CN105194737A - Tissue repair support and its preparation method and use - Google Patents
Tissue repair support and its preparation method and use Download PDFInfo
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- CN105194737A CN105194737A CN201410238853.9A CN201410238853A CN105194737A CN 105194737 A CN105194737 A CN 105194737A CN 201410238853 A CN201410238853 A CN 201410238853A CN 105194737 A CN105194737 A CN 105194737A
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Abstract
The invention provides a tissue repair support and its preparation method and use. The tissue repair support comprises a porous tridimensional network structure composed of a composite fiber layer and a hydrophilic material. The composite fiber layer comprises composite fibers composed of an adhesion factor and a hydrophobic synthesis material. The tissue repair support is used for meninge repair, spinal meninge repair, a tissue engineering support material, artificial skin, a wound accessory material, a biomembrane, a wound cladding material, a haemostasis material, a postoperative antistick material or a beauty treatment material.
Description
Technical field
The present invention relates to a kind of biomedical articles and preparation thereof, particularly a kind of tissue recovery support, Preparation Method And The Use.
Background technology
Tissue recovery support is very common in clinical position, and the reparation of such as tendon patch, ligament repair, hernia repair sheet, duramater reparation sheet, female pelvic floor functional disorder disease patch, blood vessel, suspender belt patch, skin repairing, fistula repairing, nerve trachea repairing, Cranial defect are repaired.
Because current material itself and pore structure have certain defective, with tissue component units, all there is very large diversity, especially hydrophobic synthetic material adopts general preparation method, the multi-pore structure of substantially not good similar extracellular matrix, be difficult to the characteristic complying with tissue anatomical structure, be difficult to form good tissue compliance with tissue anatomical structure and stick.
Establishment mesh sheet is product most widely used in current tissue repair fibrous membrane, but often there are the following problems for this series products: 1) rough surface, quality comparatively sclerous tissues's poor compliance, the tissue compliance of difference can produce and the friction organized when applying, to patient's discomfort significantly, easily produce foreign body sensation and pain, cause the complication such as common erosion and infection; 2) immunological rejection that causes of poor biocompatibility and such material is comparatively strong, there is more postoperative complication; When it directly contacts with internal organs, organ, easily cause damage, more serious adhesion can be caused, cause serious foreign body and immunoreation, need second operation to take out, bring misery to patient, even threat to life.The electrospinning film of current research, commonly occurs that cell is difficult to slow deficiency of growing into its inside or grow into.The performance of the tissue repair fibrous membrane that existing braiding, electrospinning are formed is all not ideal enough.
Desirable tissue renovation material needs: 1. porosity is comparatively large, is convenient to cell adhesion, creeps and grow, realize tissue regeneration fast, wants to bear Intraabdominal pressure before health tissues is not fully formed; 2. can keep good dimensional stability after implant into body, namely can not shrink and be out of shape; 3. soft, the effect be convenient to molding, increased the operability of operation, reduce the sense of discomfort of patient, improve operation; 4. good biocompatibility, can transmitting tissue grow, and reaches desirable and repairs.
Summary of the invention
the problem that invention will solve
Existing tissue renovation material pore structure is simple, hole and specific surface area limited, the fibroblast (30 microns) forming cambium cannot enter material internal, and new tissue growth is slow.
Existing synthetic tissue repair materials pore structure is single, and due to a great difference with tissue cellular construction, the active force between tissue recovery support and tissue is weak, does not have the compliance of tissue anatomical structure.
Existing establishment mesh sheet product surface is coarse, quality comparatively sclerous tissues's poor compliance, the tissue compliance of difference can produce and the friction organized when applying, to patient's discomfort significantly, easily produce foreign body sensation and pain, cause the complication such as common erosion and infection.
The mechanical strength of existing hydrophobic material own is very high, but quality is partially hard and hydrophobicity is strong, material is put into solution soak, solution is difficult to enter material internal, even if in conjunction with technology such as lyophilizing, be also difficult to allow material stiffness structure, pore structure change to produce the complying with performance, paste volt performance and multiple hole structure of tissue anatomical structure.
Existing hydrophilic material is due to a lot of hydrophilic flexibility group of material itself, lack rigid radical, mechanics of materials intensity is very poor, is easy to degrade under the more fluid environment in vivo of hydrophilic group, and degraded just occurred before cambium is not also repaired can cause repairing failure.
for the scheme of dealing with problems
Tissue recovery support of the present invention, is characterized in that, described tissue recovery support comprises the porous three-dimensional network structure formed by composite fibre layer and hydrophilic material; Described composite fibre layer comprises the composite fibre be made up of adhesion factor and hydrophobic synthetic material.
Tissue recovery support of the present invention, wherein, on the surface that described porous three-dimensional network structure can be formed at described composite fibre layer and/or interior surface.
Tissue recovery support of the present invention, wherein, described porous three-dimensional network structure, by being immersed in the solution of hydrophilic material by described composite fibre layer, being taken out postlyophilization and is formed.
Tissue recovery support of the present invention, wherein, the solution of described hydrophilic material is the aqueous solution of water soluble polymer.
Tissue recovery support of the present invention, wherein, described hydrophilic material is selected from the group be made up of hydrophilic protide, cellulose family, alcohols, ethers, chitosan class, saccharide, nitrogenous class hydroaropic substance.
Tissue recovery support of the present invention, wherein, the described adhesion factor forming described composite fibre can be present in inside and/or the surface of described composite fibre.
Tissue recovery support of the present invention, wherein, described adhesion factor be selected from there is hydrophilic protide and derived material thereof, there is hydrophilic cellulose family and derived material thereof, there is hydrophilic alcohols and derived material thereof, chitosan class and derived material, saccharide and derived material thereof, one or more in nitrogenous class hydroaropic substance.
Tissue recovery support of the present invention, wherein, described hydrophobic synthetic material comprises hydrophobic polyurethane, polyethylene terephthalate, polycaprolactone, polyglycolic acid, Poly(D,L-lactide-co-glycolide, 1,3-PD polymer, polylactic acid-caprolactone copolymers, polylactic acid.
Tissue recovery support of the present invention, wherein based on the gross weight of described composite fibre layer, the content of described adhesion factor is 5 quality %-95 quality %, is preferably 10 quality %-50 quality %, more preferably 15 quality %-30 quality %.
Tissue recovery support of the present invention, is soaked in 10s in water by described tissue recovery support, and its water absorption rate can reach 100-5000%.
Tissue recovery support of the present invention, wherein, the hot strength of described tissue recovery support is greater than 1Mpa.
Tissue recovery support of the present invention, wherein, the hydrophile-lipophile balance value HLB of described tissue recovery support is 1-13.
Tissue recovery support of the present invention, wherein, the surface of described tissue recovery support and contact angle θ≤90 ° of water.
Tissue recovery support of the present invention, wherein, described tissue recovery support and the surrounding tissue brute force that rises brokenly after pasting and lying prostrate is more than 0.10kPa.
Tissue recovery support of the present invention, wherein, the cellosilk that described composite fibre layer is 10nm-100 μm by diameter is interwoven, and has cavernous structure.
Tissue recovery support of the present invention, wherein, the aperture of described composite fibre layer support is 1-100 μm, and the aperture of described tissue recovery support is 1.3-500 μm.
The purposes of tissue recovery support of the present invention, described tissue recovery support is used for meninges repairing, spinal meninges repairing, tissue engineering bracket material, artificial skin, wound dressings, biomembrane, wound clad material, hemostatic material, post-operation adhesion preventing material or beauty treatment material.
The preparation method of tissue recovery support of the present invention, it comprises:
A () prepares the step of composite fibre layer: be dissolved in homogeneous solvent by described adhesion factor and described hydrophobic synthetic material, then be prepared by one or more combinations in electrostatic spinning technique, centrifugal force spining technology, hot melt spray webbing technology, melting electrospinning, obtain the diaphragm containing composite fibre;
B () prepares the cancellated step of porous three-dimensional: be soaked in the solution of hydrophilic material by the diaphragm containing composite fibre described in preparing, carry out lyophilization after taking-up.
Preparation method of the present invention, wherein, the described solvent used in the step of composite fibre layer of preparing at least comprises a kind of fluorine kind solvent.
Preparation method of the present invention, wherein, described fluorine kind solvent comprises hexafluoroisopropanol, trifluoroethanol, trifluoroacetic acid.
the effect of invention
Tissue recovery support of the present invention has unique multiple hole gap structure, has higher porosity and specific surface area, can better promote that cell is grown into, and accelerates cambium and repairs speed.
Form good subsides by physical force and tissue anatomical structure after implantation to lie prostrate; To the quick absorption of surrounding tissue body fluid, be further connected with organization formation under biotic induce adhesive effect; Along with creeping of collagen fiber is grown into, timbering material and surrounding tissue form one, realize repairing.
The composite fibre that different materials is formed has different controllable degradation rates, the degrade space that formed after fast material degradation is more conducive to fibrocyte and grows into rapidly, slow material of degrading can prevent part material degradation too fast in the early stage and form defect, ensures repairing prerequisite completely for enough mechanical support.
The present invention adopts bionics techniques to prepare to have the bigger serface of nanofiber, is conducive to adhesion and the propagation of cell, good tissue compliance and paste the tissue recovery support of volt effect, meets the requirement of the aspects such as the biology needed for tissue recovery support, mechanics.And surface topology is also conducive to guiding cell differentiation.Light, the soft comfort more improving patient of quality, alleviates the wound of patient, is beneficial to the reparation of patient sooner.
The present invention first adopts bionics techniques to prepare the composite fibre of different materials formation, composite fibre has hydrophobic material and water wetted material simultaneously, composite fibre layer in the solution swelling rear space structure increases, then by the technology such as lyophilization fix swelling after structure and obtain porous space structure and the surface texture of new uniqueness, form various structures simultaneously and the porous three-dimensional network structure of depositing.The degraded of hydrophobic material is comparatively slow, and implanting, early stage can keep good mechanical support effect, can form the good compatibility with surrounding tissue and paste volt property, and then reach good repairing effect in conjunction with water wetted material.
Accompanying drawing explanation
A and B of Fig. 1 is respectively the present invention's being schemed by the SEM binding the composite fibre that Summing Factor hydrophobic synthetic material is formed and the SEM figure of this composite fibre after the solvent process can dissolving wherein a kind of material for composite fibre layer.
Fig. 2 is the SEM figure on composite fibre layer surface of the present invention.
Fig. 3 is cancellated SEM figure (10 μm) of porous three-dimensional on tissue recovery support surface of the present invention.
Fig. 4 is cancellated SEM figure (1 μm) of porous three-dimensional on tissue recovery support surface of the present invention.
Fig. 5 is SEM figure (20 μm) of tissue recovery support cross section of the present invention.
Detailed description of the invention
Tissue recovery support of the present invention has the porous three-dimensional network structure formed by composite fibre layer and hydrophilic material; Described composite fibre layer comprises the composite fibre be made up of adhesion factor and hydrophobic synthetic material.
< composite fibre layer >
Composite fibre layer of the present invention, comprises the composite fibre be made up of adhesion factor and hydrophobic material.
Adhesion factor of the present invention refers to when with tissue bond, tissue recovery support can be made to have good subsides volt performance and formative tissue adhesive force to tissue, comply with anatomical structure, can absorb subsequently, promote the material that surrounding tissue blood coagulation substance formation biological slime is made a concerted effort, the hydrophilic material particularly containing one or more groups in amino, imino group, nitrilo-(secondary amino), amide, polypeptide, ester group, hydroxyl, carboxyl, ether.
The biological adhesive effect power organizing adhesive force to comprise physical force and/or adhesion factor generation of the present invention.The physical force organizing adhesive force to comprise of the present invention is the subsides volt that the depression of materials adsorption tissue surface and space reach good, increase real contact area (compared with the film formed with hydrophobic material, can 60% be greater than), and the good subsides volt property of organization formation and produce and frictional force between tissue.The physical force scope organizing adhesive force to comprise of the present invention is 0.5-10N, and preferred 0.8-8N, more preferably greater than 1N.Its method of testing is: film material shape timbering material is cut into 10mm*60mm strip, surface moisture is blotted with napkin again with after water infiltration, choose 10mm*60mm strip rabbit, the superposition that two strip symmetries, two ends interlocked gently is pressed into the strip of 10mm*70mm, the maximum pull after then making superposition with measurer for pulling force test during rectangular and rabbit generation relative motion.
Biological adhesive effect power is that the various blood coagulating proteins that can absorb very soon in surrounding tissue body fluid due to adhesion factor of the present invention enter internal stent and promote that thrombinogen (prothrombin) is converted into thrombin (thrombin), Fibrinogen (fibrinogen) is converted into fibrin (fibrin), thus acceleration blood coagulation, thus form stronger bioadhesion power with surrounding tissue.Biological cohesive force can be characterized by adhesion experiment: after the anesthesia of white rabbit pentobarbital sodium, depilation, bark fetching prepares 5cm*7cm size wound surface, wound surface is even point-like oozing of blood, being pasted by 4cm*6cm size material lies prostrate in oozing of blood wound surface, adopts 40mmHg exert pressure 5min, connects tonotransducer (JN-IB-100 type) and two road physiographs (LMS-2B), then material is drawn at the uniform velocity vertically upward, calculate biological cohesive force by maximum drawbar pull crest value.Preferred biological cohesive force is greater than 1.0g/cm
2.
In addition, the material such as various albumen, nutrition that adhesion factor of the present invention can also absorb in surrounding tissue body fluid very soon enters internal stent, can accelerate the speed of growing into of collagen fiber, promotes that cambium is grown into fast.Adhesion factor of the present invention has the several functions such as bonding, hemostasis, Promotive union, suppression scar hyperplasia.
Adhesion factor instantiation of the present invention includes but not limited to: have certain hydrophilic protide and derived material, such as collagen protein, fibrin, fibroin, the peptide polymer of elastin mimicry, gelatin; There is certain hydrophilic cellulose family and derived material, such as cellulose, modified cellulose, starch, cellulose; Hydrophilic alcohol, ethers and derived material, the block polyether class such as such as polyvinyl alcohol and Polyethylene Glycol PEG, polyethers F127, polyethers P123; Chitosan class and derived material, such as chitosan, modification of chitosan; Saccharide and corresponding derivant material, such as heparin, glucosan, alginic acid, agar, chondroitin sulfate; Nitrogenous class hydroaropic substance, the such as macromolecular material such as hydrophilic polyurethane, polyvinylpyrrolidone.Wherein, the peptide polymer of chitosan, modification of chitosan, fibrin, fibroin, elastin mimicry, polyvinyl alcohol, polyvinylpyrrolidone, collagen protein and derivant, gelatin is preferably used.
Hydrophobic material of the present invention mainly comprises hydrophobic polyurethane, polyethylene terephthalate, polycaprolactone (PCL), polyglycolic acid (PGA), Poly(D,L-lactide-co-glycolide (PLGA), 1,3-PD polymer (PDO), polylactic acid-caprolactone copolymer (PLC), polylactic acid (PLA) etc.Wherein, preferably PLA, PCL, PGA, PLGA, PLC is used.
Adhesion factor of the present invention and hydrophobic material can be bonded composite fibre in any form.Can be there is inside and/or the surface of described composite fibre in adhesion factor, described composite fibre can be core texture, cross arrangement structure, the structure of one or more combining forms in block structure.These structures can by such as controlling the ratio of adhesion factor and hydrophobic synthetic material, and preparation condition etc. adjust.
The structure of composite fibre of the present invention can adopt SEM to scheme to characterize, and as shown in Figure 1, by contrasting SEM figure and the SEM figure of this composite fibre after the solvent process can dissolving wherein a kind of material of composite fibre, can obviously find out that material has composite construction.Shown in Fig. 1-A be composite fibre of the present invention SEM figure, a kind ofly can dissolve a kind of material in described composite fibre and after the solvent that do not dissolve another kind of material processes when using, the part obtaining composite fibre silk as shown if figure 1-b by dissolving another part not by the cellosilk remnants of defeated troops of dissolving.Fig. 2 is the SEM figure on composite fibre layer surface of the present invention, can obviously see staggered fiber silk structure.
Tissue recovery support hydrophilic fibre provided by the invention and hydrophobic fibre itself have different degradation rates.Can control by regulating the ratio of adhesion factor and hydrophobic synthetic material in surface to organize body fluid to enter the speed of support, and then reach the degradation rate effectively controlling rack surface and inside, prevent the too fast cambium of material degradation from also not covering the defect of formation completely and causing tissue repair failure, the degraded of balancing material simultaneously and the speed of growing into of cambium, degrading can be cambium vacating space faster after material degradation, be beneficial to more cambium substitute creep, slower material of simultaneously degrading can have certain mechanical support at the tissue repair initial stage.
In composite fibre of the present invention, the content of adhesion factor is 5 quality %-95 quality %, is preferably 10%-50%, more preferably 15%-30%.
The surface of composite fibre layer of the present invention, has very low contact angle (θ≤90 ° or θ=0, liquid is water).
The water absorption rate of composite cellulosic membrane of the present invention is greater than 100%, and wherein, water absorption rate calculates in the following way:
Water absorption rate=[(by quality before the quality after the moisture on napkin blotting material surface-diaphragm water suction after support water suction)/diaphragm quality] × 100%
In addition, the hydrophile-lipophile balance value HLB of composite fibre provided by the invention is 1-13, is preferably 1-6.HLB value is the amount of size in surfactant molecule between hydrophilic group and lipophilic group and balance of power degree, is defined as the hydrophile-lipophile balance value of surfactant.The oleophylic of surfactant or hydrophilicrty can differentiate by the size of HLB value, and HLB value is larger, and to represent hydrophilic stronger, and HLB value is less, and to represent lipophile stronger, and generally speaking HLB value is between 1 ~ 40.HLB has important references to be worth in actual applications.Lipophilic surfactant HLB is lower, and hydrophilic surfactant active HLB is higher.Hydrophilic and oleophilic turning point HLB is 10.HLB is less than 10 for lipophile, is greater than 10 for hydrophilic.
Hydrophobic synthetic material contained by tissue recovery support provided by the invention can play certain prevention protein adsorption effect and play the effect prevented with tissue adhesion.After the water suction of described adhesion factor, the swelling support that can make is more soft, increases tissue and pastes volt property, realizes complying with pasting to lie prostrate and the subsides good with organization formation lie prostrate power with tissue anatomical structure.
Composite fibre layer of the present invention can be prepared by the following method: be dissolved in homogeneous solvent by described adhesion factor and described hydrophobic material, then be prepared by one or more combinations in electrostatic spinning technique, centrifugal force spining technology, hot melt spray webbing technology, melting electrospinning, obtain the layer containing described composite fibre.Wherein preferably use electrostatic spinning technique, centrifugal force spining technology, hot melt spray webbing technology or melting electrospinning, more preferably use centrifugal force spining technology or electrostatic spinning technique.
Composite fibre layer of the present invention, can be prepared by centrifugal force spining technology.Hydrophobic synthetic material and adhesion factor can be dissolved in same solvent simultaneously form homogeneous solution, solution enters in centrifugal box-spinning machine by boost pump, adjust the fltting speed of boost pump, centrifugal pan linear velocity and spinning temperature, solution is made to be spun into fiber rapidly, taken off from receptor by film after end, solvent removed by vacuum drying.
Composite fibre layer of the present invention, can also be prepared by electrostatic spinning technique.Hydrophobic material and adhesion factor are dissolved in same solvent simultaneously and form homogeneous solution, solution enters in electrostatic spinning machine by boost pump, the parameter such as fltting speed, transfer roller speed, spinning voltage, receiving range of adjustment boost pump, solution is made to be spun into fiber rapidly, taken off from reception roller by film after end, solvent removed by vacuum drying.
When preparing composite fibre layer of the present invention, the solvent of use is at least containing a kind of fluorine kind solvent, and described fluorine kind solvent comprises hexafluoroisopropanol, trifluoroethanol, trifluoroacetic acid etc.
The composite fibre silk that described composite fibre layer is preferably 10nm ~ 100 μm by diameter is interwoven, and has cavernous structure.
< porous three-dimensional network structure >
Tissue recovery support of the present invention has the porous three-dimensional network structure formed by composite fibre layer and hydrophilic material, on the surface that this porous three-dimensional network structure is formed at described composite fibre layer and/or interior surface.
Described porous three-dimensional network structure by being soaked in the solution of hydrophilic material by the composite fibre layer prepared (film), can being carried out lyophilization after taking-up and being formed.
Can for having certain hydrophilic protide and derived material for the formation of the cancellated hydrophilic material of porous three-dimensional, such as collagen protein, fibrin, fibroin, the peptide polymer of elastin mimicry, gelatin; There is certain hydrophilic cellulose family and derived material, such as cellulose, modified cellulose, starch, cellulose; Hydrophilic alcohol, ethers and derived material, the block polyether class such as such as polyvinyl alcohol and Polyethylene Glycol PEG, polyethers F127, polyethers P123; Chitosan class and derived material, such as chitosan, modification of chitosan; Saccharide and corresponding derivant material, such as heparin, glucosan, alginic acid, agar, chondroitin sulfate; Nitrogenous class hydroaropic substance, the such as macromolecular material such as hydrophilic polyurethane, polyvinylpyrrolidone.Wherein, the peptide polymer of chitosan, modification of chitosan, fibrin, fibroin, elastin mimicry, polyvinyl alcohol, polyvinylpyrrolidone, collagen protein and derivant, gelatin is preferably used.In described solution, there is no particular limitation for the concentration of solute, when concentration is lower, can on the surface of composite fibre layer and/or interior surface formed compared with the porous three-dimensional network structure of macropore, when concentration is higher, can on the surface of composite fibre layer and/or interior surface formed compared with the porous three-dimensional network structure of aperture.The mass percent concentration of described solution is preferably 0.1%-30%, and more preferably the mass percent concentration of described solution is 0.5-10%.
For the solvent in the solution of described hydrophilic material, there is no particular limitation, can be the Conventional solvents that can dissolve above-mentioned hydrophilic material.
Immersion of the present invention comprises conventional immersion process, ultrasonic immersion or other auxiliary immersion process.Lyophilization of the present invention comprises the method adopting the similar devices processing and manufacturings such as vacuum freeze drier, freezing type drier, vacuum rake dryer, refrigerated type air dryer.
< tissue recovery support >
Tissue recovery support of the present invention, by for first to prepare composite fibre layer (film), then can be soaked in the solution of water-soluble material by described composite fibre layer (film), prepares gained after taking-up by freeze drying process.
Tissue recovery support of the present invention there is the porous three-dimensional network structure formed by composite fibre layer and hydrophilic material, or perhaps cellular loose structure, as shown in the SEM figure of Fig. 3 and Fig. 4.The cross section of described tissue recovery support as shown in Figure 5.Aperture adopts GTTTM017-2012 method to detect.
Tissue recovery support of the present invention and the surrounding tissue brute force that rises brokenly after pasting and lying prostrate is more than 0.10kpa, preferred 0.10-30kPa.The described brute force that rises brokenly is measured in the following manner: get rabbit and be cut into the circle that diameter is 3cm, then cut out the hole that diameter is 1cm in centre; Get tissue recovery support of the present invention and be cut into the disk that diameter is 3cm, paste after infiltrating and lie prostrate in rabbit surface, ensure the complete coverage hole surface of tissue recovery support; Then be sealed in pasting the rabbit having lied prostrate support the container finish that pressure transducer is housed, ensure container tightness, then even air-blowing in container, records described tissue recovery support and is by the pressure transducer numerical value backed down the brute force that rises brokenly.In addition, the physical force of tissue recovery support of the present invention and biological adhesive effect power can be measured according to the same way of described composite cellulosic membrane.
Described tissue recovery support has the rate of water absorption be exceedingly fast, and described tissue recovery support is soaked in 10s in water, and its water absorption rate can reach 100%-5000%, preferred 100%-1000%, more preferably 100%-500%.The water absorption rate account form of described tissue recovery support is identical with the water absorption rate account form of described composite cellulosic membrane.
Described tissue recovery support has very low contact angle (θ < 90 ° or θ=0, liquid is water).
Hydrophilic material total content in described tissue recovery support is 5 quality %-95 quality %.
Tissue recovery support of the present invention can be applied to meninges repairing, tissue engineering bracket material, artificial skin, wound dressings, biomembrane, wound clad material, hemostatic material, post-operation adhesion preventing material and beauty treatment material.
Embodiment
The preparation > of < tissue recovery support
Embodiment 1
PCL and collagen protein are dissolved in hexafluoroisopropanol according to mass ratio 4:1 and form the solution that total concentration is 8 quality %, then solution enters in centrifugal box-spinning machine by boost pump, boost pump fltting speed is 150g/ minute, centrifugal pan linear velocity is 6000m/min, solution is spun into fiber rapidly, spinning temperature is 50 DEG C, is taken off by film after terminating from receptor, then removes residual solvent in diaphragm and obtains composite cellulosic membrane (being designated as materials A).The composite cellulosic membrane obtained is soaked in 5min in 2 quality % aqueous gelatin solutions, takes out postlyophilization and prepare tissue recovery support (being designated as material B).
Embodiment 2
PLLA and gelatin are dissolved in hexafluoroisopropanol according to mass ratio 2:1 and form the solution that total concentration is 8 quality %, then above-mentioned solution is loaded in electrostatic spinning syringe, the speed regulating micro-injection pump is 4 mls/hour, the voltage regulating high tension generator is 23kV, and the receiving range regulating receiving system is 19 centimetres.Receiving system uses live-rollers, translational speed 15 cel of electrospinning syringe needle, receive roller rotating speed be 120 circles/point, two pins carry out electrospinning 8 hours simultaneously.After end, film is taken off from reception roller, then remove residual solvent in diaphragm, be soaked in 30min in the aqueous solution of 7 quality % gelatin, take out postlyophilization and prepare tissue recovery support E.
Embodiment 3
PLGA and chondroitin sulfate are dissolved in trifluoroacetic acid according to mass ratio 4:1 and form the solution that total concentration is 8 quality %, then above-mentioned solution is loaded in electrostatic spinning syringe, the speed regulating micro-injection pump is 5 mls/hour, the voltage regulating high tension generator is 24kV, and the receiving range regulating receiving system is 19 centimetres.Receiving system uses live-rollers, translational speed 10 cel of electrospinning syringe needle, receive roller rotating speed be 150 circles/point, two pins carry out electrospinning 8 hours simultaneously.After end, film is taken off from reception roller, then remove residual solvent in diaphragm, be soaked in 10min in the aqueous solution of 2 quality % gelatin, take out postlyophilization and prepare tissue recovery support.
Embodiment 4
PLLA and polyethers F127 is dissolved in hexafluoroisopropanol according to mass ratio 3:1 and forms the solution that total concentration is 8 quality %, then above-mentioned solution is loaded in electrostatic spinning syringe, the speed regulating micro-injection pump is 9 mls/hour, the voltage difference regulating high tension generator is 30kV, and the receiving range regulating receiving system is 23 centimetres.Receiving system uses live-rollers, translational speed 10 cel of electrospinning syringe needle, receive roller rotating speed be 200 circles/point, two pins carry out electrospinning 8 hours simultaneously.After end, film is taken off from reception roller, then remove residual solvent in diaphragm, be soaked in 5min in the aqueous solution of 3 quality % gelatin, take out postlyophilization and prepare tissue recovery support.
Embodiment 5
PLC and PEG is dissolved in hexafluoroisopropanol according to mass ratio 1:2 and forms the solution that total concentration is 8 quality %, then above-mentioned solution is loaded in electrostatic spinning syringe, the speed regulating micro-injection pump is 6 mls/hour, the voltage difference regulating high tension generator is 27kV, and the receiving range regulating receiving system is 17 centimetres.Receiving system uses live-rollers, translational speed 10 cel of electrospinning syringe needle, receive roller rotating speed be 160 circles/point, two pins carry out electrospinning 8 hours simultaneously.After end, film is taken off from reception roller, then remove residual solvent in diaphragm, be soaked in 3min in the aqueous solution of 5 quality % gelatin, take out postlyophilization and prepare tissue recovery support.
Embodiment 6
PLC and hydrophilic polyurethane are dissolved in hexafluoroisopropanol according to mass ratio 1:3 and form the solution that total concentration is 8 quality %, then above-mentioned solution is loaded in electrostatic spinning syringe, the speed regulating micro-injection pump is 6 mls/hour, the voltage difference regulating high tension generator is 26kV, and the receiving range regulating receiving system is 15 centimetres.Receiving system uses live-rollers, translational speed 10 cel of electrospinning syringe needle, receive roller rotating speed be 300 circles/point, two pins carry out electrospinning 8 hours simultaneously.After end, film is taken off from reception roller, then remove residual solvent in diaphragm, be soaked in 3min in the aqueous solution of 6 quality % gelatin, take out postlyophilization and prepare tissue recovery support.
Comparative example 1
PCL is dissolved in hexafluoroisopropanol according to 8 quality % concentration and forms solution, then solution enters in centrifugal box-spinning machine by boost pump, boost pump fltting speed is 150g/ minute, centrifugal pan linear velocity is 6000m/min, solution is spun into fiber rapidly, spinning temperature is 50 DEG C, after end, film is taken off from receptor, then residual solvent in diaphragm is removed, be soaked in 5min in 2 quality % aqueous gelatin solutions, it is inner that water cannot infiltrate diaphragm, takes out postlyophilization and prepare support (being designated as material C).
Comparative example 2
Collagen protein is dissolved in hexafluoroisopropanol according to 8 quality % concentration and forms solution, then solution enters in centrifugal box-spinning machine by boost pump, boost pump fltting speed is 150g/ minute, centrifugal pan linear velocity is 6000m/min, and solution is spun into fiber rapidly, and spinning temperature is 50 DEG C, after end, film is taken off from receptor, then remove residual solvent in diaphragm, be soaked in 15min in 3 quality % aqueous gelatin solutions, take out postlyophilization and prepare support (being designated as material D).
Table 1 individual material properties contrasts
* NA represents that power is too little, cannot detect
method of testing
Physical force: film material shape timbering material is cut into 10mm*60mm strip, surface moisture is blotted with napkin again with after water infiltration, choose 10mm*60mm strip rabbit, the superposition that two strip symmetries, two ends interlocked gently is pressed into the strip of 10mm*70mm, the maximum pull after then making superposition with measurer for pulling force test during rectangular and rabbit generation relative motion.
Biological cohesive force: white rabbit is with after pentobarbital sodium anesthesia, depilation, bark fetching prepares 5cm*7cm size wound surface, wound surface is even point-like oozing of blood, being pasted by 4cm*6cm size material lies prostrate in oozing of blood wound surface, adopts 40mmHg exert pressure 5min, connects tonotransducer (JN-IB-100 type) and two road physiographs (LMS-2B), then material is drawn at the uniform velocity vertically upward, calculate cohesive force by maximum drawbar pull crest value.
Rise brokenly brute force: get rabbit and be cut into the circle that diameter is 3cm, then cut out the hole that diameter is 1cm in centre; Get tissue recovery support of the present invention and be cut into the disk that diameter is 3cm, paste after infiltrating and lie prostrate in rabbit surface, ensure the complete coverage hole surface of tissue recovery support; Then be sealed in pasting the rabbit having lied prostrate support the container finish that pressure transducer is housed, ensure container tightness, then even air-blowing in container, records described tissue recovery support and is by the pressure transducer numerical value backed down the brute force that rises brokenly.
< zoopery >
Experimental example 1
The material C of material B obtained in embodiment 1 and comparative example 1 is cut into 5cm × 5cm, and cleaning, sterilizing, carries out artificial skin zoopery.Experimental rabbit body weight 2-2.5kg, age 6-8 month, male and female were not limit, totally 12.Be divided at random three groups (experimental group, matched groups, blank group), often organize 4.Animal, after auricular vein injection general anesthesia, is placed on special-purpose operating table, standby hair skin of back and preoperative routine disinfection by rabbit.With scalpel by skin of back full-thickness excisional, excision area is 4*4cm, makes whole bark break and damages, repair: experimental group sticks the artificial skin obtained by material B in wound site with artificial skin, and stick in wound surface, pressing 2 minutes, diaphragm pastes and lies prostrate in wound surface.Matched group sticks the artificial skin obtained by material C in wound site, material can not paste and lie prostrate in wound surface, need fix at interval of 1cm suture.Blank is formed face and is not used artificial skin treatment.Experimental group, matched group and blank elements do not cover the aseptic oily yarn of one deck and sterile gauze on wound, fix with suture and surrounding skin.
Post operation observes animal with or without normal feed, drinking-water, body temperature, the time that each group covering is separated with wound surface and other physiological activity situation, and observes the reparation situation of wound surface.Postoperative 4th, 8 week often group get 2 rabbit euthanasia, get the whole wound surface in back and contiguous normal skin, formalin is fixed, carry out HE dyeing, the newborn situation organized in light Microscopic observation corium, inflammatory reaction situation, the thickness of epidermal structure and the regeneration situation of appendages.
Blank group rabbit after surgery all wound surface all with oily yarn or gauze adhesion.Adjuvant has obscission.Wound surface is hemorrhage, has a large amount of tissue fluid to ooze out.Linearly crust after wound healing.
The out-of-flatness of matched group wound surface artificial skin, limit and wound surface not docile, and there is the phenomenon tilting and come off; There is slight red and swollen heat in wound, there is mild infection phenomenon, have small amount tissue fluid to ooze out, wound and gauze have a small amount of adhesion.In a small amount of rectangle crust and relatively large linear crust after wound healing.
Experimental group postoperative wound surface is dry, fits closely with artificial skin, with oily yarn and gauze without adhesion.Wound is without red and swollen warm, and animal does not occur skin allergy or infects phenomenon.Material covers lower to necrosis phenomena.Rectangular crust after wound healing.Thickened by microscopic examination healing mouth skin prickle cell layer, basal cell hyperplasia enlivens, and the horny layer of epidermal area, granular layer, stratum spinosum and basal layer are all visible.The structures such as the attached organ pipe of skin corium, sweat gland, hair follicle are visible, and visible a large amount of fibroblast is parallel to skin surface arrangement, more blood capillary proliferation therebetween, a small amount of lymphocytic infiltration in local.
Observed result shows: the subsides of experimental group volt ability, anti-infection ability, the effect preventing wound surface hemorrhage are better than matched group and blank group.Experimental group wound surface has no red and swollen, downright bad.Histological observation display has matched group by the artificial skin material that the material B of embodiment 1 is obtained to wound surface and lies prostrate performance with the unexistent subsides of blank group, more can effectively promote that skin texture regenerates.
Claims (20)
1. a tissue recovery support, is characterized in that, described tissue recovery support comprises the porous three-dimensional network structure formed by composite fibre layer and hydrophilic material; Described composite fibre layer comprises the composite fibre be made up of adhesion factor and hydrophobic synthetic material.
2. tissue recovery support according to claim 1, wherein, on the surface that described porous three-dimensional network structure can be formed at described composite fibre layer and/or interior surface.
3. tissue recovery support according to claim 1 and 2, wherein, described porous three-dimensional network structure, by being immersed in the solution of hydrophilic material by described composite fibre layer, being taken out postlyophilization and is formed.
4. tissue recovery support according to claim 3, wherein, the solution of described hydrophilic material is the aqueous solution of water soluble polymer.
5. the tissue recovery support according to any one of claim 1-4, wherein, described hydrophilic material is selected from the group be made up of hydrophilic protide, cellulose family, alcohols, ethers, chitosan class, saccharide, nitrogenous class hydroaropic substance.
6. the tissue recovery support according to any one of claim 1-5, wherein, the described adhesion factor forming described composite fibre can be present in inside and/or the surface of described composite fibre.
7. the tissue recovery support according to any one of claim 1-6, wherein, described adhesion factor be selected from there is hydrophilic protide and derived material thereof, there is hydrophilic cellulose family and derived material thereof, there is hydrophilic alcohols and derived material thereof, chitosan class and derived material, saccharide and derived material thereof, one or more in nitrogenous class hydroaropic substance.
8. the tissue recovery support according to any one of claim 1-7, wherein, described hydrophobic synthetic material comprises hydrophobic polyurethane, polyethylene terephthalate, polycaprolactone, polyglycolic acid, Poly(D,L-lactide-co-glycolide, 1,3-PD polymer, polylactic acid-caprolactone copolymers, polylactic acid.
9. the tissue recovery support according to any one of claim 1-8, wherein based on the gross weight of described composite fibre layer, the content of described adhesion factor is 5 quality %-95 quality %, is preferably 10 quality %-50 quality %, more preferably 15 quality %-30 quality %.
10. the tissue recovery support according to any one of claim 1-9, is soaked in 10s in water by described tissue recovery support, and its water absorption rate can reach 100-5000%.
11. tissue recovery supports according to any one of claim 1-10, wherein, the hot strength of described tissue recovery support is greater than 1Mpa.
12. tissue recovery supports according to any one of claim 1-11, wherein, the hydrophile-lipophile balance value HLB of described tissue recovery support is 1-13.
13. tissue recovery supports according to any one of claim 1-12, wherein, the surface of described tissue recovery support and contact angle θ≤90 ° of water.
14. tissue recovery supports according to any one of claim 1-13, wherein, described tissue recovery support and the surrounding tissue brute force that rises brokenly after pasting and lying prostrate is more than 0.10kPa.
15. tissue recovery supports according to any one of claim 1-14, wherein, the cellosilk that described composite fibre layer is 10nm-100 μm by diameter is interwoven, and has cavernous structure.
16. tissue recovery supports according to any one of claim 1-15, wherein, the aperture of described composite fibre layer support is 1-100 μm, and the aperture of described tissue recovery support is 1.3-500 μm.
The purposes of the tissue recovery support described in 17. any one of claim 1-16, described tissue recovery support is used for meninges repairing, spinal meninges repairing, tissue engineering bracket material, artificial skin, wound dressings, biomembrane, wound clad material, hemostatic material, post-operation adhesion preventing material or beauty treatment material.
The preparation method of the tissue recovery support described in 18. any one of claim 1-16, it comprises:
A () prepares the step of composite fibre layer: be dissolved in homogeneous solvent by described adhesion factor and described hydrophobic synthetic material, then be prepared by one or more combinations in electrostatic spinning technique, centrifugal force spining technology, hot melt spray webbing technology, melting electrospinning, obtain the diaphragm containing composite fibre;
B () prepares the cancellated step of porous three-dimensional: be soaked in the solution of hydrophilic material by the diaphragm containing composite fibre described in preparing, carry out lyophilization after taking-up.
19. preparation methoies according to claim 18, wherein, the described solvent used in the step of composite fibre layer of preparing at least comprises a kind of fluorine kind solvent.
20. preparation methoies according to claim 19, wherein, described fluorine kind solvent comprises hexafluoroisopropanol, trifluoroethanol, trifluoroacetic acid.
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| CN201410238853.9A CN105194737B (en) | 2014-05-30 | 2014-05-30 | A kind of tissue recovery support and its preparation method and application |
| EP14818651.3A EP3015120B1 (en) | 2013-06-28 | 2014-06-26 | Tissue repair scaffold and preparation method and purpose thereof |
| PCT/CN2014/080798 WO2014206308A1 (en) | 2013-06-28 | 2014-06-26 | Tissue repair scaffold and preparation method and purpose thereof |
| CN201480037229.0A CN105339018B (en) | 2013-06-28 | 2014-06-26 | Tissue recovery support and its production and use |
| US14/392,245 US20160175487A1 (en) | 2013-06-28 | 2014-06-26 | Tissue repair scaffold and preparation method and purpose thereof |
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