CN104288833B - A kind of containing can the composite reactive bone cement and preparation method thereof of slow release drug-loading nanoparticles - Google Patents
A kind of containing can the composite reactive bone cement and preparation method thereof of slow release drug-loading nanoparticles Download PDFInfo
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Abstract
The invention discloses a kind of containing can the composite reactive bone cement of slow release drug-loading nanoparticles, comprising can slow release drug-loading nanoparticles and calcium phosphate bone cement, and this can comprise PEG-b-PLGA polymer drug carrier and hydrophobic drug by slow release drug-loading nanoparticles.The invention also discloses that this contains can the preparation method of composite reactive bone cement of slow release drug-loading nanoparticles, by using Polyethylene Glycol PEG as initiator, high molecular polymer PEG-b-PLGA is obtained as pharmaceutical carrier through ring-opening polymerization, drug-loading nanoparticles is prepared by solvent evaporation method, mixed with calcium phosphate bone cement powder by drug-loading nanoparticles, curing molding obtains this composite reactive bone cement again.Composite reactive bone cement provided by the invention reasonably can reduce " the outburst effect " of medicine after bone cement implants, be conducive to the long-term stability of contained sustained release and patient vertebral body, can be used as the composite tissue engineering support of Bone Defect Repari and treatment integration.
Description
Technical field
The invention belongs to orthopaedics field of tissue engineering technology, be specifically related to a kind of containing can the composite reactive bone cement and preparation method thereof of slow release drug-loading nanoparticles.
Background technology
In recent years, the Cranial defect caused due to industrial accident, vehicle accident, orthopaedic disease etc. presents high sickness rate, in addition China just moves towards aging society at present, and the orthopaedics degenerative disease relevant to aging population constantly increases, and causes the demand of bone reparing biological material increasing.
Bone cement is a kind of bone renovating material for bone surgery, and the more Bone Defect Repari drug release carrier of research mainly contains the strutting system of Porous Hydroxyapatite Ceramic (HAP) and polymer microsphere and calcium phosphate bone cement (CPC) compound both at home and abroad at present.Porous Hydroxyapatite Ceramic (HAP) microsphere or support good biocompatibility, but fragility is large, intensity difference, be difficult to moulding.The strutting system of polymer microsphere and calcium phosphate bone cement (CPC) compound, because calcium phosphate bone cement is converted into hydroxyapatite at room temperature self-curing, can be arbitrarily moulding, biocompatibility is good, be the Bone Defect Repari drug release carrier commonly used clinically, medicine be loaded into wherein implant into body Cranial defect position, bone tissue restoration and reconstruction can not only be promoted, also by the slow releasing of medicine, reach some disease for the treatment of as the object of infection, cancer.
But the porosity of calcium phosphate bone cement firming body compared with low, aperture is little; new bone is difficult to grow into; although can hole diameter enlargement by adding sodium bicarbonate, sodium chloride crystal grain or sucrose etc.; but lack aperture between micropore to link up; bone conduction effect is not good, and porosity increase can cause the strength of materials sharply to decline.The strutting system of polymer microsphere and calcium phosphate bone cement compound is degraded and is slowly unfavorable for bone quickly-healing simultaneously, and drug release rate is uncontrollable.And at present a lot of cancer therapy drug and antibiotic medicine due to poorly water-soluble, preparation more difficult and its application in clinical is extremely restricted, therefore, the successful exploitation with the calcium phosphate bone cement of drug slow release function jumps onto a new step by making existing material property.
Summary of the invention
For solving the problem, on the one hand, the invention provides a kind of containing can the composite reactive bone cement of slow release drug-loading nanoparticles, comprising can slow release drug-loading nanoparticles and calcium phosphate bone cement, describedly can comprise PEG-b-PLGA polymer drug carrier and hydrophobic drug by slow release drug-loading nanoparticles.
Preferably, the mass ratio of described PEG-b-PLGA polymer drug carrier and described hydrophobic drug is 5 ~ 100:1; More preferably, the mass ratio of described PEG-b-PLGA polymer drug carrier and described hydrophobic drug is 10 ~ 50:1.
Preferably, the weight average molecular weight of described PEG-b-PLGA polymer drug carrier is 8,000 ~ 60,000; More preferably, the weight average molecular weight of described PEG-b-PLGA polymer drug carrier is 10,000 ~ 30,000.
Preferably, described hydrophobic drug comprises paclitaxel, Docetaxel, amycin, cisplatin, 5-fluorouracil, camptothecine, ibuprofen, ofloxacin, erythromycin, cefazolin sodium, clindamycin or gentamycin.
Preferably, described can the mass ratio of slow release drug-loading nanoparticles and described calcium phosphate bone cement be 0.25 ~ 2:100; More preferably, described can the mass ratio of slow release drug-loading nanoparticles and described calcium phosphate bone cement be 0.5 ~ 1:100.
Preferably, described can slow release drug-loading nanoparticles comprise can slow release drug-carrying nanometer particle, can slow release medicine carrying gel or can Sustained Release Drug-Carried.
On the other hand, present invention also offers a kind of containing can the preparation method of composite reactive bone cement of slow release drug-loading nanoparticles, comprise the steps:
(1) drug-loading nanoparticles is prepared:
By lactide, Acetic acid, hydroxy-, bimol. cyclic ester, Polyethylene Glycol mixing, and then add catalyst and obtain reaction raw material; By described reaction raw material under the protection of protective gas, under 140 DEG C ~ 170 DEG C oil bath heating, carry out ring-opening polymerization, obtain polymerizate; Be dissolved in the first organic solvent after cooling by described polymerizate, then add precipitant, filtration is precipitated, and gained is precipitated vacuum drying, obtains PEG-b-PLGA polymer drug carrier;
By described PEG-b-PLGA polymer drug carrier and hydrophobic drug mixing, be dissolved in the second organic solvent and form solution, under agitation, described solution is added in polyethylene glycol 1000 vitamin E succinic acid ester aqueous solution, ultrasonic disperse in ice bath, carry out centrifugal again after described second organic solvent of decompression volatilization removing, then be resuspended in deionized water by centrifugal gained precipitation, lyophilization obtains can slow release drug-loading nanoparticles;
(2) preparation is containing can the composite reactive bone cement of slow release drug-loading nanoparticles:
What step (1) obtained can mix with calcium phosphate bone cement powder by slow release drug-loading nanoparticles, mixes well, then adds liquid-phase curing agent, obtain slurry, described slurry is placed in mould, sealing, curing molding, obtains containing can the composite reactive bone cement of slow release drug-loading nanoparticles.
Preferably, in described reaction raw material, the mass fraction of described lactide is 30% ~ 80%, and the mass fraction of Acetic acid, hydroxy-, bimol. cyclic ester is 10% ~ 40%, and the mass fraction of Polyethylene Glycol is 5% ~ 30%, and the mass fraction of catalyst is 0.1% ~ 1%.
Preferably, the weight average molecular weight of described Polyethylene Glycol is 1000 ~ 5000.
Preferably, described catalyst comprises stannous octoate, stannous iso caprylate, organic guanidine, metallic zinc, tributyltin chloride, ferric acetyl acetonade, zinc lactate, nano zine oxide, taurine, ethanol ferrum, normal propyl alcohol ferrum, isopropyl alcohol ferrum or n-butyl alcohol ferrum.
Preferably, the protection of described protective gas is by after repeatedly vacuum outgas, then carries out protective gas displacement and obtain; More preferably, after described vacuum outgas, then carry out the process of protective gas displacement, in triplicate.
Preferably, described protective gas comprises nitrogen or noble gas.
Preferably, the response time of described polyreaction is 6 ~ 18 hours; Preferred, the response time of described polyreaction is 8 ~ 12 hours.
Preferably, described first organic solvent comprises dichloromethane, chloroform, oxolane or DMF.For solvent polymerization product, this polymerizate comprises polymer and unreacted lactide monomer, glycolide monomer and the Polyethylene Glycol of polyreaction generation.
Preferably, described precipitant comprises one or more the combination in ether, normal heptane, petroleum ether, methanol and ethanol.Object is that the polymer that polyreaction is generated forms precipitation, then passes through to filter and be dissolved in unreacted lactide monomer, glycolide monomer and Polyethylene Glycol in the first organic solvent.
Preferably, vacuum drying 12 ~ 48 hours at being deposited in 30 DEG C ~ 50 DEG C described in.
Preferably, the weight average molecular weight of described PEG-b-PLGA polymer drug carrier is 8,000 ~ 60,000; More preferably, the weight average molecular weight of described PEG-b-PLGA polymer drug carrier is 10,000 ~ 30,000.
Preferably, the mass ratio of described PEG-b-PLGA polymer drug carrier and hydrophobic drug is 5 ~ 100:1; More preferably, the mass ratio of described PEG-b-PLGA polymer drug carrier and hydrophobic drug is 10 ~ 50:1.
Preferably, described hydrophobic drug comprises paclitaxel, Docetaxel, amycin, cisplatin, 5-fluorouracil, camptothecine, ibuprofen, ofloxacin, erythromycin, cefazolin sodium, clindamycin or gentamycin.
Preferably, the mass concentration of described polyethylene glycol 1000 vitamin E succinic acid ester aqueous solution is 0.03% ~ 1%.
Preferably, described second organic solvent comprises dichloromethane, chloroform, ethyl acetate or carbon tetrachloride etc.
Preferably, the time of described ultrasonic disperse is 80 ~ 300s; More preferably, the time of described ultrasonic disperse is 100 ~ 150s.
Preferably, described centrifugal rotating speed is 15000 ~ 20000rpm.
Preferably, the described centrifugal time is 10 ~ 20min.
Preferably, described can slow release drug-loading nanoparticles comprise can slow release drug-carrying nanometer particle, can slow release medicine carrying gel or can Sustained Release Drug-Carried.
Preferably, described liquid-phase curing agent comprises at least one in Na2HPO4 solution and citric acid solution.
Preferably, described can the mass ratio of slow release drug-loading nanoparticles and described calcium phosphate bone cement powder be 0.25 ~ 2:100; Preferred, described can the mass ratio of slow release drug-loading nanoparticles and described calcium phosphate bone cement powder be 0.5 ~ 1:100.
Preferably, described slurry is placed in mould, sealing, in 37 DEG C, in 100% humidity environment, curing molding obtains containing can the composite reactive bone cement of slow release drug-loading nanoparticles.
The present invention using Polyethylene Glycol PEG as initiator, a kind of biodegradable high molecular polymer PEG-b-PLGA is synthesized by ring-opening polymerization, re-use polyethylene glycol 1000 vitamin E succinic acid ester TPGS as emulsifying agent, prepare drug-loading nanoparticles by solvent evaporation method.By this high molecular polymer PEG-b-PLGA as pharmaceutical carrier, the drug-loading nanoparticles drug loading prepared is high, size is homogeneous, stable performance, and degradability is good, achieves the object of induction of bone growth.Poly lactic-co-glycolic acid (polylactic-co-glycolicacid, PLGA) be by polylactic acid (polylacticacid, PLA) be polymerized according to a certain percentage with polyglycolic acid (polyglycolicacid, PGA).As the improvement of polylactic acid PLA, PLGA had both remained its advantage---and good biocompatibility, biodegradability and mechanics processability, optimize again degradation speed and the mechanical property of material.And with peg-PLGA, can reduce and be absorbed by reticuloendothelial system and mononuclear phagocyte, thus there is macrocyclic characteristic, increase hydrophilic and the biocompatibility of material.Solve PLGA simultaneously and can produce acidic materials initiation inflammation in degradation process, the problem that too high levels causes calcium phosphate bone cement not solidify.Because containing hydrophilic molecules Polyethylene Glycol in polymer P EG-b-PLGA, the hydrophilic of whole pharmaceutical carrier can be increased, therefore it is more faster than discharging medicine by traditional drug-loading nanoparticles PLGA that the drug-loading nanoparticles that prepared by polymer P EG-b-PLGA discharges medicine, therapeutic effect can be met, and with CPC has better compatibility, in addition, PEG is human non-toxic, and nothing accumulates.Medicine carrying granule is made emulsion oil-in-water, solves cancer therapy drug and antibiotic medicine due to the problem of poorly water-soluble, preparation difficulty, be conducive to drug absorption.
Use the inventive method to prepare containing can the composite reactive bone cement of slow release nanometer drug particles, it can mix with constant weight ratio with calcium phosphate bone cement by slow release drug-loading nanoparticles, reasonably can not only reduce after bone cement implants " the outburst effect " of anticarcinogen or antibiotic medicine, be conducive to contained sustained release, the sufficient time is provided to Bone Defect Repari, and be conducive to the long-term stability of the possible patient vertebral body of long-dated survival, can be used as the composite tissue engineering support of Bone Defect Repari and treatment integration.
Accompanying drawing explanation
In order to be illustrated more clearly in the embodiment of the present invention or technical scheme of the prior art, be briefly described to the accompanying drawing used required in embodiment or description of the prior art below, apparently, accompanying drawing in the following describes is only some embodiments of the present invention, for those of ordinary skill in the art, under the prerequisite not paying creative work, other accompanying drawing can also be obtained according to these accompanying drawings.
Fig. 1 be embodiment of the present invention polymer P EG-b-PLGA hydrogen nuclear magnetic resonance spectrogram (
1hNMR);
Fig. 2 is the field emission scanning electron microscope collection of illustrative plates (FESEM) of embodiment of the present invention carrying docetaxel PEG-b-PLGA nano-particle;
Fig. 3 is particle diameter and the particle size distribution of embodiment of the present invention drug-loading nanoparticles;
Fig. 4 is the Zeta potential of embodiment of the present invention drug-loading nanoparticles;
Fig. 5 is the drug release patterns in vitro of embodiment of the present invention drug-loading nanoparticles and commodity docetaxel;
Fig. 6 is that embodiment of the present invention drug-loading nanoparticles, docetaxel and normal saline are to the cytotoxicity experiment result of human osteosarcoma U-20S cell at 24 hours;
Fig. 7 is that embodiment of the present invention drug-loading nanoparticles, docetaxel and normal saline are to the cytotoxicity experiment result of human osteosarcoma U-20S cell at 48 hours;
Fig. 8 is that embodiment of the present invention drug-loading nanoparticles, docetaxel and normal saline are to the cytotoxicity experiment result of human osteosarcoma U-20S cell at 72 hours;
Fig. 9 is the scanning electron microscope (SEM) photograph that the embodiment of the present invention contains the complex calcium phosphate bone cement of drug-loading nanoparticles.
Detailed description of the invention
To be clearly and completely described technical scheme of the present invention below.Obviously, described embodiment is only the present invention's part embodiment, instead of whole embodiments.Based on the embodiment in the present invention, those of ordinary skill in the art, not making the every other embodiment obtained under creative work prerequisite, belong to the scope of protection of the invention.
embodiment 1
Containing can the preparation method of active bone cement of slow release drug-loading nanoparticles, comprise the steps:
(1) nano-particle of carrying docetaxel is prepared:
By mass percentage take the lactide monomer of 30%, the glycolide monomer of 40% and 29.9% Polyethylene Glycol (M
w=5000) mix, put into polymerization pipe, then add mass percentage be 0.1% tributyltin chloride obtain reaction raw material; Carry out inflated with nitrogen after vacuumize degassing, in triplicate, make to form nitrogen protection environment in polymerization pipe; By polymerization pipe tube sealing, under 150 DEG C of oil bath heating, carry out ring-opening polymerization, react and obtain polymerizate after 6 hours; The polymerizate of gained is dissolved in the first organic solvent dichloromethane after cooling, this polymerizate comprises the polymer that polyreaction generates, unreacted lactide monomer, glycolide monomer and Polyethylene Glycol, then add the polymer that excessive methanol makes polyreaction generate and precipitate; Cross to filter to be dissolved in dichloromethane and be precipitated after unreacted lactide, glycolide monomer and Polyethylene Glycol, gained is deposited in 40 DEG C of vacuum dryings 24 hours, obtain the PEG-b-PLGA polymer drug carrier that weight average molecular weight is 12000.
Fig. 1 is the hydrogen nuclear magnetic resonance spectrogram (1HNMR) of polymer P EG-b-PLGA: methyl (CH in the lactide in PLGA segment
3) signal of hydrogen and methine (CH) hydrogen appears at 1.62ppm (d peak) and 5.21ppm (a peak) place respectively, the signal peak being arranged in 4.62 ~ 4.82ppm (b peak) place then belongs to the methylene (CH of Acetic acid, hydroxy-, bimol. cyclic ester
2) proton.Polyethylene Glycol PEG methylene (CH
2) absworption peak appear at 3.65ppm (c peak) place, this is the characteristic peak of Polyethylene Glycol.Can illustrate that PEG-b-PLGA polymer drug carrier synthesizes successfully from the result of the nmr spectrum of Fig. 1.
The PEG-b-PLGA polymer drug carrier (Mw=12000) and the 1mg Docetaxel that accurately take the above-mentioned preparation of 100mg are dissolved in 8mL second organic solvent dichloromethane, and mix homogeneously forms solution; Under agitation, this solution being joined 120mL mass concentration is in the TPGS aqueous solution of 0.03%, with 25w power ultrasonic dispersion 120s under condition of ice bath, forms emulsion oil-in-water; Again through decompression volatilization removing dichloromethane, under the rotating speed of 20000rpm, centrifugal 15min is precipitated subsequently; By the above-mentioned precipitation of deionized water wash three times, with except emulsion breaker TPGS and free medicine, then be resuspended in 10mL deionized water by gained precipitation, lyophilization obtains the PEG-b-PLGA nanoparticle product of carrying docetaxel.
Fig. 2 is the scanning electron microscope result (FESEM) of carrying docetaxel PEG-b-PLGA nano-particle: as can be seen from the figure, Nanoparticle Size is more homogeneous, spherical in shape, and particle diameter is greatly about about 200nm.
Fig. 3 is particle diameter and the particle size distribution of drug-loading nanoparticles: as can be seen from the figure, particle size distribution is narrower, and particle diameter, greatly about about 200nm, demonstrates the observed result of scanning electron microscope further.
Fig. 4 is the Zeta potential of drug-loading nanoparticles: the Zeta potential of nanoparticle is about-16mV, and the absolute value of surface charge is higher, and between granule, repulsive interaction is comparatively strong, thus stablizes at decentralized photo camber.
Fig. 5 is the drug release patterns in vitro of drug-loading nanoparticles and commodity docetaxel: the medicament slow release curve measuring nanoparticle by dialysis, this embodiment gained drug-carrying nanometer particle of 5mg is scattered in 1mL release medium PBST (phosphate Tween buffer) solution (by 8.5gNaCl, 2.2gNa
2hPO
4, 0.3gNaH
2pO
4, 1.0g tween 80 and 1000mL deionized water composition, obtain through autoclaving) in, formation suspension.Nano-particle suspension is placed in bag filter, seals bag mouth.Airtight bag filter puts into 50mL centrifuge tube, adds 15mLPBST, is placed in water bath with thermostatic control shaking table in 37 DEG C, vibrates under 120rpm rotating speed.In a certain time interval, from centrifuge tube, taking out 10mL solution for analyzing, supplementing the fresh PBST of equivalent simultaneously in centrifuge tube.2mL dichloromethane extraction is added, aqueous phase discarded in the sample collected.The sample that extraction obtains is through being dissolved in 5mL mobile phase acetonitrile: water (50/50, V/V), passes into N
2dichloromethane is volatilized, until solution becomes clarification, frit, then adds mobile phase, is settled to 10mL.Through 0.45 μm of membrane filtration before mobile phase uses, and supersound process.Each sample introduction 20 μ L, the peak area of HPLC working sample.Calculate the amount of the Docetaxel of each release, according to Plotting data drug-loading nanoparticles In-vitro release curves, acquired results is shown in Fig. 5.
HPLC checks that condition is as follows:
Chromatographic column is C-18 post (YMC, 150mm*4.6mm, 5 μm)
Mobile phase: acetonitrile: water (50:50, V/V)
Flow velocity: 1.0mL/min
Column temperature: 30 DEG C
Ultraviolet detection wavelength is 227nm
The linear equation of Docetaxel standard curve is:
A=21.59*C (R
2=0.9998) wherein A is chromatogram peak area; C is sample concentration.
As can be known from Fig. 5, the PEG-b-PLGA nano-particle release profiles of carrying docetaxel is biphase release characteristic and adjoint initial " burst effect ".(An Wante medicine (AventisPharma) contrasts, and can find out that the drug release rate of PEG-b-PLGA nano-particle is moderate, easily meet clinical requirement to adopt widely used Docetaxel preparation docetaxel Taxotere clinically.
Fig. 6 ~ Fig. 8 is drug-loading nanoparticles, docetaxel and normal saline to human osteosarcoma U-20S cell at 24 hours, the cytotoxicity experiment result of 48 hours and 72 hours: adopt 3-(4, 5-dimethylthiazole-2)-2, 5-diphenyltetrazolium bromide bromine salt (MMT) sends out the cytotoxicity measuring this nanoparticle, by human osteosarcoma U-20S cell (ATCC, Rockville, MD) be inoculated in 96 porocyte culture plates, after cell culture 24h is adherent, discard outmoded culture medium, rinse once with PBS (phosphate buffer), add testing sample drug-loading nanoparticles, positive control (docetaxel, An Wante medicine), negative control (normal saline) cultivates 24h respectively, 48h, 72h.After predefined time intervals, discard outmoded culture medium, rinse once with PBS, every hole adds 100 μ L containing the cell culture medium of MTT1mg/mL, 37 DEG C hatch 4h after, discard MTT, every hole adds the dimethyl sulfoxide (DMSO) of 100 μ L, cultivate 2h for dark 37 DEG C, vibration 10min, measures the absorbance of 570nm wavelength by microplate reader.
Result shows, drug-loading nanoparticles has obvious toxicity to osteosarcoma cell, and cytotoxicity is greater than commercial Docetaxel preparation docetaxel.In addition, MTT experiment result illustrates that the toxicity of drug-carrying nanometer particle to U-20S cell has time and concentration dependent.
(2) preparation is containing can the composite reactive bone cement of slow release drug-loading nanoparticles:
The nano-particle of carrying docetaxel step (1) prepared mixes with the mass ratio of 0.25:100 with calcium phosphate bone cement powder, mixes well; Add the Na that mass concentration is 2%
2hPO
4solution, as liquid-phase curing agent, obtains slurry, inserts in glass tubing by this slurry, and two seals; Put into constant temperature 37 DEG C, 100% humidity environment, treat that slurry is shaped completely, after stable, take out, namely obtain containing can the composite reactive bone cement of slow release drug-loading nanoparticles.
Fig. 9 is the scanning electron microscope (SEM) photograph of the complex calcium phosphate bone cement containing drug-loading nanoparticles: as can be seen from the figure, be distributed in medicine carrying uniform particles on calcium phosphate bone cement hole wall, and the inside also has a lot of hole, these holes can continue to maintain the function of material as support, can grow into for cell adhesion and new organization and space is provided, accelerated material is degraded, and accelerates skeleton reparation.
embodiment 2
Containing can the preparation method of active bone cement of slow release drug-loading nanoparticles, comprise the steps:
(1) nano-particle of amycin is carried in preparation:
By mass percentage take the lactide monomer of 35%, the glycolide monomer of 29% and 35% Polyethylene Glycol (Mw=5000) mixing, put into polymerization pipe, then add mass percentage be 1% tributyltin chloride obtain reaction raw material; Carry out inflated with nitrogen after vacuumize degassing, in triplicate, make to form nitrogen protection environment in polymerization pipe; By polymerization pipe tube sealing, under 150 DEG C of oil bath heating, carry out ring-opening polymerization, react and obtain polymerizate after 8 hours; The polymerizate of gained is dissolved in the first organic solvents, chloroform after cooling, this polymerizate comprises the polymer that polyreaction generates, unreacted lactide monomer, glycolide monomer and Polyethylene Glycol, then add the polymer that excessive methanol makes polyreaction generate and precipitate; Cross to filter to be dissolved in dichloromethane and be precipitated after unreacted lactide, glycolide monomer and Polyethylene Glycol, gained is deposited in 30 DEG C of vacuum dryings after 24 hours, obtain the PEG-b-PLGA polymer drug carrier that weight average molecular weight is 8,000.
The PEG-b-PLGA polymer drug carrier (Mw=8,000) and the 20mg amycin that accurately take the above-mentioned preparation of 100mg are dissolved in 8mL second organic solvent chloroform, and mix homogeneously forms solution; Under agitation, this solution being joined 120mL mass concentration is in the TPGS aqueous solution of 0.05%, with 25w power ultrasonic dispersion 80s under condition of ice bath, forms emulsion oil-in-water; Again through decompression volatilization removing dichloromethane, under the rotating speed of 15000rpm, centrifugal 10min is precipitated subsequently; By the above-mentioned precipitation of deionized water wash three times, with except emulsion breaker TPGS and free medicine, then be resuspended in 10mL deionized water by gained precipitation, lyophilization must carry the PEG-b-PLGA nanoparticle product of amycin.
(2) preparation is containing can the composite reactive bone cement of slow release drug-loading nanoparticles:
The nano-particle carrying amycin step (1) prepared mixes with the mass ratio of 2:100 with calcium phosphate bone cement powder, mixes well; Add the Na that mass concentration is 2%
2hPO
4solution, as liquid-phase curing agent, obtains slurry, inserts in glass tubing by this slurry, and two seals; Put into constant temperature 37 DEG C, 100% humidity environment, treat that slurry is shaped completely, after stable, take out, namely obtain containing can the composite reactive bone cement of slow release drug-loading nanoparticles.
embodiment 3
Containing can the preparation method of active bone cement of slow release drug-loading nanoparticles, comprise the steps:
(1) nano-particle of 5-fluorouracil is carried in preparation:
By mass percentage take the lactide monomer of 50%, the glycolide monomer of 40% and 9% Polyethylene Glycol (Mw=2000) mixing, put into polymerization pipe, then add mass percentage be 1% stannous iso caprylate obtain reaction raw material; Carry out inflated with nitrogen after vacuumize degassing, in triplicate, make to form nitrogen protection environment in polymerization pipe; By polymerization pipe tube sealing, under 170 DEG C of oil bath heating, carry out ring-opening polymerization, react and obtain polymerizate after 12 hours; The polymerizate of gained is dissolved in the first organic solvent tetrahydrofuran through cooling, this polymerizate comprises the polymer that polyreaction generates, unreacted lactide monomer, glycolide monomer and Polyethylene Glycol, then add the polymer that excess diethyl ether makes polyreaction generate and precipitate; Cross to filter to be dissolved in dichloromethane and be precipitated after unreacted lactide, glycolide monomer and Polyethylene Glycol, gained is deposited in 40 DEG C of vacuum dryings after 48 hours, obtain the PEG-b-PLGA polymer drug carrier that weight average molecular weight is 36100.
The PEG-b-PLGA polymer drug carrier (Mw=36100) and the 10mg5-fluorouracil that accurately take the above-mentioned preparation of 100mg are dissolved in 8mL second organic solvent ethyl acetate, and mix homogeneously forms solution; Under agitation, this solution being joined 120mL mass concentration is in the TPGS aqueous solution of 0.1%, with 25w power ultrasonic dispersion 100s under condition of ice bath, forms emulsion oil-in-water; Again through decompression volatilization removing dichloromethane, under the rotating speed of 20000rpm, centrifugal 15min is precipitated subsequently, by the above-mentioned precipitation of deionized water wash three times, to remove emulsion breaker TPGS and free medicine, be resuspended in 10mL deionized water by gained precipitation again, lyophilization must carry the PEG-b-PLGA nanoparticle product of 5-fluorouracil.
(2) preparation is containing can the composite reactive bone cement of slow release drug-loading nanoparticles:
The nano-particle carrying 5-fluorouracil step (1) prepared mixes with the mass ratio of 0.5:100 with calcium phosphate bone cement powder, mixes well; Add mass concentration be the sodium citrate aqueous solution of 2% as liquid-phase curing agent, obtain slurry, slurry inserted in glass tubing, two seal; Put into constant temperature 37 DEG C, 100% humidity environment, treat that slurry is shaped completely, after stable, take out, namely obtain containing can the composite reactive bone cement of slow release drug-loading nanoparticles.
embodiment 4
Containing can the preparation method of active bone cement of slow release drug-loading nanoparticles, comprise the steps:
(1) nano-particle of camptothecine is carried in preparation:
By mass percentage take the lactide monomer of 50%, the glycolide monomer of 40% and 9% Polyethylene Glycol (Mw=2000) mixing, put into polymerization pipe, then add mass percentage be 1% stannous iso caprylate obtain reaction raw material; Carry out inflated with nitrogen after vacuumize degassing, in triplicate, make to form nitrogen protection environment in polymerization pipe; By polymerization pipe tube sealing, under 170 DEG C of oil bath heating, carry out ring-opening polymerization, react and obtain polymerizate after 12 hours; The polymerizate of gained is dissolved in the first organic solvent dichloromethane through cooling, this polymerizate comprises the polymer that polyreaction generates, unreacted lactide monomer, glycolide monomer and Polyethylene Glycol, then add the polymer that excess diethyl ether makes polyreaction generate and precipitate; Cross to filter to be dissolved in dichloromethane and be precipitated after unreacted lactide, glycolide monomer and Polyethylene Glycol, gained is deposited in 50 DEG C of vacuum dryings after 48 hours, obtain the PEG-b-PLGA polymer drug carrier that molecular weight is 36100.
The PEG-b-PLGA polymer drug carrier (Mw=36100) and the 5mg camptothecine that accurately take the above-mentioned preparation of 100mg are dissolved in 8mL second organic solvent dichloromethane, and mix homogeneously forms solution; Under agitation, this solution being joined 120mL mass concentration is in the TPGS aqueous solution of 1%, with 25w power ultrasonic dispersion 150s under condition of ice bath, forms emulsion oil-in-water; Again through decompression volatilization removing dichloromethane, under the rotating speed of 20000rpm, centrifugal 15min is precipitated subsequently, by the above-mentioned precipitation of deionized water wash three times, to remove emulsion breaker TPGS and free medicine, be resuspended in 10mL deionized water by gained precipitation again, lyophilization must carry the PEG-b-PLGA nanoparticle product of camptothecine.
(2) preparation is containing can the composite reactive bone cement of slow release drug-loading nanoparticles:
The nano-particle carrying camptothecine step (1) prepared mixes with the mass ratio of 1:100 with calcium phosphate bone cement powder, mixes well; Slurry, as liquid-phase curing agent, is inserted in glass tubing by the sodium citrate solution adding mass concentration 2%, and two seals; Put into constant temperature 37 DEG C, 100% humidity environment, treat that slurry is shaped completely, after stable, take out, namely obtain containing can the composite reactive bone cement of slow release drug-loading nanoparticles.
embodiment 5
Containing can the preparation method of active bone cement of slow release drug-loading nanoparticles, comprise the steps:
(1) nano-particle of ibuprofen is carried in preparation:
By mass percentage get the lactide monomer of 80%, the glycolide monomer of 14% and 5.8% Polyethylene Glycol (Mw=2000) mixing, put into polymerization pipe, then add mass percentage be 0.2% stannous iso caprylate obtain reaction raw material; Carry out inflated with nitrogen after vacuumize degassing, in triplicate, make to form nitrogen protection environment in polymerization pipe; By polymerization pipe tube sealing, under 140 DEG C of oil bath heating, carry out ring-opening polymerization, react and obtain polymerizate after 18 hours; Gained polymerizate is dissolved in the first organic solvent dichloromethane through cooling, this polymerizate comprises the polymer that polyreaction generates, unreacted lactide monomer, glycolide monomer and Polyethylene Glycol, then and add the polymer that excessive oil ether makes polyreaction generate and precipitate; Cross to filter to be dissolved in dichloromethane and be precipitated after unreacted lactide, glycolide monomer and Polyethylene Glycol, gained is deposited in 40 DEG C of vacuum dryings after 12 hours, obtain the PEG-b-PLGA polymer drug carrier that molecular weight is 56800.
The PEG-b-PLGA polymer drug carrier (Mw=56800) and the 10mg ibuprofen that accurately take the above-mentioned preparation of 100mg are dissolved in 8mL second organic solvent dichloromethane, and mix homogeneously forms solution; Under agitation, this solution being joined 120mL mass concentration is in the TPGS aqueous solution of 0.03%, with 25w power ultrasonic dispersion 300s under condition of ice bath, forms emulsion oil-in-water; Again through decompression volatilization removing dichloromethane, under the rotating speed of 20000rpm, centrifugal 15min is precipitated subsequently, by the above-mentioned precipitation of deionized water wash three times, to remove emulsion breaker TPGS and free medicine, be resuspended in 10mL deionized water by gained precipitation again, lyophilization must carry the PEG-b-PLGA nanoparticle product of ibuprofen.
(2) preparation is containing can the composite reactive bone cement of slow release drug-loading nanoparticles:
The nano-particle carrying ibuprofen step (1) prepared mixes with the mass ratio of 1:100 with calcium phosphate bone cement powder, mixes well; Add the Na that mass concentration is 2%
2hPO
4slurry, as liquid-phase curing agent, is inserted in glass tubing by solution, and two seals; Put into constant temperature 37 DEG C, 100% humidity environment, treat that slurry is shaped completely, after stable, take out, namely obtain containing can the composite reactive bone cement of slow release drug-loading nanoparticles.
Use the inventive method to prepare containing can the composite reactive bone cement of slow release nanometer drug particles, reasonably can not only reduce after implanting " the outburst effect " of anticarcinogen or antibiotic medicine, be conducive to contained sustained release, the sufficient time is provided to Bone Defect Repari, and be conducive to the long-term stability of the possible patient vertebral body of long-dated survival, can be used as the composite tissue engineering support of Bone Defect Repari and treatment integration.
Above to invention has been detailed description; its object is to allow the personage being familiar with this art can understand content of the present invention and be implemented; can not limit the scope of the invention with this; the equivalence change that all spirit according to the present invention are done or modification, all should be encompassed in protection scope of the present invention.
Claims (10)
1. one kind containing can the composite reactive bone cement of slow release drug-loading nanoparticles, comprising can slow release drug-loading nanoparticles and calcium phosphate bone cement, it is characterized in that: describedly can comprise PEG-b-PLGA polymer drug carrier and hydrophobic drug by slow release drug-loading nanoparticles.
2. composite reactive bone cement as claimed in claim 1, is characterized in that: the mass ratio of described PEG-b-PLGA polymer drug carrier and described hydrophobic drug is 5 ~ 100:1.
3. composite reactive bone cement as claimed in claim 1, is characterized in that: the weight average molecular weight of described PEG-b-PLGA polymer is 8,000 ~ 60,000.
4. composite reactive bone cement as claimed in claim 1, is characterized in that: described hydrophobic drug comprises paclitaxel, Docetaxel, amycin, cisplatin, 5-fluorouracil, camptothecine, ibuprofen, ofloxacin, erythromycin, cefazolin sodium, clindamycin or gentamycin.
5., containing can the preparation method of composite reactive bone cement of slow release drug-loading nanoparticles, comprise the steps:
(1) preparation can slow release drug-loading nanoparticles:
By lactide, Acetic acid, hydroxy-, bimol. cyclic ester, Polyethylene Glycol mixing, and then add catalyst and obtain reaction raw material; By described reaction raw material under the protection of protective gas, under 140 DEG C ~ 170 DEG C oil bath heating, carry out ring-opening polymerization, obtain polymerizate; Be dissolved in the first organic solvent after cooling by described polymerizate, then add precipitant, filtration is precipitated, and gained is precipitated vacuum drying, obtains PEG-b-PLGA polymer drug carrier;
By described PEG-b-PLGA polymer drug carrier and hydrophobic drug mixing, be dissolved in the second organic solvent and form solution, under agitation, described solution is added in polyethylene glycol 1000 vitamin E succinic acid ester aqueous solution, ultrasonic disperse in ice bath, carry out centrifugal again after described second organic solvent of decompression volatilization removing, then be resuspended in deionized water by centrifugal gained precipitation, lyophilization obtains can slow release drug-loading nanoparticles;
(2) preparation is containing can the composite reactive bone cement of slow release drug-loading nanoparticles:
What step (1) obtained can mix with calcium phosphate bone cement powder by slow release drug-loading nanoparticles, mixes well, then adds liquid-phase curing agent, obtain slurry, described slurry is placed in mould, sealing, curing molding, obtains containing can the composite reactive bone cement of slow release drug-loading nanoparticles.
6. preparation method as claimed in claim 5, it is characterized in that: in described reaction raw material, the mass fraction of described lactide is 30% ~ 80%, the mass fraction of described Acetic acid, hydroxy-, bimol. cyclic ester is 10% ~ 40%, the mass fraction of described Polyethylene Glycol is 5% ~ 30%, and the mass fraction of described catalyst is 0.1% ~ 1%.
7. preparation method as claimed in claim 5, is characterized in that: described catalyst comprises stannous octoate, stannous iso caprylate, organic guanidine, metallic zinc, tributyltin chloride, ferric acetyl acetonade, zinc lactate, nano zine oxide, taurine, ethanol ferrum, normal propyl alcohol ferrum, isopropyl alcohol ferrum or n-butyl alcohol ferrum.
8. preparation method as claimed in claim 5, is characterized in that: described precipitant comprise in ether, normal heptane, petroleum ether, methanol and ethanol one or more.
9. preparation method as claimed in claim 5, is characterized in that: described hydrophobic drug comprises paclitaxel, Docetaxel, amycin, cisplatin, 5-fluorouracil, camptothecine, ibuprofen, ofloxacin, erythromycin, cefazolin sodium, clindamycin or gentamycin.
10. preparation method as claimed in claim 5, is characterized in that: the mass concentration of described polyethylene glycol 1000 vitamin E succinic acid ester aqueous solution is 0.03% ~ 1%.
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| US11331191B2 (en) | 2015-08-12 | 2022-05-17 | Howmedica Osteonics Corp. | Bioactive soft tissue implant and methods of manufacture and use thereof |
| CA2938576A1 (en) * | 2015-08-12 | 2017-02-12 | Howmedica Osteonics Corp. | Methods for forming scaffolds |
| CN105879048B (en) * | 2016-05-10 | 2018-09-21 | 苏州大学张家港工业技术研究院 | The preparation method of functional living being degradable nano particle based on polyaminoacid |
| CN106620856A (en) * | 2016-11-08 | 2017-05-10 | 上海纳米技术及应用国家工程研究中心有限公司 | Chemotherapy-thermotherapy type bone repair material on basis of nano-carriers and preparation and application of chemotherapy-thermotherapy type bone repair material |
| WO2018107243A1 (en) * | 2016-12-16 | 2018-06-21 | Field Orthopaedics Pty Ltd | A medical implant and a method of coating a medical implant |
| CN107158473B (en) * | 2017-05-08 | 2019-12-27 | 上海纳米技术及应用国家工程研究中心有限公司 | Calcium phosphate bone cement embedded with drug-loaded silica plastid and preparation method and application thereof |
| CN108553687B (en) * | 2018-02-06 | 2021-04-27 | 重庆医科大学附属永川医院 | Injection type bone filling material and preparation method and use method thereof |
| CN109054000B (en) * | 2018-07-24 | 2021-08-06 | 中山大学 | Nano drug-loading system based on polysalicylic acid and preparation method and application thereof |
| CN110170068B (en) * | 2019-05-27 | 2023-09-26 | 南通大学附属医院 | Multi-functional medical suture and preparation method thereof |
| CN113181368A (en) * | 2020-01-13 | 2021-07-30 | 上海中医药大学附属龙华医院 | Preparation method and application of biodegradable PLGA-TK-PEG nano drug carrier |
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| CN101618209A (en) * | 2008-06-30 | 2010-01-06 | 上海高科联合生物技术研发有限公司 | Local sustained release preparation for preventing and treating osteomyelitis, preparation method and application thereof |
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