CN103654999A - Neutral restoration catheter support of multilayer structure and manufacturing method of neutral restoration catheter support - Google Patents
Neutral restoration catheter support of multilayer structure and manufacturing method of neutral restoration catheter support Download PDFInfo
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- CN103654999A CN103654999A CN201310639284.4A CN201310639284A CN103654999A CN 103654999 A CN103654999 A CN 103654999A CN 201310639284 A CN201310639284 A CN 201310639284A CN 103654999 A CN103654999 A CN 103654999A
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Abstract
The invention provides a neutral restoration catheter support of a multilayer structure and manufacturing method of the neutral restoration catheter support in order to meet requirements of neural tissue engineering. The neutral restoration catheter support structurally comprises an inner pipe, a middle pipe and an outer pipe which are made of incompletely different materials, the inner pipe is made of poly-dl-lactide, the middle pipe is made of a lactic acid caprolactone copolymer, and the outer pipe is made of poly-l-lactide. The neutral catheter support is characterized in that the support has certain mechanical performance, is bendable and extrudable, is 10-15MPa in tensile strength and 5-13% of tensile deformation and can restore to an original form after being bent or extruded.
Description
Technical field
The present invention relates to a kind of Biodegradable high-molecular nerve rehabilitating tube support with multiple structure and preparation method thereof, belong to neural tissue engineering field.
Background technology
After peripheral nerve injury, damaged alternative reparation is a global problem always.Peripheral nerve defection is repaired in the autologous neural transplantation of at present clinical main employing, exist and will bring secondary damage to patient, and operating difficulty is large, patient's problems such as be difficult to return to one's perfect health but exist autologous neural transplantation.Using artificial nerve graft to repair peripheral nerve defection as support is the study hotspot in peripheral nerve field in recent years, and expectation can substitute nerve autograft reparation peripheral nerve defection, but there is no at present desirable product appearance.
Research at present thinks that rational neural recovery support should meet following requirement:
(1) the material biodegradable of support, and degradation rate should be able to regulate;
(2) support has certain mechanical performance, can provide enough growing space and mechanical strengths for nerve;
(3) there is good biocompatibility, can not affect tissue activity and function.
Large quantity research shows, Poly-L-lactic acid, poly-dl-lactide and lactic acid caprolactone copolymer, there is good biocompatibility, three kinds of materials combinations can be regulated effectively to the performance of material, Poly-L-lactic acid and poly-dl-lactide can reinforcing material intensity, and lactic acid caprolactone copolymer can reinforcing material toughness and elastic modelling quantity.
By the prepared degradable macromolecule tubular bracket of electrostatic spinning technique, can realize degradation rate should be able to regulate, as simple polymeric membrane is compared, can analog cell outer surface substrate microscopic appearance and structure, good biocompatibility, reaches the object of adhesion and the growth of cell.But because electrostatic spinning lacks necessary mechanical strength, this makes troubles to such support cell culture and implant surgery operation before implanting.Although incorporate other biological material, all can not meet enough mechanical strengths and Growth of Cells space completely.
So needing at present a kind of ability that can play supporting and space can have again loose structure provides rational three dimensions to be convenient to the plantation of organizing relevant cell and the propagation of nerve injury.
Summary of the invention
In order to overcome the deficiencies in the prior art, what the invention provides prepared by a kind of biodegradable macromolecule has nerve rehabilitating tube support of three-decker and preparation method thereof, applies to neural tissue engineering field.
A kind of nerve rehabilitating tube support of multiple structure, it is characterized in that, nerve rehabilitating tube support has inner tube, middle pipe and outward appearance pipe three-decker, the macromolecular material of its every kind structure forms different, inner tube is poly-dl-lactide, middle pipe is lactic acid caprolactone copolymer, and outer tube is Poly-L-lactic acid.
The weight average molecular weight of poly-dl-lactide component is than being Poly-L-lactic acid: dextrorotation polylactic acid=7:3, the weight average molecular weight of lactic acid caprolactone copolymer component is than being Poly-L-lactic acid: polycaprolactone=7:3.
The internal diameter of nerve rehabilitating tube support is 2～4 millimeters, and external diameter is 2.2～4.2 millimeters, and length is 10～200 millimeters.
Interior tube thickness is 0.6～0.8 millimeter, and the thickness of middle pipe is 0.1～0.2 millimeter, and the thickness of outer tube is 0.1～0.4 millimeter.
The average fibre diameter of middle pipe is 240～450 millimeters, and the average fibre diameter of outer tube is 1.1～1.4 microns.
A preparation method for the nerve rehabilitating tube support of multiple structure, is characterized in that, comprises the steps:
(1) poly-dl-lactide is dissolved in dichloromethane, heating condensing reflux dissolve, and form solution A;
(2) by the dissolved solution of lactic acid caprolactone copolymer in dichloromethane and N-dimethylformamide mixed solvent, form solution B;
(3) by the dissolved solution of Poly-L-lactic acid in dichloromethane and N-dimethylformamide mixed solvent in, form solution C;
(4) with bar-shaped mould, insert in above-mentioned solution A, after appropriateness stirs, vertically take out at once, be upside down in ventilation and place, room temperature volatilizees more than 6 hours, obtains inner tube;
(5) build jet deck I, syringe needle is No. 5 straight angle syringe needles, and needle point and receiving system distance are 10 centimetres; The solution B obtaining in step (2) is packed in syringe, inner tube and bar-shaped mould are vertically fixed on metal rotation device, electrostatic spinning is collected in rotation;
(6) build jet deck II, syringe needle is No. 10 straight angle syringe needles, and needle point and receiving system distance are 10 centimetres; The solution C obtaining in step (3) is packed in syringe, inner tube and bar-shaped mould are vertically fixed on metal rotation device, electrostatic spinning is collected in rotation;
(7) lyophilization is processed 12 hours, and support is soaked 10 minutes in 40 ℃ of warm water through bubble, can support is separated with bar-shaped mould.
In solution A, solution B and solution C, the concentration range of macromolecular material is followed successively by 10～25%, 1～1.5% and 1～1.5%; In solution B and solution C, the volume ratio of dichloromethane and N-dimethylformamide is 7:3.
Bar-shaped mould diameter is 2～3 millimeters.
While adopting in electrostatic spinning technique preparation tubular construction, injection rate is 0.4mL/h, 0.5～1 milliliter of injection volume.
While adopting electrostatic spinning technique to prepare outer tube structure, injection rate is 0.4mL/h, 0.5～1 milliliter of injection volume.
Nerve rehabilitating tube support provided by the invention material used is biological Polymer materialspreparation that can be degradable, has good biocompatibility, and the relative rate of increase of cell (RGR) is 1 grade; Support has the independently three layers of tubular structure in interior China and foreign countries, and inner tube is solid Poly-L-lactic acid, strengthens the mechanical strength of support, can keep recovering original form after support extruding or Bending Deformation; The fiber of the difference size of middle pipe and outer tube provides necessary space for cell attachment and growth.
Preparation technology provided by the invention is simple, and without special large-scale instrument, with low cost, preparation time is short.
Accompanying drawing explanation
Fig. 1 is the perspective view of the nerve rehabilitating tube support prepared of the present invention.1 is outer tube, and 2 is middle pipe, and 3 is inner tube.
Fig. 2 is the three-decker schematic diagram of the nerve rehabilitating tube support prepared of the present invention.1 is the micrometer fibers structure of outer tube, 2 nanofibrous structures that are middle pipe, 3 solid constructions that are inner tube.
The specific embodiment
Embodiment 1:
Step is as follows:
(1) take in the dichloromethane that 3g poly-dl-lactide is dissolved in 20ml, heating condensing reflux dissolve 1 hour, form solution A;
(2) take in the dichloromethane and N-dimethylformamide (volume ratio is 7:3) mixed solvent that 0.3g lactic acid caprolactone copolymer is dissolved in 10ml, heating condensing reflux dissolve 1 hour, form solution B;
(3) take in the dichloromethane and N-dimethylformamide (volume ratio is 7:3) mixed solvent that 0.6g Poly-L-lactic acid is dissolved in 10ml, heating condensing reflux dissolve 1 hour, form solution C;
(4) with the stainless steel bar that diameter is 3mm, insert in above-mentioned solution A, after appropriateness stirs, vertically take out at once, be upside down in ventilation and place, room temperature volatilizees more than 6 hours, obtains inner tube;
(5) build jet deck I, syringe needle is No. 5 straight angle syringe needles, and needle point and receiving system distance are 10cm.The solution B obtaining in step (2) is packed in syringe, inner tube and stainless steel bar are vertically fixed on metal rotation device, the rotation of the speed of 50rpm.Injection rate is 0.4mL/h, and injection volume 0.5mL obtains the middle pipe that nanofiber forms.
(6) build jet deck II, syringe needle is No. 10 straight angle syringe needles, and needle point and receiving system distance are 10cm.The solution C obtaining in step (3) is packed in syringe, inner tube and stainless steel bar are vertically fixed on metal rotation device, the rotation of the speed of 50rpm.Injection rate is 0.4mL/h, and injection volume 0.7mL obtains the outer tube that micrometer fibers forms.
(7) 12h is processed in lyophilization, and support is soaked to 10min through bubble in 40 ℃ of warm water, can support is separated with stainless steel bar.
The nerve rehabilitating tube support of gained has inner tube, the tube and the outer tube three-decker, is hollow tubular structures.Stent length is 200mm, can be crooked and stretch, there is stronger toughness and enabling capabilities, and its hot strength is 15MPa; Stretch-proof shape becomes 13%.Adopt vernier caliper measurement, its interior diameter is about 3.1mm, and external diameter is about 4.3mm.Through sem observation, interior tube thickness is about 0.8mm, and the thickness of middle pipe is about 0.15mm, and the thickness of outer tube is about 0.25mm; The average fibre diameter of outer tube is 1.1 μ m, and the average fibre diameter of middle pipe is 430nm.
Embodiment 2:
Step is as follows:
(1) take in the dichloromethane that 2g poly-dl-lactide is dissolved in 20ml, heating condensing reflux dissolve 1 hour, form solution A;
(2) take in the dichloromethane and N-dimethylformamide (volume ratio is 7:3) mixed solvent that 0.3g lactic acid caprolactone copolymer is dissolved in 10ml, heating condensing reflux dissolve 1 hour, form solution B;
(3) take in the dichloromethane and N-dimethylformamide (volume ratio is 7:3) mixed solvent that 0.6g Poly-L-lactic acid is dissolved in 10ml, heating condensing reflux dissolve 1 hour, form solution C;
(4) with the stainless steel bar that diameter is 3mm, insert in above-mentioned solution A, after appropriateness stirs, vertically take out at once, be upside down in ventilation and place, room temperature volatilizees more than 6 hours, obtains inner tube;
(5) build jet deck I, syringe needle is No. 5 straight angle syringe needles, and needle point and receiving system distance are 10cm.The solution B obtaining in step (2) is packed in syringe, inner tube and stainless steel bar are vertically fixed on metal rotation device, the rotation of the speed of 50rpm.Injection rate is 0.4mL/h, and injection volume 1mL obtains the middle pipe that nanofiber forms.
(6) build jet deck II, syringe needle is No. 10 straight angle syringe needles, and needle point and receiving system distance are 10cm.The solution C obtaining in step (3) is packed in syringe, inner tube and stainless steel bar are vertically fixed on metal rotation device, the rotation of the speed of 50rpm.Injection rate is 0.4mL/h, and injection volume 0.2mL obtains the outer tube that micrometer fibers forms.
(7) 12h is processed in lyophilization, and support is soaked to 10min through bubble in 40 ℃ of warm water, can support is separated with stainless steel bar.
The nerve rehabilitating tube support of gained has inner tube, the tube and the outer tube three-decker, is hollow tubular structures.Stent length is 100mm, can be crooked and stretch, there is stronger toughness and enabling capabilities, and its hot strength is 12MPa; Stretch-proof shape becomes 11%.Adopt vernier caliper measurement, its interior diameter is about 3.0 mm, and external diameter is about 3.8mm.Through sem observation, interior tube thickness is about 0.6mm, and the thickness of middle pipe is about 0.1mm, and the thickness of outer tube is about 0.1mm; The average fibre diameter of outer tube is 1.1 μ m, and the average fibre diameter of middle pipe is 430nm.
Embodiment 3:
Step is as follows:
(1) take in the dichloromethane that 3g poly-dl-lactide is dissolved in 20ml, heating condensing reflux dissolve 1 hour, form solution A;
(2) take in the dichloromethane and N-dimethylformamide (volume ratio is 7:3) mixed solvent that 0.2g lactic acid caprolactone copolymer is dissolved in 10ml, heating condensing reflux dissolve 1 hour, form solution B;
(3) take in the dichloromethane and N-dimethylformamide (volume ratio is 7:3) mixed solvent that 0.8g Poly-L-lactic acid is dissolved in 10ml, heating condensing reflux dissolve 1 hour, form solution C;
(4) with the stainless steel bar that diameter is 2mm, insert in above-mentioned solution A, after appropriateness stirs, vertically take out at once, be upside down in ventilation and place, room temperature volatilizees more than 6 hours, obtains inner tube;
(5) build jet deck I, syringe needle is No. 5 straight angle syringe needles, and needle point and receiving system distance are 10cm.The solution B obtaining in step (2) is packed in syringe, inner tube and stainless steel bar are vertically fixed on metal rotation device, the rotation of the speed of 50rpm.Injection rate is 0.4mL/h, and injection volume 0.5mL obtains the middle pipe that nanofiber forms.
(6) build jet deck II, syringe needle is No. 10 straight angle syringe needles, and needle point and receiving system distance are 10cm.The solution C obtaining in step (3) is packed in syringe, inner tube and stainless steel bar are vertically fixed on metal rotation device, the rotation of the speed of 50rpm.Injection rate is 0.4mL/h, and injection volume 0.7mL obtains the outer tube that micrometer fibers forms.
(7) 12h is processed in lyophilization, and support is soaked to 10min through bubble in 40 ℃ of warm water, can support is separated with stainless steel bar.
The nerve rehabilitating tube support of gained has inner tube, the tube and the outer tube three-decker, is hollow tubular structures.Stent length is 100mm, can be crooked and stretch, there is stronger toughness and enabling capabilities, and its hot strength is 10MPa; Stretch-proof shape becomes 11%.Adopt vernier caliper measurement, its interior diameter is about 2.0 mm, and external diameter is about 4.2mm.Through sem observation, interior tube thickness is about 0.8mm, and the thickness of middle pipe is about 0.1mm, and the thickness of outer tube is about 0.3mm; The average fibre diameter of outer tube is 1.4 μ m, and the average fibre diameter of middle pipe is 240nm.
Embodiment 4:
Step is as follows:
(1) take in the dichloromethane that 5g poly-dl-lactide is dissolved in 20ml, heating condensing reflux dissolve 1 hour, form solution A;
(2) take in the dichloromethane and N-dimethylformamide (volume ratio is 7:3) mixed solvent that 0.2g lactic acid caprolactone copolymer is dissolved in 10ml, heating condensing reflux dissolve 1 hour, form solution B;
(3) take in the dichloromethane and N-dimethylformamide (volume ratio is 7:3) mixed solvent that 0.8g Poly-L-lactic acid is dissolved in 10ml, heating condensing reflux dissolve 1 hour, form solution C;
(4) with the stainless steel bar that diameter is 3mm, insert in above-mentioned solution A, after appropriateness stirs, vertically take out at once, be upside down in ventilation and place, room temperature volatilizees more than 6 hours, obtains inner tube;
(5) build jet deck I, syringe needle is No. 5 straight angle syringe needles, and needle point and receiving system distance are 10cm.The solution B obtaining in step (2) is packed in syringe, inner tube and stainless steel bar are vertically fixed on metal rotation device, the rotation of the speed of 50rpm.Injection rate is 0.4mL/h, and injection volume 1mL obtains the middle pipe that nanofiber forms.
(6) build jet deck II, syringe needle is No. 10 straight angle syringe needles, and needle point and receiving system distance are 10cm.The solution C obtaining in step (3) is packed in syringe, inner tube and stainless steel bar are vertically fixed on metal rotation device, the rotation of the speed of 50rpm.Injection rate is 0.4mL/h, and injection volume 1mL obtains the outer tube that micrometer fibers forms.
(7) 12h is processed in lyophilization, and support is soaked to 10min through bubble in 40 ℃ of warm water, can support is separated with stainless steel bar.
The nerve rehabilitating tube support of gained has inner tube, the tube and the outer tube three-decker, is hollow tubular structures.Stent length is 100mm, can be crooked and stretch, there is stronger toughness and enabling capabilities, and its hot strength is 11MPa; Stretch-proof shape becomes 10%.Adopt vernier caliper measurement, its interior diameter is about 3.2 mm, and external diameter is about 4.4mm.Through sem observation, interior tube thickness is about 0.6mm, and the thickness of middle pipe is about 0.2mm, and the thickness of outer tube is about 0.4mm; The average fibre diameter of outer tube is 1.4 μ m, and the average fibre diameter of middle pipe is 240nm.
Embodiment 5:
Nerve rehabilitating tube support prepared in embodiment 1 ~ 4 is carried out to Study on biocompatibility.
Nerve rehabilitating tube support prepared in embodiment 1 ~ 4 is cut into the segment that length is 0.5cm left and right with scalpel.Ultraviolet sterilization is placed on respectively in 48 orifice plates after processing.Every hole inoculates respectively 10
5individual HMS cell, cultivates in cell culture incubator 24 hours, with CCK-8(days roots of cytoactive test kit, China) carry out cytoactive test, the OD value under the optical filter of 450nm wavelength has been reacted cell gross activity.10
5individual HMS cell is seeded in behind a hole, do not have the cell of support and equal number in incubator, cultivate 24 hours be matched group.Calculate the relative rate of increase of cell (RGR) and use following formula:
RGR (%)=(sample OD/ contrasts OD) * 100%
The relative rate of increase of cell and toxic reaction classification are in Table 1.
The relative rate of increase of table 1 nerve rehabilitating tube support cell and toxic reaction classification
Note: toxic reaction classification: the toxic reaction classification of material is as follows according to RGR: 0 grade is RGR >=100%; 1 grade is 80% ~ 99%; 2 grades is 50% ~ 79%; 3 grades is 30% ~ 49%; 4 grades is 0% ~ 29%
As seen from the above table, the relative rate of increase RGR(% of the cell of the support in embodiment 1 ~ embodiment 4) be all I level, biocompatibility reaction is good.
Claims (10)
1. the nerve rehabilitating tube support of a multiple structure, it is characterized in that, nerve rehabilitating tube support has inner tube, middle pipe and outward appearance pipe three-decker, the macromolecular material of its every kind structure forms different, inner tube is poly-dl-lactide, middle pipe is lactic acid caprolactone copolymer, and outer tube is Poly-L-lactic acid.
2. the nerve rehabilitating tube support of multiple structure according to claim 1, it is characterized in that, the weight average molecular weight of poly-dl-lactide component is than being Poly-L-lactic acid: dextrorotation polylactic acid=7:3, the weight average molecular weight of lactic acid caprolactone copolymer component is than being Poly-L-lactic acid: polycaprolactone=7:3.
3. the nerve rehabilitating tube support of multiple structure according to claim 1, is characterized in that, the internal diameter of nerve rehabilitating tube support is 2～4 millimeters, and external diameter is 2.2～4.2 millimeters, and length is 10～200 millimeters.
4. the nerve rehabilitating tube support of multiple structure according to claim 1, is characterized in that, interior tube thickness is 0.6～0.8 millimeter, and the thickness of middle pipe is 0.1～0.2 millimeter, and the thickness of outer tube is 0.1～0.4 millimeter.
5. the nerve rehabilitating tube support of multiple structure according to claim 1, is characterized in that, the average fibre diameter of middle pipe is 240～450 millimeters, and the average fibre diameter of outer tube is 1.1～1.4 microns.
6. according to the preparation method of the nerve rehabilitating tube support of multiple structure described in claim 1-5 any one, it is characterized in that, comprise the steps:
(1) poly-dl-lactide is dissolved in dichloromethane, heating condensing reflux dissolve, and form solution A;
(2) by the dissolved solution of lactic acid caprolactone copolymer in dichloromethane and N-dimethylformamide mixed solvent, form solution B;
(3) by the dissolved solution of Poly-L-lactic acid in dichloromethane and N-dimethylformamide mixed solvent in, form solution C;
(4) with bar-shaped mould, insert in above-mentioned solution A, after appropriateness stirs, vertically take out at once, be upside down in ventilation and place, room temperature volatilizees more than 6 hours, obtains inner tube;
(5) build jet deck I, syringe needle is No. 5 straight angle syringe needles, and needle point and receiving system distance are 10 centimetres; The solution B obtaining in step (2) is packed in syringe, inner tube and bar-shaped mould are vertically fixed on metal rotation device, electrostatic spinning is collected in rotation;
(6) build jet deck II, syringe needle is No. 10 straight angle syringe needles, and needle point and receiving system distance are 10 centimetres; The solution C obtaining in step (3) is packed in syringe, inner tube and bar-shaped mould are vertically fixed on metal rotation device, electrostatic spinning is collected in rotation;
(7) lyophilization is processed 12 hours, and support is soaked 10 minutes in 40 ℃ of warm water through bubble, can support is separated with bar-shaped mould.
7. the preparation method of the nerve rehabilitating tube support of multiple structure according to claim 6, is characterized in that, in solution A, solution B and solution C, the concentration range of macromolecular material is followed successively by 10～25%, 1～1.5% and 1～1.5%; In solution B and solution C, the volume ratio of dichloromethane and N-dimethylformamide is 7:3.
8. the preparation method of the nerve rehabilitating tube support of multiple structure according to claim 6, is characterized in that, bar-shaped mould diameter is 2～3 millimeters.
9. the preparation method of the nerve rehabilitating tube support of multiple structure according to claim 6, is characterized in that, while adopting in electrostatic spinning technique preparation tubular construction, injection rate is 0.4mL/h, 0.5～1 milliliter of injection volume.
10. the preparation method of the nerve rehabilitating tube support of multiple structure according to claim 6, is characterized in that, while adopting electrostatic spinning technique to prepare outer tube structure, injection rate is 0.4mL/h, 0.5～1 milliliter of injection volume.
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| 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 |
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| CN106963991A (en) * | 2017-03-02 | 2017-07-21 | 天津索玛科技有限公司 | A kind of modified degradable support and preparation method thereof |
| CN109124822A (en) * | 2018-06-25 | 2019-01-04 | 上海理工大学 | A kind of aorta pectoralis support ring pre-bending shaping mould |
| WO2020136656A1 (en) * | 2018-12-27 | 2020-07-02 | Yeda Research And Development Co. Ltd. | Nanocomposites comprising biodegradable polymers and inorganic nanoparticles, methods of preparation and uses thereof |
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| CN102961783A (en) * | 2012-04-20 | 2013-03-13 | 南开大学 | Construction method of anticoagulant artificial blood vessel scaffold material |
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| CN109124822A (en) * | 2018-06-25 | 2019-01-04 | 上海理工大学 | A kind of aorta pectoralis support ring pre-bending shaping mould |
| WO2020136656A1 (en) * | 2018-12-27 | 2020-07-02 | Yeda Research And Development Co. Ltd. | Nanocomposites comprising biodegradable polymers and inorganic nanoparticles, methods of preparation and uses thereof |
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| CN103654999B (en) | 2016-07-20 |
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