CN102166378A - Tissue regeneration guiding membrane and preparation method thereof - Google Patents
Tissue regeneration guiding membrane and preparation method thereof Download PDFInfo
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- CN102166378A CN102166378A CN2011100068208A CN201110006820A CN102166378A CN 102166378 A CN102166378 A CN 102166378A CN 2011100068208 A CN2011100068208 A CN 2011100068208A CN 201110006820 A CN201110006820 A CN 201110006820A CN 102166378 A CN102166378 A CN 102166378A
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Abstract
The invention relates to a tissue regeneration guiding membrane and a preparation method thereof, aiming at solving the technical problems that the membrane is limited in thickness, poor in cell growth effect, and limited in strength in the prior art. The tissue regeneration guiding membrane has a layer structure. The tissue regeneration guiding membrane sequentially has the arrangement sequence of an irregularly-configured fiber layer with irregularly-configured fibers, a parallelly-configured fiber layer with parallelly-configured fibers and a nettly-configured fiber layer with nettly-configured fibers, wherein each fiber layer comprises aliphatic polyester. The tissue regeneration guiding membrane can preferably meet the clinical requirement for guiding the tissue regeneration and repair.
Description
Technical field
The present invention relates to a kind of medical device and preparation method thereof, is a kind of guide tissue regeneration film and preparation method thereof specifically.
Background technology
The guide tissue regeneration technology is used for making the paradenlal tissue regeneration of being lost in the periodontal disease development the earliest and produces new-attachment of periodontium.Its principle is to cover periodontal bone forfeiture position and exposed root of the tooth position with barrier film, utilizes membranous physical barriers effect, and the root that stops gingival epithelium cell and connective tissue is the paradenlal tissue regeneration space for the creativity to intyrusive.
But the guide tissue regeneration film of absorbability and bioresorbable does not have similar histology and clinical repair effect on as the mechanicalness isolation barrier.But with the politef be representative can not absorb guide tissue regeneration film because it can not be degraded in vivo voluntarily, need second operation to take out, not only increased patient's misery, also strengthened the probability of postoperative infection.And, owing to be difficult to composite bio-active material and antibiotic etc., can not can only play simple barrier action by resorbable membrane.
The biological development that can absorb guide tissue regeneration film has improved above-mentioned deficiency comprehensively.Have the natural biological degradation material of excellent biology performance such as collagen, gelatin, chitosan etc., have the synthesising biological degradation material of excellent mechanical performances such as polylactic acid, polylactic acid-glycolic guanidine-acetic acid, polyamide etc., and have the inorganic constituents of bone induced growth ability such as nanometer hydroxyapatite etc., all be used to the preparation that biology can absorb guide tissue regeneration film.But, the membrane material of one-component, single layer structure, be to satisfy guide tissue regeneration film will have the requirement that biocompatibility, mechanical stability, bone are induced generative capacity, epithelial cell and connective tissue barrier simultaneously, therefore only in structure with form and exist the guide tissue regeneration film of gradient function variation could satisfy clinical requirement better.
Chinese invention patent CN1216653C discloses a kind of preparation method that is used for the nanocrystalline hydroxyapatite/collagen/polylactic acid-glycolic guanidine-acetic acid composite film material of guide tissue regeneration, and it is coarse that prepared film has one side, the ganoid construction features of one side.
Chinese invention patent CN1319604C discloses a kind of double layer composite collagen base guide tissue regeneration material and preparation method thereof, is prepared into the double-layer composite material with compacted zone and weaker zone structure with collagen and hyaluronic acid or its sodium salt.
Chinese invention patent CN100408115C discloses a kind of biological material film and preparation method with loose structure, it is characterized in that between the both side surface of medical polyamide composition or medical polyamide/nano osteoid apatite composite parts film with the aperture gradual change be distributed with the hole of running through mutually, the one side of film is that the aperture is the dense face of the tiny hole of 0.01～30um, and another side is that the aperture is the loose face of macropore of 30～300um.
The loose porous side of these membrane materials can help tissue regeneration towards defective region, and a fine and close side can stop epithelial cell and connective tissue to enter defective region towards surrounding tissue.Membrane structure although it is so to be can satisfying the clinical requirement of guide tissue regeneration film, but based on the network of fibers characteristic of n cell epimatrix, from bionical viewpoint, electrostatic spinning technique is obtaining application aspect the guide tissue regeneration film of preparation multiple structure.
Among the open CN101584885A of Chinese invention patent, a certain proportion of natural biologic material, degradable polymer, nanometer hydroxyapatite mixture are dissolved in the specific solvent, the method that by the layer upon layer electrostatic spinning, with the aluminium foil is receptor has prepared three layers of gradient composite membrane, the membrane material one side is coarse loose, the one-sided smooth densification.But in this patent of invention was open, because the variation of electric field intensity, the thickness of the film of layer upon layer electrostatic spinning preparation can be restricted; In addition, be the non-woven fibre film that receptor is collected with the aluminium foil, though the porosity height, often because of fiber accumulations density height has suppressed the effect that cell enters growth, and intensity is also very limited.
Summary of the invention
The present invention is exactly a technical problem bad for the effect that the thickness that solves the film that exists in the prior art is restricted, cell enters growth and that intensity is limited, and a kind of guide tissue regeneration film and preparation method thereof is provided.
For this reason, the invention provides a kind of guide tissue regeneration film, it has layer structure, it puts in order and is followed successively by: fiber is parallel arrangement fibrous layer that the random random fibrous layer of arranging of arranging, fiber be parallel arrangement and fiber and is grid that grid the arranges fibrous layer of arranging, and all contains aliphatic polyester in the above-mentioned fibrous layer.
Optimized technical scheme of the present invention is that the random fibrous layer of arranging also contains natural polymer; The grid fibrous layer of arranging also contains natural polymer; The grid fibrous layer of arranging also contains nanometer hydroxyapatite; Film thickness is 0.05～2mm, and more preferably thickness is 0.1～0.5mm; Aliphatic polyester is a kind of in Poly-L-lactic acid, polycaprolactone, polylactic acid-glycolic guanidine-acetic acid, polylactic acid-caprolactone, the polylactic acid-glycolic guanidine-acetic acid-caprolactone; Natural polymer is a kind of in collagen or the gelatin; Fibrolaminar number of plies n satisfies 3≤n≤30, and the random fibrous layer number of plies x that arranges, parallel arrangement fibrous layer number of plies y and the grid fibrous layer number of plies z that arranges satisfies 1≤x, y, z≤10; The fibrolaminar shop of the parallel arrangement of plural number layer angle is 0～180 °.
The present invention provides a kind of method of guide tissue regeneration film simultaneously, and it contains and has the following steps:
(1) aliphatic polyester being dissolved in solvent, stirring under the room temperature and obtained the solution A that concentration is 0.05～0.4g/ml in 6～24 hours, above-mentioned solution A is carried out electrostatic spinning, is receiving system with the stainless steel drum, receives the parallel arrangement fibrous layer;
(2) natural polymer is dissolved in solvent, stirred under the room temperature 6～24 hours, obtain the solution B that concentration is 0.05～0.4g/ml, above-mentioned solution B is mixed with solution A, obtain the solution C that the solution total concentration is 0.05～0.4g/ml, the weight ratio of aliphatic polyester and natural polymer is 90/10～10/90 in this solution C, and this solution C is carried out electrostatic spinning, be receiving system with aluminium foil and copper mesh respectively, receive random arrange fibrous layer and the grid fibrous layer of arranging;
(3) nanometer hydroxyapatite is scattered in solvent, obtaining content behind the ultra-sonic dispersion is 0.1～0.5g/ml suspension, get above-mentioned suspension and add solution A or solution C, the weight ratio that obtains polymer and nanometer hydroxyapatite is 100/0～70/30 solution, above-mentioned solution is carried out electrostatic spinning, with the copper mesh is receiving system, receives the grid fibrous layer of arranging;
(4) fibrous membrane that step (1) is obtained carries out drawing-off along the fiber alignment direction, drafting force 200～500g, 50～100 ℃ of drawing temperatures, degree of draft 100～300%;
(5) fibrous membrane that step (2), (3), (4) are obtained places vacuum drying oven, 25～35 ℃ of constant temperature, and vacuum is lower than under the 40Pa, dry 12～48h;
(6) with the fibrous layer that obtains in the step (5) by the parallel arrangement fibrous layer in the centre, random arrange fibrous layer and the grid order of fibrous layer in both sides of arranging superposes, after being the natural polymer solution immersion of 0.001～0.02g/ml with concentration then, lyophilization obtains guide tissue regeneration film.
Optimized technical scheme of the present invention is, solvent is a kind of in trifluoroethanol or the hexafluoroisopropanol; Aliphatic polyester is a kind of in Poly-L-lactic acid, polycaprolactone, polylactic acid-glycolic guanidine-acetic acid, polylactic acid-caprolactone, the polylactic acid-glycolic guanidine-acetic acid-caprolactone; Natural polymer is a kind of in collagen or the gelatin; Used copper mesh aperture is 100～550 μ m in step (2) and (3).
Guide tissue regeneration film provided by the invention, be followed successively by to a side from a side: the random fibrous layer of arranging towards defective region towards surrounding tissue, can be towards surrounding tissue, highdensity fiber accumulations has interception to the growth that enters of epithelium or connective tissue, helps the attaching growth of epithelium or fibroblast etc. simultaneously; The parallel arrangement fibrous layer can guarantee the mechanical strength of film, plays growing into to damaged place such as stoping epithelium or fibroblast simultaneously; The grid fibrous layer of arranging can be towards defective region, and loose fiber architecture zone between its grid helps the attaching growth of bone or periodontal ligament cell.Specifically shown in table 1 and accompanying drawing 4,5,6,7.
Table 1: the mechanical property with Poly-L-lactic acid fibrous membrane of different fiber architecture compares (n=10, p＜0.05)
The present invention adopts compound biodegradable aliphatic polyester and the natural polymer main component as the guide tissue regeneration film both sides of gradient function structure, the structure that hydrophobicity degradable aliphatic polyester composition helps after the fibrous membrane chance water chance heat is kept, and the hydrophilic natural macromolecule composition helps improving biocompatibility and cellular affinity.
Grid of the present invention is arranged in the fibrous layer, and the adding of nanometer hydroxyapatite composition has improved the inductivity to osteanagenesis.
The present invention is by the layer combination of different fibrous layers shop, can realize independent control to the individual feature layer, can make guide tissue regeneration film have isotropic high mechanical properties by multi-angle (0～180 °) the shop layer of biodegradable aliphatic polyester fibrous membrane that multi-layer fiber is arranged in parallel.
A plurality of fibrous layers that the present invention will spread layer by the natural polymer aqueous solution carry out compoundly, and interlayer is combined, and has increased biocompatibility simultaneously.
The present invention is by selecting the different biodegradable aliphatic polyesters of forming, absorption rate in the degradation rate of regulation and control gradient function structure guide tissue regeneration film and the body.
The present invention utilizes electrostatic spinning technique to prepare at composition respectively and the fibrous membrane that there are differences on arranging, again by the compound mode of shop layer, not only be easier to realize the gradient function structure of guide tissue regeneration film, also make the control that more is easy to get of the thickness of guide tissue regeneration film and mechanical property, somatomedin and antibiotic introducing and sustained release are easier to realize, can satisfy the clinical needs that guide tissue regeneration is repaired better.
Below in conjunction with accompanying drawing, with the specific embodiment content of the present invention is elaborated, but the present invention is not limited to following these examples, do not breaking away under the above-mentioned technological thought situation of the present invention, various replacements and change according to ordinary skill knowledge and customary means are made all should comprise within the scope of the invention.
Description of drawings
Fig. 1 be the fiber random arrangement of electrostatic spinning of the present invention preparation Poly-L-lactic acid/gelatin (50/50, the w/w) stereoscan photograph of composite cellulosic membrane (JSM 4700 type scanning electron microscopies, NEC);
Fig. 2 is the stereoscan photograph (JSM 4700 type scanning electron microscopies, NEC) of the Poly-L-lactic acid fibrous membrane that is arranged in parallel of the fiber of electrostatic spinning of the present invention preparation;
Fig. 3 is the stereoscan photograph (JSM 4700 type scanning electron microscopies, NEC) that the fiber of electrostatic spinning of the present invention preparation is Poly-L-lactic acid/nanometer hydroxyapatite composite cellulosic membrane that grid arranges;
Fig. 4 is a periodontal ligament cell after cultivating 7 days on the Poly-L-lactic acid fibrous layer of fiber random arrangement, laser confocal microscope observation of cell stress fiber photo (from left to right: by fibrolaminar table and in, slice thickness 1.18 μ m);
Fig. 5 is a periodontal ligament cell after cultivating 7 days on the parallel Poly-L-lactic acid fibrous layer of 100% hot drawing-off, laser confocal microscope observation of cell stress fiber photo (from left to right: by the table of fibrous membrane and in, slice thickness 1.17 μ m);
Fig. 6 is that (a:x 200 for the SEM morphologic observation photo of periodontal ligament cell after cultivating 3 days on the grid type Poly-L-lactic acid fibrous layer; B:x 500; C:x 2000);
Fig. 7 is a periodontal ligament cell after cultivating 7 days on the grid type Poly-L-lactic acid nano fibrous membrane, laser confocal microscope observe the nonwoven zone cell stress fiber photo (from left to right: by the table of fibrous membrane and in, slice thickness 5 μ m).
The specific embodiment
Embodiment 1
1, Poly-L-lactic acid (the Shandong medical device research is produced, down with) is dissolved in trifluoroethanol (Shanghai chemical reagent company limited is produced, analytical pure, down with), stirring at room 12 hours obtains the solution A that concentration is 10% (g/ml);
2, gelatin (Sigma-Aldrich produces, down together) is dissolved in trifluoroethanol, stirring at room 12 hours, obtaining concentration is 10% (g/ml) solution B;
3, solution A and B equal proportion are mixed, obtain solution C, the weight ratio of Poly-L-lactic acid and gelatin is 1/1, polymer total concentration 10% (g/ml);
4, getting solution A and carry out electrostatic spinning, is receiving system with the stainless steel drum, cylinder coiling speed 12m/s, and voltage 12kV, receiving range 15cm, flow velocity 0.8mL/h obtains the Poly-L-lactic acid fibrous membrane that fiber is arranged in parallel, spinning 10h, the about 100 μ m of thickness;
5, getting solution C and carry out electrostatic spinning, is receiving system with the aluminium foil, voltage 12kV, and receiving range 20cm, flow velocity 0.4mL/h obtains the Poly-L-lactic acid/gelatin-compounded fibrous membrane of fiber random arrangement, spinning 10h, the about 50 μ m of thickness;
6, getting solution C and carry out electrostatic spinning, is receiving system with the copper mesh of 400 μ m mesh, voltage 12kV, and receiving range 20cm, flow velocity 0.4mL/h obtains fiber and is Poly-L-lactic acid/gelatin-compounded fibrous membrane that grid is arranged, spinning 10h, the about 50 μ m of thickness;
7, the fibrous membrane that step 4,5,6 is obtained places vacuum drying oven, 30 ℃ of constant temperature, and vacuum is lower than under the 12Pa, dry 24h;
8, the Poly-L-lactic acid fibrous membrane that is arranged in parallel of the fiber that step 4 is obtained carries out drawing-off along the fiber alignment direction, drafting force 300g, 100 ℃ of drawing temperatures, degree of draft 100%;
9, the fibrous membrane with step 8 drawing-off superposes the fibrous membrane that step 5,6 is obtained successively through the dry back of step 7, floods with 0.1% (g/ml) aqueous gelatin solution, and lyophilization obtains the compound fibrous membrane of gradient functionization, the about 200 μ m of thickness.
The used used accessory of electrostatic spinning apparatus following (following examples are identical) among the present invention: it is Baoding Lange constant flow pump of TS2-60 that medical injection pump is selected model for use; 30kV, 3mA product that the high direct voltage electrostatic generator selects for use east, Tianjin civilian high voltage power supply factory to produce; The high speed rotary-drum receiving system is self-control, and specification is 0～7000rpm.
Embodiment 2
1, polylactic acid-glycolic guanidine-acetic acid (the Shandong medical device research is produced, down together) is dissolved in hexafluoroisopropanol, stirring at room 24 hours obtains the solution A that concentration is 40% (g/ml);
2, gelatin is dissolved in hexafluoroisopropanol, stirring at room 24 hours, obtaining concentration is 40% (g/ml) solution B;
3,3ml solution A and 1ml solution B are mixed, obtain solution C, the weight ratio of polylactic acid-glycolic guanidine-acetic acid and gelatin is 3/1, polymer total concentration 40% (g/ml);
4, a certain amount of nanometer hydroxyapatite is scattered in hexafluoroisopropanol, 400W ultra-sonic dispersion 5min obtains pasty state suspension D, nanometer hydroxyapatite content 50% (g/ml);
5, get 10ml solution A and 2ml pasty state suspension D mixing and obtain solution E, the weight ratio of polymer and nanometer hydroxyapatite is 4/1;
6, getting solution A and carry out electrostatic spinning, is receiving system with the stainless steel drum, cylinder coiling speed 12m/s, voltage 12kV, receiving range 15cm, flow velocity 0.5mL/h, obtain the polylactic acid-glycolic guanidine-acetic acid fibrous membrane that fiber is arranged in parallel, spinning 10h, the about 400 μ m of thickness;
7, getting solution C and carry out electrostatic spinning, is receiving system with the aluminium foil, voltage 12kV, and receiving range 20cm, flow velocity 0.5mL/h obtains the polylactic acid-glycolic guanidine-acetic acid/gelatin-compounded fibrous membrane of fiber random arrangement, spinning 10h, the about 400 μ m of thickness;
8, getting solution E and carry out electrostatic spinning, is receiving system with the copper mesh of 400 μ m mesh, voltage 12kV, receiving range 20cm, flow velocity 0.5mL/h obtains fiber and is polylactic acid-glycolic guanidine-acetic acid/nanometer hydroxyapatite composite cellulosic membrane that grid is arranged, spinning 10h, the about 400 μ m of thickness;
9, the fibrous membrane that step 6,7,8 is obtained places vacuum drying oven, 35 ℃ of constant temperature, and vacuum is lower than under the 12Pa, dry 48h;
10, the polylactic acid-glycolic guanidine-acetic acid fibrous membrane that is arranged in parallel of the fiber that step 6 is obtained carries out drawing-off along the fiber alignment direction, drafting force 500g, 70 ℃ of drawing temperatures, degree of draft 200%;
11, the fibrous membrane with step 8 drawing-off superposes successively through the dry back of step 7 with 5,6 fibrous membranes that obtain, and floods with 2% (g/ml) aqueous gelatin solution, and lyophilization obtains the compound fibrous membrane of gradient functionization, thickness 1mm.
Embodiment 3
1, polycaprolactone (the Shandong medical device research is produced, down together) is dissolved in trifluoroethanol, stirring at room 6 hours obtains the solution A that concentration is 5% (g/ml);
2, gelatin is dissolved in trifluoroethanol, stirring at room 6 hours, obtaining concentration is 5% (g/ml) solution B;
3, solution A and B equal proportion are mixed, obtain solution C, the weight ratio of polycaprolactone and gelatin is 1/1, polymer total concentration 5% (g/ml);
4, a certain amount of nanometer hydroxyapatite is scattered in trifluoroethanol, 400W ultra-sonic dispersion 5min obtains pasty state suspension D, nanometer hydroxyapatite content 50% (g/ml);
5, get 10ml solution C and 0.2ml pasty state suspension D mixing and obtain solution F, the weight ratio of polymer and nanometer hydroxyapatite is 5/1;
6, getting solution A and carry out electrostatic spinning, is receiving system with the stainless steel drum, cylinder coiling speed 12m/s, and voltage 12kV, receiving range 15cm, flow velocity 0.4mL/h obtains the polycaprolactone fibrous membrane that fiber is arranged in parallel, spinning 10h, the about 20 μ m of thickness;
7, getting solution C and carry out electrostatic spinning, is receiving system with the aluminium foil, voltage 12kV, and receiving range 20cm, flow velocity 0.4mL/h obtains the polycaprolactone/gelatin-compounded fibrous membrane of fiber random arrangement, spinning 10h, the about 20 μ m of thickness;
8, getting solution F and carry out electrostatic spinning, is receiving system with the copper mesh of 100 μ m mesh, voltage 12kV, receiving range 20cm, flow velocity 0.4mL/h obtains fiber and is polycaprolactone/gelatin/nanometer hydroxyapatite composite cellulosic membrane that grid is arranged, spinning 10h, the about 20 μ m of thickness;
9, the fibrous membrane that step 6,7,8 is obtained places vacuum drying oven, 25 ℃ of constant temperature, and vacuum is lower than under the 40Pa, dry 12h;
10, the polycaprolactone fibrous membrane that is arranged in parallel of the fiber that step 6 is obtained carries out drawing-off along the fiber alignment direction, drafting force 200g, 50 ℃ of drawing temperatures, degree of draft 300%;
11, the fibrous membrane with step 8 drawing-off superposes successively through the dry back of step 7 with 5,6 fibrous membranes that obtain, and floods with 0.5% (g/ml) aqueous gelatin solution, and lyophilization obtains the compound fibrous membrane of gradient functionization, thickness 0.05mm.
Embodiment 4
1, polylactic acid-glycolic guanidine-acetic acid-caprolactone (the Shandong medical device research is produced, down together) is dissolved in trifluoroethanol, stirring at room 24 hours obtains the solution A that concentration is 30% (g/ml);
2, gelatin is dissolved in trifluoroethanol, stirring at room 12 hours, obtaining concentration is 20% (g/ml) solution B;
3,2ml solution A and 3ml solution B are mixed, obtain solution C, the weight ratio of poly-polylactic acid-glycolic guanidine-acetic acid-caprolactone and gelatin is 1/1, polymer total concentration 24% (g/ml);
4, a certain amount of nanometer hydroxyapatite is scattered in trifluoroethanol, 400W ultra-sonic dispersion 5min obtains pasty state suspension D, nanometer hydroxyapatite content 50% (g/ml);
5, get 10ml solution C and 0.5ml pasty state suspension D mixing and obtain solution F, the weight ratio of polymer and nanometer hydroxyapatite is 10/1;
6, getting solution A and carry out electrostatic spinning, is receiving system with the stainless steel drum, cylinder coiling speed 12m/s, voltage 15kV, receiving range 18cm, flow velocity 0.8mL/h, obtain polylactic acid-glycolic guanidine-acetic acid-caprolactone fibrous membrane that fiber is arranged in parallel, spinning 10h, the about 250 μ m of thickness;
7, getting solution C and carry out electrostatic spinning, is receiving system with the aluminium foil, voltage 15kV, and receiving range 20cm, flow velocity 0.4mL/h obtains the polylactic acid-glycolic guanidine-acetic acid-caprolactone/gelatin-compounded fibrous membrane of fiber random arrangement, spinning 20h, the about 150 μ m of thickness;
8, get solution F and carry out electrostatic spinning, copper mesh with 400 μ m mesh is a receiving system, voltage 15kV, receiving range 20cm, flow velocity 0.4mL/h, obtain fiber and be polylactic acid-glycolic guanidine-acetic acid-caprolactone/gelatin/nanometer hydroxyapatite composite cellulosic membrane that grid is arranged, spinning 20h, the about 150 μ m of thickness;
9, the fibrous membrane that step 6,7,8 is obtained places vacuum drying oven, 30 ℃ of constant temperature, and vacuum is lower than under the 12Pa, dry 24h;
10, polylactic acid-glycolic guanidine-acetic acid-caprolactone fibrous membrane of being arranged in parallel of the fiber that step 6 is obtained carries out drawing-off along the fiber alignment direction, drafting force 200g, 70 ℃ of drawing temperatures, degree of draft 100%;
11, the fibrous membrane with step 8 drawing-off superposes successively through the dry back of step 7 with 5,6 fibrous membranes that obtain, and floods with 0.5% (g/ml) aqueous gelatin solution, and lyophilization obtains the compound fibrous membrane of gradient functionization, thickness 650 μ m.
Embodiment 5
1, polylactic acid-caprolactone is dissolved in trifluoroethanol, stirring at room 12 hours obtains the solution A that concentration is 10% (g/ml);
2, collagen is dissolved in trifluoroethanol, stirring at room 12 hours, obtaining concentration is 10% (g/ml) solution B;
3, solution A and B equal proportion are mixed, obtain solution C, the weight ratio of polylactic acid-caprolactone and collagen is 1/1, polymer total concentration 10% (g/ml);
4, a certain amount of nanometer hydroxyapatite is scattered in trifluoroethanol, 400W ultra-sonic dispersion 5min obtains pasty state suspension D, nanometer hydroxyapatite content 50% (g/ml);
5, get 10ml solution C and 0.2ml pasty state suspension D mixing and obtain solution F, the weight ratio of polymer and nanometer hydroxyapatite is 10/1;
6, getting solution A and carry out electrostatic spinning, is receiving system with the stainless steel drum, cylinder coiling speed 12m/s, voltage 12kV, receiving range 15cm, flow velocity 0.8mL/h, obtain polylactic acid-glycolic guanidine-acetic acid-caprolactone fibrous membrane that fiber is arranged in parallel, spinning 10h, the about 100 μ m of thickness;
7, getting solution C and carry out electrostatic spinning, is receiving system with the aluminium foil, voltage 12kV, and receiving range 20cm, flow velocity 0.4mL/h obtains the polylactic acid-caprolactone/collagen composite fiber film of fiber random arrangement, spinning 10h, the about 50 μ m of thickness;
8, getting solution F and carry out electrostatic spinning, is receiving system with the copper mesh of 400 μ m mesh, voltage 12kV, receiving range 20cm, flow velocity 0.4mL/h obtains fiber and is polylactic acid-caprolactone/collagen/nanometer hydroxyapatite composite cellulosic membrane that grid is arranged, spinning 10h, the about 50 μ m of thickness;
9, the fibrous membrane that step 6,7,8 is obtained places vacuum drying oven, 30 ℃ of constant temperature, and vacuum is lower than under the 12Pa, dry 24h;
10, polylactic acid-caprolactone fibrous membrane of being arranged in parallel of the fiber that step 6 is obtained carries out drawing-off along the fiber alignment direction, drafting force 300g, 100 ℃ of drawing temperatures, degree of draft 100%;
11, the fibrous membrane with step 8 drawing-off superposes the fibrous membrane that step 5,6 is obtained successively through the dry back of step 7, floods with 0.5% (g/ml) collagenic aqueous solution, and lyophilization obtains the compound fibrous membrane of gradient functionization, thickness 200 μ m.
Embodiment 6
1, the polylactic acid-glycolic guanidine-acetic acid-caprolactone of fiber random arrangement/gelatin-compounded fibrous membrane preparation method is with embodiment 4;
2, polylactic acid-glycolic guanidine-acetic acid-caprolactone fibrous membrane preparation method of being arranged in parallel of fiber is with embodiment 4;
3, fiber is polylactic acid-glycolic guanidine-acetic acid-caprolactone/gelatin/nanometer hydroxyapatite composite cellulosic membrane that grid arranges with embodiment 4;
4, polylactic acid-glycolic guanidine-acetic acid-caprolactone/gelatin-compounded fibrous membrane with 4 layers of fiber random arrangement, polylactic acid-glycolic guanidine-acetic acid-caprolactone fibrous membrane that 4 layers of fiber is arranged in parallel and 4 layers of fiber are polylactic acid-glycolic guanidine-acetic acid-caprolactone/gelatin/nanometer hydroxyapatite composite cellulosic membrane that grid arranges and superpose successively, wherein polylactic acid-glycolic guanidine-acetic acid-caprolactone fibrous membrane of being arranged in parallel of four layers of fiber is respectively with 0,45,90,135 ° of four directions are spread layer, flood with 0.5% (g/ml) aqueous gelatin solution, lyophilization obtains the compound fibrous membrane of gradient functionization, the about 2mm of thickness.
Embodiment 7
1, the Poly-L-lactic acid of fiber random arrangement/gelatin-compounded fibrous membrane preparation method is with embodiment 1;
2, the Poly-L-lactic acid fibrous membrane preparation method that is arranged in parallel of fiber is with embodiment 1;
3, fiber is Poly-L-lactic acid/gelatin-compounded fibrous membrane that grid arranges with embodiment 1;
4, the acid fiber by polylactic film that the polylactic acid/gelatin-compounded fibrous membrane of 10 layers of fiber random arrangement, 10 layers of fiber are arranged in parallel and 10 layers of fiber are polylactic acid/gelatin-compounded fibrous membrane that grid arranges and superpose successively, wherein the acid fiber by polylactic film that is arranged in parallel of 10 layers of fiber is spread layer with 0,45,90,135 ° of four direction respectively, flood with 0.5% (g/ml) aqueous gelatin solution, lyophilization obtains the compound fibrous membrane of gradient functionization, thickness 2mm.
Claims (15)
1. guide tissue regeneration film, it is characterized in that, it has layer structure, it puts in order and is followed successively by: fiber is parallel arrangement fibrous layer that the random random fibrous layer of arranging of arranging, fiber be parallel arrangement and fiber and is grid that grid the arranges fibrous layer of arranging, and all contains aliphatic polyester in the above-mentioned fibrous layer.
2. guide tissue regeneration film according to claim 1 is characterized in that the described random fibrous layer of arranging also contains natural polymer.
3. guide tissue regeneration film according to claim 1 is characterized in that the described grid fibrous layer of arranging also contains natural polymer.
4. according to claim 1 or 3 described guide tissue regeneration films, it is characterized in that the described grid fibrous layer of arranging also contains nanometer hydroxyapatite.
5. guide tissue regeneration film according to claim 1 and 2 is characterized in that its thickness is 0.05～2mm.
6. guide tissue regeneration film according to claim 3 is characterized in that its thickness is 0.1～0.5mm.
7. guide tissue regeneration film according to claim 1 is characterized in that described aliphatic polyester is a kind of in Poly-L-lactic acid, polycaprolactone, polylactic acid-glycolic guanidine-acetic acid, polylactic acid-caprolactone, the polylactic acid-glycolic guanidine-acetic acid-caprolactone.
8. according to claim 2 or 3 described guide tissue regeneration films, it is characterized in that described natural polymer is a kind of in collagen or the gelatin.
9. guide tissue regeneration film according to claim 1 is characterized in that fibrolaminar total number of plies n satisfies 3≤n≤30, and the random fibrous layer number of plies x that arranges, parallel arrangement fibrous layer number of plies y and the grid fibrous layer number of plies z that arranges satisfies 1≤x, y, z≤10.
10. guide tissue regeneration film according to claim 1 is characterized in that the fibrolaminar shop of the parallel arrangement layer angle of plural number is 0～180 °.
11. prepare the method for the described guide tissue regeneration film of claim 1, it is characterized in that containing and have the following steps:
(1) aliphatic polyester being dissolved in solvent, stirring under the room temperature and obtained the solution A that concentration is 0.05～0.4g/ml in 6～24 hours, above-mentioned solution A is carried out electrostatic spinning, is receiving system with the stainless steel drum, receives the parallel arrangement fibrous layer;
(2) natural polymer is dissolved in solvent, stirred under the room temperature 6～24 hours, obtain the solution B that concentration is 0.05～0.4g/ml, above-mentioned solution B is mixed with solution A, obtain the solution C that the solution total concentration is 0.05～0.4g/ml, the weight ratio of aliphatic polyester and natural polymer is 90/10～10/90 in this solution C, and this solution C is carried out electrostatic spinning, be receiving system with aluminium foil and copper mesh respectively, receive random arrange fibrous layer and the grid fibrous layer of arranging;
(3) nanometer hydroxyapatite is scattered in solvent, obtaining nanometer hydroxyapatite content behind the ultra-sonic dispersion is 0.1～0.5g/ml suspension, get above-mentioned suspension and add solution A or solution C, the weight ratio that obtains polymer and nanometer hydroxyapatite is 100/0～70/30 solution, above-mentioned solution is carried out electrostatic spinning, with the copper mesh is receiving system, receives the grid fibrous layer of arranging;
(4) fibrous membrane that step (1) is obtained carries out drawing-off along the fiber alignment direction, drafting force 200～500g, 50～100 ℃ of drawing temperatures, degree of draft 100～300%;
(5) fibrous membrane that step (2), (3), (4) are obtained places vacuum drying oven, 25～35 ℃ of constant temperature, and vacuum is lower than under the 40Pa, dry 12～48h;
(6) with the fibrous layer that obtains in the step (5) by the parallel arrangement fibrous layer in the centre, random arrange fibrous layer and the grid order of fibrous layer in both sides of arranging superposes, after being the natural polymer solution immersion of 0.001～0.02g/ml with concentration then, lyophilization obtains guide tissue regeneration film.
12. the preparation method of guide tissue regeneration film according to claim 11 is characterized in that described solvent is trifluoroethanol or hexafluoroisopropanol.
13. the preparation method of guide tissue regeneration film according to claim 11 is characterized in that described aliphatic polyester is a kind of in Poly-L-lactic acid, polycaprolactone, polylactic acid-glycolic guanidine-acetic acid, polylactic acid-caprolactone, the polylactic acid-glycolic guanidine-acetic acid-caprolactone.
14. the preparation method of guide tissue regeneration film according to claim 11 is characterized in that natural polymer is a kind of in collagen or the gelatin.
15. the preparation method of guide tissue regeneration film according to claim 11 is characterized in that copper mesh aperture used in described step (2) and (3) is 100～550 μ m.
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