CN101366982A - Double braid nerve trachea and preparation method thereof - Google Patents
Double braid nerve trachea and preparation method thereof Download PDFInfo
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- CN101366982A CN101366982A CN 200810200387 CN200810200387A CN101366982A CN 101366982 A CN101366982 A CN 101366982A CN 200810200387 CN200810200387 CN 200810200387 CN 200810200387 A CN200810200387 A CN 200810200387A CN 101366982 A CN101366982 A CN 101366982A
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Abstract
The invention provides a double-layer knitting nerve duct having the characteristics that the double-layer knitting nerve duct consists of an outer layer tube wall knitted by polyglycollide-lactide yarns and an inner layer tube wall made from chitin non-woven fabrics, and also provides a method for preparing the double-layer knitting nerve duct having the characteristics that the method comprises the following steps of: attaching the chitin non-woven fabrics on a core bar and knitting the polyglycollide-lactide yarns on an outer layer; forming the polyglycollide-lactide yarns for 2 and 5 minutes at a temperature of between 90 and 105 DEG C, cleaning the polyglycollide-lactide yarns with anhydrous alcohol, and forming the polyglycollide-lactide yarns for 1 and 2 minutes at a temperature of between 90 and 100 DEG C; taking down a polyglycollide-lactide knitting layer and a chitin non-woven fabric layer from the core bar at the same time, and freeze-dry the polyglycollide-lactide knitting layer for 2 and 3 hours at a temperature of between 50 below zero and 40 DEG C. The double-layer knitting nerve duct has the advantages of long-distance bridging defect, good quality and fast speed for nerve regeneration and good mechanical property, and contributes to regulation of a pH value in the body.
Description
Technical field
The present invention relates to a kind of double braid nerve trachea with and preparation method thereof, belong to bio-medical material and tissue engineering technique field.
Background technology
Nerve trachea is the support that is used to guide the injured nerve growth with biological or abiotic prefabricated one-tenth tubular structure.It bridges between the neural broken ends of fractured bone, is used for repairing nerve damage.At present, be used for the biodegradable absorbing material of artificial nerve catheter both at home and abroad and be mainly polylactic acid (PLA), chitosan (chitosan), collagen (collagen), polyglycolic acid (PGA), polyphosphazene (polyphosphazenes) and their copolymer etc., reason is that these material sources are extensive, low price; Biodegradable, to nervous tissue's nonhazardous effect.In addition, along with people to the cell of peripheral nerve regeneration and the further investigation of molecular mechanism, people recognize that gradually nerve trachea not only will be as the occasional passage of neuranagenesis, the more important thing is to have the biologic activity that promotes axon regeneration.Research focus about the peripheral nerve conduit is the biological activity that strengthens conduit at present, and its main method is to add life active substance in nerve trachea, improves the microenvironment of neuranagenesis, improves axon regeneration speed.Concrete grammar comprises that (1) is implanted into schwann cell at nerve trachea, and (2) conduit contains the neurotrophic factor that can slowly discharge; (3) in conduit, add the extracellular matrix composition; (4) the above-mentioned two or more method of use in conjunction improves the activity of nerve trachea.
Super high molecular weight poly-dl-lactide (PDLLA) is the most widely used a kind of degradation material of present peripheral nerve tissue engineering, has excellent biological compatibility.(Chinese science B collects in " organizational project and peripheral nerve reparation-III. gather d; 1-lactic acid pipe is induced neural research of repairing " in king's body states etc., 2001,31 (2), propose 109-144) to repair the damaged sciatic bridge joint of rat 10mm with the polylactic acid pipe, the result shows: the polylactic acid pipe is the bridge joint neurologic defect effectively, in time degrades and absorbs, and neural surrounding tissue is not had rejection.Simultaneously it had both helped the regeneration of neural axon, can alleviate the influence that surrounding tissue is repaired nerve again, thereby for the reparation of neurologic defect provides preferably " microenvironment ", reached and repair close repairing effect with neural transplantation.But polylactic acid exists the difficult control of degradation rate, degraded back acid product can cause cell injury and aseptic inflammation as tube material, and therefore single poly-lactic acid material can not well meet the demands, and often needs more compound other materials.
Kuang Yong etc. are at " chitin and chitosan are in the experimentation [J] of the schwann cell compatibility " (Chinese reconstruction surgical magazine, 1998,12:90-93) point out in the literary composition that chitin and chitosan have excellent biological compatibility to the schwann cell of In vitro culture, chitosan is better than chitin to the compatibility of schwann cell.Gu Xiaosong etc. " the artificial organ nerve graft is repaired the damaged experimentation [J] of Canis familiaris L. sciatic nerve " (natural science progress, 2002,12 (4): propose 381-386) with chitosan make porous, be convenient to the conduit that mass exchange and blood vessel are grown into, be placed with the polyglycolic acid support that is beneficial to schwann cell and the orderly guiding growth of nervous process in the tube chamber.The problem that this material exists at present is that fragility is higher, easy cracked subsiding when tube wall is thin.If tube wall is blocked up, then can prolongs soak time, and may produce the local compression effect regenerating nerve.
In order to allow injured nerve carry out material and information exchange with the external world, nerve trachea high infiltration in recent years, porous has been subjected to attention.Li Qingfeng etc. " different permeability chitosan biomembrane composite conduits are repaired the experimentation [J] of peripheral nerve defection " (shanghai Medicine, 2000,23 (7): propose 390-392), by (more than the 10 μ m of aperture), (aperture 10 μ m are following) and the 3 kinds of chitosan-based conduits of atresia of half permeability of studying full permeability, what the result thought half permeability is best.Can not only allow erythrocyte, oxygen, micromolecule nutrient substance to pass through, intercept unfavorable composition and entered in the conduit, its local microenvironment is beneficial to neural regeneration.In addition, the somebody prepares multiwalled collagem membrane pipe, and mechanical strength is improved a lot, and the PGA pipe etc. that is filled with collagen sponge.And there is experiment to point out that poly butyric salt can be used for the neurologic defect of longer distance.
At present, studies show that much that the Biodegradable macromolecule can freely design the structure of nerve trachea, and obtain good neural repairing effect.Lundborg is at " Bioartificial nerve gratis:A prototype " (Scand J Plast Rencostr Hand Surg, 1996, the notion of " inherent support " has been proposed 30:105), some Polymer Synthesizing fiber filaments are inserted the nerve trachea intracavity, form inherent support, play the effect of stable matrix bridge.Subsequently, Hadlock is in " A tissue-engineered conduit for peripheral nerve repair " (Arch Otolaryngol Head Neck Surg, 1998; 124:1081) imagination that has proposed " tube-in-tube " in the literary composition is again come analog neuron bundle structure, to solve sensory nerve bundle and the involutory problem of motor tract.At home, Dai Chuanchang is at " with the experimentation of Method of Tissue Engineering bridge joint peripheral nerve defection " (Chinese surgical magazine, 2000, structure polyglycolic acid support has been proposed 38:388-390), this support has the ribbon rope of parallel longitudinal arrangement, spatial configuration, schwann cell is climbed after the full ribbon rope, repairs the damaged scabies of rat 15mm sciatic nerve with it and is bordering on nerve autograft.The somebody has studied non-tubular shape, non-fibrous structure.The paper of Toba " Regeneration of canine peroneal nerve with the use of a polyglycolicacid-collagen tube filled with laminin-soaked collagen sponge:a comparative study ofcollagen sponge and collagen fibers as filling materials for nerve conduits[J] " (JBiomed Mater, 2001,58 (6): propose 622-630) to make the composite conduit with polyglycolic acid and collagen, spongy collagen that respectively laminin was soaked and fibrous collagen filling are wherein.
At present, domestic Donghua University has realized making the artificial nerve trachea of degradability with braider: the plastotype that (1) is good; (2) easy and simple to handle; (3) in the neuranagenesis process, can keep shape and not subside, can degrade after regeneration is finished; (4) tube wall has certain permeability, is beneficial to the conveying of neurotrophic factor and the exchange of metabolite, promotes neural growth.The Zhang Junfeng of Donghua University etc. further investigate its braiding structure and yarn selection etc., selected suitable yarn raw material, and research have drawn best braiding structure; Liu Guohua etc. have done further investigation to the tensile mechanical properties and the biocompatibility that weave used polymeric polyglycolide-polylactide (PGLA) material, and a series of experiments of process show, Weaving type PGLA degradable nerve regeneration conduit has possessed necessary mechanical property and degradation property, and successful in the in vivo test of rat and Canis familiaris L.; Lou Lin etc. continue to select for use the PGLA material to weave, and mode such as plasma treatment is improved the performance of nerve trachea.
On this basis, but also there is tangible deficiency in braid nerve trachea aspect the damaged distance of nerve trachea bridge joint, the quality that improves neuranagenesis and the speed increasing.Because neural chemotactic and contact guidance are two big principles of neuranagenesis, the neurologic defect distance is long more, and contact guidance is just difficult more, thus long apart from damaged be an insoluble problem.Above deficiency does not exist only in braid nerve trachea, but the difficult problem that whole tissue engineering nerve conduit field faces jointly.How designing the nerve trachea model that can impel the quick effective regeneration of peripheral nerve, is the vital task that a lot of experts and scholars face.
Summary of the invention
The purpose of this invention is to provide a kind of braid nerve trachea, but the damaged distance of its bridge joint, and the neuranagenesis quality is good, speed is fast.
Another object of the present invention provides the preparation method of above-mentioned braid nerve trachea.
In order to achieve the above object, technical scheme of the present invention provides a kind of double braid nerve trachea, it is characterized in that, the inner layer tube wall that outer wall that is woven into by the polymeric polyglycolide-polylactide yarn and chitin non-woven fabrics constitute is formed.
The present invention also provides the preparation method of above-mentioned double braid nerve trachea, it is characterized in that, concrete steps are:
Step 1. invests the chitin non-woven fabrics and forms inner layer tube wall on the plug, and the polymeric polyglycolide-polylactide yarn is knitted to its outer outer wall that forms;
Step 2. is cleaned with dehydrated alcohol at 90～105 ℃ of typing 2～5min, at 90～100 ℃ of typing 1～2min;
The inner layer tube wall that outer wall that step 3. is woven into polymeric polyglycolide-polylactide and chitin non-woven fabrics constitute was backed off from plug simultaneously, at-50--40 ℃ lyophilization 2-3 hour.
The present invention has overcome the shortcoming and defect of existing braid nerve trachea, and a kind of thinking and method of new preparation braid nerve trachea is provided.The outer tube wall of double braid nerve trachea provided by the invention has permeability preferably, can prevent the intrusion of fibrous connective tissue, also is beneficial to the exchange of pipe ectendotrophy material and metabolite; Internal layer has passage preferably, and these passages not only can the directional guide axon growth, also can promote neuranagenesis by presetting competent cell; Endothecium structure also provides preferably carrier, provides adapt circumstance for mass exchange for the adhesion of cell and growth.By the cell in vitro complication experiment, tentative confirmation double braid nerve trachea of the present invention has cell compatibility preferably, helps the adhesion and the growth of cell.
The present invention has the following advantages:
1. the chitin non-woven fabrics is as the internal layer of nerve trachea, and its excellent surface texture helps neural adhesion growth, but can increase the quality and the speed of the damaged distance of nerve trachea bridge joint, raising neuranagenesis;
2. mechanical property is good;
3. help regulating intravital pH value (chitin be beneficial in and the acidic materials that produce in degraded of PGLA), under similarity condition, pH value becomes 7 by original 5.8.
The specific embodiment
Specify the present invention below in conjunction with embodiment.
Embodiment 1
A kind of double braid nerve trachea, the inner layer tube wall that outer wall that is woven into by the polymeric polyglycolide-polylactide yarn and chitin non-woven fabrics constitute is formed.
Above-mentioned double braid nerve trachea is prepared by following method: the chitin non-woven fabrics is invested form inner layer tube wall on the plug, the polymeric polyglycolide-polylactide yarn is knitted to its outer outer wall that forms; At 90 ℃ of typing 2min, clean with dehydrated alcohol, at 90 ℃ of typing 1min; The inner layer tube wall that outer wall that polymeric polyglycolide-polylactide is woven into and chitin non-woven fabrics constitute was backed off from plug simultaneously ,-50 ℃ of lyophilizations 2 hours.
Embodiment 2
A kind of double braid nerve trachea, the inner layer tube wall that outer wall that is woven into by the polymeric polyglycolide-polylactide yarn and chitin non-woven fabrics constitute is formed.
Above-mentioned double braid nerve trachea is prepared by following method: the chitin non-woven fabrics is invested form inner layer tube wall on the plug, the polymeric polyglycolide-polylactide yarn is knitted to its outer outer wall that forms; At 105 ℃ of typing 5min, clean with dehydrated alcohol, at 100 ℃ of typing 2min; The inner layer tube wall that outer wall that polymeric polyglycolide-polylactide is woven into and chitin non-woven fabrics constitute was backed off from plug simultaneously ,-40 ℃ of lyophilizations 3 hours.
Embodiment 3
A kind of double braid nerve trachea, the inner layer tube wall that outer wall that is woven into by the polymeric polyglycolide-polylactide yarn and chitin non-woven fabrics constitute is formed.
Above-mentioned double braid nerve trachea is prepared by following method: the chitin non-woven fabrics is invested form inner layer tube wall on the plug, the polymeric polyglycolide-polylactide yarn is knitted to its outer outer wall that forms; At 100 ℃ of typing 3.5min, clean with dehydrated alcohol, at 95 ℃ of typing 1.5min; The inner layer tube wall that outer wall that polymeric polyglycolide-polylactide is woven into and chitin non-woven fabrics constitute was backed off from plug simultaneously ,-45 ℃ of lyophilizations 2.5 hours.
The performance test of the double braid nerve trachea of embodiment 1-3 is as follows:
1. the measurement of porosity and comparison: the notion suitable with porosity is " relative density ", and it is the porous body apparent density and the ratio of corresponding fine and close material density.In experiment, adopt following formula to calculate porosity:
K is a porosity in the formula, ρ
rBe relative density (nondimensional decimal), ρ
*Apparent density (g/cm for nerve trachea
3), ρ
SActual density (g/cm for nerve trachea
3).
Wherein, the actual density of nerve trachea adopts pycnometric method density measurement method to obtain.
The porosity of double braid nerve trachea sees the following form among conventional monolayers braid nerve trachea and the embodiment 1-3:
By data in the table as can be seen, from actual density, the actual density of braid nerve trachea of the present invention is less than in the past monolayer Weaving type nerve trachea, and the reduction of conduit density helps strengthening the oxygen permeability in the cell growth process.From porosity, the porosity of two kinds of materials is suitable, and all about 80% (because the existence of error in the experiment can not conclude from small difference that porosity is any better) all satisfies the requirement of cell growth in the nerve trachea.Realized simultaneously, when reducing conduit density, do not influenced the porosity of conduit.Solved the contradiction between former studies mesopore rate and the conduit density.
2. tensile property is relatively:
Experimental apparatus: WDW-20 microcomputer control universal testing machine, draw speed: 20mm/min, clamping spacing: 20mm, specimen length: 60mm.If directly be clamped in conduit on the chuck,, the conduit both sides easily produce stress concentration because being crushed in the clamping port, therefore the hardboard of band recess in the middle of at first adhering in the grip block both sides, each inserts one section rod iron at the conduit two ends, guarantee that conduit clamps and do not produce distortion at clamp mouth part, concentrate influence to eliminate stress the conduit brute force.
The tensile property of double braid nerve trachea sees the following form among conventional monolayers braid nerve trachea and the embodiment 1-3:
Because carrying out coating, do not handle double braid nerve trachea of the present invention, therefore powerful less.But extension at break increases.The present invention not only makes the adding of internal layer help the adhesion and the neural growth of cell, and has saved chitosan slurry coating operation.
3. twisting property relatively
Experimental apparatus: reverse strength tester, constant predetermined amount: 90 degree, torsional mode: decide under the angle measurement standard atmosphere condition (20 ℃ ± 2 ℃, RH 65% ± 2%) and test.
The twisting property of double braid nerve trachea sees the following form among conventional monolayers braid nerve trachea and the embodiment 1-3:
The antitorque commentaries on classics performance of double braid nerve trachea of the present invention is suitable with the monolayer nerve trachea.Reason is that the chitosan slurry coating of monolayer nerve trachea has strengthened the antitorque commentaries on classics performance of conduit, and the adding of the internal layer of double braid nerve trachea of the present invention has reached same effect.But the adding of chitin non-woven fabrics internal layer for the adhesion of cell and neural growth provide carrier, more helps neural reparation than monolayer braid nerve trachea.
4. degraded percentage rate and compression performance change
According to data, there are some conduits slight collapse phenomenon in experimentation, to occur, had a strong impact on neural growth.Therefore, be necessary the radial compression changes of properties of conduit in degradation process studied.PGLA degraded back is acid, can influence intravital acid-base balance, therefore need measure for the pH value after the nerve trachea degraded.
Weight-loss ratio and pH value among conventional monolayers braid nerve trachea and the embodiment 1-3 after around the double braid nerve trachea external degradation see the following form:
As can be seen from the table, double-deck nerve trachea of the present invention has solved the sour environment influence that reparation causes to nerve that polymeric polyglycolide-polylactide produces well in degraded.
Compression performance among conventional monolayers braid nerve trachea and the embodiment 1-3 in the double braid nerve trachea external degradation experimentation changes (elastic recovery loss rate %) and sees the following form:
Wherein, compression performance changes the following experiment parameter of employing in the test: 25 ℃ of temperature, and humidity 65%, the number of occurrence 5 times, compression speed 6mm/min, compression distance are 50% of conduit initial diameter.
As can be seen from the table, after the last fortnight degraded, the elastic recovery loss rate of monolayer braid nerve trachea is less, and reason is that the last fortnight mainly is the chitosan serosity degraded that is used for coating, and the mechanical property loss of conduit itself is imitated little.But the beginning of the 3rd week, the elastic recovery loss rate of monolayer nerve trachea has tangible rising, and the elastic resilience of double braid nerve trachea of the present invention demonstrates its remarkable advantages.
Claims (2)
1. a double braid nerve trachea is characterized in that, the inner layer tube wall that outer wall that is woven into by the polymeric polyglycolide-polylactide yarn and chitin non-woven fabrics constitute is formed.
2. a kind of preparation method of double braid nerve trachea according to claim 1 is characterized in that concrete steps are:
Step 1. invests the chitin non-woven fabrics and forms inner layer tube wall on the plug, and the polymeric polyglycolide-polylactide yarn is knitted to its outer outer wall that forms;
Step 2. is cleaned with dehydrated alcohol at 90～105 ℃ of typing 2～5min, at 90～100 ℃ of typing 1～2min;
The inner layer tube wall that outer wall that step 3. is woven into polymeric polyglycolide-polylactide and chitin non-woven fabrics constitute was backed off from plug simultaneously, at-50--40 ℃ lyophilization 2-3 hour.
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Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101829356A (en) * | 2010-05-06 | 2010-09-15 | 东华大学 | Tissue engineering artificial tendon scaffold enhanced body and preparation method thereof |
| CN104382672A (en) * | 2014-11-14 | 2015-03-04 | 东华大学 | Double-layer multichannel nerve conduit and preparation method thereof |
Family Cites Families (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN100479785C (en) * | 2005-04-07 | 2009-04-22 | 首都医科大学北京神经科学研究所 | Producing method for double-layer artificial nerve catheter |
| CN1730117B (en) * | 2005-07-15 | 2011-04-27 | 东华大学 | Process for preparing peripheral nerve regeneration conduit |
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Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101829356A (en) * | 2010-05-06 | 2010-09-15 | 东华大学 | Tissue engineering artificial tendon scaffold enhanced body and preparation method thereof |
| CN101829356B (en) * | 2010-05-06 | 2013-06-05 | 东华大学 | Tissue engineering artificial tendon scaffold enhanced body and preparation method thereof |
| CN104382672A (en) * | 2014-11-14 | 2015-03-04 | 东华大学 | Double-layer multichannel nerve conduit and preparation method thereof |
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