CN1100572C - Chitosan catheter for repair of nerve - Google Patents
Chitosan catheter for repair of nerve Download PDFInfo
- Publication number
- CN1100572C CN1100572C CN 99123745 CN99123745A CN1100572C CN 1100572 C CN1100572 C CN 1100572C CN 99123745 CN99123745 CN 99123745 CN 99123745 A CN99123745 A CN 99123745A CN 1100572 C CN1100572 C CN 1100572C
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- solution
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- chitosan
- gelatin
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- 229920001661 Chitosan Polymers 0.000 title claims abstract description 28
- 210000005036 nerve Anatomy 0.000 title abstract description 11
- 230000008439 repair process Effects 0.000 title abstract description 5
- 238000002360 preparation method Methods 0.000 claims abstract description 26
- HEMHJVSKTPXQMS-UHFFFAOYSA-M Sodium hydroxide Chemical compound [OH-].[Na+] HEMHJVSKTPXQMS-UHFFFAOYSA-M 0.000 claims abstract description 21
- 108010010803 Gelatin Proteins 0.000 claims abstract description 19
- 229920000159 gelatin Polymers 0.000 claims abstract description 19
- 239000008273 gelatin Substances 0.000 claims abstract description 19
- 235000019322 gelatine Nutrition 0.000 claims abstract description 19
- 235000011852 gelatine desserts Nutrition 0.000 claims abstract description 19
- QTBSBXVTEAMEQO-UHFFFAOYSA-N Acetic acid Chemical compound CC(O)=O QTBSBXVTEAMEQO-UHFFFAOYSA-N 0.000 claims abstract description 15
- 238000000034 method Methods 0.000 claims abstract description 10
- SXRSQZLOMIGNAQ-UHFFFAOYSA-N Glutaraldehyde Chemical compound O=CCCCC=O SXRSQZLOMIGNAQ-UHFFFAOYSA-N 0.000 claims abstract description 5
- 230000001537 neural effect Effects 0.000 claims description 13
- 239000011521 glass Substances 0.000 claims description 12
- 238000003756 stirring Methods 0.000 claims description 10
- 238000000465 moulding Methods 0.000 claims description 8
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Chemical compound O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims description 7
- 238000004108 freeze drying Methods 0.000 claims description 6
- 238000007710 freezing Methods 0.000 claims description 5
- 230000008014 freezing Effects 0.000 claims description 5
- 239000007788 liquid Substances 0.000 claims description 5
- 239000012153 distilled water Substances 0.000 claims description 4
- 238000011010 flushing procedure Methods 0.000 claims description 4
- BHTJEPVNHUUIPV-UHFFFAOYSA-N pentanedial;hydrate Chemical compound O.O=CCCCC=O BHTJEPVNHUUIPV-UHFFFAOYSA-N 0.000 claims description 4
- 230000008569 process Effects 0.000 claims description 4
- KDXKERNSBIXSRK-RXMQYKEDSA-N D-lysine Chemical compound NCCCC[C@@H](N)C(O)=O KDXKERNSBIXSRK-RXMQYKEDSA-N 0.000 claims description 3
- 230000006837 decompression Effects 0.000 claims description 3
- 239000008367 deionised water Substances 0.000 claims description 3
- 229910021641 deionized water Inorganic materials 0.000 claims description 3
- 238000001914 filtration Methods 0.000 claims description 3
- 230000007935 neutral effect Effects 0.000 claims description 3
- 229950000845 politef Drugs 0.000 claims description 3
- 238000007789 sealing Methods 0.000 claims description 3
- 230000006641 stabilisation Effects 0.000 claims description 3
- 238000011105 stabilization Methods 0.000 claims description 3
- 238000013461 design Methods 0.000 claims description 2
- 238000001035 drying Methods 0.000 claims description 2
- 239000000463 material Substances 0.000 abstract description 10
- 108010039918 Polylysine Proteins 0.000 abstract 1
- 230000009982 effect on human Effects 0.000 abstract 1
- 230000002503 metabolic effect Effects 0.000 abstract 1
- 229920000656 polylysine Polymers 0.000 abstract 1
- 231100000331 toxic Toxicity 0.000 abstract 1
- 230000002588 toxic effect Effects 0.000 abstract 1
- 210000004126 nerve fiber Anatomy 0.000 description 6
- 229920000954 Polyglycolide Polymers 0.000 description 3
- 230000006196 deacetylation Effects 0.000 description 3
- 238000003381 deacetylation reaction Methods 0.000 description 3
- 238000002474 experimental method Methods 0.000 description 3
- 239000002207 metabolite Substances 0.000 description 3
- 229920000747 poly(lactic acid) Polymers 0.000 description 3
- 239000004633 polyglycolic acid Substances 0.000 description 3
- 239000004626 polylactic acid Substances 0.000 description 3
- 238000002390 rotary evaporation Methods 0.000 description 3
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 description 2
- 241001465754 Metazoa Species 0.000 description 2
- 230000006378 damage Effects 0.000 description 2
- 230000000694 effects Effects 0.000 description 2
- 210000002950 fibroblast Anatomy 0.000 description 2
- 230000006870 function Effects 0.000 description 2
- 238000004519 manufacturing process Methods 0.000 description 2
- 230000004048 modification Effects 0.000 description 2
- 238000012986 modification Methods 0.000 description 2
- 230000035479 physiological effects, processes and functions Effects 0.000 description 2
- 229920000729 poly(L-lysine) polymer Polymers 0.000 description 2
- 238000011084 recovery Methods 0.000 description 2
- 210000004116 schwann cell Anatomy 0.000 description 2
- 230000001954 sterilising effect Effects 0.000 description 2
- 238000004659 sterilization and disinfection Methods 0.000 description 2
- 230000000638 stimulation Effects 0.000 description 2
- 206010020880 Hypertrophy Diseases 0.000 description 1
- 102000015336 Nerve Growth Factor Human genes 0.000 description 1
- 108010025020 Nerve Growth Factor Proteins 0.000 description 1
- 208000028389 Nerve injury Diseases 0.000 description 1
- 241000700157 Rattus norvegicus Species 0.000 description 1
- 230000002378 acidificating effect Effects 0.000 description 1
- 230000009471 action Effects 0.000 description 1
- 230000036982 action potential Effects 0.000 description 1
- 238000001467 acupuncture Methods 0.000 description 1
- 239000003519 biomedical and dental material Substances 0.000 description 1
- 210000004204 blood vessel Anatomy 0.000 description 1
- 230000002490 cerebral effect Effects 0.000 description 1
- 238000010382 chemical cross-linking Methods 0.000 description 1
- 238000006243 chemical reaction Methods 0.000 description 1
- 150000001875 compounds Chemical class 0.000 description 1
- 230000002950 deficient Effects 0.000 description 1
- 230000000593 degrading effect Effects 0.000 description 1
- 239000003814 drug Substances 0.000 description 1
- 230000007831 electrophysiology Effects 0.000 description 1
- 238000002001 electrophysiology Methods 0.000 description 1
- 238000011156 evaluation Methods 0.000 description 1
- 239000000835 fiber Substances 0.000 description 1
- 150000004676 glycans Chemical class 0.000 description 1
- 238000010562 histological examination Methods 0.000 description 1
- 238000001727 in vivo Methods 0.000 description 1
- 238000011534 incubation Methods 0.000 description 1
- 230000005012 migration Effects 0.000 description 1
- 238000013508 migration Methods 0.000 description 1
- 230000008764 nerve damage Effects 0.000 description 1
- 229940053128 nerve growth factor Drugs 0.000 description 1
- 231100000957 no side effect Toxicity 0.000 description 1
- 230000008520 organization Effects 0.000 description 1
- 210000000578 peripheral nerve Anatomy 0.000 description 1
- 229920001282 polysaccharide Polymers 0.000 description 1
- 239000005017 polysaccharide Substances 0.000 description 1
- 230000001172 regenerating effect Effects 0.000 description 1
- 238000011160 research Methods 0.000 description 1
- 210000003497 sciatic nerve Anatomy 0.000 description 1
- 230000035807 sensation Effects 0.000 description 1
- 229920002379 silicone rubber Polymers 0.000 description 1
- 239000004945 silicone rubber Substances 0.000 description 1
- 238000001356 surgical procedure Methods 0.000 description 1
- 230000001228 trophic effect Effects 0.000 description 1
- 238000000108 ultra-filtration Methods 0.000 description 1
- 239000003643 water by type Substances 0.000 description 1
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- Materials For Medical Uses (AREA)
Abstract
The present invention belongs to biomedical materials which are mainly composed of the components with the proportion by weight: 100 portions of chitosan, 5 to 10 portions of gelatin, and a tiny amount of glutaraldehyde, acetic acid, sodium hydroxide and polylysine. A method for preparing conduits used for nerve repair of the present invention comprises the preparation of original solution for preparing conduits and conduit preparation. The present invention is natural, biodegradable and novel materials manufactured for nerve repair without toxic or side effect on human bodies by a metabolic product.
Description
The invention belongs to fields such as bio-medical material, clinical medical neurosurgery, department of cerebral surgery and preclinical medicine.Be used for the conduit that peripheral nerve is repaired, incipient stage is to make of silicone rubber, this material is not degraded, after the nerve reparation, this conduit is still deposited in vivo as foreign body, excites nerve, and must take out through second operation, and second operation damages inevitably, excites nerve, and brings great misery and extra financial burden to the patient.This material is abandoned in the nerve reparation.But the polylactic acid of degrading with the dirt thing again, polyglycolic acid etc. were as the material of nerve rehabilitating tube afterwards, though this material is biodegradable, the neural reparation do not need to do second operation again,, polylactic acid, polyglycolic acid are the materials of synthetic on the one hand, complex process, the cost height, on the other hand, the metabolite of these two kinds of materials is acidic materials, body there is stimulation, still is in animal experiment stage so far, still have very big distance apart from clinical experiment.
The objective of the invention is to overcome present neural repair widely used polylactic acid, polyglycolic acid expense height in the research, metabolite has defectives such as stimulation to body, but develops degraded, that its metabolite has no side effect to human body, the manufacturing nerve repair material of a kind of natural dirt thing.
A kind of conduit that is used for neural reparation that the present invention proposes is characterized in that mainly being made up of chitosan, gelatin, glutaraldehyde, acetic acid, sodium hydroxide, poly-D-lysine, and its part by weight is: 100 parts of chitosans: 5～10 parts in gelatin, all the other trace.
Each component of the present invention technical parameter require as follows:
1) chitosan: deacetylation is greater than 75%, and molecular weight (representing with viscosity usually) is big as far as possible, and is general
More than 150mPaS.
2) gelatin, Sigma company product
3) glutaraldehyde, homemade analytical pure
4) acetic acid, homemade analytical pure
5) sodium hydroxide, homemade analytical pure
6) poly-D-lysine, homemade analytical pure
The present invention proposes a kind of method of making above-mentioned chitosan catheter, it is characterized in that, comprises that the preparation and the conduit of preparation conduit original solution prepares two parts, specifically may further comprise the steps:
One, preparation conduit original solution preparation:
1. chitosan is dissolved in the acetum, fully stirs silk cover filtering.
2. gelatin is dissolved in the deionized water, is made into gelatin solution.
3. gelatin solution is slowly added in the chitosan solution, stir, drip NaHCO
3Solution is transferred pH to 5.1-6.0, drips glutaraldehyde water solution, stirred for several minute.
4. mid-more than 12 hours at the room temperature shaking table, eliminate the bubble in the solution.
Two, the conduit preparation comprises two kinds of schemes:
Get the Glass rod of setting diameter, with the rotating speed rotation plug of 20～50rpm.At the uniform velocity vertically insert solution, leave standstill a moment treat the spin stabilization of solution with Glass rod after, Glass rod is lifted out liquid level with the speed of 1～5mm/s, place horizontal level, still at the uniform velocity rotate with the speed of 20～50rpm, so that solution is evenly distributed on Glass rod, said process is repeated in the air-dry back of room temperature, reaches 300-500 μ m to pipe thickness and takes off.
Utilize politef to be processed into mould and prepare conduit, in the chitosan solution adding mould with molding, bubble is eliminated in decompression, fills it up with sealing; Put into freezing a few hours under-50～-80 ℃ the temperature then, treat freezing fully after, take out, open the mould closed port, place the freezer dryer lyophilization, the moisture in the molding solution distributes fully; Mould plug and conduit are together taken out again, place that NaOH solution is handled, typing, the reuse distilled water flushing is to neutral, will be enclosed within conduit on the plug at last and be placed in the baking oven in dry, and heat-treat, and temperature is higher than 80 ℃, but is no more than 120 ℃.Take out plug at last, get conduit.
Characteristics of the present invention be preferred natural marine organisms polysaccharide one chitosan as stock, its deacetylation is about 75%-90%, molecular weight is big as much as possible.By chemical crosslinking modification, modification, be dissolved in acetic acid solution, utilize rotary evaporation method and freeze-drying to be prepared into wall thickness 0.3-1mm, the above conduit of internal diameter 1-5mm, length 20mm, conduit has certain elasticity and intensity.Catheter wall can stop the intrusion of outside organization, schwann cell, good hypertrophy, the migration environment of fibroblast can be provided again, simultaneously fibroblast, the excretory various nerve growth factor of schwann cell and trophic factors are played the effect of gathering, make it reach the needed concentration of neuranagenesis.
Fig. 1 is the mould example structure sketch map in the inventive method
The embodiment that a kind of preparation of the present invention is used for the neural chitosan catheter of repairing is described in detail as follows:
One, the preparation of preparation conduit original solution
1. deacetylation is that 85% chitosan is dissolved in 2% (v/v) acetum, and the final concentration that makes chitosan is 2% (m/v), fully stirs, and 200 mesh sieve thin,tough silk filter.
2. gelatin is dissolved in 50 ℃ of deionized waters, is made into 10% gelatin solution (m/v).
3. gelatin solution is slowly added in the chitosan solution stirring, Dropwise 5 %NaHCO
3Solution is transferred pH to 5.1-6.0, drips 0.25% glutaraldehyde water solution, fully stirs, and the proportion control of chitosan and gelatin is about 19: 1.The final concentration of glutaraldehyde is not higher than 0.00125%, stirs 30 minutes.
4. in 37 ℃ of shaking tables, spend the night, eliminate the bubble in the solution.
Two, conduit preparation
The Glass rod of cut-off footpath 1.3mm is with the rotating speed rotation plug of 30rpm.At the uniform velocity vertically insert solution, leave standstill after 1min treats the spin stabilization of solution with Glass rod, Glass rod is lifted out liquid level with the speed of 1mm/s, place horizontal level, still at the uniform velocity rotate with the speed of 30rpm, so that solution is evenly distributed on Glass rod, said process is repeated in the air-dry back of room temperature, reaches 0.2-0.3mm to pipe thickness and takes off.
Utilize politef to be processed into mould, mould structure comprises as shown in Figure 1: sleeve 1, place the plug 2 in the sleeve, and be fixed at the nut 3,4 at sleeve two ends.Be the space of the chitosan solution of ccontaining desire molding between sleeve and the plug, the length of conduit is by length (length that the also has plug 2 accordingly) decision of sleeve 1.Catheter diameter is by the external diameter decision of plug 2, and the external diameter of conduit is by the internal diameter decision of sleeve 1.
With nut 3, sleeve 1 and plug 2 fixing after, add the chitosan solution 5 of desiring molding by the other end, bubble is eliminated in decompression, fills it up with, with nut 4 sealings.
Put into freezing 5h under-70 ℃ the temperature then, take out, twist remove nut 4 after, place the freezer dryer lyophilization, the moisture in the molding solution distributes fully; Plug 2 and conduit are together taken out again, place NaOH solution to handle, finalize the design, the reuse distilled water flushing is to neutral, and the conduit that will be enclosed within last on the plug is placed on drying in the baking oven, and heat-treats 100 ℃ of temperature.Take out plug at last, get conduit.
Using method:
1, conduit is washed, is dipped to neutrality with distilled water
2, reuse is 75% alcohol-pickled 10 minutes
3, the Hanks liquid with sterilization soaks, washes and remove residual ethanol
4, preparation 0.1% Poly-L-Lysine Solution, the ultrafiltration sterilization, with above-mentioned through Hanks liquid soak, after the flushing
Conduit insert in the Poly-L-Lysine Solution of being joined to soak to take out behind the 2h and use.
The effect experiment of present embodiment:
The sciatic nerve of Wistar rat is cut off, the people causes the model of the neural gaps of 0.5cm as nerve injury, with we the preparation chitosan catheter the damage neural near-end and far-end between " bridging ", after eight months, histological examination, the regenerating nerve fiber is compared with the normal control nerve fiber, very big similarity is arranged, the nerve fiber regular shape, sub-circular, diameter chap, the nerve fiber that has good in a large number myelinization, the obvious beam splitting of nerve fiber has the blood vessel of a large amount of associations, because the time is long not enough, be that with normal neural difference nerve fiber quantity is many, the beam splitting number is many, and not as obviously normal, fibre diameter is inhomogeneous; The electrophysiology evaluation shows that the nervous physiology function is recovered to a great extent, for no other reason than that fall short of recovery time, compares with normal nerve, and the nerve trunk compound action potential of reparation is longer incubation period, and conduction velocity is slower, and amplitude of action is lower; Animal limb sensation and motion conditions are observed, and repairing the batter toe has certain grip, the acupuncture significant reaction, though still towing of walking, degree obviously reduces.Show that it is effective that chitosan catheter is used for neural the reparation, but the recovery fully of nervous physiology function needs the longer time.
Claims (4)
1, a kind of conduit that is used for neural reparation is characterized in that mainly being made up of chitosan, gelatin, glutaraldehyde, acetic acid, sodium hydroxide, poly-D-lysine, and its part by weight is: 100 parts of chitosans: gelatin 5-10 part, all the other trace.
2, the preparation method that is used for the neural conduit of repairing as claimed in claim 1 is characterized in that, comprises that the preparation and the conduit of preparation conduit original solution prepares two parts, specifically may further comprise the steps:
One, preparation conduit original solution preparation:
1) chitosan is dissolved in the acetum, fully stirs silk cover filtering;
2) gelatin is dissolved in the deionized water, is made into gelatin solution;
3) gelatin solution is slowly added in the chitosan solution, stir, drip NaHCO
3Solution is transferred pH to 5.1-6.0, drips glutaraldehyde water solution, stirred for several minute;
4) mid-more than 12 hours at the room temperature shaking table, eliminate the bubble in the solution;
Two, conduit preparation:
1) get the Glass rod of setting diameter, the rotating speed rotation plug with 20～50rpm at the uniform velocity vertically inserts solution;
2) leave standstill treat the spin stabilization of solution a moment with Glass rod after, Glass rod is lifted out liquid level with the speed of 1～5mm/s, place horizontal level, still at the uniform velocity rotate, so that solution distributes on Glass rod all with the speed of 20～50rpm;
3) said process is repeated in the air-dry back of room temperature, reaches 300-500 μ m to pipe thickness and takes off.
3, the preparation method that is used for the neural conduit of repairing as claimed in claim 1 is characterized in that, comprises that the preparation and the conduit of preparation conduit original solution prepares two parts, specifically may further comprise the steps:
One, preparation conduit original solution preparation:
1) chitosan is dissolved in the acetum, fully stirs silk cover filtering;
2) gelatin is dissolved in the deionized water, is made into gelatin solution;
3) gelatin solution is slowly added in the chitosan solution, stir, drip NaHCO
3Solution is transferred pH to 5.1-6.0, drips glutaraldehyde water solution, stirred for several minute;
4) mid-more than 12 hours at the room temperature shaking table, eliminate the bubble in the solution;
Two, conduit preparation:
1) utilize politef to be processed into mould and prepare conduit, in the chitosan solution adding mould with molding, bubble is eliminated in decompression, fills it up with sealing;
2) put into freezing a few hours under-50～-80 ℃ the temperature then, treat freezing fully after, take out, open the mould closed port, place the freezer dryer lyophilization, the moisture in the molding solution distributes fully;
3) mould plug and conduit are together taken out again, place NaOH solution to handle, finalize the design, the reuse distilled water flushing is to neutral, and the conduit that will be enclosed within last on the plug is placed on drying in the baking oven, and heat-treats, and temperature is higher than 80 ℃, but is no more than 120 ℃.Take out plug at last, get conduit.
4, the preparation mould that is used for the neural conduit of repairing as claimed in claim 3 its, it is characterized in that, comprising: sleeve, place the plug in the sleeve, be fixed at the nut at sleeve two ends; The space of the chitosan solution of promising ccontaining desire molding between this sleeve and the plug.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN 99123745 CN1100572C (en) | 1999-11-19 | 1999-11-19 | Chitosan catheter for repair of nerve |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN 99123745 CN1100572C (en) | 1999-11-19 | 1999-11-19 | Chitosan catheter for repair of nerve |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN1262961A CN1262961A (en) | 2000-08-16 |
| CN1100572C true CN1100572C (en) | 2003-02-05 |
Family
ID=5282954
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN 99123745 Expired - Fee Related CN1100572C (en) | 1999-11-19 | 1999-11-19 | Chitosan catheter for repair of nerve |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN1100572C (en) |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1802434B (en) * | 2003-06-05 | 2011-03-23 | 索尼株式会社 | Immobilization support, process for producing the same, electrode, process for producing the same, electrode reaction utilizing apparatus and process for producing the same |
Families Citing this family (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN100417666C (en) * | 2006-09-07 | 2008-09-10 | 重庆大学 | Lysine-chitosan resin and preparation method thereof |
| CN107412854A (en) * | 2017-07-05 | 2017-12-01 | 北京大清生物技术股份有限公司 | A kind of tissue engineering bracket material for CO2 laser weld and preparation method thereof |
| CN111016081B (en) * | 2019-12-16 | 2021-12-14 | 天新福（北京）医疗器材股份有限公司 | Mold and method for preparing multichannel nerve conduit |
| CN114848922B (en) * | 2022-04-19 | 2023-01-10 | 浙江大学 | Composite conduit material doped with mechanoluminescence material and preparation method thereof |
-
1999
- 1999-11-19 CN CN 99123745 patent/CN1100572C/en not_active Expired - Fee Related
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1802434B (en) * | 2003-06-05 | 2011-03-23 | 索尼株式会社 | Immobilization support, process for producing the same, electrode, process for producing the same, electrode reaction utilizing apparatus and process for producing the same |
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| Publication number | Publication date |
|---|---|
| CN1262961A (en) | 2000-08-16 |
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