CN105079883B - A kind of multi-stage nano fiber composite medicine-carried periodontium material and preparation method thereof - Google Patents
A kind of multi-stage nano fiber composite medicine-carried periodontium material and preparation method thereof Download PDFInfo
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Abstract
The invention belongs to medical material tech field, discloses a kind of multi-stage nano fiber composite medicine-carried periodontium material and preparation method thereof.Described multi-stage nano fiber composite medicine-carried periodontium material is made up of the PLA that mass fraction is 50%~99% and blended fiber, composite fibre or acid fiber by polylactic/chitosan microball composite construction that 1%~50% chitosan is formed；Its preparation method is：Polylactic acid fiber membrane or support are prepared first, then recombination chitosan fiber or chitosan microball in polylactic acid fiber membrane or support；Or acid fiber by polylactic and chitin fiber formation blended fiber or composite fibre are prepared simultaneously, obtain multi-stage nano fiber composite medicine-carried periodontium material.In preparation process, fat-soluble, water-soluble and nanoparticulate drug composition be could be incorporated on composite the material of the present invention, and for adding from now on, multi-medicament will be modified or function improvement makes great sense.
Description
Technical field
The invention belongs to Degradable Biomedical Materials technical field, and in particular to a kind of multi-stage nano fiber composite can carry
Medicine periodontium material and preparation method thereof.
Background technology
From after PGA (polyglycolic acid) is used for Orthopedic Clinical by Rokkane in 1984 first, the absorbable material of biodegradation
The experiment and clinical research of material obtain fast development, and obtain certain achievement.This kind of material obtains in orthopaedic trauma at present
Develop rapidly.Periodontium is in the moist microenvironment in oral cavity, constantly oozing out for level in gingival sulcus fluid, is largely caused a disease in periodontium
Bacterium, which has many factors, causes the treatment of periodontitis and the treatment of wound bone to have significant difference, the main function of wound bone material
It is to promote knitting, therefore, the artificial substituting material for periodontal surgery is higher than the material requirements of wound bone.
The artificial substituting material commonly used in periodontal surgery has bone meal and GTR (guide tissue regeneration) film, in periodontosis,
Periodontium Cranial defect is once formed, it may be difficult to returns to original degree, organization healing is often formed after periodontal surgery in addition
Epithelium regeneration heals, and this will improve the recurrence rate of periodontosis.Therefore, how induction material successfully loaded by periodontium material
Material, antibacterial medicines and antibiotics composition, so as to promote periodontium Cranial defect to recover, suppress Periodontal Pathogens growth simultaneously
The recurrence rate of periodontosis is reduced, is that researcher in this field needs to solve the problems, such as.
The content of the invention
In order to solve the shortcomings that above prior art and weak point, primary and foremost purpose of the invention is to provide a kind of multistage
The compound medicine-carried periodontium material of nanofiber.
Another object of the present invention is to provide the preparation of above-mentioned multi-stage nano fiber composite medicine-carried periodontium material
Method.
The object of the invention is achieved through the following technical solutions：
A kind of multi-stage nano fiber composite medicine-carried periodontium material, by the PLA that mass fraction is 50%~99%
(PLA) and 1%~50% chitosan (CS) formed blended fiber, composite fibre or acid fiber by polylactic/chitosan microball it is multiple
Structure is closed to form；The form of described blended fiber is that acid fiber by polylactic and chitin fiber mutually intert or are layered in one
Rise；The form of the composite fibre is that acid fiber by polylactic and chitin fiber form island, core-skin, bias or composite junction arranged side by side
Structure；The form of the acid fiber by polylactic/chitosan microball composite construction is that chitosan microball is distributed in PLA fibre with graininess
Dimension table face.
Preferably, the weight average molecular weight of the PLA is 1 × 104~9 × 107G/mol, the weight average molecular weight of chitosan
For 1 × 104~9 × 107g/mol。
Described multi-stage nano fiber composite medicine-carried periodontium material also includes carrying medicine composition, described load medicine composition
Medicine composition or nano particle are carried including water solubility, the described water-soluble medicine composition that carries is carried on chitin fiber or chitosan microball
Inside or surface；Described nano particle is carried on inside or the table of chitin fiber, chitosan microball or acid fiber by polylactic
Face.
Preferably, it is described it is water-soluble carry medicine composition be fibroblast growth factor, tetracycline, Amoxicillin, first nitre file,
At least one of Liuwei Dihuang Wan, Puerarin, hedysarum polybotys saccharide and iron lactoprotein；Described nano particle refer to Nano Silver and
At least one of ZnO.
The preparation method of above-mentioned multi-stage nano fiber composite medicine-carried periodontium material, including following preparation process：
Prepare polylactic acid fiber membrane or support first, then in polylactic acid fiber membrane or support recombination chitosan fiber or
Chitosan microball, or acid fiber by polylactic and chitin fiber formation blended fiber or composite fibre are prepared simultaneously, obtain multistage and receive
Rice fiber composite medicine-carried periodontium material.
Above-mentioned periodontium material also loading nano silvery by the following method：Nano Silver deionized water suspension is blown and beaten through air gun
Uniform and ultrasonic disperse, directly it is added dropwise in material surface, air-dries, obtain multi-stage nano fiber composite loading nano silvery periodontium
Material.
The polylactic acid fiber membrane or the preparation method of support are as follows：
PLA is dissolved in solvent, the PLA solution that mass fraction is 5~40% is made into, then passes through Static Spinning
Silk method, film split method, meltblown, centrifugal spinning, freeze-drying, liquid cause split-phase method or flash method obtain polylactic acid fiber membrane or
Support.
Preferably, described solvent refers to that chloroform, dichloromethane, dimethylformamide, dimethyl acetamide, dimethyl are sub-
Mixing more than one or both of sulfone, hexafluoroisopropanol, six cyclopentadiene and tetrahydrofuran.
The preparation method of recombination chitosan fiber or chitosan microball in polylactic acid fiber membrane or support is as follows：
Dissolve chitosan in acetic acid, trifluoroacetic acid, trifluoroethanol and the hexafluoro isopropyl that mass fraction is 30%~90%
In one or both of alcohol solvent mixed above, the chitosan solution that mass fraction is 5~30% is made into, then passes through electrostatic
Spinning, electrostatic spraying, electrostatic dipping or dipping freeze-drying polylactic acid fiber membrane or rack surface formed chitin fiber or
Chitosan microball.
Acid fiber by polylactic is prepared while described and chitin fiber forms the preparation method of blended fiber or composite fibre
It is as follows：
Dissolve chitosan in acetic acid, trifluoroacetic acid, trifluoroethanol and the hexafluoro isopropyl that mass fraction is 30%~90%
In one or both of alcohol solvent mixed above, the chitosan solution that mass fraction is 5~30% is made into, then by chitosan
Solution obtains mixed emulsion with PLA solution in the presence of surfactant；Again by mixed emulsion by method of electrostatic spinning,
Film splits method, meltblown, centrifugal spinning, freeze-drying, liquid and causes split-phase method or flash method to obtain PLA/CS blended fibers or multiple
Condensating fiber.
Described chitosan solution is additionally added water-soluble load medicine composition or nano particle in process for preparation and dissolves and pass through
It is uniformly dispersed, obtains chitosan-loaded drug solns.
Preferably, described surfactant refers to tween, sapn, dodecyl sodium sulfate, lauryl sodium sulfate, poly-
It is more than one or both of vinyl alcohol (PVA), polyvinylpyrrolidone, carboxymethyl cellulose (CMC) and polyacrylic acid mixed
Close.
Preferably, the specific preparation process of the mixed emulsion is：1%~10% matter is first added into the solvent of PLA
The surfactant of fraction is measured, stirring and emulsifying forms emulsification system, then adds chitosan solution, poly- breast is added after stirring and emulsifying
Acid, dissolve, homogenizing, deaeration, obtain mixed emulsion；The solvent of the PLA refers to chloroform, dichloromethane, dimethyl formyl
It is more than one or both of amine, dimethyl acetamide, dimethyl sulfoxide (DMSO), hexafluoroisopropanol, six cyclopentadiene and tetrahydrofuran
Mixing.
The present invention preparation method and resulting product has the following advantages that and beneficial effect：
(1) in the preparation process of periodontium material of the invention, water soluble drug can be loaded in chitosan solution, fat
Soluble drug can load on the shaping that nanofiber is carried out in PLA material, and nano particle such as Nano Silver can directly be sprayed onto material
Surface, fat-soluble, water-soluble and nanoparticulate drug composition could be incorporated on composite, and it for adding a variety of medicines from now on
Thing is modified or function improvement makes great sense, and possesses vast potential for future development；
(2) structure of multi-stage nano fiber composite medicine-carried periodontium material of the invention and the diameter of parodontium collagen
And form is similar, wound bone material is different from, is regenerated available for promotion organization after periodontal surgery, improves operative effect；
(3) multi-stage nano fiber composite medicine-carried periodontium material of the invention is bicomponent structural, is unfavorable for cell
The PLA of adhesion directly contacts with gingiva tissue；Contacted beneficial to the chitosan of cell propagation with bone tissue, beneficial to skeletonization；
(4) present invention is modified using chitosan to PLA, elastic properties of materials step-down, it is sufficient to maintain periodontium reparation
Space, degradation speed then obtains appropriate proportioning to meet reality by changing using PLA molecular weight and regulation CS dosage
The requirement that border uses；
(5) preparation method of the invention is combined using a variety of forming techniques, and products therefrom effectively overcomes simple quiet
The defects of fibrofelt obtained by electrical spinning method is uneven.
Embodiment
With reference to embodiment, the present invention is described in further detail, but the implementation of the present invention is not limited to this.
Embodiment 1
PLA (PLA, Mw=3 × 105G/mol after) being dried in vacuo (80 DEG C, 10h), chloroform is used to be configured to for solvent
The mass fraction of PLA is 20% solution, and magnetic agitation 5h, standing and defoaming 4h obtain PLA solution；The PLA solution that will be prepared
PLA film is prepared using method of electrostatic spinning, electrospinning conditions are：The distance between receiver board and syringe needle 12cm, in 15kV voltages
Lower PLA solution carries out electrospinning with 0.5ml/h flow velocity, and the tunica fibrosa that electrospinning is obtained is dried in vacuo, and obtains pure PLA film；
Chitosan (CS, Mw=8 × 104G/mol concentration) is used to be configured to the CS solution that mass fraction is 3% for 90% acetic acid solvent,
Then CS solution is sprayed onto in pure PLA film in a manner of 0.6ml/h flow velocity is by electrostatic spraying, received in electrostatic spray process
Plate is 10cm with syringe needle distance, and the voltage between syringe needle and receiver board is 20kV, is dried in vacuo again, that is, obtains CS-PLA films；Will
Nano Silver deionized water suspension is blown and beaten uniformly and through ultrasonic disperse through air gun, is directly added dropwise on CS-PLA films surface, is air-dried, obtain
Multi-stage nano fiber composite loads silver-colored periodontium material.
The fibre diameter of PLA is 300~500nm in periodontium material obtained by the present embodiment, and CS is distributed with graininess
On PLA film surface, its diameter range is 400~800nm, and its structure is acid fiber by polylactic/chitosan microball composite construction；PLA
The collagenous fibres diameter of fiber and periodontal membrane tissue can simulate periodontal close to (diameter of collagenous fibres about 200~1500nm)
The microenvironment of theca cell growth, the structure are different from wound bone material (tradition wound bone material simulation bone structure, for more preferable mould
Intend bone tissue, be often prepared to the loose structure of similar cancellous bone.Spinning fibre structure and parodontium prepared by the present invention is fine
Dimension is close, suitable paradenlal tissue regeneration).In the cell propagation obtained by the present embodiment on periodontium material (using from big
The bone marrow stroma stem cell of mouse, numbering RAWMX-01001) it is significantly faster than that (i.e. cell is directly inoculated in cell to blank control group
Culture plate), cell mineralization experiments (using the bone marrow stroma stem cell from rat, numbering RAWMX-01001) display ore deposit
It is more obvious to change tubercle.Therefore, periodontium material obtained by the present embodiment can meet the requirement for inducing periodontium growth.
Embodiment 2
PLA (PLA, Mw=9 × 107G/mol after) being dried in vacuo (80 DEG C, 10h), using chloroform:Dimethylformamide
Weight ratio is 1:1 mixed solvent is solvent, and the mass fraction for being configured to PLA is 14% solution, stirs 5h, is stood de-
Bubble 4h obtains PLA solution；The PLA solution prepared is prepared into PLA film using method of electrostatic spinning, electrospinning conditions are：Receiver board
The distance between syringe needle 12cm, PLA solution carries out electrospinning with 0.5ml/h flow velocity under 15kV voltages, and electrospinning is obtained
Tunica fibrosa is dried in vacuo, and obtains pure PLA film；Chitosan (CS Mw=7 × 104G/mol 90% hexafluoroisopropanol) is used：
Trifluoroacetic acid (1:1) the CS solution that mass fraction is 5% is configured to, CS solution is then passed through into electrostatic with 0.6ml/h flow velocity
The mode of spraying is sprayed onto the two sides of pure PLA film, and the distance between receiver board and syringe needle 10cm in electrostatic spray process, syringe needle is with connecing
It is 20kV to receive the voltage between plate, the vacuum drying of gained sample, produces CS-PLA composite membranes, the structure of composite membrane is sandwich knot
Structure, wherein CS are in PLA both sides, using PLA as intermediate layer；Nano Silver deionized water suspension is blown and beaten uniformly and through ultrasound through air gun
It is scattered, directly it is added dropwise in CS-PLA composite film surfaces, air-dries, obtains multi-stage nano fiber composite and load silver-colored periodontium material.
The fibre diameter of PLA is 300~500nm in periodontium material obtained by the present embodiment, and CS is distributed with graininess
On PLA film surface, its diameter range is 500~900nm, and its structure is acid fiber by polylactic/chitosan microball composite construction；PLA
The collagenous fibres diameter of fiber and periodontal membrane tissue can simulate periodontal close to (diameter of collagenous fibres about 200~1500nm)
The microenvironment of theca cell growth, the structure are different from wound bone material (tradition wound bone material simulation bone structure, for more preferable mould
Intend bone tissue, be often prepared to the loose structure of similar cancellous bone.Spinning fibre structure and parodontium prepared by the present invention is fine
Tie up close to suitable paradenlal tissue regeneration).(use in the cell propagation obtained by the present embodiment on periodontium material and derive from rat
Bone marrow stroma stem cell, numbering RAWMX-01001) be significantly faster than that blank control group (i.e. cell be directly inoculated in cell training
Support plate), cell mineralization experiments (using the bone marrow stroma stem cell from rat, numbering RAWMX-01001) display mineralising
Tubercle is more obvious.Therefore, periodontium material obtained by the present embodiment can meet the requirement for inducing periodontium growth.
Embodiment 3
PLA (PLA, Mw=2.4 × 106G/mol after) being dried in vacuo (80 DEG C, 10h), using chloroform:Dimethyl formyl
Amine weight ratio is 1:1 mixed solvent is solvent, and the mass fraction for being configured to PLA is 24% solution, stirs 5h, stands
Deaeration 4h obtains PLA solution；Chitosan (CS Mw=1 × 104G/mol quality point) is configured to using 90% trifluoroacetic acid solution
Number is 5% CS solution；Using double needle pattern, i.e. a syringe needle carries out the electrostatic spinning of PLA solution, and another syringe needle is carried out
The electrostatic spraying of CS solution.The distance between receiver board and syringe needle 12cm, PLA solution is with 0.5ml/h stream under 15kV voltages
Speed carries out electrospinning, vacuum drying, obtains pure PLA film；CS solution is 12cm with syringe needle distance with 0.6ml/h flow velocity, receiver board,
Voltage is molded for 20kV between syringe needle and receiver board, and sample is dried in vacuo, that is, obtains CS-PLA composite membranes, gained composite membrane knot
Structure is the core-sheath composite structure that CS and PLA mutually mix, and wherein CS is sandwich layer, and PLA is cortex；By Nano Silver deionized water suspension
Through air gun piping and druming uniformly and through ultrasonic disperse, directly it is added dropwise in CS-PLA composite film surfaces, air-dries, obtain multi-stage nano fiber and answer
Close and load silver-colored periodontium material.
The fibre diameter of PLA is 300~500nm in periodontium material obtained by the present embodiment, CS distribution of fiber diameters
Scope is 600~800nm, the collagenous fibres diameter of PLA fibers and periodontal membrane tissue close to (diameter of collagenous fibres about 200~
1500nm), the microenvironment of periodontal ligament cell growth can be simulated, the structure is different from (the tradition wound bone material simulation of wound bone material
Bone structure, in order to more preferably simulate bone tissue, often it is prepared to the loose structure of similar cancellous bone.Spinning prepared by the present invention
Silk fiber structure and periodontal membrane fiber are close to suitable paradenlal tissue regeneration).Thin on periodontium material obtained by the present embodiment
Born of the same parents' propagation (using the bone marrow stroma stem cell from rat, numbering RAWMX-01001) is significantly faster than that blank control group (i.e.
Cell is directly inoculated in Tissue Culture Plate), (using the bone marrow stroma stem cell from rat, numbering is cell mineralization experiments
RAWMX-01001) show that Mineral nodules are more obvious.Therefore, periodontium material obtained by the present embodiment can meet to induce tooth
The requirement of all tissue growths.
Embodiment 4
PLA (PLA, Mw=8 × 105G/mol after) being dried in vacuo (80 DEG C, 10h), dimethylformamide is used to be molten
Agent, the mass fraction for being configured to PLA is 28% solution, stirs 5h, standing and defoaming 4h obtains PLA solution；Chitosan (CS
Mw=4 × 104G/mol) using 90% mass fraction hexafluoroisopropanol and trifluoroacetic acid mixed solvent (hexafluoroisopropanol and
The volume ratio of trifluoroacetic acid is 1:3) the CS solution that mass fraction is 5% is configured to, and 2wt% first nitre is added in CS solution
File is uniformly dissolved to obtain CS load drug solns；Using double needle pattern is used, i.e. a syringe needle carries out the electrostatic spinning of PLA solution,
Another syringe needle carries out the electrostatic spraying of CS solution, and using method of electrostatic spinning prepared by PLA solution into PLA film, receiver board and syringe needle
The distance between 15cm, PLA solution carries out electrospinning with 0.5ml/h flow velocity under 25kV voltages, is dried in vacuo, obtained pure
PLA film；It is 12cm with syringe needle distance that CS, which carries drug solns with 0.6ml/h flow velocity, receiver board, and voltage is between syringe needle and receiver board
20kV is molded, and sample is dried in vacuo, that is, obtains multi-stage nano fiber composite load first nitre tooth grinding week organization material, resulting materials
Structure is the island composite construction that CS and PLA mutually mix, and wherein CS is island phase, and PLA is marine facies.
The fibre diameter of PLA is 500-700nm in periodontium material obtained by the present embodiment, CS distribution of fiber diameters
Scope is 500-800nm, the collagenous fibres diameter of PLA fibers and periodontal membrane tissue close to (diameter of collagenous fibres about 200~
1500nm), the microenvironment of periodontal ligament cell growth can be simulated, the structure is different from (the tradition wound bone material simulation of wound bone material
Bone structure, in order to more preferably simulate bone tissue, often it is prepared to the loose structure of similar cancellous bone.Spinning prepared by the present invention
Silk fiber structure and periodontal membrane fiber are close to suitable paradenlal tissue regeneration).Thin on periodontium material obtained by the present embodiment
Born of the same parents' propagation (using the bone marrow stroma stem cell from rat, numbering RAWMX-01001) is significantly faster than that blank control group (i.e.
Cell is directly inoculated in Tissue Culture Plate), (using the bone marrow stroma stem cell from rat, numbering is cell mineralization experiments
RAWMX-01001) show that Mineral nodules are more obvious.Therefore, periodontium material obtained by the present embodiment can meet to induce tooth
The requirement of all tissue growths.
Embodiment 5
PLA (PLA, Mw=3 × 106G/mol after) being dried in vacuo (80 DEG C, 10h), using dimethylformamide:Hexafluoro
The weight ratio of isopropanol is 1:2 mixed solvent is solvent, and the mass fraction for preparing PLA is 50% solution, stirs 5h,
Standing and defoaming 4h obtains PLA solution；Chitosan (CS Mw=3 × 105G/mol) using 60% mass fraction hexafluoroisopropanol and
(volume ratio of hexafluoroisopropanol and trifluoroacetic acid is 1 to the mixed solvent of trifluoroacetic acid:3) it is molten to be made into the CS that mass fraction is 3%
Liquid, and the ZnO of addition 2% is uniformly dispersed to obtain CS load drug solns in CS solution；Using using double needle pattern, i.e. a pin
Head carries out the electrostatic spinning of PLA solution, and another syringe needle carries out the electrostatic spraying of CS solution, PLA solution is used into electrostatic spinning
Method prepares PLA film, and the distance between receiver board and syringe needle 15cm, PLA solution is carried out with 0.5ml/h flow velocity under 25kV voltages
Electrospinning, pure PLA film is obtained after vacuum drying；It is 12cm with syringe needle distance that CS, which carries drug solns with 0.6ml/h flow velocity, receiver board,
Voltage is molded for 20kV between syringe needle and receiver board, and sample is dried in vacuo, that is, obtains multi-stage nano fiber composite loading ZnO tooth
All organization materials, gained structure of composite membrane are the core-sheath composite structure that CS and PLA mutually mix, and wherein CS is sandwich layer, and PLA is skin
Layer.
The fibre diameter of PLA is 600-1200nm in periodontium material obtained by the present embodiment, CS distribution of fiber diameters
Scope is 500-800nm, the collagenous fibres diameter of PLA fibers and periodontal membrane tissue close to (diameter of collagenous fibres about 200~
1500nm), the microenvironment of periodontal ligament cell growth can be simulated, the structure is different from (the tradition wound bone material simulation of wound bone material
Bone structure, in order to more preferably simulate bone tissue, often it is prepared to the loose structure of similar cancellous bone.Spinning prepared by the present invention
Silk fiber structure and periodontal membrane fiber are close to suitable paradenlal tissue regeneration).Thin on periodontium material obtained by the present embodiment
Born of the same parents' propagation (using the bone marrow stroma stem cell from rat, numbering RAWMX-01001) is significantly faster than that blank control group (i.e.
Cell is directly inoculated in Tissue Culture Plate), (using the bone marrow stroma stem cell from rat, numbering is cell mineralization experiments
RAWMX-01001) show that Mineral nodules are more obvious.Therefore, periodontium material obtained by the present embodiment can meet to induce tooth
The requirement of all tissue growths.
Embodiment 6
PLA (PLA, Mw=6 × 105G/mol after) being dried in vacuo (80 DEG C, 10h), using dimethyl sulfoxide (DMSO):Dichloromethane
The weight ratio of alkane is 1:2 mixed solvent is solvent, and the mass fraction for preparing PLA is 14% solution, stirs 5h, stands
Deaeration 4h obtains PLA solution；PLA solution is prepared into PLA film, the distance between receiver board and syringe needle using centrifugation method of electrostatic spinning
20cm, PLA solution is carried out electrospinning, pure PLA is obtained after vacuum drying with 450r/min, 15ml/h flow velocity under 35kV voltage
Film；Chitosan (CS Mw=2.6 × 105G/mol the hexafluoroisopropanol of 60% mass fraction) is used to be configured to quality point for solvent
Number is 3% CS solution, CS solution is adhered in pure PLA film using electrostatic impregnation technology, voltage 5kV, impregnates solid-to-liquid ratio
For 1:15, dip time 5min, are then dried in vacuo again, that is, obtain CS-PLA films, i.e. acid fiber by polylactic and chitosan is fine
Dimension forms eccentric structure；Nano Silver deionized water suspension is blown and beaten uniformly and through ultrasonic disperse through air gun, is directly added dropwise in CS-PLA
Film surface, air-dry, obtain multi-stage nano fiber composite and load silver-colored periodontium material.
The fibre diameter of PLA is 300-500nm in periodontium material obtained by the present embodiment, CS distribution of fiber diameters
Scope is 400-800nm, the collagenous fibres diameter of PLA fibers and periodontal membrane tissue close to (diameter of collagenous fibres about 200~
1500nm), the microenvironment of periodontal ligament cell growth can be simulated, the structure is different from (the tradition wound bone material simulation of wound bone material
Bone structure, in order to more preferably simulate bone tissue, often it is prepared to the loose structure of similar cancellous bone.Spinning prepared by the present invention
Silk fiber structure and periodontal membrane fiber are close to suitable paradenlal tissue regeneration).Thin on periodontium material obtained by the present embodiment
Born of the same parents' propagation (using the bone marrow stroma stem cell from rat, numbering RAWMX-01001) is significantly faster than that blank control group (i.e.
Cell is directly inoculated in Tissue Culture Plate), (using the bone marrow stroma stem cell from rat, numbering is cell mineralization experiments
RAWMX-01001) show that Mineral nodules are more obvious.Therefore, periodontium material obtained by the present embodiment can meet to induce tooth
The requirement of all tissue growths.
Embodiment 7
PLA (PLA, Mw=6 × 105G/mol after) being dried in vacuo (80 DEG C, 10h), using dimethyl sulfoxide (DMSO):Dichloromethane
The weight ratio of alkane is 1:2 mixed solvent is solvent, and the mass fraction for preparing PLA is 14% solution, stirs 5h, stands
Deaeration 4h obtains PLA solution；PLA solution is prepared into PLA film, the distance between receiver board and syringe needle using centrifugation method of electrostatic spinning
20cm, solution is carried out electrospinning, pure PLA film is obtained after vacuum drying with 450r/min, 15ml/h flow velocity under 35kV voltage；
Chitosan (CS Mw=2.6 × 105G/mol the hexafluoroisopropanol aqueous solution of 60% mass fraction) is used to be configured to quality for solvent
Fraction is 3% CS solution, then 5wt% Amoxicillin medicine is added into CS solution, obtains CS and carries drug solns, CS is carried into medicine
Solution is adhered in pure PLA film using freeze drying process after dipping, that is, obtains multi-stage nano fiber composite load Amoxicillin
Periodontium material.
The fibre diameter of PLA is 300-500nm in periodontium material obtained by the present embodiment, and CS is distributed with graininess
On PLA film surface, its diameter distribution is 400-800nm, and the collagenous fibres diameter of PLA fibers and periodontal membrane tissue approaches
(diameter of collagenous fibres about 200~1500nm), can simulate the microenvironment of periodontal ligament cell growth, and the structure is different from wound bone
(tradition wound bone material simulation bone structure, in order to more preferably simulate bone tissue, is often prepared to the porous of similar cancellous bone to material
Structure.Spinning fibre structure prepared by the present invention and periodontal membrane fiber are close to suitable paradenlal tissue regeneration).In the present embodiment institute
Obtain the cell propagation on periodontium material (using the bone marrow stroma stem cell from rat, numbering RAWMX-01001)
Blank control group (i.e. cell is directly inoculated in Tissue Culture Plate) is significantly faster than that, cell mineralization experiments are (using from rat
Bone marrow stroma stem cell, numbering RAWMX-01001) show that Mineral nodules are more obvious.Therefore, periodontal group obtained by the present embodiment
Knit the requirement that material can meet to induce periodontium growth.
Embodiment 8
PLA (PLA, Mw=1 × 104G/mol after) being dried in vacuo (80 DEG C, 10h), PLA fibers are prepared using meltblown
Framework material, framework material is dried in vacuo to obtain pure PLA scaffold material；Chitosan (CS Mw=9 × 107G/mol) use
The hexafluoroisopropanol aqueous solution of 90% mass fraction is the CS solution that solvent is configured to that mass fraction is 1%, passes through electrostatic spraying
Mode CS solution is sprayed onto in pure PLA scaffold material with 0.6ml/h flow velocity, receiver board and syringe needle distance are 10cm, syringe needle
Voltage between receiver board is 20kV, is dried in vacuo again, that is, it is shelf layer to obtain PLA fibers, and miscellaneous plant CS fibers is more in layer
The level compound medicine-carried periodontium material of nanofiber.
The fibre diameter of PLA is 800-1400nm in periodontium material obtained by the present embodiment, CS distribution of fiber diameters
Scope is 700-1200nm, the collagenous fibres diameter of PLA fibers and periodontal membrane tissue close to (diameter of collagenous fibres about 200~
1500nm), the microenvironment of periodontal ligament cell growth can be simulated, the structure is different from (the tradition wound bone material simulation of wound bone material
Bone structure, in order to more preferably simulate bone tissue, often it is prepared to the loose structure of similar cancellous bone.Spinning prepared by the present invention
Silk fiber structure and periodontal membrane fiber are close to suitable paradenlal tissue regeneration).Thin on periodontium material obtained by the present embodiment
Born of the same parents' propagation (using the bone marrow stroma stem cell from rat, numbering RAWMX-01001) is significantly faster than that blank control group (i.e.
Cell is directly inoculated in Tissue Culture Plate), (using the bone marrow stroma stem cell from rat, numbering is cell mineralization experiments
RAWMX-01001) show that Mineral nodules are more obvious.Therefore, periodontium material obtained by the present embodiment can meet to induce tooth
The requirement of all tissue growths.
Embodiment 9
In dimethyl sulfoxide (DMSO):The weight ratio of dichloromethane is 1:2 in the mixed solvent adds 1wt% sorbester p18,
After 3000r/min is emulsified 30 minutes at a high speed, it is that solvent is configured to matter to add with the hexafluoroisopropanol aqueous solution of 60% mass fraction
Measure the chitosan (CS.M that fraction is 3%w=2.6 × 105G/mol) solution, 2h is emulsified at a high speed in 5000r/min, adds 14wt%
PLA (PLA, the M of (80 DEG C, 10h) after vacuum dryingw=6 × 105G/mol), 5h is stirred, 4h is stood and obtains PLA/CS mixing
Emulsion；5wt% tetracycline medicine is added wherein in CS solution process for preparation, forms medicine-carried system；By PLA/CS mixed emulsions
PLA/CS composite cellulosic membranes are prepared using method of electrostatic spinning, the distance between receiver board and syringe needle 20cm, spun under 35kV voltages
Silk shaping, obtain PLA/CS core-skin multi-stage nanos fiber composite load tetracycline periodontal organization material.
The fibre diameter of PLA/CS core-skin composite fibers is 600-1500nm, CS in periodontium material obtained by the present embodiment
With irregular structure distribution in fibrous outer surfaces, the collagenous fibres diameter of PLA fibers and periodontal membrane tissue is close to (collagenous fibres
Diameter about 200~1500nm), can simulate periodontal ligament cell growth microenvironment, the structure be different from wound bone material (tradition
Bone material simulation bone structure is created, in order to more preferably simulate bone tissue, is often prepared to the loose structure of similar cancellous bone.This hair
Bright prepared spinning fibre structure and periodontal membrane fiber are close to suitable paradenlal tissue regeneration).In periodontal group obtained by the present embodiment
The cell propagation knitted on material is significantly faster than that (using the bone marrow stroma stem cell from rat, numbering RAWMX-01001)
Blank control group (i.e. cell is directly inoculated in Tissue Culture Plate), cell mineralization experiments (use the bone marrow matrix from rat
Stem cell, numbering RAWMX-01001) show that Mineral nodules are more obvious.Therefore, periodontium material obtained by the present embodiment can
To meet the requirement of induction periodontium growth.
Embodiment 10
In dimethyl acetamide:The weight ratio of dichloromethane is 1:2 in the mixed solvent adds 1wt% polysorbate60,
3000r/min after emulsification 30 minutes, is added with the mixture of the hexafluoroisopropanol of 90% mass fraction and trifluoroethanol at a high speed
(volume ratio of hexafluoroisopropanol and trifluoroethanol is 1 to the aqueous solution:1) it is that mixed solvent is configured to the shell that mass fraction is 3% and gathered
Sugar (CS.Mw=3 × 105G/mol) solution, 2h is emulsified at a high speed in 5000r/min, after addition 14wt% vacuum drying (80 DEG C,
PLA (PLA, M 10h)w=6 × 105G/mol), 5h is stirred, 4h is stood and obtains PLA/CS mixed emulsions；Wherein CS solution is matched somebody with somebody
ZnO medicines are added during system, form medicine-carried system；PLA/CS mixed emulsions are prepared into PLA/CS using centrifugation method of electrostatic spinning
Composite cellulosic membrane, the distance between receiver board and syringe needle 20cm, centrifugal rotation speed 400r/min, it is spun under 35kV voltages
Type, obtain PLA/CS core-skin multi-stage nano fiber composite loading ZnO periodontium materials.
The fibre diameter of PLA/CS core-skin composite fibers is 800-1800nm, CS in periodontium material obtained by the present embodiment
Part is separated, PLA fibers and parodontium in fibrous outer surfaces, part with irregular structure distribution individually into threadiness with PLA
The collagenous fibres diameter of tissue can simulate periodontal ligament cell growth close to (diameter of collagenous fibres about 200~1500nm)
Microenvironment, the structure is different from wound bone material, and (tradition wound bone material simulates bone structure, in order to more preferably simulate bone tissue, often
It is prepared to the loose structure of similar cancellous bone.Spinning fibre structure prepared by the present invention and periodontal membrane fiber are close to suitable tooth
All regenerations).(marrow mesenchymal stem from rat is used in the cell propagation obtained by the present embodiment on periodontium material
Cell, numbering RAWMX-01001) it is significantly faster than that blank control group (i.e. cell is directly inoculated in Tissue Culture Plate), cell ore deposit
It is more obvious to change experiment (using the bone marrow stroma stem cell from rat, numbering RAWMX-01001) display Mineral nodules.
Therefore, periodontium material obtained by the present embodiment can meet the requirement for inducing periodontium growth.
Embodiment 11
The 10wt% PVA aqueous solution (PVA mass concentrations are 15%) is added in chloroform solvent, 3000r/min is high
After speed emulsification 30 minutes, addition is with the chitosan (CS.M that trifluoroacetic acid is that solvent is configured to that mass fraction is 15%w=1.4 ×
104G/mol) solution, 2h is emulsified at a high speed in 5000r/min, add the PLA of (80 DEG C, 10h) after 20wt% vacuum drying
(PLA, Mw=4 × 105G/mol), 5h is stirred, 4h is stood and obtains PLA/CS mixed emulsions；Wherein added in CS solution process for preparation
Iron lactoprotein, form medicine-carried system；PLA/CS mixed emulsions are molded using freeze-drying, PLA/CS islands shape multistage is obtained and receives
Rice fiber composite load iron lactoprotein periodontium material.
The fibre diameter of PLA/CS islands shape composite fibre is 1000- in periodontium material obtained by the present embodiment
1600nm, CS part are separated, the structure in fibrous outer surfaces, part with irregular structure distribution individually into threadiness with PLA
Being different from wound bone material, (tradition wound bone material simulation bone structure, in order to more preferably simulate bone tissue, is often prepared to similar pine
The loose structure of matter bone.Spinning fibre structure prepared by the present invention and periodontal membrane fiber are close to suitable paradenlal tissue regeneration).
(using the bone marrow stroma stem cell from rat, numbering is cell propagation obtained by the present embodiment on periodontium material
RAWMX-01001 blank control group (i.e. cell is directly inoculated in Tissue Culture Plate)) is significantly faster than that, cell mineralization experiments (use
From the bone marrow stroma stem cell of rat, numbering RAWMX-01001) show that Mineral nodules are more obvious.Therefore, this implementation
Example gained periodontium material can meet the requirement for inducing periodontium growth.
Embodiment 12
In chloroform:The weight ratio of tetrahydrofuran is 1:The 10wt% PVA aqueous solution (PVA mass is added in 1 solvent
Concentration is that 20%), for 3000r/min at a high speed after emulsification 30 minutes, addition is that solvent is configured to mass fraction and is with trifluoroacetic acid
15% chitosan (CS.Mw=1.8 × 105G/mol) solution, in 5000r/min, emulsification 2h, addition 10wt% vacuum are done at a high speed
PLA (PLA, the M of (80 DEG C, 10h) after dryw=9 × 107G/mol), 5h is stirred, 4h is stood and obtains PLA/CS mixed emulsions；Its
Hedysarum polybotys saccharide is added in middle CS solution process for preparation, forms medicine-carried system；PLA/CS mixed emulsions are split into melt-blown shaping using film
PLA/CS islands shape multi-stage nano fiber composite load hedysarum polybotys saccharide periodontium material is obtained, meltblown forming temperature is 180
DEG C, spinning pressure 2MPa, 80 DEG C of environment temperature.
The fibre diameter of PLA/CS islands shape composite fibre is 800-1500nm in periodontium material obtained by the present embodiment,
CS parts are separated, the periodontium material in fibrous outer surfaces, part with irregular structure distribution individually into threadiness with PLA
Material can simulate the microenvironment of periodontal ligament cell growth, and the collagenous fibres diameter of PLA fibers and periodontal membrane tissue is close to (collagen is fine
The diameter of dimension about 200~1500nm), the structure is different from wound bone material (tradition wound bone material simulation bone structure, in order to more
Bone tissue is simulated well, is often prepared to the loose structure of similar cancellous bone.Spinning fibre structure and periodontal prepared by the present invention
Membrane fiber is close to suitable paradenlal tissue regeneration).In the cell propagation obtained by the present embodiment on periodontium material (using deriving from
The bone marrow stroma stem cell of rat, numbering RAWMX-01001) it is significantly faster than that (i.e. cell is directly inoculated in carefully blank control group
Born of the same parents' culture plate), cell mineralization experiments (using the bone marrow stroma stem cell from rat, numbering RAWMX-01001) display
Mineral nodules are more obvious.Therefore, periodontium material obtained by the present embodiment can meet the requirement for inducing periodontium growth.
Embodiment 13
In six cyclopentadiene:The weight ratio of dichloromethane is 1:2 in the mixed solvent adds 1wt% dodecyl sulphate
Sodium, after 3000r/min is emulsified 30 minutes at a high speed, it is that solvent is configured to add with the hexafluoroisopropanol aqueous solution of 60% mass fraction
Mass fraction is 3% chitosan (CS.Mw=2.6 × 105G/mol) solution, 2h is emulsified at a high speed in 5000r/min, is added
PLA (PLA, the M of (80 DEG C, 10h) after 14wt% vacuum dryingw=6 × 105G/mol), 5h is stirred, 4h is stood and obtains PLA/
CS mixed emulsions；5wt% Puerarin solution is added wherein in CS solution process for preparation, forms medicine-carried system；PLA/CS is mixed
Close emulsion and PLA/CS composite cellulosic membranes, the distance between receiver board and syringe needle 20cm, in 35kV electricity are prepared using method of electrostatic spinning
Spinning moulding is depressed, obtains PLA/CS core-skin multi-stage nanos fiber composite load Puerarin periodontium material.
The fibre diameter of PLA/CS core-skin composite fibers is 600-1500nm, CS in periodontium material obtained by the present embodiment
With irregular structure distribution in fibrous outer surfaces, the periodontium material can simulate the microenvironment of periodontal ligament cell growth,
The structure is different from wound bone material, and (tradition wound bone material simulation bone structure, in order to more preferably simulate bone tissue, is often prepared to
The loose structure of similar cancellous bone.Spinning fibre structure prepared by the present invention and periodontal membrane fiber close to suitable periodontium again
It is raw).(using the bone marrow stroma stem cell from rat, compiled in the cell propagation obtained by the present embodiment on periodontium material
Number it is RAWMX-01001) it is significantly faster than that blank control group (i.e. cell is directly inoculated in Tissue Culture Plate), cell mineralization experiments
(using the bone marrow stroma stem cell from rat, numbering RAWMX-01001) display Mineral nodules are more obvious.Therefore,
Periodontium material obtained by the present embodiment can meet the requirement for inducing periodontium growth.
Embodiment 14
1wt% lauryl sodium sulfate is added in dimethylacetamide solvent, 3000r/min is emulsified 30 minutes at a high speed
Afterwards, it is chitosan that solvent is configured to that mass fraction is 3% to add with the hexafluoroisopropanol aqueous solution of 60% mass fraction
(CS.Mw=2.6 × 105G/mol) solution, 2h is emulsified at a high speed in 5000r/min, after addition 14wt% vacuum drying (80 DEG C,
PLA (PLA, M 10h)w=5 × 105G/mol), 5h is stirred, 4h is stood and obtains PLA/CS mixed emulsions；Wherein CS solution is matched somebody with somebody
5wt% fibroblast growth factor is added during system, forms medicine-carried system；PLA/CS mixed emulsions are used into Static Spinning
Silk method prepares PLA/CS composite cellulosic membranes, the distance between receiver board and syringe needle 20cm, the spinning moulding under 25kV voltages, obtains
PLA/CS parallel type multi-stage nanos fiber composite loads fibroblast growth factor periodontium material.
The fibre diameter of PLA/CS parallel composite fibers is 600-1500nm in periodontium material obtained by the present embodiment,
CS and PLA form bilateral fibre structure, are partly present in PLA fiber peripherals with free state, and the periodontium material can be with
The microenvironment of periodontal ligament cell growth is simulated, the structure is different from wound bone material and (tradition wound bone material simulation bone structure, is
More preferable simulation bone tissue, often it is prepared to the loose structure of similar cancellous bone.Spinning fibre structure prepared by the present invention with
Periodontal membrane fiber is close to suitable paradenlal tissue regeneration).In the cell propagation obtained by the present embodiment on periodontium material (using next
Come from the bone marrow stroma stem cell of rat, numbering RAWMX-01001) it is significantly faster than that (i.e. cell is directly inoculated with blank control group
In Tissue Culture Plate), cell mineralization experiments (use the bone marrow stroma stem cell from rat, numbering RAWMX-01001)
Show that Mineral nodules are more obvious.Therefore, periodontium material obtained by the present embodiment can meet to induce periodontium growth
It is required that.
Embodiment 15
1wt% Tween 80 is added in chloroform solvent, 3000r/min after emulsification 30min, is added with 60% mass at a high speed
The hexafluoroisopropanol aqueous solution of fraction is the chitosan (CS.M that the mass fraction that solvent is configured to is 1%w=9 × 107g/mol)
Solution, 2h is emulsified at a high speed in 5000r/min, add PLA (PLA, the M of (80 DEG C, 10h) after 14wt% vacuum dryingw=5 ×
105G/mol), 5h is stirred, 4h is stood and obtains PLA/CS mixed emulsions；5wt% Six-elements are added wherein in CS solution process for preparation
Yellow ball suspension, form medicine-carried system；PLA/CS mixed emulsions are prepared into PLA/CS composite membranes using flash method shaping, obtain PLA/
CS parallel type multi-stage nanos fiber composite loads Liuwei Dihuang Wan periodontium material.
The fibre diameter of PLA/CS parallel composite fibers is 800-2000nm in periodontium material obtained by the present embodiment,
CS and PLA form bilateral fibre structure, are partly present in PLA fiber peripherals with free state, and the periodontium material can be with
The microenvironment of periodontal ligament cell growth is simulated, the structure is different from wound bone material and (tradition wound bone material simulation bone structure, is
More preferable simulation bone tissue, often it is prepared to the loose structure of similar cancellous bone.Spinning fibre structure prepared by the present invention with
Periodontal membrane fiber is close to suitable paradenlal tissue regeneration).In the cell propagation obtained by the present embodiment on periodontium material (using next
Come from the bone marrow stroma stem cell of rat, numbering RAWMX-01001) it is significantly faster than that (i.e. cell is directly inoculated with blank control group
In Tissue Culture Plate), cell mineralization experiments (use the bone marrow stroma stem cell from rat, numbering RAWMX-01001)
Show that Mineral nodules are more obvious.Therefore, periodontium material obtained by the present embodiment can meet to induce periodontium growth
It is required that.
Above-described embodiment is the preferable embodiment of the present invention, but embodiments of the present invention are not by above-described embodiment
Limitation, other any Spirit Essences without departing from the present invention with made under principle change, modification, replacement, combine, simplification,
Equivalent substitute mode is should be, is included within protection scope of the present invention.
Claims (4)
1. a kind of preparation method of multi-stage nano fiber composite medicine-carried periodontium material, it is characterised in that including following preparation
Step：
(1) preparation of polylactic acid fiber membrane or support：PLA is dissolved in solvent, it is 5~40% to be made into mass fraction
PLA solution, method, meltblown, centrifugal spinning, freeze-drying, liquid are then split by method of electrostatic spinning, film and cause split-phase method
Or flash method obtains polylactic acid fiber membrane or support；Described solvent refers to chloroform, dichloromethane, dimethylformamide, diformazan
Mixing more than one or both of yl acetamide, dimethyl sulfoxide (DMSO), hexafluoroisopropanol, six cyclopentadiene and tetrahydrofuran；
(2) recombination chitosan micro-sphere in polylactic acid fiber membrane or support：Dissolve chitosan in mass fraction for 30%~
In one or both of 90% acetic acid, trifluoroacetic acid, trifluoroethanol and hexafluoroisopropanol solvent mixed above, quality is made into
Fraction is 5~30% chitosan solution, then forms chitosan in polylactic acid fiber membrane or rack surface by electrostatic spraying
Microballoon；
(3) loading nano silvery：Nano Silver deionized water suspension blows and beats uniformly simultaneously ultrasonic disperse through air gun, is directly added dropwise in step
(2) resulting materials surface, air-dry, obtain multi-stage nano fiber composite loading nano silvery periodontium material；
In gained multi-stage nano fiber composite loading nano silvery periodontium material, the quality of PLA and chitosan between the two
Fraction is 50%~99% PLA and 1%~50% chitosan.
2. a kind of preparation method of multi-stage nano fiber composite medicine-carried periodontium material according to claim 1, its
It is characterised by：The weight average molecular weight of the PLA is 1 × 104~9 × 107G/mol, the weight average molecular weight of chitosan for 1 ×
104~9 × 107g/mol。
3. a kind of preparation method of multi-stage nano fiber composite medicine-carried periodontium material according to claim 1, its
It is characterised by：Described chitosan solution is additionally added water-soluble load medicine composition or nano particle dissolving or scattered in process for preparation
Uniformly, chitosan-loaded drug solns are obtained；The described water-soluble medicine composition that carries is fibroblast growth factor, tetracycline, A Moxi
At least one of woods, first nitre file, Liuwei Dihuang Wan, Puerarin, hedysarum polybotys saccharide and iron lactoprotein；Described nano particle refers to
At least one of Nano Silver and ZnO.
A kind of 4. multi-stage nano fiber composite medicine-carried periodontium material, it is characterised in that：It is any by claims 1 to 3
Method described in is prepared.
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| WO2017139775A1 (en) * | 2016-02-12 | 2017-08-17 | Tricol Biomedical, Inc. | Chitosan superfine fiber systems |
| CN106390208A (en) * | 2016-10-09 | 2017-02-15 | 华南理工大学 | Three-dimensional support material containing hierarchical porous structures and preparation and application |
| CN109529125B (en) * | 2018-12-28 | 2023-01-24 | 佛山科学技术学院 | Method for preparing biological tissue engineering scaffold by solvent spraying |
| WO2020185714A1 (en) * | 2019-03-08 | 2020-09-17 | Optimed Technology, Inc. | Functionalized prosthetic interfaces for the prevention and treatment of dental conditions |
| CN110344126A (en) * | 2019-06-18 | 2019-10-18 | 浙江理工大学 | A kind of centrifugal spinning preparation method of porous cellulose acetate micro/nano-fibre film |
| CN111110922B (en) * | 2019-12-25 | 2020-10-27 | 四川大学 | Periodontal biological module for 3D biological printing and construction method and application thereof |
| CN111705377A (en) * | 2020-06-17 | 2020-09-25 | 北京天助畅运医疗技术股份有限公司 | Preparation method of microporous core-shell structure degradable nanofiber |
| CN112353780A (en) * | 2020-11-12 | 2021-02-12 | 盐城工学院 | Drug sustained and controlled release platform system with double nano composite structures |
| CN112962220B (en) * | 2021-02-02 | 2022-10-14 | 武汉理工大学 | Polypyrrole chitosan drug-loaded fiber membrane for artificial nerve conduit and preparation method thereof |
| CN113633822B (en) * | 2021-09-14 | 2022-08-23 | 青岛大学 | Polymer nanofiber/microparticle photosolder composite microsphere and preparation method and application thereof |
| CN115814170B (en) * | 2023-01-30 | 2024-03-01 | 安徽医科大学 | Preparation method and application of trans-scale bionic composite prosthesis for guiding periodontal tissue regeneration |
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