CN102085393A - Biodegradable nerve conduit with bilayer structure and preparation method thereof - Google Patents
Biodegradable nerve conduit with bilayer structure and preparation method thereof Download PDFInfo
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Abstract
The invention relates to a biodegradable nerve conduit with a bilayer structure and a preparation method thereof. The biodegradable nerve conduit comprises a reticular monofilament liner which is made of polyglycollide-lactide (PLGA) and used as a framework structure and a nano fiber felt which is made of a biodegradable material and used as an outer layer structure. The preparation method comprises the following steps of: (1) preparing PLGA monofilaments by adopting a fused spinning method, stretching for orientating, and then weaving into a reticular liner conduit; (2) dissolving the biodegradable material into an organic solvent to prepare a spinning stock solution with a mass fraction of 3-20 percent; and (3) carrying out electrostatic spinning by using the spinning stock solution on the surface of the reticular liner conduit by taking the reticular liner conduit as a receiving device to obtain the biodegradable nerve conduit. The biodegradable nerve conduit has good biocompatibility, proper degradation time and higher mechanical strength and can promote the regeneration and the restoration of damaged nerves. The preparation method has the advantages of simpleness of operation, low cost, environmental protection, high economic benefit and good application prospect.
Description
Technical field
The invention belongs to the field of biodegradable nerve trachea, particularly a kind of have double-deck biodegradable nerve trachea and preparation method thereof.
Background technology
The 19th-century end is at first taken off bone calcium by Cluck by employing and has been made the bone pipe and come bridge joint neurologic defect position, carries out nerve reparation, and the research that utilizes conduit to repair neurologic defect is constantly carried out.Comprise that utilization that Weiss proposed in nineteen forty-four do not have the notion that the method for stitching thread conduitization is come repairing nerve damage, and the artificial neuron implantation technique that grows up gradually of the later stage seventies 20th century, promptly adopt biomaterial to prepare nerve trachea, and the microenvironment of in conduit cavity, building nerve growth, guiding also promotes neuranagenesis.In recent years, along with the continuous research and the exploration of tissue engineering technique research, engineered neural transplantation becomes the ideal scheme of repairing peripheral nerve injury.But up to the present, the progress of nerve reparation is still slowly, for how obtaining ideal neuranagenesis form, and further improves the recovery capability of function of nervous system, become the direction that the various countries scholar constantly makes great efforts.
Perineural damage roughly can be divided into two classes: between the N/D damage and the broken ends of fractured bone damaged two classes are arranged between the broken ends of fractured bone.For N/D damage between the broken ends of fractured bone mainly is to adopt operation methods such as adventitia coincide, the bundle film the coincide reparation that directly coincide at present.But,, thereby can make regenerating tissues not arrive original position and lose function because the hypertrophy of its severed nerve colloid and neural peripheral connective tissue all can hinder the growth forward of regenerating nerve fiber for damaged damage is arranged between the broken ends of fractured bone.Therefore being the main difficult problem that present medical circle faces to the perineural reparation that damage is arranged between the broken ends of fractured bone, also is various countries scientist's research focus.
In recent years, development along with tissue engineering technique, adopt the ultimate principle and the method for tissue engineering, according to the biological characteristics of neuranagenesis, preparation has the carrier mass of good biocompatibility and repairs perineural damage to become main direction of studying as nerve trachea.Repair peripheral nerve injury by the nerve trachea bridge joint, the conduit bridge joint peripheral nerve injury broken ends of fractured bone made from biology or non-biological material exactly is for the neuranagenesis that damages provides regeneration room.Its advantage is: on the one hand, because neuranagenesis does not have penetrance, in growth course if run into obstacle then the growth meeting stop at once, nerve trachea can prevent the intrusion of connective tissue on every side and the formation of cicatrix for neuranagenesis provides a clog-free passage; On the other hand, nerve trachea helps keeping endogenous and ectogenic neurotrophic factor, somatomedin etc. can promote the excitor substance of synapse growth.That is to say that the mechanism of utilizing the method for nerve trachea to repair peripheral nerve injury is exactly to be that neuranagenesis is built suitable " microenvironment ".In the last few years, the research worker of various countries had been invented various types of nerve tracheas, mainly contained non-nervous tissue's conduit, non-biodegradation nerve trachea and biodegradable nerve trachea, simply was described below:
People such as Dellon [Dellon et al., Bioabsorbable surgical device for treating nerve defects, U.S.Pat.No.4870966] invented softness, the netted nerve trachea of porous degradable of the knitting or weaving method preparation of a kind of usefulness.The catheter design rolling is curved, in order to avoid damage inner regenerative system when needs are crooked.Catheter interior is very smooth, and neural axon is in the optimum growh environment.Li[Li, Multi-layered, semi-permeable conduit for nerve regeneration, U.S.Pat.No.4,963,146; Li, Multi-layered, semi-permeable conduit for nerve regeneration comprised of type 1 collagen, its method of manufacture and a method of nerve regeneration using said conduit, U.S.Pat.No.5,026,381] Fa Ming nerve trachea tube wall is made up of the semi-permeable type i collagen of multilamellar, bore hole size it is said that these holes optionally allow the neuron factor to pass through by stoping the fibroid cell in the tube wall between 0.006～5 micron.Nichols[Nichols, Nerve regeneration conduit, U.S.Pat.No.5019087] also invented similar nerve trachea, tube wall is made up of type i collagen and laminin, it is said the growth that proteic adding can accelerator nerve.
Also this has made huge contribution to domestic research worker on the one hand at the development nerve trachea: people such as Ao Qiang [Ao Qiang etc., a kind of nervous tissue engineering tubular bracket and preparation method thereof: 200410009259.9[P] .2004-06-25] invented a kind of nervous tissue engineering tubular bracket and preparation method thereof, this nervous tissue's engineering tubular bracket is by the chitosan tube wall and have axial multichannel biogenetic derivation filling substrate and form, and each interchannel has the micropore that is interconnected, can prevent the intrusion of fibrous connective tissue, also be beneficial to the exchange of pipe ectendotrophy material and metabolite.It is 1～5mm semipermeability chitosan hollow pipe that this method at first prepares internal diameter, in pipe, pour into biological macromolecule solns such as chitosan, collagen or gelatin then, utilize particular manufacturing craft and Freeze Drying Technique, preparation has the tubular bracket that nerve injury is repaired that can be used for of 7～50 axial passages.The multi-channel nerve conduit that gained has biomimetic features helps cell adhesion, migration and guiding aixs cylinder oriented growth, is applicable to the reparation and the regeneration of nerve injury.This preparation technology is easy and simple to handle, and a kind of thinking and method of new preparation nervous tissue engineering rack is provided.People such as Zhang Jing [Zhang Jing etc. can promote the composite collagen nerve trachea and hollow wet spinning manufacturing process: the 200410099205.6[P thereof of neuranagenesis] .2004-12-29] a kind of composite collagen nerve trachea and hollow wet spinning manufacturing process thereof that promotes neuranagenesis disclosed.By collagen protein, the composite collagen spinning solution that chitosan and porogen are formed makes the composite collagen nerve trachea by one-step method hollow wet spinning manufacturing process.Spinning technique combines with hole forming technology in this method, and disposal molding hollow semi permeability nerve trachea has reduced the complexity of catheter fabrication.Be easy to spin out the nerve trachea of different apertures, porosity, diameter and cross sectional shape, be fit to the environment that various types of cells adheres to growth, promote neural regeneration thereby create.Xu Guofeng [Xu Guofeng, biological nerve duct: 200510120792.7[P] .2005-12-20] a kind of biological nerve duct and preparation method thereof disclosed, it is formed by removing the thin-walled body that animal membrane material that antigen handles makes through the crosslinked fixing and active reagent of no-aldehyde fixative and strong hydrogen bonding reagent and being fixed on the helical mount that elongated slip coiled coil that the animal membrane material of being handled by preceding method on the tube wall is cut into is bonded.Advantage of the present invention is: it is made by the natural biologic material that with the animal tissue is raw material, and non-immunogenicity can not cause rejection, has the favorable tissue compatibility, can induce division, propagation and the migration of neurocyte, promotes the regeneration of nervous tissue.It can guarantee nervous tissue's regeneration requisite space passage, can see through the nutrient substance and the blood vessel of growing into again, the nutritional need of supply nervous tissue growth, for good microenvironment is created in the reproducibility reparation of nervous tissue, after the Regeneration and Repair of nervous tissue is finished, it can be degraded and absorbed, not as the existence of foreign body forever.People such as Wu Qiong [Wu Qiong etc., a kind of nerve conduit stent and preparation method thereof: 200910001598.5[P] .2009-01-15] a kind of nerve conduit stent is provided, it is made of polyhydroxyalkanoate, and the inwall of described nerve conduit stent is fine and close small pore-like structure, and the aperture is below the 10 μ m; Outer wall is loose macroporous structure, and the aperture is 30-100 μ m.This nerve conduit stent adopts and earlier polyhydroxyalkanoate series polymer and organic solvent is mixed and made into uniform solution, mixed proportion is that every gram polyhydroxyalkanoate series polymer adds the 8-20ml organic solvent, the water-soluble granular of different-diameter will be added again in the above-mentioned solution, after evenly stirring, dip in pole shape mould and to get mixture; After treating that solution solidifies fully, the stripping porogen obtains loose structure.People such as Fan Cunyi [Fan Cunyi etc., a kind of nerve trachea and its production and use: 200810208123.9[P] .2008-12-29] a kind of nerve trachea and its production and use disclosed, described nerve trachea is made of " shell-core " structure nano fiber, the sandwich layer of described " shell-core " structure nano fiber contains bioactive ingredients, and the shell of described " shell-core " structure nano fiber contains Biodegradable material; The internal diameter of described nerve trachea is 0.5-5.0mm, and external diameter is 1.0-6.0mm; Described bioactive ingredients is a neurotrophic factor; Described Biodegradable material is selected from following one or more: lactic acid-polyglycolic acid copolymer (PLGA), polylactic acid one caprolactone copolymer P (LLA-CL), poly-trihydroxy-butyric acid (PHB), or poly phosphate (PPE).But the nerve trachea in its invention does not have framing structure, and its mechanical property can be affected.
Summary of the invention
Technical problem to be solved by this invention provides a kind of double-deck biodegradable nerve trachea and preparation method thereof that has, this nerve trachea biocompatibility is good, have suitable degradation time and higher mechanical strength, can promote the injured nerve Regeneration and Repair; Preparation method is simple to operate, and cost is low, and is environmentally friendly, and the economic benefit height has a good application prospect.
A kind of double-deck biodegradable nerve trachea that has of the present invention comprises: netted monofilament liner of making as the polymeric polyglycolide-polylactide PLGA of framing structure and the nanofiber mats of making as the degradable biomaterial of layer structure.
Described degradable biomaterial is one or more in polymeric polyglycolide-polylactide PLGA, polylactic acid PLA, poly butylene succinate PBS, polycaprolactone (PCL), the fibroin albumen.
Described nerve trachea length is 10～100mm, and internal diameter is 0.5～5mm, and wall thickness is 0.02～1.5mm.
A kind of preparation method with double-deck biodegradable nerve trachea of the present invention comprises:
(1) adopts melt spinning method to make the PLGA monofilament, be woven into netted liner conduit behind the stretch orientation;
(2) degradable biomaterial is dissolved in the organic solvent, makes mass fraction and be 3～20% spinning solution;
(3) with the netted liner conduit in the step (1) as receiving system, the spinning solution in the step (2) is carried out electrostatic spinning at netted liner catheter surface, promptly get and have double-deck biodegradable nerve trachea.
Organic solvent in the described step (2) is oxolane, N, one or more in dinethylformamide DMF, acetone, chloroform, formic acid, the trifluoroacetic acid.
Electrostatic spinning process parameter in the described step (3) is that the flow of boost pump is 1.3mL/h, and plus high-pressure is 7～10kv, and receiving range is 18cm.
The cladding material of nerve trachea is the three-dimensional net structure of biodegradable high-specific surface area, high porosity, can be as the support of organizational project.The intensity and the flexibility of the fiber number by adjusting the PGLA monfil, the adjustable nerve trachea of count of reticulate vessel; Can regulate and control the structural parameters such as wall thickness, density, voidage, bore hole size of nerve trachea by the spinning technology parameter of regulating outer electrostatic spinning felt.The present invention can be applied to the reparation of peripheral nervous system.
Beneficial effect
(1) nerve trachea tube wall of the present invention is a kind of structure that possesses high-specific surface area, high porosity, three-dimensional network shape that adopts the coaxial electrostatic spinning method to be prepared from, and still reproduces neural tissue engineering bracket when playing guiding, carrying the nutrient effect;
(2) liner of nerve trachea of the present invention is the PLGA monofilament that adopts melt spinning method to be prepared from, because high-molecular weight PLGA not only has excellent biological compatibility and suitable degradation time, also have higher mechanical strength, be suitable as the framing structure of nerve trachea;
(3) nerve trachea of the present invention can be erected a bridge block at be badly damaged fracture and the very difficult neural two ends of sewing up by surgical operation, plays the effect that promotes the injured nerve Regeneration and Repair;
(4) preparation method of the present invention is simple to operate, and cost is low, and is environmentally friendly, the economic benefit height.
Description of drawings
The liner reticulate vessel of Fig. 1 for adopting the high molecular degradable fiber to make;
Fig. 2 is prepared nerve trachea outward appearance.
The specific embodiment
Below in conjunction with specific embodiment, further set forth the present invention.Should be understood that these embodiment only to be used to the present invention is described and be not used in and limit the scope of the invention.Should be understood that in addition those skilled in the art can make various changes or modifications the present invention after the content of having read the present invention's instruction, these equivalent form of values fall within the application's appended claims institute restricted portion equally.
Embodiment 1
(1) the PGLA monofilament that melt spinning is made is woven into the netty circular tube that internal diameter is 2mm, and the rotation of being inserted in the electrostatic spinning machine after the fixed length typing receives on the pipe, and rotary speed is 4000rpm;
(2) PLA is dissolved in oxolane/DMF solution (volume ratio of oxolane: DMF=3: 1), makes the PLA mass percent after the stirring and be PLA/ oxolane/DMF solution of 6%; Fibroin albumen is dissolved in the trifluoroacetic acid solution, makes fibroin albumen quality percentage ratio after the stirring and be 10% spinning solution;
(3) above-mentioned two kinds of spinning solutions are imported respectively in two syringe pumps, two syringe pumps have the shower nozzle in inside and outside two same axle center, carry out electrostatic spinning, and the flow of adjusting boost pump is 1.3mL/h, and plus high-pressure is 10kv, and receiving range is 18cm;
(4) after electrostatic spinning finishes, take off nerve trachea, use cobalt 60 illumination-based disinfections, pack, nerve trachea length is 50mm, and wall thickness is 1mm.
Embodiment 2
(1) the PGLA monofilament that melt spinning is made is woven into the netty circular tube that internal diameter is 2mm, and the rotation of being inserted in the electrostatic spinning machine after the fixed length typing receives on the pipe, and rotary speed is 4000rpm;
(2) PBS/PCL (mass ratio 1: 4) is dissolved in the chloroformic solution, makes the solute mass fraction after the stirring and be 10% PBS/PCL/ chloroformic solution; Gelatin is dissolved in the formic acid solution, makes the gelatin mass fraction after the stirring and be 10% spinning solution;
(3) two kinds of spinning solutions are imported respectively in two syringe pumps, two syringe pumps have the shower nozzle in inside and outside two same axle center, carry out electrostatic spinning, and the flow of adjusting boost pump is 1.3mL/h, and plus high-pressure is 7.53kv, and receiving range is 18cm;
(4) after electrostatic spinning finishes, take off nerve trachea, use cobalt 60 illumination-based disinfections, pack, nerve trachea length is 10mm, and wall thickness is 0.02mm.
Embodiment 3
(1) the PGLA monofilament that melt spinning is made is woven into the netty circular tube that internal diameter is 0.5mm, and the rotation of being inserted in the electrostatic spinning machine after the fixed length typing receives on the pipe, and rotary speed is 4000rpm;
(2) PBS/PLA (mass ratio 1: 4) is dissolved in the chloroformic solution, makes the solute mass fraction after the stirring and be 10% PBS/PLA/ chloroformic solution; Gelatin is dissolved in the formic acid solution, makes the gelatin mass fraction after the stirring and be 10% spinning solution;
(3) two kinds of spinning solutions are imported respectively in two syringe pumps, two syringe pumps have the shower nozzle in inside and outside two same axle center, carry out electrostatic spinning, and the flow of adjusting boost pump is 1.3mL/h, and plus high-pressure is 8kv, and receiving range is 18cm;
(4) after electrostatic spinning finishes, take off nerve trachea, use cobalt 60 illumination-based disinfections, pack, nerve trachea length is 100mm, and wall thickness is 1.5mm.
Embodiment 4
(1) the PGLA monofilament that melt spinning is made is woven into the netty circular tube that internal diameter is 5mm, and the rotation of being inserted in the electrostatic spinning machine after the fixed length typing receives on the pipe, and rotary speed is 4000rpm;
(2) PBS/PLA (mass ratio 1: 4) is dissolved in the chloroformic solution, makes the solute mass fraction after the stirring and be 10% PBS/PLA/ chloroformic solution; Gelatin is dissolved in the formic acid solution, makes the gelatin mass fraction after the stirring and be 10% spinning solution;
(3) two kinds of spinning solutions are imported respectively in two syringe pumps, two syringe pumps have the shower nozzle in inside and outside two same axle center, carry out electrostatic spinning, and the flow of adjusting boost pump is 1.3mL/h, and plus high-pressure is 8kv, and receiving range is 18cm;
(4) after electrostatic spinning finishes, take off nerve trachea, use cobalt 60 illumination-based disinfections, pack, nerve trachea length is 80mm, and wall thickness is 0.5mm.
Claims (6)
1. one kind has double-deck biodegradable nerve trachea, comprising: netted monofilament liner of making as the polymeric polyglycolide-polylactide PLGA of framing structure and the nanofiber mats of making as the degradable biomaterial of layer structure.
2. a kind of double-deck biodegradable nerve trachea that has according to claim 1, it is characterized in that: described degradable biomaterial is one or more in polymeric polyglycolide-polylactide PLGA, polylactic acid PLA, poly butylene succinate PBS, polycaprolactone (PCL), the fibroin albumen.
3. a kind of double-deck biodegradable nerve trachea that has according to claim 1, it is characterized in that: described nerve trachea length is 10～100mm, and internal diameter is 0.5～5mm, and wall thickness is 0.02～1.5mm.
4. preparation method with double-deck biodegradable nerve trachea comprises:
(1) adopts melt spinning method to make the PLGA monofilament, be woven into netted liner conduit behind the stretch orientation;
(2) degradable biomaterial is dissolved in the organic solvent, makes mass fraction and be 3～20% spinning solution;
(3) with the netted liner conduit in the step (1) as receiving system, the spinning solution in the step (2) is carried out electrostatic spinning at netted liner catheter surface, promptly get and have double-deck biodegradable nerve trachea.
5. a kind of preparation method according to claim 4 with double-deck biodegradable nerve trachea, it is characterized in that: the organic solvent in the described step (2) is oxolane, N, one or more in dinethylformamide DMF, acetone, chloroform, formic acid, the trifluoroacetic acid.
6. a kind of preparation method according to claim 4 with double-deck biodegradable nerve trachea, it is characterized in that: the electrostatic spinning process parameter in the described step (3) is that the flow of boost pump is 1.3mL/h, plus high-pressure is 7～10kv, and receiving range is 18cm.
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Application publication date: 20110608 |