CN104644560A - Hyaluronic acid modified hectorite amide nanoparticle and preparation and application of hyaluronic acid modified hectorite amide nanoparticle - Google Patents
Hyaluronic acid modified hectorite amide nanoparticle and preparation and application of hyaluronic acid modified hectorite amide nanoparticle Download PDFInfo
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Abstract
The invention relates to a hyaluronic acid-modified amide hectorite nanoparticle and a preparation and an application of the hyaluronic acid modified hectorite amide nanoparticle. The hyaluronic acid modified hectorite amide nanoparticle is (3-amino propyl) ethoxydimethylsilane modified hectorite; and the mass fraction of the hyaluronic acid in the nanoparticle is 17.17%-19.09%. The preparation method comprises the following steps: modifying a hectorite surface with (3-amino propyl) ethoxydimethylsilane on to obtain hectorite amide; and through covalent action of amino and carboxyl, grafting the hectorite amide surface with hyaluronic acid, thus obtaining the hyaluronic acid modified hectorite amide nanoparticle. The stability and the biocompatibility of the nanoparticle are improved; meanwhile, the hyaluronic acid-modified hectorite amide nanoparticle has specific targeting action on cancer cells with high expression for a CD44 receptor, and can be applied to targeted delivery of anti-cancer drugs; and the hyaluronic acid-modified hectorite amide nanoparticle is simple in preparation method, mild in reaction condition and easy to operate, and has an industrial implementation prospect.
Description
Technical field
The invention belongs to nano drug-carrying Material Field, particularly a kind of hyaluronic acid decorated lithium amide saponite nano-particle and Synthesis and applications thereof.
Background technology
In the numerous disease progression of the mankind, there is the major disease of serious threat human life health---tumor.In the chemotherapy process of tumor, the key affecting its therapeutic effect is that can cancer therapy drug arrive tumor cell and have certain drug level, thus reaches the object suppressing the propagation of cancer cell even to kill cancerous cell.But a large amount of drug molecules makes therapeutic effect be extremely restricted because having the defects such as poorly water-soluble, cell membrane permeater ate be low.Therefore, build the drug delivery system with targeted drug conveying function and become the key solving this difficult problem.
Along with the development of nanotechnology and the appearance of a large amount of novel nano-material, some functionalized nano materials have been used as drug delivery carrier and have demonstrated great advantage.In numerous anti-cancer medicament carrier systems, just becoming current study hotspot using nanoclay hectorite as the medicine-carried system of platform.Hectorite (Laponite, LAP) is a kind of mineral belonging to smectite race, is dispersed in water and can forms flat crystal.Hectorite there is special space sandwich and surface with a large amount of negative charges, can bag medicine carrying thing effectively, in load and the conveying of hydrophobic drug Itraconazole, antitumor drug amycin etc., embody advantage.But find suitable carrier and be also nowhere near, many cancer therapy drugs can not pass cell membrane, even if medicine is loaded onto tumor cell surface by pharmaceutical carrier, still lack effective mechanism of internalization.Along with going deep into of studying tumor cell, researcher finds that tumor cell has the receptor of a series of overexpression, if the part relevant to receptor or antibody to be connected to medicine or pharmaceutical carrier surface, the drug targeting of institute's load can be transported to specific tissue and tumor cell by receptor mediated endocytosis, the specificity of this method and all very high to the affinity of tumor cell, substantially increases targeting and the transport efficacy of medicine.
The higher polysaccharides that hyaluronic acid (HA) is made up of unit D-Glucose aldehydic acid and N-acetyl-glucosamine, be the important component of vertebrates body fluid and tissue, play an important role in some bioprocesss are as repair in trauma, cell proliferation, cell adhesion and cell migration etc.In addition, HA is also a kind of conventional tumor targeted molecular, it can with the combination of the CD44 receptor-specific of some tumor cell surface high expresseds, applied to the aspects such as gene therapy, molecular imaging and drug delivery widely.Compared with the targeting agents such as micromolecule folic acid, hyaluronan molecule is a kind of macromole with good aqueous solubility, modify and can not only give the target function of carrier to tumor cell to the surface of carrier, colloidal stability and the biocompatibility of carrier can also be improved, show unique advantage.
Therefore, utilize the own structural characteristics load antitumor drug doxorubicin hydrochloride of LAP, giving carrier targeting by the mode of finishing HA again, by being expected to the antitumous effect, the reduction toxic and side effects that improve amycin, there is important theory significance and practical value.
Summary of the invention
Technical problem to be solved by this invention is to provide a kind of hyaluronic acid decorated lithium amide saponite nano-particle and Synthesis and applications thereof, this lithium amide saponite nano-particle improves stability and the biocompatibility of nano-particle, to the cancerous cell of CD44 receptor high expressed, there is specific targeting simultaneously, can be applicable to targeted cancer therapy drug; Preparation method is simple, and reaction condition is gentle, is easy to operation, has the prospect of industrialized implementation.
A kind of hyaluronic acid decorated lithium amide saponite nano-particle of the present invention, described lithium amide saponite nano-particle is the hectorite that (3-aminopropyl) dimethylethoxysilane is modified, and in nano-particle, hyaluronic mass fraction is 17.17-19.09%.
The preparation method of a kind of hyaluronic acid decorated lithium amide saponite nano-particle of the present invention, comprising:
(1) with dissolution with solvents hyaluronic acid HA, add activator and stir, activated carboxyl 2-4h, obtains HA solution;
(2) configure hectorite LAP dispersion liquid, get (3-aminopropyl) dimethylethoxysilane and join in LAP dispersion liquid, then 50 ~ 60 DEG C of standing 14-18h, dialysis, obtained lithium amide saponite LM solution;
(3) get the HA solution of gained in (1), add in step (2) gained LM solution, after stirring reaction 70-74h, dialysis, finally obtains hyaluronic acid decorated lithium amide saponite nano-particle LM-HA.
Solvent in described step (1) is ultra-pure water; The concentration of HA solution is 25-27mg/mL.
Activator in described step (1) is 1-ethyl-(3-dimethylaminopropyl) phosphinylidyne diimine EDC and N-hydroxysuccinimide NHS, and the mol ratio of EDC, NHS and carboxylic acid is 5:5:1.
The concentration of the LAP dispersion liquid in described step (2) is 8-10mg/mL, LAP is 2-3:1 with the mass ratio of (3-aminopropyl) dimethylethoxysilane.
The mol ratio of LM and the HA in described step (3) is 1:1.25-1:1.5.
Stirring in described step (1) and (3) is magnetic agitation, and mixing speed is 100-150r/min.
Dialysis in described step (2) and step (3) is distill water dialysis, and bag filter molecular cut off is 8000-14000, and dialysis time is 3d.
The application of a kind of hyaluronic acid decorated lithium amide saponite nano-particle of the present invention, amycin DOX aqueous solution is added in hyaluronic acid decorated lithium amide saponite nano-particle aqueous solution, stir, centrifugal, wash and disperse, obtaining the hyaluronic acid decorated lithium amide saponite nano-particle LM-HA/DOX of load amycin; Wherein, the concentration of amycin DOX aqueous solution is 1-2mg/mL, and hyaluronic acid decorated lithium amide saponite nano-particle and the mass ratio of amycin are 4:1-2:1.
Described stirring is magnetic agitation, and mixing time is 12-24h, and mixing speed is 100-150r/min; Described centrifugation rate is 5000r/min, and centrifugation time is 10min.
In the LM-HA/DOX obtained, LM-HA nano-particle is 67.2% to the load efficiency of DOX.
The present invention adopts silane coupler to modify LAP surface and obtains lithium amide saponite, then obtains hyaluronic acid decorated lithium amide saponite LM-HA by chemical bonding HA.This carrier not only by hyaluronic targeting by tumor cell initiatively picked-up, can extend carrier circulation time in vivo simultaneously, improve drug release characteristics.LM-HA carrier of the present invention is used for antitumor drug load, has good Targeting Effect, can be used as desirable targeted nano carrier to the tumor cell of CD44 receptor high expressed, realizes load and the targeted of medicine, gene or diagnosis molecular probe.
The LM-HA nano-particle that the present invention uses the method such as thermogravimetric analysis, infrared analysis to characterize the present invention to prepare, ultraviolet-visible spectrum test can verify the successful load of medicine.Check toxicity and the targeting of the cervical cancer cell (Hela cell) of LM-HA/DOX effects on surface CD44 receptor high expressed of the present invention with MTT cytotoxicity method, flow cytometry and laser confocal microscope simultaneously.Concrete test result is as follows:
(1) thermogravimetric analysis
Can find out at accompanying drawing 1, within the scope of 200-600 DEG C, before modifying, the weight loss of nano-particle (LAP) is 5.09%.After modification, the weight loss of LM and LM-HA is 10.59% and 22.93 respectively.Weightlessness in this temperature range mainly comes from the thermal decomposition of organic molecule, and therefore the weightlessness of LM and LM-HA increases proves that silane coupled agent molecule and hyaluronan molecule all successfully modify hectorite surface.
(2) infrared analysis
In the infrared test result shown in accompanying drawing 2,1260cm
-1place is the stretching vibration of Si-O key in LM, proves LAP success amination.The FT-IR collection of illustrative plates of LM-HA, the characteristic peak of HA appears at 1615cm
-1(amino-compound I vibration peak), 1413cm
-1(the midplane extrusion vibration peak of carboxyl), 1152cm
-1(the skeletal vibration peak of acetal) and 1043cm
-1(C-O stretching vibration peak), proves that HA is successfully connected on LM.
(3) ultraviolet-visible spectrum test
In the ultraviolet-visible spectrum test result shown in accompanying drawing 3, before medicine carrying, LM-HA does not all have obvious absworption peak in 200 to 800nm wave-length coverages.After load DOX, material demonstrates an obvious absworption peak near 480nm.According to bibliographical information, this absworption peak is the ultraviolet characteristic absorption peak of DOX.Therefore, the hectorite nano-particle of finishing still can load antitumor drug DOX.By UV-quantitative analysis, be under the condition of LM-HA:DOX=3:1 at rate of charge, the medicine carrying efficiency of LM-HA is 67.2%.
(4) extracorporeal releasing test
Accompanying drawing 4 is the cumulative in vitro release profiles of LM-HA/DOX.In 168 hours, it is high that DOX adds up release ratio Cumulative release amount in the neutral environment of pH=7.4 in the weak acid environment of pH=5.4 from LM-HA/DOX.This illustrates that the medicine-carried system based on hectorite all has pH response, namely the rate of release in the weak acid environment similar with tumor tissues is fast, preparation is high, and rate of release is slow in the neutral environment of normal structure, preparation is low, illustrates that this system effectively can reduce the release of DOX in vivo at normal structure place in cyclic process, medicine is concentrated on the release of tumor tissues position, decrease the toxicity of DOX normal tissue, improve the inhibition to tumor cell malignant proliferation.
(5) biocompatibility and antitumous effect evaluation
For the biocompatibility of evaluation carrier, select the cervical cancer cell Hela cell of CD44 receptor high expressed as model, by the cell viability after the LM-HA nano-particle Dual culture of MTT colorimetric method for determining Hela cell and variable concentrations, as Fig. 5 a.Compared with PBS matched group, LM-HA does not have significant difference (cell survival rate is all more than 88%) to the survival rate of cell within the scope of experimental concentration 1.5 to 96 μ g/mL, shows that material has good biocompatibility.In order to verify the antitumous effect of drug-loading nanoparticles, by MTT colorimetric method for determining with the vigor of the Hela cell of simple DOX and the LM-HA/DOX Dual culture 24h of variable concentrations, as Fig. 5 b.Under identical drug level condition, the cytoactive through the Hela of LM-HA/DOX process is starkly lower than the cytoactive of the Hela of pure medicine DOX process.This result shows that targeting material LM-HA/DOX of the present invention has good biocompatibility and is better than the antitumous effect of naked medicine.
(6) cytophagy experiment
The targeting of the nano-particle prepared for model cell evaluation with the cervical cancer cell Hela cell of CD44 receptor high expressed, with concentration be simultaneously the Hela cell of the pure HA Dual culture 2h of 1mM in contrast.
The confocal microscopy of Hela cell respectively after LM-HA/DOX, HA treated+LM-HA/DOX and PBS and pure DOX process, as Fig. 6.The wherein dyed display blue-fluorescence of nucleus, DOX has red fluorescence.The Hela cell contrasting pure DOX and LM-HA/DOX process finds, the Hela cell of LM-HA/DOX process has stronger red fluorescence, shows that the medicine wrapped up through carrier is more easily by cellular uptake.Meanwhile, compared with the Hela cell of HA treated+LM-HA/DOX process, the Hela cell of LM-HA/DOX process has stronger red fluorescence, shows LM-HA/DOX more easily by Hela cellular uptake that CD44 expresses.The flow cytomery of Hela cell after LM-HA/DOX, HA treated+LM-HA/DOX and PBS and pure DOX process, as Fig. 7.Compared with the Hela cell of pure DOX process (Fig. 7 b), the Hela cell (Fig. 7 d) of LM-HA/DOX process has the fluorescent value significantly strengthened, and illustrates that the medicine wrapped up through carrier is more easily by cellular uptake.Hela cell after free hyaluronic acid Dual culture and LM-HA/DOX cultivate (Fig. 7 c), then the fluorescence intensity in cell obviously reduces.This illustrates that the picked-up of cell to medicine carrying microgranule obviously reduces, and LM-HA/DOX enters the approach of cell mainly by the CD44 receptor of Hela cell surface high expressed and the direct specific recognition reaction of the hyaluronic acid of carrier surface.Confocal microscopy picture and Flow cytometry data absolutely prove that hyaluronic acid decorated carrier material significantly improves the specific binding capacity of drug delivery system, promote that cancerous cell is to the picked-up of medicine carrying complex, imparts the characteristic of medicine-carried system active targeting tumor.
beneficial effect
Invention increases stability and the biocompatibility of nano-particle, to the cancerous cell of CD44 receptor high expressed, there is specific targeting simultaneously, can be applicable to targeted cancer therapy drug; Preparation method is simple, and reaction condition is gentle, is easy to operation, has the prospect of industrialized implementation.
Accompanying drawing explanation
Fig. 1 is the thermogravimetric analysis figure of LAP, LM and LM-HA;
Fig. 2 is the infrared structure analysis of LM, LM-HA and HA;
Fig. 3 is the uv absorption spectra of LM-HA, LM-HA/DOX and DOX;
Fig. 4 is the cumulative release curve that DOX discharges from LM-HA/DOX under condition of different pH;
Fig. 5 is that mtt assay tests the cell viability of Hela cell (a) after the LM-HA process 24h of variable concentrations obtained; (b) cell viability after DOX and the LM-HA/DOX process 24h of variable concentrations (* p < 0.05, * * p < 0.005, * * * p<0.001);
Fig. 6 is the Laser Scanning Confocal Microscope picture (concentration of medicine group DOX is 0.5 μ g/mL) after Hela cell and PBS, DOX, LM-HA/DOX, HA-treated+LM-HA/DOX Dual culture 4h;
Fig. 7 is Hela cell and (a) PBS, (b) DOX, (c) LM-HA/DOX, and the flow cytometer fluorescence intensity displacement diagram after (d) HA-treated+LM-HA/DOX Dual culture 4h, (e) is the average fluorescent strength of cell in above-mentioned each group of material.
Detailed description of the invention
Below in conjunction with specific embodiment, set forth the present invention further.Should be understood that these embodiments are only not used in for illustration of the present invention to limit the scope of the invention.In addition should be understood that those skilled in the art can make various changes or modifications the present invention, and these equivalent form of values fall within the application's appended claims limited range equally after the content of having read the present invention's instruction.
Embodiment 1
(1) hyaluronic acid (HA) (being purchased from Dong Yuan bio tech ltd, Zhenjiang) of 518mg is dissolved with the ultra-pure water of 20mL, add 1-ethyl-(3-dimethylaminopropyl) phosphinylidyne diimine (EDC) that 1mL concentration is 16.9mg/mL and stir 0.5h, then adding 1mL concentration is that 10.1mg/mL N-hydroxysuccinimide (NHS) activates 3h;
(2) LAP powder is got, be dissolved in ultra-pure water, obtained concentration is the dispersion liquid of 10mg/mL, getting 40 μ L (3-aminopropyl) vibration limit, dimethylethoxysilane limit is added dropwise in 10mL LAP dispersion liquid, then 50 DEG C of standing 16h, by product with 8000-14000 bag filter at distill water dialysis 72h, obtained amidized hectorite (LM) solution.
(3) the HA solution of activation is made into the solution that concentration is 2mg/mL.Get 1mL and add the EDC that 1mL concentration is 2.2mg/mL wherein, then add the NHS solution that 1mL concentration is 1.32mg/mL, magnetic agitation 3h.Slowly add the lithium amide saponite LM aqueous solution that 1mL concentration is 4mg/mL, magnetic agitation reacts 3 days.After reaction terminates, the LM-HA solution obtained all being transferred to molecular cut off size is in the bag filter of 8000-14000.Using distill water dialysis 72h.After dialysis terminates, by transferred product in bag filter to 15mL centrifuge tube, 4 DEG C of preservations.
Embodiment 2
Get the LM-HA aqueous solution that 1mL concentration is 3mg/mL, add the DOX aqueous solution that 1mL concentration is 1mg/mL, at the condition lower magnetic force stirring reaction 24h of lucifuge, after reaction terminates, solution is transferred to respectively in 15mL centrifuge tube, centrifugal under rotating speed is 5000rpm condition (10min), the precipitate with deionized water obtained washs 3 times, can obtain drug-loading nanoparticles LM-HA/DOX.
Embodiment 3
LM-HA/DOX is dispersed into the buffer of pH=7.4 and pH=5.4 the solution that concentration is the 1mg/mL concentration of DOX (in the LM-HA/DOX) respectively, get 1mL put into bag filter fix, be placed in the container of the buffer containing the different pH of 9mL, be placed in 37 DEG C of shaking tables and vibrate.30min, 1h, 2h, 4h, 6h, 9h and 12h spot sampling started, gets a sample every 24h later.Get the outer liquid 1mL of bag filter at every turn, then outside bag filter, add corresponding buffer solution 1mL.Measure the dialysis solution that takes out at 480nm place absorbance, calculate the release profiles that DOX under external condition of different pH discharges from LM-HA/DOX.(Fig. 4)
Embodiment 4
Collect exponential phase Hela cell, actual conditions is: Hela cell adopts DMEM culture medium, and culture medium is all containing 10% hyclone, and 1% is dual anti-.Incubator temperature is 37 DEG C, and titanium dioxide concentration is 5%.Get the certain density PBS solution of preparation, and to spend the night sterilization with ultra-vioket radiation.Then LM-HA and the nano granule suspension of 1.5,3.0,6.0,12,24,48 and 96 μ g/mL is respectively at superclean bench fresh culture compound concentration.Hela cell seeding after 96 orifice plates respectively with fresh culture (concentration is 1.5,3.0,6.0,12,24,48 and 96 μ g/mL) the Dual culture 24h at 37 DEG C containing LM-HA nano-particle.Then, 20 μ L MTT (5mg/mL) are added in cultivation plate hole, after continuing to cultivate 4h at 37 DEG C, discard culture fluid, and add 200 μ L DMSO, vibrate after crystalline solid being dissolved completely in 15 minutes, use enzyme-linked immunosorbent assay instrument to measure the light absorption value of each hole solution at 570nm place, and calculate the vigor of cell according to this value.The impact of the material on cell proliferation of variable concentrations with buffer PBS for matched group compares.(Fig. 5 a)
Only relevant with the antitumor drug DOX of its load in order to prove the antitumor drug effect of synthesized LM-HA/DOX complex.Preparation is containing the material of different DOX concentration (0.1,0.2,0.5,1.0,2.0,4.0 and 8.0 μ g/mL) LM-HA/DOX and pure DOX.Hela cell seeding after 96 orifice plates respectively with fresh culture (containing DOX concentration 0.1,0.2,0.5,1.0,2.0,4.0 and 8.0 μ g/mL) the Dual culture 24h at 37 DEG C containing LM-HA/DOX and pure DOX nano-particle.Then, 20 μ L MTT (5mg/mL) are added in cultivation plate hole, after continuing to cultivate 4h at 37 DEG C, discard culture fluid, and add 200 μ L DMSO, vibrate after crystalline solid being dissolved completely in 15 minutes, use enzyme-linked immunosorbent assay instrument to measure the light absorption value of each hole solution at 570nm place, and calculate the vigor of cell according to this value.The impact of the material on cell proliferation of variable concentrations with buffer PBS for matched group compares.(Fig. 5 b)
Embodiment 5
Collect exponential phase Hela cell, being positioned over by coverslip in 12 porocyte culture plates and soaking 12h with fresh DMEM medium, after then discarding culture medium, every hole supplements 1.0mL fresh culture and inoculates 5 × 10
4individual Hela cell, incubated overnight cell, culture medium is discarded after adherent, cell more respectively with containing PBS, material LM-HA/DOX, culture medium Dual culture 4h at 37 DEG C of HA treated+LM-HA/DOX and pure DOX (DOX concentration is 0.5 μ g/mL) nano-particle, then three times are cleaned with PBS, then use that 2.5% glutaraldehyde (0.5mL) fixes 15 minutes, DAPI (0.8mL) dyes 8 minutes successively, finally coverslip is positioned on microscope slide, by the pattern of oily mirror (63 ×) observation of cell.(Fig. 6) LM-HA/DOX, HA treated+LM-HA/DOX and pure DOX is mixed with two kinds of nano granule suspensions that DOX concentration is 0.5 μ g/mL respectively with fresh cell medium.Hela cell is first with 2 × 10
5the density in/hole is planted in 12 orifice plates, after incubated overnight cell attachment, first discard culture fluid, then cell more respectively with LM-HA/DOX, culture medium solution (DOX concentration is 0.5 μ g/mL) the Dual culture 4h at 37 DEG C of HA treated+LM-HA/DOX and pure DOX nano-particle, and with the cell of PBS process as a control group.After Dual culture, cell PBS cleans three times, more centrifugal with trypsinization, abandons supernatant, by cell suspension in 1mL PBS.By the average fluorescent strength after flow cytomery cytophagy nano-particle.(Fig. 7)
Claims (10)
1. a hyaluronic acid decorated lithium amide saponite nano-particle, it is characterized in that: described lithium amide saponite nano-particle is the hectorite that (3-aminopropyl) dimethylethoxysilane is modified, and in nano-particle, hyaluronic mass fraction is 17.17-19.09%.
2. a preparation method for hyaluronic acid decorated lithium amide saponite nano-particle, comprising:
(1) with dissolution with solvents hyaluronic acid HA, add activator and stir, activated carboxyl 2-4h, obtains HA solution;
(2) configure hectorite LAP dispersion liquid, get (3-aminopropyl) dimethylethoxysilane and join in LAP dispersion liquid, then 50 ~ 60 DEG C of standing 14-18h, dialysis, obtained lithium amide saponite LM solution;
(3) get the HA solution of gained in (1), add in step (2) gained LM solution, after stirring reaction 70-74h, dialysis, finally obtains hyaluronic acid decorated lithium amide saponite nano-particle LM-HA.
3. the preparation method of a kind of hyaluronic acid decorated lithium amide saponite nano-particle according to claim 2, is characterized in that: the solvent in described step (1) is ultra-pure water; The concentration of HA solution is 25-27mg/mL.
4. the preparation method of a kind of hyaluronic acid decorated lithium amide saponite nano-particle according to claim 2, it is characterized in that: the activator in described step (1) is 1-ethyl-(3-dimethylaminopropyl) phosphinylidyne diimine EDC and N-hydroxysuccinimide NHS, and the mol ratio of EDC, NHS and carboxyl is 5:5:1.
5. the preparation method of a kind of hyaluronic acid decorated lithium amide saponite nano-particle according to claim 2, it is characterized in that: the concentration of the LAP dispersion liquid in described step (2) is 8-10mg/mL, LAP is 2-3:1 with the mass ratio of (3-aminopropyl) dimethylethoxysilane.
6. the preparation method of a kind of hyaluronic acid decorated lithium amide saponite nano-particle according to claim 2, is characterized in that: the mol ratio of LM and the HA in described step (3) is 1:1.25-1:1.5.
7. the preparation method of a kind of hyaluronic acid decorated lithium amide saponite nano-particle according to claim 2, is characterized in that: the stirring in described step (1) and (3) is magnetic agitation, and mixing speed is 100-150r/min.
8. the preparation method of a kind of hyaluronic acid decorated lithium amide saponite nano-particle according to claim 2, it is characterized in that: the dialysis in described step (2) and step (3) is distill water dialysis, bag filter molecular cut off is 8000-14000, and dialysis time is 3d.
9. the application of a hyaluronic acid decorated lithium amide saponite nano-particle as claimed in claim 1, it is characterized in that: amycin DOX aqueous solution is added in hyaluronic acid decorated lithium amide saponite nano-particle aqueous solution, stir, centrifugal, wash and disperse, obtaining the hyaluronic acid decorated lithium amide saponite nano-particle LM-HA/DOX of load amycin; Wherein, the concentration of amycin DOX aqueous solution is 1-2mg/mL, and hyaluronic acid decorated lithium amide saponite nano-particle and the mass ratio of amycin are 4:1-2:1.
10. the application of a kind of hyaluronic acid decorated lithium amide saponite nano-particle according to claim 9, it is characterized in that: described stirring is magnetic agitation, mixing time is 12-24h, and mixing speed is 100-150r/min; Described centrifugation rate is 5000r/min, and centrifugation time is 10min.
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| CN109045305A (en) * | 2018-07-31 | 2018-12-21 | 东华大学 | A kind of preparation method of the hectorite nano particle of TPGS modification |
| CN109045305B (en) * | 2018-07-31 | 2021-12-10 | 东华大学 | Preparation method of TPGS-modified hectorite nanoparticles |
| CN110013559A (en) * | 2019-05-14 | 2019-07-16 | 东华大学 | A kind of extra small ferrum nano material of double-metal hydroxide-of HA targeting and its preparation and application |
| CN110013559B (en) * | 2019-05-14 | 2021-08-31 | 东华大学 | HA-targeted bimetallic hydroxide-ultra-small iron nano material and preparation and application thereof |
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