Two, background technology:
Tissues such as people's skeleton because of disease or wound etc. causes tissue defect, need artificial material to repair or rescue.This class repair materials comprises metal, inorganic material, macromolecular material etc.
The bone renovating material intensity height that rustless steel or titanium alloy are made, cost is lower, and frequency of utilization is very high, but stress shielding may occur, may cause bone resorption, and knitting is delayed, and histocompatibility is not so good, also can cause inflammatory reaction etc.
Hydroxyapatite, tricalcium phosphate are the bioceramices of using always, and the calcium phosphate bone cement also is usually used in hard tissue repair, and it has better biocompatibility, and hydroxyapatite also has the induced osteogenesis effect, but quality is too crisp.Someone is compound with bone morphogenetic protein (BMP) and hydroxyapatite, and this material can be accelerated knitting, but fragility is big, and this class material biodegradation rate is slower.
In macromolecular material, the composite bone cement of being made up of methyl methacrylate etc. also is to use very wide hard tissue repairing material at present.Bone cement has better biocompatibility, but fragility is big, the hardness height, and material is not firm with combining of body tissue, can not be absorbed by body.
Another kind of hard tissue repairing material is to absorb macromolecular material, mainly is the polyesters macromolecule.The granular bone repair materials that absorbable polyester family macromolecule material is made is applied clinically.In order to accelerate knitting, report also arranged with skeletal growth factor and macromolecular material are compound.The result shows that bone morphogenetic protein (BMP) can induce bone synthetic with polylactic acid-based high molecular composite.Because macromolecular material is progressively degraded in body and is produced acid micromolecular compound, the non-bacterial inflammatory reaction may occur.
In order to improve mechanical property that can absorb hard tissue repairing material etc., there is the people that hydroxyapatite (HAP) or tricalcium phosphate are mixed with the polyesters macromolecular material, this material can degraded and absorbed.In this inorganic organic polymer composite, add bone morphogenetic protein (BMP) and can accelerate the differentiation and the growth of osteocyte, induce bone synthetic, accelerate knitting.The processing technique more complicated of present this class tissue renovation material.
Can absorb polymer fiber material as reinforcing material in order to improve the intensity of bone renovating material, can to adopt, make the bone renovating material with higher-strength, what adopt usually is the thicker fibrous material of diameter.
In addition, a kind of guiding bone renovating material is with the tricalcium phosphate particle and can absorbs the compound porous film material of making of macromolecular material, the bridge formation formula covers position to be repaired, bone stump is filled with absorbable material, the guiding osteocyte is grown along coverlay, but the pore structure of the porous film material that methods such as lyophilization make is difficult to control.So it is simple to be necessary to develop a class processing technique, material hole size and cell are close or littler, and the fibrous material diameter is thinner, is easy to degraded and absorbed, as required the tissue renovation material of designing material composition, shape, structure.
Three, summary of the invention:
1, technical problem
The purpose of this invention is to provide a kind of bioresorbable good, can carry out cutting, reliable in structure according to the physiological structure of defect, simple tissue renovation material of processing technique and preparation method thereof.
2, technical scheme
Absorbable superfine fibre tissue remedial materials of the present invention, this material comprise the synthos ultrafine particle and can absorb the high molecular superfine fiber material that high molecular superfine fiber is 1: 2~9: 1 with the ratio of the quality of synthos ultrafine particle.Because fiber is piled up the loose structure of formation rule regularly.The synthos ultrafine particle is sintering or unsintered hydroxyapatite (Ca
10(PO
4)
6(OH)
2), tricalcium phosphate (Ca
3(PO
4)
2), calcium pyrophosphate (Ca
2P
2O
7),, calcium hydrogen phosphate (CaHPO
4), dicalcium phosphate dihydrate (CaHPO
4.2H
2The mixture of one or more synthos O), its granularity is less than 5 microns.
The high molecular superfine fiber material is can absorb synthetic high polymer maybe can absorb natural macromolecular ultrafine fibrously, and fibre diameter is between 20 nanometers to 3 micron.Can absorb synthetic high polymer is poly--L-lactic acid, poly--(D, L)-and a kind of or following at least two kinds of monomeric copolymers in lactic acid, polyglycolic acid, polycaprolactone, poly-butyrolactone, poly-valerolactone, poly-anhydride, poly--a-amino acid: L-lactic acid, D, L-lactic acid, hydroxyacetic acid, caprolactone, butyrolactone, valerolactone, aminoacid, or two or more blend wherein, high molecular weight average molecular weight is 10,000-1,500,000.Can absorb natural polymer and be a kind of or their derivant or two or more blend wherein in chitosan, chitosan derivatives, hyaluronic acid, derivatives of hyaluronic acids, chondroitin sulfate, collagen, carrageenin, alginate, gelatin, glucosan, fibroin, keratoprotein, the albumin.
Absorbable superfine fibre tissue remedial materials can also contain one or more following trace mineral supplements: xenogenesis bone morphogenetic protein or people's gene reorganization bone morphogenetic protein (BMP), BMP-1, BMP-2, BMP-3, BMP-4, BMP-5, BMP-6, BMP-7, BMP-8, growth/differentiation factor GDF-5, GDF-6, GDF-7, or transforming growth factor-beta (TGF-β), insulin like growth factor, platelet derived growth factor, fibroblast growth factor, antibiotic, immunosuppressant, antibacterial, hormone, vitamin, aminoacid, polypeptide, protein, enzyme, the gross mass of additive is less than 5% of the repair materials gross mass.
The preparation method of absorbable superfine fibre tissue remedial materials is: one or more macromolecular materials are disperseed or be dissolved in and become clear solution or mixture in the solvent, synthos particle, additive etc. are dispersed in the above-mentioned solution, obtain the electrospinning mixture, add in the storage tank then, be connected to one or more injector heads by syringe pump and conduit, what be connected with one or more injector heads can be single electrospinning mixture or multiple electrospinning mixture.Injector head connects high voltage electric field, and voltage is 1 kilovolt~120 kilovolts.Accept partial earthing, in electric field, carry out electrostatic spinning or EFI according to pre-designed shape then, obtain the high molecular superfine fiber material of phosphoric acid calcium salt particle, additive, through post-treatment operations such as cutting or heat treatments, or mix with additive, obtain the tissue renovation material of membranaceous, block, tubulose, column or other different shape.High voltage electric field can be high pressure positive electric field, high pressure negative electric field, high-voltage alternating electric field.
3, beneficial effect
(1) the present invention compared with prior art has following advantage:
(2) tissue renovation material provided by the invention is the composite of membranaceous, block, the tubulose, column or other shape that contain the high molecular superfine fiber of ultrafine particle.
(3) tissue renovation material provided by the invention has the simple feature of processing technique.Adopt macromolecule to mix electrospinning technology with the calcium phosphate particle and obtain the superfine fibre composite, through cutting, machining such as laminate, obtain tissue renovation material, the clinical reparation that is used for tissue defect can obtain good effect.
(4) the present invention can absorb macromolecule, synthos mix electrospinning and post-treatment makes porous organization's repair materials, have higher intensity and permeability.
(5) the present invention is dispersed in active additive in the solvent and carries out electrospinning with macromolecule, guarantees that material is not subjected to the effect of extreme conditions such as High Temperature High Pressure, and the additive activity is unaffected.
(6) the present invention's super-fine fiber material that macromolecule, synthos etc. is made by mould or machining or heat treatment, can make the repair materials that adapts with the tissue physiology anatomical structure.
Four, the specific embodiment
Absorbable superfine fibre tissue remedial materials of the present invention comprises the synthos ultrafine particle and can absorb the high molecular superfine fiber material that high molecular superfine fiber is 1: 2~9: 1 with the ratio of the quality of synthos ultrafine particle.Because fiber is piled up the loose structure of formation rule regularly.The synthos ultrafine particle is sintering or unsintered hydroxyapatite (Ca
10(PO
4)
6(OH)
2), tricalcium phosphate (Ca
3(PO
4)
2), calcium pyrophosphate (Ca
2P
2O
7),, calcium hydrogen phosphate (CaHPO
4), dicalcium phosphate dihydrate (CaHPO
4.2H
2The mixture of one or more synthos O), its granularity is less than 5 microns.
The high molecular superfine fiber material is can absorb synthetic high polymer maybe can absorb natural macromolecular ultrafine fibrously, and fibre diameter is between 20 nanometers to 3 micron.Can absorb synthetic high polymer is poly--L-lactic acid, poly--(D, L)-and a kind of or following at least two kinds of monomeric copolymers in lactic acid, polyglycolic acid, polycaprolactone, poly-butyrolactone, poly-valerolactone, poly-anhydride, poly--a-amino acid: L-lactic acid, D, L-lactic acid, hydroxyacetic acid, caprolactone, butyrolactone, valerolactone, aminoacid, or two or more blend wherein, high molecular weight average molecular weight is 10,000-1,500,000.Can absorb natural polymer and be a kind of or their derivant or two or more blend wherein in chitosan, chitosan derivatives, hyaluronic acid, derivatives of hyaluronic acids, chondroitin sulfate, collagen, carrageenin, alginate, gelatin, glucosan, fibroin, keratoprotein, the albumin.
Absorbable superfine fibre tissue remedial materials can also contain one or more following trace mineral supplements: xenogenesis bone morphogenetic protein or people's gene reorganization bone morphogenetic protein (BMP), BMP-1, BMP-2, BMP-3, BMP-4, BMP-5, BMP-6, BMP-7, BMP-8, growth/differentiation factor GDF-5, GDF-6, GDF-7, or transforming growth factor-beta (TGF-β), insulin like growth factor, platelet derived growth factor, fibroblast growth factor, antibiotic, immunosuppressant, antibacterial, hormone, vitamin, aminoacid, polypeptide, protein, enzyme, the gross mass of additive is less than 5% of the repair materials gross mass.
The preparation method of absorbable superfine fibre tissue remedial materials is: one or more macromolecular materials are disperseed or be dissolved in and become clear solution or mixture in the solvent, synthos particle, additive etc. are dispersed in the above-mentioned solution, obtain the electrospinning mixture, add in the storage tank then, be connected to one or more injector heads by syringe pump and conduit, what be connected with one or more injector heads can be single electrospinning mixture or multiple electrospinning mixture.Injector head connects high voltage electric field, and voltage is 1 kilovolt~120 kilovolts.Accept partial earthing, in electric field, carry out electrostatic spinning or EFI according to pre-designed shape then, obtain the high molecular superfine fiber material of phosphoric acid calcium salt particle, additive, through post-treatment operations such as cutting or heat treatments, or mix with additive, obtain the tissue renovation material of membranaceous, block, tubulose, column or other different shape.High voltage electric field can be high pressure positive electric field, high pressure negative electric field, high-voltage alternating electric field.
Tissue renovation material proposed by the invention can be used for various tissue defects reparation, substitute, the bone that causes as wound is damaged, it is damaged that infection, deformity, tumor etc. cause.The hard tissue repairing material of suitable shape and size can be used for the hard tissue repair of operations such as plastic surgery, oral surgery, neurosurgery, craniomaxillofacial surgery.Material of the present invention also is a tissue engineering bracket material.
Embodiment 1
The preparation of hard tissue repairing material: can absorb synthesized polymer material and can adopt poly--L-lactic acid, poly--(D, L)-and a kind of or following at least two kinds of monomeric copolymers in lactic acid, polyglycolic acid, polycaprolactone, poly-butyrolactone, the poly-valerolactone: L-lactic acid, (D, L)-lactic acid, hydroxyacetic acid, caprolactone, butyrolactone, valerolactone, or their blend.
For example, 1.5 grammes per square metre average molecular weights are about 130,000 poly (l-lactic acid) and are dissolved in the 10ml acetone, be made into the macromolecular solution of 10% (mass fraction), stirred 24 hours, agglomerating tricalcium phosphate (Ca
3(PO
4)
2) the 0.2g powder body adds in the 5ml acetone, ultra-sonic dispersion mixed with above-mentioned macromolecular solution after 20 minutes, add in the storage tank then, be connected to injector head by syringe pump and conduit, injector head connects the high pressure positive electric field, and distance is controlled at about 5cm solution flow rate 6ml/h between injector head and reception thing, voltage is 10 kilovolts, carries out electrospinning.Obtain fiber film material,, obtain multi-layer film material after cutting is superimposed through vacuum drying.0.5 gram multi-layer film material was soaked 15 minutes in containing 10 milliliters of buffer solns of 1 milligram of bone morphogenetic protein(BMP), obtain active bone repair material.Embodiment 2
The preparation of hard tissue repairing material: can adopt the synthetic hydroxyapatite (Ca of low temperature without agglomerating synthos
10(PO
4)
6(OH)
2), tricalcium phosphate (Ca
3(PO
4)
2), calcium pyrophosphate (Ca
2P
2O
7),, calcium hydrogen phosphate (CaHPO
4), dicalcium phosphate dihydrate (CaHPO
4.2H
2O) a kind of in or the mixture of two or more synthos wherein.
For example, be that 130,000 poly (l-lactic acid)s are dissolved in the 5ml acetone with a certain amount of weight average molecular weight, be made into the macromolecular solution of 10% (mass fraction), will be without agglomerating tricalcium phosphate (Ca
3(PO
4)
2) 0.2g adds in the 5ml acetone, ultra-sonic dispersion mixed with above-mentioned macromolecular solution after 20 minutes, add in the storage tank then, be connected to injector head by syringe pump and conduit, injector head connects the high pressure positive electric field, and distance is controlled at about 5cm solution flow rate 6ml/h between injector head and reception thing, voltage is about 10kv, carries out electrospinning.Through vacuum drying, promptly obtain the porous hard tissue renovation material behind the oxirane disinfection.
Embodiment 3
With 1 grammes per square metre average molecular weight is that 130,000 poly-(L-lactic acid-co-D, L-lactic acid) are dissolved in the 10ml acetone, is made into the macromolecular solution of 10% (mass fraction), will be without agglomerating tricalcium phosphate powder body (Ca
3(PO
4)
2) 0.2g adds in the 5ml acetone, ultra-sonic dispersion mixed with above-mentioned macromolecular solution after 20 minutes, add in the storage tank then, be connected to injector head by syringe pump and conduit, injector head connects the high pressure positive electric field, and distance is controlled at about 5cm solution flow rate 6ml/h between injector head and reception thing, voltage is about 10kv, carries out electrospinning.Through vacuum drying, promptly obtain the porous hard tissue renovation material behind the oxirane disinfection.
Embodiment 4
With 1 grammes per square metre average molecular weight is that 130,000 poly-(L-lactic acid-co-D, L-lactic acid) are dissolved in the 10ml acetone, is made into the macromolecular solution of 10% (mass fraction), will be without agglomerating hydroxyapatite (Ca
10(PO
4)
6(OH)
2) 0.2g adds in the 5ml acetone, ultra-sonic dispersion mixed with above-mentioned macromolecular solution after 20 minutes, add in the storage tank then, be connected to 3 injector heads by syringe pump and conduit, injector head connects the high pressure positive electric field, and distance is controlled at about 5cm between injector head and reception thing, solution flow rate 6ml/h, voltage is about 10kv, carries out electrospinning, obtains the tubular tissue repair materials after vacuum drying, coiling, cutting.
Embodiment 5
With 1 grammes per square metre average molecular weight is that 150,000 polycaprolactones are dissolved in the 10ml oxolane, is made into the macromolecular solution of 10% (mass fraction), will be without agglomerating tricalcium phosphate powder body (Ca
3(PO
4)
2) 0.2g adds in the 1ml oxolane, mix with above-mentioned macromolecular solution, add in the storage tank then, be connected to injector head by syringe pump and conduit, injector head connects the high pressure negative electric field, and distance is controlled at about 5cm solution flow rate 6ml/h between injector head and reception thing, voltage is about 10kv, carries out electrospinning.Through vacuum drying, promptly obtain the porous hard tissue renovation material behind the oxirane disinfection.
Embodiment 6
With 1 grammes per square metre average molecular weight is that 130,000 poly (l-lactic acid)s are dissolved in the 5ml acetone, is made into the macromolecular solution of 10% (mass fraction), will be without agglomerating tricalcium phosphate (Ca
3(PO
4)
2) 0.2g adds in the 5ml acetone, ultra-sonic dispersion mixed with above-mentioned macromolecular solution after 20 minutes, add in the storage tank then, be connected to injector head by syringe pump and conduit, injector head connects the high pressure negative electric field, and distance is controlled at about 5cm solution flow rate 6ml/h between injector head and reception thing, voltage is about 10kv, carries out electrospinning.The fibrous membrane that obtains obtains multi-layer film material, oxirane disinfection after vacuum drying, superimposed, cutting.Multi-layer film material was soaked in bone marrow 15 minutes, obtain active bone repair material.
Embodiment 7
With 1 grammes per square metre average molecular weight is that 130,000 poly (l-lactic acid)s are dissolved in the 5ml acetone, is made into the macromolecular solution of 10% (mass fraction), will be without agglomerating tricalcium phosphate (Ca
3(PO
4)
2) 0.2g adds in the 5ml acetone, ultra-sonic dispersion mixed with above-mentioned macromolecular solution after 20 minutes, add in the storage tank then, be connected to injector head by syringe pump and conduit, injector head connects the high pressure negative electric field, and distance is controlled at about 10cm solution flow rate 6ml/h between injector head and reception thing, voltage is about 10kv, carries out electrospinning.The fibrous membrane that obtains obtains multi-layer film material, oxirane disinfection through vacuum drying after cutting is superimposed.Multi-layer film material was soaked 15 minutes in the big enzyme cellulose solution of celebrating, obtain bone renovating material.
Embodiment 8
With 0.5 grammes per square metre average molecular weight is that the polyoxyethylene of 130,000 poly (l-lactic acid)s and 0.5 gram-molecular weight 100,000 is dissolved in the 10ml dimethyl formamide, is made into the macromolecular solution of 10% (mass fraction), will be without agglomerating tricalcium phosphate powder body (Ca
3(PO
4)
2) 0.2g adds in the 2ml dimethyl formamide, ultra-sonic dispersion mixed with above-mentioned macromolecular solution after 20 minutes, add in the storage tank then, be connected to injector head by syringe pump and conduit, injector head connects the high pressure negative electric field, and distance is controlled at about 15cm solution flow rate 6ml/h between injector head and reception thing, voltage is about 13kv, carries out electrospinning.The fibrous membrane that obtains obtains multi-layer film material through vacuum drying after cutting is superimposed.
Embodiment 9
With being dissolved in the 5ml hexafluoroisopropanol of 0.2 gram bovine collagen, be made into the macromolecular solution of 4% (mass fraction), will be without agglomerating tricalcium phosphate powder body (Ca
3(PO
4)
2) 0.05g adds in the above-mentioned solution, ultra-sonic dispersion 20 minutes adds in the storage tank then, be connected to injector head by syringe pump and conduit, injector head connects the high pressure negative electric field, and distance is controlled at about 5cm between injector head and reception thing, solution flow rate 6ml/h, voltage are about 10kv, carry out electrospinning.The fibrous membrane that obtains obtains multi-layer film material through vacuum drying after cutting is superimposed.
Embodiment 10
With 5 grammes per square metre average molecular weights is that 130,000 poly (l-lactic acid)s are dissolved in the 45ml acetone, is made into the macromolecular solution of 10% (mass fraction), will be without agglomerating tricalcium phosphate (Ca
3(PO
4)
2) lg adds in the 5ml acetone, ultra-sonic dispersion mixed with above-mentioned macromolecular solution after 20 minutes, add in the storage tank then, be connected to 6 injector heads by syringe pump and conduit, injector head connects the high pressure negative electric field, and distance is controlled at about 15cm solution flow rate 18ml/h between injector head and reception thing, voltage is about 13kv, carries out electrospinning.The fibrous membrane that obtains is through vacuum drying, and cutting is superimposed, through hot pressing, obtains bulk material.
Embodiment 11
With 1 grammes per square metre average molecular weight is that 130,000 poly (l-lactic acid)s are dissolved in the 10ml acetone, is made into the macromolecular solution of 10% (mass fraction), will be without agglomerating tricalcium phosphate (Ca
3(PO
4)
2) lg adds in the solution, ultra-sonic dispersion adds syringe after 20 minutes then, is connected to an injector head by conduit, injector head connects the high pressure negative electric field, injector head with receive between thing apart from being controlled at about 15cm, solution flow rate 6ml/h, voltage are about 12kv.With being dissolved in the 5ml hexafluoroisopropanol of 0.2 gram bovine collagen, be made into the macromolecular solution of 4% (mass fraction), will be without agglomerating tricalcium phosphate powder body (Ca
3(PO
4)
2) 0.05g adds in the above-mentioned solution, ultra-sonic dispersion 20 minutes adds in the syringe then, be connected to another injector head by conduit, injector head connects the high pressure negative electric field, and distance is controlled at about 15cm between injector head and reception thing, solution flow rate 6ml/h, voltage are about 15kv.Carry out common-battery together and spin, obtain composite cellulosic membrane.Through vacuum drying, obtain multi-layer film material after cutting is superimposed.