CN103751851A - Preparation method of inorganic/organic multi-drug controlled release composite nano fiber scaffold - Google Patents
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Abstract
The invention relates to a preparation method of inorganic/organic multi-drug controlled release composite nano fiber scaffold, which includes the steps of preparing drug loading nanoparticle with particle size of 5-500 nm, preparing mixed solution of high molecular polymer and drug, adding multiple drug loading nanoparticles into the polymer solution, ultrasonically stirring, and carrying out electrostatic spinning to obtain inorganic/organic multi-drug controlled release composite nano fiber scaffold with diameter of 50-800 nm. The preparation method provided by the invention has the advantages of mild reaction conditions, simple operation, and easy experimental apparatus, and is in favor or large-scale production; the obtained multi-drug controlled release composite nano fiber scaffold has good stability and good biocompatibility and biodegradability, and can be stored for a long period of time, and the degradation product is non-toxic. The inorganic/organic multi-drug controlled release composite nano fiber scaffold can be widely applied in the fields of tissue engineering, biomedical engineering, drug therapy and the like.
Description
Technical field
The invention belongs to many medicine controlled releasings composite nano fiber scaffold field, particularly a kind of preparation method of inorganic/organic many medicine controlled releasings composite nano fiber scaffold.
Background technology
Along with scientific and technical progress, the development of pharmaceutics has entered brand-new epoch.In order to control better drug dose, to improve utilization ratio of drug and reduce poisonous side effect of medicine, effectively treat disease and can alleviate again patient suffering, the future development that various kinds of drug preparation is discharging towards storage-stable, control.In medicine controlled releasing system, except medicine itself, pharmaceutical carrier is also being played the part of important role, and the form of medicine controlled release carrier mainly comprises nanoparticle and nanofiber etc.Medicine, bioactie agent etc. are entered to the inside of nanoparticle or fiber by the mode of blend, absorption or parcel, can prepare novel medicine controlled release carrier.Medicine-carried nano particles or nanofiber can effectively be controlled the action time of drug release, prolong drug, see through biological barrier targeted medicine, set up new route of administration etc., and this demonstrates the incomparable advantage of other delivery systems aspect medicine controlled releasing.
Desirable drug controlled release system must possess following two kinds of advantages: be to reduce administration number of times or administration frequency on the one hand, alleviate patient suffering; Be the stability that can increase medicine on the other hand, reduce the half-life, extend action time, improve therapeutic effect.And single medicine slow-releasing system exists problems, as in organizational project, the object of tissue is successfully repaired in the very difficult realization of monofactor or medicine, often needs to use collaborative the multiple factor, and effect just can be significantly like this; In clinical, the use of curative drug is also like this, some disease is some inflammation or tumor especially, experiencing after initial effectively treatment, still recurrence unavoidably, this one of them very important reason is exactly that antibacterial or tumor cell have produced drug resistance to single curative drug, and this just means the effect that rational therapeutic alliance can be brought into play to greatest extent medicine and kills harmful cell or antibacterial, and this is that single medicine slow-releasing system cannot be realized.Many drug controlled release systems can have been given play to best drug effect, not only can deliver fat-soluble medicine, also can deliver water soluble drug; Both can load curative drug, also loading functional medicine simultaneously, these need are by the pharmaceutical carrier of reasonably arranging in pairs or groups.The structure of multiple medicines thing medicine-carried system not only depends on the selection of pharmaceutical carrier, also depends on preparation method.Electrostatic spinning technique (abbreviation Static Spinning) is a kind of method of preparing submicron or nanofiber, there is the advantages such as simple and efficient, with low cost, structure is controlled, can prepare the slow releasing carrier of medication that fibre diameter is little, specific surface area is large, porosity is high, in recent years get more and more people's extensive concerning, as Chinese patent (application number 03137309) discloses a kind of method and apparatus that utilizes electrostatic spinning to prepare tissue engineering bracket material; Chinese patent (application number 200610038663) discloses a kind of water soluble anti-cancer medicine slow-release fiber preparation and preparation method thereof etc.But these patents all just utilize single nanofiber to deliver a kind of medicine, seldom there is the report of three kinds or multi-medicament slow-releasing system.Therefore, utilize electrostatic spinning technique that multi-medicament or medicine-carried nano particles and medicine are prepared into many medicine slow release stents, this support can, according to the different mechanism of morbidity and the different qualities of medicine, be realized the synergism of multiple medicines thing, has very large application prospect.
Summary of the invention
Technical problem to be solved by this invention is to provide a kind of preparation method of inorganic/organic many medicine controlled releasings composite nano fiber scaffold, and the method reaction condition gentleness is easy and simple to handle; Experimental provision is simple and easy, is beneficial to production in enormous quantities; The multiple medicines thing controlled release nanometer fibrous framework good stability obtaining, can preserve for a long time, and have good biocompatibility and biodegradability, and its catabolite is nontoxic; Can be widely used in the fields such as organizational project, biomedical engineering, Drug therapy.
The preparation method of a kind of inorganic/organic many medicine controlled releasings composite nano fiber scaffold of the present invention, comprising:
(1) prepare respectively two kinds or more of medicine-carried nano particles;
Nanoparticle can be bought from other companies, also can be synthetic according to prior art.With regard to synthetic, different nanoparticles have different preparation technologies.Take mesoporous silicon dioxide nano particle that is in the news and Poly(D,L-lactide-co-glycolide nanoparticle as example:
Sub-preparation technology is as follows for medicine carrying mesoporous silicon dioxide nano particle: step 1: prepare mesoporous silicon nanoparticle.The sodium hydroxide of the cetyl trimethyl ammonium bromide of 1 gram and 0.28 gram is dissolved in 480 ml deionized water, as for 80 ℃, stir 1 hour, dropwise add the tetraethyl orthosilicate of 5 milliliters, centrifugal stir 20 hours in 80 ℃ of environment after, water and washing with alcohol, after collection, as for adding in the mixed solution that contains 5 milliliters of hydrochloric acid and 500 milliliters of dehydrated alcohol, at 120 ℃, reflux 24 hours, after centrifugal drying, make mesoporous silicon nanoparticle.Step 2: medicine loads take anticarcinogen amycin as example, amycin is mixed with to the solution of 10 mg/ml, adds 1 gram of mesoporous silicon nanoparticle having prepared in amycin solution, evacuation repeatedly, through fully stirring, centrifugal collection, makes medicine carrying mesoporous silicon nanoparticle;
The preparation technology of Poly(D,L-lactide-co-glycolide medicine-carried nanospheres: the PLGA of preparing 1 milligram of amycin and 10 milligrams is dissolved in 1 milliliter of dichloromethane and is made into PLGA solution, it is in 1% poly-vinyl alcohol solution that PLGA solution is joined to mass fraction, within ultrasonic 2 minutes, form just emulsion, again the first emulsion of preparation is joined in the poly-vinyl alcohol solution that 10 milliliters of mass fractions are 0.2%, form multiple emulsion, stir 4 hours, after centrifugal and lyophilization, make medicine carrying PLGA nanoparticle,
(2) polymer material solution that preparation mass fraction is 5～50%, and add another kind of medicine by 0.5～10% of front a kind of polymer quality;
(3) the two kinds or more of medicine-carried nano particles of being prepared by step (1) add respectively in the polymeric medicine mixed solution of step (2), through ultrasonic, stir, be prepared into even suspension, then carry out electrostatic spinning, obtain inorganic/organic many medicine controlled releasings composite nano fiber scaffold; Wherein, nanoparticle is polymer quality 1～20%.
Nanoparticle in described step (1) is one or more in inorganic non-metallic nanoparticle, organic polymer nanoparticle, metal nanoparticle; Nano-carrier structure is one or several in spheroid, vesicle, liquid pearl; Diameter of carrier is 5～500 nanometers.
Described inorganic non-metallic nanoparticle is one or several in mesoporous silicon nanoparticle, hydroxyapatite nano particle, mesopore bioactive glass, CNT, Graphene; Organic polymer nanoparticle is one or more in poly lactic-co-glycolic acid Nano microsphere, Polylactide Sustained Release Nanospheres, liposome, gelatin nanosphere, microgel; Metal nanoparticle is one or several in ferroso-ferric oxide, golden nanometer particle, Nano silver grain, zinc oxide.
Medicine in described step (1) and (2) is one or more in somatomedin, gene, enzyme, curative drug, antibiotic, vitamin, fluorescent dye.
Described somatomedin is one or several in phosphoric acid sphingol, VEGF, basic fibroblast growth factor, epidermal growth factor; Gene is one or more that disturb in little RNA, messenger RNA, plasmid DNA; Enzyme is one or more in protease, lipase, catalase, amylase; Curative drug is amycin, dexamethasone, lomustine, TANSHINONES, carmustine, dexrazoxane, fludarabine phosphate, capecitabine, paclitaxel, replaces one or more in lucky Austria, oxaliplatin, docetaxel, vinorelbine, elemene, hydroxy camptothecin, fluorescein sodium; Antibiotic is one or more in penicillins, cephalosporins, sulfonamides, glycopeptide class, steroidal anti-inflammatory medicine, NSAID (non-steroidal anti-inflammatory drug); Vitamin is one or more in vitamin A, vitamin B, vitamin C, vitamin D, vitamin E; Fluorescent dye is one or more in Fluorescein isothiocyanate, rhodamine B, Nile red.
Polymeric material in described step (2) is the one in natural material, synthesized polymer material.
Described natural material is animal gelatin, fibroin albumen, collagen protein, fibrin, chitosan, cellulose, chitin, glucosan, chitinous one; Synthesized polymer material is the one of polylactic acid, Poly(D,L-lactide-co-glycolide, polyglycolic acid, polycaprolactone.
Electrostatic spinning process parameter in described step (3) is: voltage is 10-22kV, and fltting speed is 0.3-5ml/h, and receiving range is 14-20cm, and temperature is 20-30 ℃, and humidity is 30-50%.
The present invention mainly utilizes Static Spinning technology to carry out blending and prepare the nano fiber scaffold that is loaded with multiple medicines thing carrying the nanoparticle of different pharmaceutical and macromolecular material, can be used as tissue engineering bracket and drug sustained release system.
beneficial effect
(1) the present invention is easy and simple to handle; Experimental provision is simple and easy, is beneficial to production in enormous quantities;
(2) the short and mild condition of preparation time of the present invention, can not cause release in advance and the inactivation of the bioactive ingredients being carried in microcarrier;
(3) many medicine controlled releasings inorganic/organic nano fibrous framework that prepared by the present invention, can realize many drug synergisms, improves therapeutic effect;
(4) many medicine controlled releasings inorganic/organic nano fibrous framework good stability that prepared by the present invention, can preserve for a long time, and have good biocompatibility and biodegradability.
Accompanying drawing explanation
Fig. 1 is the TEM photo of embodiment 1 products therefrom;
Fig. 2 is the TEM photo of embodiment 3 products therefroms;
Fig. 3 is the SEM photo of embodiment 1 products therefrom;
Fig. 4 is the SEM photo of embodiment 3 products therefroms.
The specific embodiment
Below in conjunction with specific embodiment, further set forth the present invention.Should be understood that these embodiment are only not used in and limit the scope of the invention for the present invention is described.In addition should be understood that those skilled in the art can make various changes or modifications the present invention after having read the content of the present invention's instruction, these equivalent form of values fall within the application's appended claims limited range equally.
Embodiment 1
Be loaded with mesoporous silicon oxide/hydroxyapatite/polylactic acid composite nano fiber scaffold of amycin, hydroxy camptothecin and paclitaxel, for treatment of cancer.
(1) prepare mesoporous silicon dioxide nano particle, to its load amycin;
(2) prepare hydroxyapatite, to its load hydroxy camptothecin;
(3) the hexafluoroisopropanol solution 10ml of the polylactic acid that configuration quality volume ratio is 1:10, and add the third drug taxol by polylactic acid quality 1%;
(4) medicine-carried nano particles of being prepared by step (1), (2), join respectively in the solution of step (3) preparation, wherein, carry the amount of amycin mesoporous silicon oxide with respect to 15% of polylactic acid, add the amount of carrying hydroxy camptothecin hydroxyapatite with respect to 15% of polylactic acid quality.
(5) with syringe, draw this solution, regulation voltage is 18kV, and fltting speed is 3ml/h, receiving range is 15cm, temperature is 25 ℃, and humidity is 30%, makes mesoporous silicon/hydroxyapatite/polylactic acid composite nano fiber scaffold of load amycin, hydroxy camptothecin and paclitaxel three medicines.The charging ratio of this composite nano fiber scaffold amycin, hydroxy camptothecin and paclitaxel is respectively 1.5%, 1.8% and 0.01%, and average fibre diameter is 480nm, and hot strength is 1.4MPa, and elastic modelling quantity is 26.4MPa.
Embodiment 2
Be loaded with the liposome/mesoporous silicon oxide/gelatin-compounded nano fiber scaffold of plasmid DNA, dexamethasone and S1P, for bone tissue restoration.
(1) membrane process is prepared liposome, to its load plasmid DNA;
(2) prepare mesoporous silicon dioxide nano particle, to its load S1P;
(3) the isinglass aqueous solution 10ml that preparation mass volume ratio is 9:10, and in this solution, add another kind of medicine dexamethasone by the quality of isinglass 5%;
(4) the medicament-carried nano body of being prepared by step (1), (2), join respectively in the solution of step (3) preparation, wherein, add the liposome that carries plasmid DNA with respect to 10% of gelatin quality, add the mesoporous silicon amount of carrying S1P with respect to 10% of gelatin quality;
(5) with syringe, draw the mixing material in (4), regulation voltage is 14kV, and fltting speed is 0.3ml/h, receiving range is 15cm, temperature is 30 ℃, and humidity is 50%, makes the three medicine composite gelatin fibrous frameworks that contain plasmid DNA, dexamethasone and S1P;
(6) gained nano fiber scaffold stifling crosslinked 24h in 5% genipin alcohol vapour, the effect that both can reach sterilization degerming can also be cross-linked, and makes gained nanofiber change hydrophobicity support into from original Superhydrophilic.The charging ratio of plasmid DNA, dexamethasone and S1P is respectively 0.5%, 4.5% and 0.01%, and average fibre diameter is 750nm.
Embodiment 3
PLGA nanosphere/mesoporous silicon/fibroin albumen the composite nano fiber scaffold that is loaded with vitamin A, vitamin C and vitamin E, can be used for skin repair.
(1) double emulsion is prepared polylactic-co-glycolic acid nanosphere, and loads vitamin A;
(2) prepare mesoporous silicon dioxide nano particle, to its load vitamin E;
(3) preparation quality percent by volume is 20% silk fibroin water solution 10ml, and in this solution, adds medicine vitamin C by fibroin albumen quality 5%;
(4) medicine-carried nano particles of being prepared by step (1), (2), join respectively in the solution of step (3) preparation, wherein, add and carry vitamin A PLGA nanosphere with respect to 5% of the quality of fibroin albumen, add and carry ground vitamin E mesoporous silicon with respect to 10% of fibroin albumen quality;
(5) with syringe, draw the mixing material in (4), regulation voltage is 15kV, and fltting speed is 0.3ml/h, receiving range is 15cm, temperature is 25 ℃, and humidity is 35%, makes the Multivitamin composite filament fibroin fibrous framework that contains vitamin A, vitamin C and vitamin E;
(6) gained nano fiber scaffold stifling crosslinked 24h in 75% alcohol vapour, the effect that both can reach sterilization degerming can also be cross-linked, and makes gained nanofiber change hydrophobicity support into from original Superhydrophilic.The charging ratio of vitamin A, vitamin C and vitamin E is respectively 0.8%, 1% and 0.5%, and average fibre diameter is 750nm.
Claims (8)
1. a preparation method for inorganic/organic many medicine controlled releasings composite nano fiber scaffold, comprising:
(1) prepare respectively two kinds or more of medicine-carried nano particles;
(2) polymer material solution that preparation mass fraction is 5～50%, and add another kind of medicine by 0.5～10% of front a kind of polymer quality;
(3) the two kinds or more of medicine-carried nano particles of being prepared by step (1) add respectively in the polymeric medicine mixed solution of step (2), through ultrasonic, stir, be prepared into even suspension, then carry out electrostatic spinning, obtain inorganic/organic many medicine controlled releasings composite nano fiber scaffold; Wherein, nanoparticle is polymer quality 1～20%.
2. the preparation method of a kind of inorganic/organic many medicine controlled releasings composite nano fiber scaffold according to claim 1, is characterized in that: the nanoparticle in described step (1) is one or more in inorganic non-metallic nanoparticle, organic polymer nanoparticle, metal nanoparticle; Nano-carrier structure is one or several in spheroid, vesicle, liquid pearl; Diameter of carrier is 5～500 nanometers.
3. the preparation method of a kind of inorganic/organic many medicine controlled releasings composite nano fiber scaffold according to claim 2, is characterized in that: described inorganic non-metallic nanoparticle is one or several in mesoporous silicon nanoparticle, hydroxyapatite nano particle, mesopore bioactive glass, CNT, Graphene; Organic polymer nanoparticle is one or more in poly lactic-co-glycolic acid Nano microsphere, Polylactide Sustained Release Nanospheres, liposome, gelatin nanosphere, microgel; Metal nanoparticle is one or several in ferroso-ferric oxide, golden nanometer particle, Nano silver grain, zinc oxide.
4. the preparation method of a kind of inorganic/organic many medicine controlled releasings composite nano fiber scaffold according to claim 1, is characterized in that: the medicine in described step (1) and (2) is one or more in somatomedin, gene, enzyme, curative drug, antibiotic, vitamin, fluorescent dye.
5. the preparation method of a kind of inorganic/organic many medicine controlled releasings composite nano fiber scaffold according to claim 4, is characterized in that: described somatomedin is one or several in phosphoric acid sphingol, VEGF, basic fibroblast growth factor, epidermal growth factor; Gene is one or more that disturb in little RNA, messenger RNA, plasmid DNA; Enzyme is one or more in protease, lipase, catalase, amylase; Curative drug is amycin, dexamethasone, lomustine, TANSHINONES, carmustine, dexrazoxane, fludarabine phosphate, capecitabine, paclitaxel, replaces one or more in lucky Austria, oxaliplatin, docetaxel, vinorelbine, elemene, hydroxy camptothecin, fluorescein sodium; Antibiotic is one or more in penicillins, cephalosporins, sulfonamides, glycopeptide class, steroidal anti-inflammatory medicine, NSAID (non-steroidal anti-inflammatory drug); Vitamin is one or more in vitamin A, vitamin B, vitamin C, vitamin D, vitamin E; Fluorescent dye is one or more in Fluorescein isothiocyanate, rhodamine B, Nile red.
6. the preparation method of a kind of inorganic/organic many medicine controlled releasings composite nano fiber scaffold according to claim 1, is characterized in that: the polymeric material in described step (2) is the one in natural material, synthesized polymer material.
7. the preparation method of a kind of inorganic/organic many medicine controlled releasings composite nano fiber scaffold according to claim 6, is characterized in that: described natural material is animal gelatin, fibroin albumen, collagen protein, fibrin, chitosan, cellulose, chitin, glucosan, chitinous one; Synthesized polymer material is the one of polylactic acid, Poly(D,L-lactide-co-glycolide, polyglycolic acid, polycaprolactone.
8. the preparation method of a kind of inorganic/organic many medicine controlled releasings composite nano fiber scaffold according to claim 1, it is characterized in that: the electrostatic spinning process parameter in described step (3) is: voltage is 10-22kV, fltting speed is 0.3-5ml/h, receiving range is 14-20cm, temperature is 20-30 ℃, and humidity is 30-50%.
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