanoscale electrospun fiber membrane, in prepared electrospun fiber membrane table
Face grafting I-type collagen is modified, and obtains modified Poly(D,L-lactide-co-glycolide PLGA film；
(2) by submucous layer of small intestine cell tissue be soaked in the Triton X-100 being 0.2% by volumetric concentration-
In the de-cell liquid that 100 (Triton X-100) and the ammoniacal liquor that volumetric concentration is 0.03% are mixed, after the de- cell of vibration-
Carry out cryogenic freezing vacuum drying treatment at a temperature of 40 DEG C, obtain small intestine acellular matrix SIS film；
(3) according to urethra caliber require, the Poly(D,L-lactide-co-glycolide PLGA film that will be obtained by step (1) and
The small intestine acellular matrix SIS film that step (2) obtains makes the rectangle of rule, obtains internal layer 1 with outward with 9/0 nylon line suture
Layer 2.
The aseptic groin fat taking out new zealand white rabbit, collagenase method isolates rabbit fat mesenchymal stem cell
, and carry out stationary culture in vitro culture (ADMSCs)；To the fat mesenchymal stem cell (ADMSCs) after culture using disease slowly
The method of poison transfection builds overexpression VEGF (VEGF) 165, obtains VEGF (VEGF) 165
The fat mesenchymal stem cell of gene transfection；Fat mesenchymal stem cell (ADMSCs) after culture is in the blood platelet of 20ng/ml
Induce 2 weeks under derivative growth factor (PDGF-BB) conventional culture conditions, be divided into smooth muscle cell；Using liquid phase culture side
Method is inoculated between the fat of VEGF (VEGF) 165 gene transfection on the internal layer 1 being obtained by above-mentioned preparation method
Mesenchymal stem cells, inoculate the smooth muscle cell being converted by fat mesenchymal stem cell on outer layer 2；Outer layer 2 is located at the outer of internal layer 1
On side；Described internal layer 1 be tubular structure, by two length be 30mm, width be 3mm OBL polylactic acid-glycolic
Acetic acid copolymer PLGA film and two length are 30mm, width is 3mm OBL small intestine acellular matrix SIS film
Longitudinally spaced arrangement is stitched together in the form of sheets, synthesizes tubulose around catheter external latasuture；Outer layer 2 by two length be 30mm,
Width be 3mm OBL Poly(D,L-lactide-co-glycolide PLGA be 30mm with two length, width be 3mm rectangle
Arrange and suture and make strip in the small intestine acellular matrix SIS horizontal interval of shape, outer layer 2 is spirally wound on the appearance of internal layer 1
On face.
Fig. 5 is the electron microscope of the Poly(D,L-lactide-co-glycolide PLGA film obtaining；Fig. 6 is that the small intestine obtaining takes off carefully
The electron microscope of cytoplasmic matrix SIS film.
Embodiment 2
A kind of preparation method of degradable composite tubular urethra rack, this preparation method comprises the following steps：
(1) by macromolecular material polylactic acid PLA and polyglycolic acid PGA in molar ratio 75:25 ratio mixing, using quiet
Electrospinning is obtained Poly(D,L-lactide-co-glycolide PLGA nanoscale electrospun fiber membrane, and thickness is 0.4mm, and porosity is
90%, a diameter of 300-450nm of Electrospun, aperture is 60-100 μm；In prepared electrospinning fibre film surface grafting type i collagen egg
It is modified in vain, obtain modified Poly(D,L-lactide-co-glycolide PLGA film；
(2) by submucous layer of small intestine cell tissue be soaked in the Triton X-100 being 0.2% by volumetric concentration-
In the de-cell liquid that 100 (Triton X-100) and the ammoniacal liquor that volumetric concentration is 0.03% are mixed, after the de- cell of vibration-
Carry out cryogenic freezing at a temperature of 42 DEG C to process and vacuum drying treatment, obtain small intestine acellular matrix SIS film；
(3) according to urethra caliber require, the Poly(D,L-lactide-co-glycolide PLGA film that will be obtained by step (1) and
The small intestine acellular matrix SIS film that step (2) obtains is cut into the rectangle of rule, obtains internal layer 1 He with 9/0 nylon line suture
Outer layer 2.
The aseptic groin fat taking out new zealand white rabbit, collagenase method isolates rabbit fat mesenchymal stem cell
, and carry out stationary culture in vitro culture (ADMSCs)；To the fat mesenchymal stem cell (ADMSCs) after culture using disease slowly
The method of poison transfection builds overexpression VEGF (VEGF) 165, obtains VEGF (VEGF) 165
The fat mesenchymal stem cell of gene transfection；Fat mesenchymal stem cell (ADMSCs) after culture is in the blood platelet of 20ng/ml
Induce 2 weeks under derivative growth factor (PDGF-BB) conventional culture conditions, be divided into smooth muscle cell；Using liquid phase culture side
Method is inoculated between the fat of VEGF (VEGF) 165 gene transfection on the internal layer 1 being obtained by above-mentioned preparation method
Mesenchymal stem cells, inoculate the smooth muscle cell being converted by fat mesenchymal stem cell on outer layer 2；Outer layer 2 is located at the outer of internal layer 1
On side；Described internal layer 1 be tubular structure, by two length be 30mm, width be 3mm OBL polylactic acid-glycolic
Acetic acid copolymer PLGA film and two length are 30mm, width is 3mm OBL small intestine acellular matrix SIS film
Longitudinally spaced arrangement is stitched together in the form of sheets, synthesizes tubulose around catheter external latasuture；Outer layer 2 by two length be 30mm,
Width be 3mm OBL Poly(D,L-lactide-co-glycolide PLGA be 30mm with two length, width be 3mm rectangle
Arrange and suture and make strip in the small intestine acellular matrix SIS horizontal interval of shape, outer layer 2 is spirally wound on the appearance of internal layer 1
On face.
Embodiment 3
A kind of preparation method of degradable composite tubular urethra rack, this preparation method comprises the following steps：
(1) by macromolecular material polylactic acid PLA and polyglycolic acid PGA in molar ratio 85:15 ratio mixing, using quiet
Electrospinning is obtained Poly(D,L-lactide-co-glycolide PLGA nanoscale electrospun fiber membrane, in prepared electrospun fiber membrane table
Face grafting I-type collagen is modified, and obtains modified Poly(D,L-lactide-co-glycolide PLGA film；
(2) by submucous layer of small intestine cell tissue be soaked in the Triton X-100 being 0.2% by volumetric concentration-
In the de-cell liquid that 100 (Triton X-100) and the ammoniacal liquor that volumetric concentration is 0.03% are mixed, after the de- cell of vibration-
Carry out cryogenic freezing vacuum drying treatment at a temperature of 50 DEG C, obtain small intestine acellular matrix SIS film；
(3) according to urethra caliber require, the Poly(D,L-lactide-co-glycolide PLGA film that will be obtained by step (1) and
The small intestine acellular matrix SIS film that step (2) obtains is cut into the rectangle of rule, obtains internal layer 1 He with 9/0 nylon line suture
Outer layer 2.
The aseptic groin fat taking out new zealand white rabbit, collagenase method isolates rabbit fat mesenchymal stem cell
, and carry out stationary culture in vitro culture (ADMSCs)；To the fat mesenchymal stem cell (ADMSCs) after culture using disease slowly
The method of poison transfection builds overexpression VEGF (VEGF) 165, obtains VEGF (VEGF) 165
The fat mesenchymal stem cell of gene transfection；Fat mesenchymal stem cell (ADMSCs) after culture is in the blood platelet of 20ng/ml
Induce 2 weeks under derivative growth factor (PDGF-BB) conventional culture conditions, be divided into smooth muscle cell；Using liquid phase culture side
Method is inoculated between the fat of VEGF (VEGF) 165 gene transfection on the internal layer 1 being obtained by above-mentioned preparation method
Mesenchymal stem cells, inoculate the smooth muscle cell being converted by fat mesenchymal stem cell on outer layer 2；Outer layer 2 is located at the outer of internal layer 1
On side；Described internal layer 1 be tubular structure, by two length be 30mm, width be 3mm OBL polylactic acid-glycolic
Acetic acid copolymer PLGA film and two length are 30mm, width is 3mm OBL small intestine acellular matrix SIS film
Longitudinally spaced arrangement is stitched together in the form of sheets, synthesizes tubulose around catheter external latasuture；Outer layer 2 by two length be 30mm,
Width be 3mm OBL Poly(D,L-lactide-co-glycolide PLGA be 30mm with two length, width be 3mm rectangle
Arrange and suture and make strip in the small intestine acellular matrix SIS horizontal interval of shape, outer layer 2 is spirally wound on the appearance of internal layer 1
On face.
Claims (7)
1. a kind of degradable composite tubular urethra rack is it is characterised in that this degradable composite tubular urethra rack is
A kind of degradable tissue engineering bracket, including internal layer and outer layer, internal layer is tubulose, and outer layer is located on the outer surface of internal layer, outward
Layer is wrapped on the outer surface of internal layer by spiral way；Described internal layer and outer layer all using degradable macromolecular material and
Acellular matrix is made；Described make internal layer and the degradable high polymer material of outer layer adopts poly lactic-co-glycolic acid copolymerization
Thing PLGA film, described acellular matrix adopts small intestine acellular matrix SIS film；Poly(D,L-lactide-co-glycolide
Interconnect between PLGA film and small intestine acellular matrix SIS film；Described a kind of degradable composited pipe
The preparation method of shape urethra rack, comprises the following steps：
(1) macromolecular material polylactic acid PLA and polyglycolic acid PGA are pressed 50:50～85:15 mixed in molar ratio, using quiet
Electrospinning is obtained Poly(D,L-lactide-co-glycolide PLGA nanoscale electrospun fiber membrane, in prepared electrospun fiber membrane table
Face grafting I-type collagen is modified, and obtains modified Poly(D,L-lactide-co-glycolide PLGA film；
(2) submucous layer of small intestine cell tissue is soaked in the Triton X-100-100 being 0.2% by volumetric concentration
In the de-cell liquid that (Triton X-100) and the ammoniacal liquor that volumetric concentration is 0.03% are mixed, -40 after the de- cell of vibration
DEG C～-50 DEG C at a temperature of carry out cryogenic freezing vacuum drying treatment, obtain small intestine acellular matrix SIS film；
(3) according to the requirement of urethra caliber, the Poly(D,L-lactide-co-glycolide PLGA film that will be obtained by step (1) and step
(2) the small intestine acellular matrix SIS film obtaining makes the rectangle of rule, is closed with linear slit and obtains internal layer and outer layer.
2. a kind of degradable composite tubular urethra rack according to claim 1 is it is characterised in that described poly- breast
It is combined by linear slit between acid-co-glycolic acid PLGA film and small intestine acellular matrix SIS film；Described is poly-
Poly lactic coglycolic acid PLGA film and small intestine acellular matrix SIS film are all in strip structure.
3. a kind of degradable composite tubular urethra rack according to claim 2 is it is characterised in that described internal layer
On Poly(D,L-lactide-co-glycolide PLGA film and small intestine acellular matrix SIS film horizontal positioned, longitudinally spaced arrangement
It is stitched together in the form of sheets, synthesize tubulose around catheter external latasuture.
4. a kind of degradable composite tubular urethra rack according to claim 2 is it is characterised in that described outer layer
On Poly(D,L-lactide-co-glycolide PLGA film and small intestine acellular matrix SIS film level be intervally arranged and be sewn to
It is in strip together.
5. a kind of degradable composite tubular urethra rack according to claim 1 preparation method it is characterised in that
Described line adopts nylon wire.
6. a kind of degradable composite tubular urethra rack according to claim 1 preparation method it is characterised in that
The mol ratio of described polylactic acid PLA and polyglycolic acid PGA is 75:25.
7. a kind of degradable composite tubular urethra rack according to claim 1 preparation method it is characterised in that
Done using the fat mesenchymal that liquid phase cultural method inoculates the transfection of VEGF (VEGF) 165 gene on internal layer
Cell, inoculates the smooth muscle cell being converted by fat mesenchymal stem cell on outer layer.
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| CN107596447A (en) * | 2017-10-11 | 2018-01-19 | 广州新诚生物科技有限公司 | A kind of biological medicinal membrane with tissue repair function and preparation method thereof |
| CN108853579A (en) * | 2018-07-27 | 2018-11-23 | 吉林大学 | A kind of preparation method of artificial tubular biomaterial |
| CN110680961B (en) * | 2019-11-12 | 2021-07-27 | 东南大学 | Preparation method of multilayer spiral urethral tissue engineering scaffold |
| CN113244021A (en) * | 2021-05-12 | 2021-08-13 | 太原理工大学 | Support for preventing external auditory canal stenosis through 3D printing acellular matrix and preparation method |
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| CN204618494U (en) * | 2014-12-09 | 2015-09-09 | 金华市人民医院 | A kind of degradable composite tubular urethra rack |
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| FI942170A (en) * | 1993-06-15 | 1994-12-16 | Esa Viherkoski | Tubular device for holding the urethra open |
| WO2004110315A1 (en) * | 2003-06-16 | 2004-12-23 | Nanyang Technological University | Polymeric stent and method of manufacture |
| CN101433735B (en) * | 2007-11-13 | 2013-06-12 | 北京大清生物技术有限公司 | Method for preparing SIS tissue repair material |
| CN101987050A (en) * | 2009-08-06 | 2011-03-23 | 浙江普洛医药科技有限公司 | Degradable double-layer compound ureteral stent tube |
| CN101810883B (en) * | 2009-12-25 | 2013-09-11 | 上海太亨科贸有限公司 | Bio-derived material with high tissue compatibility and long acting anti-infection as well as preparation method and application thereof |
| CN102475910A (en) * | 2010-11-22 | 2012-05-30 | 大连创达技术交易市场有限公司 | Esophageal tissue constructed by tissue engineering |
| CN102274491A (en) * | 2011-07-13 | 2011-12-14 | 中国人民解放军第四军医大学 | Stem cell combination growth factor injection for promoting angiogenesis of ischemic tissue, and its preparation method and application method |
| CN102488926B (en) * | 2011-12-16 | 2013-10-30 | 东华大学 | Tissue engineering scaffold for urethra reconstruction and preparation method thereof |
| CN102895702B (en) * | 2012-11-09 | 2014-02-26 | 四川大学华西医院 | Composite manual bile duct and preparation method thereof |
| CN103877622B (en) * | 2014-03-26 | 2016-04-20 | 中山大学 | A kind of Electrospun nano-fibers-ECM coupled biomaterial and its preparation method and application |
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| CN204618494U (en) * | 2014-12-09 | 2015-09-09 | 金华市人民医院 | A kind of degradable composite tubular urethra rack |
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