A kind of electroactive nano fiber scaffold material and preparation method thereofTechnical field
The present invention relates to tissue regeneration and material for repairing and preparation method thereof, particularly to a kind of electricityActive nano fiber scaffold material and preparation method thereof.
Background technology
Artificial bone repair materials is owing to wide material sources, performance are controlled and low cost, at bone renovating materialIn occupy critical role.But, presently used artificial material still can not meet clinical demand,Its main cause is that artificial material can not be as biological tissue, with physiological stress or biochemical stimulationMake respective reaction, the most do not possess biological reactivity, make permanent replacement of human body hard tissue defect repairMultiple theory receives challenge greatly.Therefore, change permanent replacement and repair idea, give full play to machineBody self-regeneration potentiality, exploitation there is biological activity and biological reactivity material to become current human body hardThe Main Trends of The Development of tissue renovation material.
Osseous tissue forms the electricity environment relied on, and the electroactive Bionic Design for bone renovating material carriesSupply important evidence.Electroactive for making embedded material have, researcher is by conventional piezoelectric pottery metatitanic acidBarium (BaTiO3) or potassium-sodium niobate (KNN) etc. be applied to technical field of biological material.But simple pottery materialThere is the high shortcoming being difficult to processing of fragility in material, thus is usually combined into macromolecule polymer materialType is used for Bone Defect Repari.
Such as, the Chinese invention patent of Publication No. CN 104208754A discloses a kind of piezoelectricity workProperty bone-repairing composite material and preparation method thereof, wherein, material is by the pottery with nucleocapsid structureParticulate filler and polymeric matrix are constituted, and the ceramic particle uniform filling with nucleocapsid structure is dispersed in instituteState in polymeric matrix；The nucleome of the ceramic particle filler with nucleocapsid structure is ceramic particle, potteryThe Surface coating of porcelain granule has Coated with Organic Matter layer.
This composite forming method maintains good electrology characteristic, pliability and clinical operability.But in terms of material structure, it is fine and close real wall-shaped material, maintain cell growth microenvironment withAnd the three-dimensional net structure feature of the aspect such as nutrition transmission and natural extracellular matrix is not quite similar, byThe Bone Defect Repari effect that this causes also remains to be further improved.
Summary of the invention
The present invention is contemplated to solve that current material structure is to be improved, Bone Defect Repari effect needs into oneWalk perfect technical problem, it is provided that a kind of electroactive nano fiber scaffold with three-dimensional net structureMaterial and preparation method thereof.
To this end, the present invention provides a kind of electroactive nano fiber scaffold material, material is by surface modificationCeramic particle filler and Poly-L-lactic acid, Poly(D,L-lactide-co-glycolide, polycaprolactone inOne or more polymeric matrixs are constituted, ceramic particle filling surface one layer of lemon of cladding of surface modificationLemon acid sodium, electroactive nano fiber scaffold material is three-dimensional net structure.
Preferably, the material of ceramic particle filler be Barium metatitanate., in barium strontium titanate and potassium-sodium niobateOne or more.
Preferably, electroactive nano fiber scaffold material is micro/nano fibrous membrane material, wherein said electricityActive nano fibre diameter is 100nm～600nm, and the thickness of fibrous membrane is 50 μm～500 μm,A diameter of 50nm～200nm of ceramic particle filler.
Preferably, in electroactive nano fiber scaffold material, the ceramic particle filler of surface modification accounts forThe mass percent of electroactive nano fiber scaffold material is 1wt.%～20wt.%, and remaining is polymerizationThing matrix.
Meanwhile, the present invention provides the preparation method of a kind of electroactive nano fiber scaffold material, surfaceThe preparation process of modified ceramic particle filler uses aqua-solution method to complete；Polymeric matrix and surfaceThe composite molding process of modified ceramic particle filler is completed by method of electrostatic spinning.
Preferably, the preparation process of the ceramic particle filler of surface modification includes that sodium citrate solution is joinedSystem, nano-particle dispersion in sodium citrate solution include that ultrasonic agitation process, nano-particle are outstandingThe mixing of supernatant liquid and polymer solution and the electrostatic spinning process of mixed liquor.
Preferably, the method that the present invention provides, comprise the steps: that (1) weighs sodium citrate and putsIn 2ml organic solvent trifluoroethanol, it is configured to the sodium citrate solution that concentration is 0.02g/mL,Take ceramic particle to be placed in sodium citrate solution, carry out at sonic oscillation, stirring at ambient temperatureManage each 30min, alternately 3～5 times so that the ceramic particle filler of surface modification is in a solventForm homodisperse suspension；(2) weigh 1g polymer, add 8mL organic solvent trifluoroEthanol, stirring 3h～8h makes it be completely dissolved, obtains polymer solution；(3) by step (1)Middle gained suspension adds in step (2) resulting polymers solution, and stirring 6h～12h makes surfaceModified ceramic particle uniform filling is dispersed in polymeric matrix, obtains mixed liquor；(4) will stepSuddenly, in (3) gained mixed liquor injects injector for medical purpose, syringe is placed in electrostatic spinning apparatus, quietThe device that receives of Electrospun is the one in flat board, copper mesh or roller, and electrostatic potential is 10kV～20KV, spinning flow velocity is 0.2mL/h～1.0mL/h, syringe needle and the distance received between device be 10cm～30cm, makes solvent volatilize completely in gained nano fibrous membrane is placed in vacuum drying oven, obtains electricity and livesProperty nano fiber scaffold material.
The invention have the benefit that
(1) present invention is by carrying out surface modification to nano-ceramic particle, effectively reduces nanometerThe reunion of grain, improves the interface between ceramic particle and polymer, it is achieved that ceramic particle is poly-Dispersed in compound matrix, improves the electric property of composite, is made up of nanofiberNano fiber scaffold material maintain good three-dimensional net structure characteristic.
(2) timbering material obtained by the present invention and skeletonization relevant cell have good affinity,Osteoblastic adhesion and propagation can be promoted；After et al. Ke, visible material has good tissue phaseCapacitive.
(3) electroactive nano fiber scaffold material provided by the present invention is tied at macro property and microcosmicGood performance it is respectively provided with, during Bone Defect Repari, for adhesion, the life of skeletonization relevant cell on structureLong and differentiation provides microenvironment preferably；Additionally, this preparation method can make bone renovating material haveThere is good biological reactivity, contribute to improving Bone Defect Repari effect further.
Accompanying drawing explanation
Fig. 1 is that the scanning electron microscope of nano fiber scaffold material described in the embodiment of the present invention 1 is shoneSheet；
Fig. 2 is that barium titanate nano ceramic particle described in the embodiment of the present invention 1 divides in PLLA fiberThe transmission electron microscope photo of cloth；
Fig. 3 is the X-ray diffractogram of nano fiber scaffold material described in the embodiment of the present invention 1；
Fig. 4 is the electric property figure (nanometer of nano fiber scaffold material described in the embodiment of the present invention 1The dielectric constant of fiber scaffold material is 1.15 ± 0.02)；
Fig. 5 is the external osteoblasts cultivation 1 of nano fiber scaffold material described in the embodiment of the present invention 1It electron scanning micrograph；
Fig. 6 is the internal reparation defect of skull 4 of nano fiber scaffold material described in the embodiment of the present invention 1The light microscopic photo of all tissue slices；
Fig. 7 is the morphology observation after mesenchymal stem cells MSCs growth on two kinds of membrane materials 5 days(A is BTO/P (VDF-TrFE) composite casting film；B is BTO/PLLA composite nano fiberFilm)；
Fig. 8 is mesenchymal stem cells MSCs proliferation activity detection on two kinds of membrane materials(*p<0.05)。
Detailed description of the invention
The invention provides a kind of electroactive nano fiber scaffold material and preparation method thereof, below knotThe present invention will be further described to close the drawings and specific embodiments.
Embodiment 1
(1) weighing sodium citrate to be placed in 2ml organic solvent trifluoroethanol, being configured to concentration is 0.02The sodium citrate solution of g/mL, takes 0.07g BaTiO3Ceramic particle is placed in sodium citrate solution,Carry out sonic oscillation, each 30min of stir process at ambient temperature, alternately 3 times so that tableThe BaTiO that face is modified3Ceramic particle filler (BNP@sodium citrate) is formed in a solvent and uniformly dividesThe suspension dissipated；
(2) weigh 1g polymer P LLA, add 8mL organic solvent trifluoroethanol, stir 6hMake it be completely dissolved, obtain polymer PLLA solution；
(3) gained suspension in step (1) is added step (2) resulting polymers PLLA moltenIn liquid, stirring 10h makes the BaTiO of surface modification3Ceramic particle filler (BNP@sodium citrate)It is dispersed in polymer P LLA matrix, obtains mixed liquor；
(4) step (3) gained mixed liquor is injected in injector for medical purpose, syringe is placed in quietElectric spinning device, the reception device of electrostatic spinning is flat board, and electrostatic potential is 13kV, spinning flow velocityIt is 15cm for 0.8mL/h, syringe needle and the distance received between device.Gained nano fibrous membrane is placed inMake solvent volatilize completely in vacuum drying oven, obtain a kind of electroactive nano fiber scaffold material.
Barium titanate nano pottery is contained by the electroactive nano fiber scaffold material of above step gainedGranule and polymer P LLA, it is micro/nano fibrous membrane material, and thickness is 200 μm, nanofiberA diameter of 400nm.Nano-ceramic particle quality in electroactive nano fiber scaffold material containsAmount is 7wt.%.
Embodiment 2
(1) weighing sodium citrate to be placed in 2ml organic solvent trifluoroethanol, being configured to concentration is 0.02The sodium citrate solution of g/mL, takes 0.05g BaTiO3Ceramic particle is placed in sodium citrate solution,Carry out sonic oscillation, each 30min of stir process at ambient temperature, alternately 4 times so that tableThe BaTiO that face is modified3Ceramic particle filler (BNP@sodium citrate) is formed in a solvent and uniformly dividesThe suspension dissipated；
(2) weigh 1g polymer P LLA, add 8mL organic solvent trifluoroethanol, stir 5hMake it be completely dissolved, obtain polymer PLLA solution；
(3) gained suspension in step (1) is added step (2) resulting polymers PLLA moltenIn liquid, stirring 12h makes the BaTiO of surface modification3Ceramic particle filler (BNP@sodium citrate)It is dispersed in polymer P LLA matrix, obtains mixed liquor；
(4) step (3) gained mixed liquor is injected in injector for medical purpose, syringe is placed in quietElectric spinning device, the reception device of electrostatic spinning is flat board, and electrostatic potential is 15kV, spinning flow velocityIt is 16cm for 1.0mL/h, syringe needle and the distance received between device.Gained nano fibrous membrane is placed inMake solvent volatilize completely in vacuum drying oven, obtain a kind of electroactive nano fiber scaffold material.
Barium titanate nano pottery is contained by the electroactive nano fiber scaffold material of above step gainedGranule and polymer P LLA, it is micro/nano fibrous membrane material, and thickness is 300 μm, nanofiberA diameter of 300nm.Nano-ceramic particle quality in electroactive nano fiber scaffold material containsAmount is 5wt.%.
Embodiment 3
(1) weighing sodium citrate to be placed in 2ml organic solvent trifluoroethanol, being configured to concentration is 0.02The sodium citrate solution of g/mL, takes 0.01g BaTiO3Ceramic particle is placed in sodium citrate solution,Carry out sonic oscillation, each 30min of stir process at ambient temperature, alternately 4 times so that tableThe BaTiO that face is modified3Ceramic particle filler (BNP@sodium citrate) is formed in a solvent and uniformly dividesThe suspension dissipated；
(2) weigh 1g polymer P LLA, add 8mL organic solvent trifluoroethanol, stir 8hMake it be completely dissolved, obtain polymer PLLA solution；
(3) gained suspension in step (1) is added step (2) resulting polymers PLLA moltenIn liquid, stirring 8h makes the BaTiO of surface modification3Ceramic particle filler (BNP@sodium citrate)It is dispersed in polymer P LLA matrix, obtains mixed liquor；
(4) step (3) gained mixed liquor is injected in injector for medical purpose, syringe is placed in quietElectric spinning device, the reception device of electrostatic spinning is flat board, and electrostatic potential is 15kV, spinning flow velocityIt is 12cm for 1.0mL/h, syringe needle and the distance received between device.Gained nano fibrous membrane is placed inMake solvent volatilize completely in vacuum drying oven, obtain a kind of electroactive nano fiber scaffold material.
Barium titanate nano pottery is contained by the electroactive nano fiber scaffold material of above step gainedGranule and polymer P LLA, it is micro/nano fibrous membrane material, and thickness is 150 μm, nanofiberA diameter of 200nm.Nano-ceramic particle quality in electroactive nano fiber scaffold material containsAmount is 1wt.%.
Embodiment 4
(1) weighing sodium citrate to be placed in 2ml organic solvent trifluoroethanol, being configured to concentration is 0.02The sodium citrate solution of g/mL, takes 0.1g potassium sodium niobate ceramic granule and is placed in sodium citrate solution,Carry out sonic oscillation, each 30min of stir process at ambient temperature, alternately 5 times so that tableThe potassium sodium niobate ceramic granular filler (KNN@sodium citrate) of face modification is formed uniformly in a solventScattered suspension；
(2) weigh 1g polymer P LLA, add 8mL organic solvent trifluoroethanol, stir 7hMake it be completely dissolved, obtain polymer PLLA solution；
(3) gained suspension in step (1) is added step (2) resulting polymers PLLA moltenIn liquid, stirring 12h makes potassium sodium niobate ceramic granular filler (the KNN@citric acid of surface modificationSodium) it is dispersed in polymer P LLA matrix, obtain mixed liquor；
(4) step (3) gained mixed liquor is injected in injector for medical purpose, syringe is placed in quietElectric spinning device, the reception device of electrostatic spinning is copper mesh, and electrostatic potential is 18kV, spinning flow velocityIt is 18cm for 0.5mL/h, syringe needle and the distance received between device.Gained nano fibrous membrane is placed inMake solvent volatilize completely in vacuum drying oven, obtain a kind of electroactive nano fiber scaffold material.
Contain potassium-sodium niobate nanometer by the electroactive nano fiber scaffold material of above step gained to make potteryPorcelain granule and polymer P LLA, it is micro/nano fibrous membrane material, and thickness is 300 μm, NanowireA diameter of 450nm of dimension.Nano-ceramic particle quality in electroactive nano fiber scaffold materialContent is 10wt.%.
Embodiment 5
(1) weigh sodium citrate to be placed in 2ml organic solvent (such as trifluoroethanol), be configured to denseDegree is the sodium citrate solution of 0.02g/mL, takes 0.2g barium-strontium titanate ceramic granule and is placed in citric acidIn sodium solution, carry out sonic oscillation, each 30min of stir process at ambient temperature, alternately 3Secondary so that the barium-strontium titanate ceramic granular filler (BST@sodium citrate) of surface modification is in a solventForm homodisperse suspension；
(2) weigh 1g polymer P LLA, add 8mL organic solvent trifluoroethanol, stir 8hMake it be completely dissolved, obtain polymer PLLA solution；
(3) gained suspension in step (1) is added step (2) resulting polymers PLLA moltenIn liquid, stirring 12h makes the barium-strontium titanate ceramic granular filler (BST@sodium citrate) of surface modificationIt is dispersed in polymer P LLA matrix, obtains mixed liquor；
(4) step (3) gained mixed liquor is injected in injector for medical purpose, syringe is placed in quietElectric spinning device, the reception device of electrostatic spinning is roller, and electrostatic potential is 18kV, spinning flow velocityIt is 20cm for 0.3mL/h, syringe needle and the distance received between device.Gained nano fibrous membrane is placed inMake solvent volatilize completely in vacuum drying oven, obtain a kind of electroactive nano fiber scaffold material.
Contain barium strontium titanate nanometer by the electroactive nano fiber scaffold material of above step gained to make potteryPorcelain granule and polymer P LLA, it is micro/nano fibrous membrane material, and thickness is 400 μm, NanowireA diameter of 500nm of dimension.Nano-ceramic particle quality in electroactive nano fiber scaffold materialContent is 20wt.%.
Comparative example 6
The electroactive nano fiber scaffold material that the present invention uses electrostatic spinning technique to prepare is being tiedIn three-dimensional network spline structure on structure, the host material cultivated as cell, can be not only cellGrowth provides three dimensional matrix microenvironment, makes cell present three dimensional growth state, and cell can be grown into materialSpace is internal also organically combines (as shown in Figure 7) with material, and the transmission of beneficially nutritional labeling,Thus significantly improve cell proliferation and tissue repair.And for tradition reality wall two dimensional surface material,Cell is showed only as flat growth conditions, and its proliferation activity is less than on nano fiber scaffold materialOsteoblast activity (as shown in Figure 8).