CN114081956A - Polyelectrolyte multilayer film-calcium carbonate nano-drug carrier and preparation method and application thereof - Google Patents
Polyelectrolyte multilayer film-calcium carbonate nano-drug carrier and preparation method and application thereof Download PDFInfo
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Abstract
The polyelectrolyte multilayer film-calcium carbonate nano-drug carrier comprises a core and a shell, wherein the core is nano-sized calcium carbonate particles with a porous structure, the calcium carbonate nanoparticles are used as a supporting core, and polyelectrolyte chitosan/sodium alginate with good biocompatibility is modified on the surfaces of the calcium carbonate nanoparticles by a layer-by-layer self-assembly method to form an ultrathin film structure with good biocompatibility and the functions of protecting the deformation of the core and slowly releasing drugs. According to the invention, by electrostatically adsorbing gold nanoparticles on the polyelectrolyte multilayer film structure, the therapeutic drugs carried by the nano-drug carrier can be effectively controlled and rapidly released under the action of near-infrared laser photo-thermal, diseases can be treated by means of the controllable release of local photo-thermal and chemotherapeutic drugs, and the toxic and side effects on organisms are reduced.
Description
Technical Field
The invention relates to biological medicine, in particular to a polyelectrolyte multilayer film-calcium carbonate nano-drug carrier, a preparation method and application thereof.
Background
Cancer has been a major public health problem worldwide for centuries, threatening human life. However, conventional treatment modalities, including surgical resection, chemotherapy, and radiation therapy, have certain limitations. At present, surgery cannot completely remove all tumor tissues, resulting in a high recurrence rate, chemotherapy often lacks selectivity for cancer cells and causes severe side effects on normal organs, and radiation therapy often damages healthy cells and tissues in the vicinity of cancer cells and causes problems of multidrug resistance of cells. Cancer is a serious health hazard, so that people are eager to find new alternative therapies with enhanced efficacy and reduced adverse effects.
In recent years, nanoparticle-based Drug Delivery Systems (DDS) have attracted increasing attention. Nanoparticles (NPs) have been studied for their high drug loading, tailored release characteristics and various functions. In order to enhance the therapeutic efficacy of the drug. Many novel drug delivery strategies have emerged. Wherein, the polymer drug-loaded nanoparticle has certain advantages. Advantages of polymer drug-loaded nanoparticles include not only effective drug encapsulation but also controlled drug release, wherein drug release can be achieved by external stimuli as well as by the internal microenvironment (e.g., pH and temperature) of the biological tissue itself.
The preparation of polyelectrolyte multilayer film drug-loaded nanoparticles by layer-by-layer self-assembly technology (LBL) has become one of the most common methods for developing multifunctional drug delivery due to its advantages of biodegradability, simple operation process, etc. The layer-by-layer self-assembly technology is firstly researched by Iler et al, and is proposed by Decher et al in 1966, the method is a method for forming polyelectrolyte supermolecule multilayer assembly by utilizing electrostatic attraction and complexation between polyanion and polycation, and multilayer ultrathin organic thin films or hybrid films with different properties are prepared by continuously depositing a dilute polyelectrolyte solution with positive and negative charges onto a charged solid template. Which utilizes charge-charge interactions between the template and the polyelectrolyte monolayer to form a multi-layered nanostructure held together by electrostatic forces. The formation of the LBL system is mainly due to electrostatic interactions, hydrogen bonding, hydrophobic interactions and van der waals forces. LBL has several advantages: (1) the performance of the coating is accurately controlled; (2) environment-friendly, mild in condition and low in manufacturing cost; (3) versatility of being useful for all available surface coatings; (4) obtaining a uniform film with controllable thickness; (5) loading and controlled release of biomolecules/drugs. Polyelectrolyte capsules were introduced in 1998 and have attracted considerable attention as potential drug carriers, and nano-and micron-sized capsules based on LBL polyelectrolyte multilayer films have not only scientific but also technical value, since they have potential application prospects in the fields of medicine, drug delivery, artificial cells or viruses, catalysis, and the like.
Calcium carbonate (CaCO)3) Is one of the most abundant minerals in nature and has three polymorphic forms; calcite, aragonite and vaterite. And calcium carbonate particles in the form of vaterite (CaCO)3) Because of its biocompatibility, large surface area and its ability to rapidly decompose under mild conditions (pH below 6.5). This pH sensitivity opens new possibilities for targeted delivery, as the microenvironment in tumors is usually more acidic than in normal tissues. The high porosity of the vaterite polycrystal determines the high drug loading, which has led to interest in incorporating various bioactive substances, such as proteins, drugs and DNA. Since vaterite is the most unstable phase in calcium carbonate, which gradually recrystallizes to the stable calcite phase after incubation in water-based solutions, such loaded particles release the drug without any external influence. At the same time, the material exhibits high stability in the dry state, enabling long-term storage of the carrier under standard conditions. Biocompatibility testing of the vaterite submicron particles showed no evidence of cytotoxicity, with no effect on in vitro viability or metabolic activity.
Thus, CaCO3The biodegradability, pH sensitivity, and simplicity and cheapness of the manufacturing techniques of the particles provide a prospect for their biomedical applications.
Photothermal therapy (PTT) is a topical treatment used to cure solid cancers or tumors in different parts of the body. During photothermal therapy, the photosensitizer is excited by using electromagnetic radiation and ordinary infrared radiation. The photosensitizer functions to absorb light or energy and further convert it into heat, further causing local hyperthermia, causing ablation of the tumor. Gold Nanoparticles (GNPs) are small gold particles with a diameter of 1nm to 100nm, and GNPs with controllable shape and size are synthesized by different physical methods (microwave, laser ablation, uv irradiation) and chemical methods.
Reference documents:
[1] international cancer research institute: 1930 ten thousand Cancer patients are newly added in 2020 world, 1000 ten thousand Cancer patients are removed (2020), International Agency for Research on Cancer 2020,12
[2]Zehui He,Yongtai Zhang,Nianping Feng.Cell membrane-coated nanosized active targeted drug delivery systems homing to tumor cells:A review[J].Materials Science and Engineering,2020,106(110298):0928-493.
[3]W.Yu,X.He,Z.Yang,X.Yang,et al.Sequentially responsive biomimetic nanoparticles with optimal size in combination with checkpoint blockade for cascade synergetic treatment of breast cancer and lung metastasis[J].Biomaterials,2019,217,119309.
[4]D.Duan,H.Liu,M.Xu,et al,Size-controlled synthesis of drug-loaded zeolitic imidazolate framework in aqueous solution and size effect on their cancer theranostics in vivo,ACS Appl.Mater.Interfaces.,2018,10,42165-42174.
[5]Gao C,Wu Z,Lin Z,et al.Polymeric Capsule Cushioned Leukocyte Membrane Vesicles as a Biomimetic Delivery Platform.Nanoscale,2016,8:3548-3554.
[6]Aryal S,Hu C M,Fang R H,et al.Erythrocyte Membrane-cloaked Polymeric Nanoparticles for Controlled Drug Loading and Release.Nanomedicine,2013,8(8):1271-1280
[7]P.Hraber,C.Kuiken,K.Yusim,Evidence for Human Leukocyte Antigen Heterozygote Advantage Against Hepatitis C Virus Infection.Hepatology,2007,46(6):1713-1721.
[8] (iii) construction of Yanluo, Yanhui, Huyu, Zhao Wei Zhi, (J) inner Mongolia petrochemical, (2021, 47 (04)) 3-6, layer-by-layer self-assembled porous membrane and functionalized coating.
[9] Zhang Yi Ping. design of M cell targeting starch-based layer-by-layer self-assembled microcapsules and targeting controlled release delivery mechanism research [ D ]. university of southern China, 2020.
[10] Nano (Shifeta Naemi Tonateni), layer by layer self-assembly method, a biomimetic rigid biocompatible polyelectrolyte/calcium carbonate film [ D ]. china university of geology (beijing), 2019.
Disclosure of Invention
The invention aims to perform functional design on polyelectrolyte multilayer film nano-particles based on a layer-by-layer self-assembly technology to prepare the polyelectrolyte multilayer film nano-drug-carrying particles with drug-carrying and photothermal treatment functions. So as to overcome the defects of low water solubility, low bioavailability, poor chemical stability and the like of a plurality of medicaments, thereby obtaining better clinical curative effect.
In order to achieve the above purpose, the technical scheme of the invention is as follows:
the preparation method of the polyelectrolyte multilayer film-calcium carbonate nano-drug carrier comprises the following steps:
preparing calcium carbonate nanoparticles;
calcium carbonate nano particles are used as a template core, and are assembled on a template layer by layer through layer-by-layer self-assembly by utilizing the electrostatic adsorption effect between polyanion and polycation, and the steps are repeated for a certain number of times to obtain the polyelectrolyte multilayer film;
soaking the polyelectrolyte multilayer film in a proper solution for crosslinking for a period of time;
gold nanoparticles with a photothermal function are modified on the middle layer of the polyelectrolyte multilayer film by an electrostatic adsorption method to obtain the polyelectrolyte multilayer film-calcium carbonate nano-drug carrier.
Further, the preparation method of the calcium carbonate nanoparticles comprises the following steps:
when the temperature of the system is 40 ℃, the volume ratio of 1: 1 of 0.01-0.1M NaCO3Adding glycerol in the volume ratio of 1: 1 of 0.01-0.1M CaCl2Putting the mixed solution into an ultrasonic machine for ultrasonic treatment for 30 seconds, stirring for 5 minutes at the rotating speed of 500-1000rpm/min by virtue of a magnetic stirring platform to obtain a product, and carrying out fractional centrifugation and collection on the product.
Further, during self-assembly, chitosan with positive charges and sodium alginate with negative charges are used as polyelectrolyte multilayer film synthesis components to synthesize the polyelectrolyte multilayer film adsorbed composite nanoparticles.
Further, the preparation method of the chitosan solution comprises the following steps:
0.2-0.5g of water-soluble chitosan was weighed and dissolved in 2.5M NaCl solution to prepare 100mL of solution for use.
Further, the preparation method of the sodium alginate solution comprises the following steps:
0.2-0.5g of sodium alginate is weighed and dissolved in 100mL of NaCl solution with the concentration of 2.5M to prepare sodium alginate solution for later use.
Further, the method for preparing the polyelectrolyte multilayer film comprises the following steps:
weighing a certain amount of dry calcium carbonate nano particles as a template, alternately adsorbing chitosan and sodium alginate on the surfaces of calcium carbonate porous particles through layer-by-layer self-assembly, washing the calcium carbonate particles with deionized water twice before transferring the calcium carbonate particles from one solution to another solution, and removing the excess polyelectrolyte solution which is not bonded on the surfaces to ensure the uniformity of the membrane; repeating for 4 times to form 4 double-layer chitosan/sodium alginate composite membranes to obtain the polyelectrolyte multilayer membrane.
Further, the synthesis method of the gold nanoparticles comprises the following steps:
heating a 0.01% chloroauric acid solution to boiling, heating 1% sodium citrate and stirring for 15min to recover to room temperature while stirring, finally, centrifuging at 12000rpm for 5min to collect particles, and washing with deionized water for 3 times to obtain gold nanoparticles with a rod-like structure, wherein the gold nanoparticles have negative charges in the solution.
The invention also provides the polyelectrolyte multilayer film-calcium carbonate nano-drug carrier synthesized by the method.
The invention also provides application of the polyelectrolyte multilayer film-calcium carbonate nano-drug carrier in preparation of anti-cancer drugs.
Furthermore, the drug is a fat-soluble or water-soluble broad-spectrum anti-cancer drug wrapped by the polyelectrolyte multilayer film-calcium carbonate nano drug carrier.
Compared with the prior art, the invention has the beneficial technical effects that:
the invention prepares a new polyelectrolyte multilayer film n-calcium carbonate nano drug carrier, the particle size of calcium carbonate nano particles is controllable (10-10000nm), the calcium carbonate nano particles have surface porous structure, and can regulate and control the encapsulation of fat-soluble/water-soluble broad-spectrum anticancer drugs, thus effectively reducing the toxic and side effects of the dissolution-drug delivery through organic solvent in the traditional treatment; through the modification protection effect of the polyelectrolyte multilayer film, the porous structure of calcium carbonate can be effectively maintained, and the amount of the effective drug carried by the calcium carbonate can reach a focus area. The carbonate inner core can stably exist in neutral body fluid such as blood, and the polyelectrolyte multilayer film (CHI/ALG) n-calcium carbonate nano-drug carrier is degraded in an acidic environment in cancer cells after entering a tumor area, and rapidly releases the drug under the stress of carbon dioxide air pressure, so that a high-efficiency treatment effect is generated, and the problems that the drug carrier is slowly released and is easy to generate drug resistance are solved; meanwhile, the modification of the polyelectrolyte multilayer film by the photosensitizer and the gold nanoparticles can synergistically treat diseases through photothermal effect and drug release in a near-infrared region range, and the disease treatment efficiency is improved.
Drawings
FIG. 1 is CaCO3SEM images of nanoparticles;
FIG. 2 is an SEM image of a polyelectrolyte multilayer film;
FIG. 3 is an SEM image of polyelectrolyte multilayer film adsorbed on the surface of drug-loaded calcium carbonate;
FIG. 4a is an SEM image of calcium carbonate nanoparticles unprotected by a polyelectrolyte multilayer film; FIG. 4b is an SEM image of calcium carbonate particles modified with a polyelectrolyte multilayer film;
FIG. 5 is a graph of the ultraviolet absorption spectrum of photosensitizer-gold nanoparticles;
FIG. 6 is a transmission electron micrograph of photosensitizer-gold nanoparticles;
FIG. 7 shows the comparison of drug loading for different templates;
FIG. 8 shows the results of comparative cytotoxicity tests;
FIG. 9 shows the results of in vitro drug efficacy evaluation;
FIG. 10 is a graph of the results of a comparison of the natural permeation rate and the photo-thermal induced release rate, where a is CaCO3B is (CHI/ALG)4@CaCO3。
Detailed Description
Examples
Synthesis of polyelectrolyte multilayer film-calcium carbonate nano-drug carrier
(1) The preparation method of the calcium carbonate nano particles adopts a hydration method:
at a system temperature of 40 ℃, the volume ratio (V: V) of the components is 1: 1 of (0.01-0.1) M NaCO3Adding glycerol in the volume ratio (V: V) of 1: 1 of (0.01-0.1) M CaCl2Putting the mixed solution into an ultrasonic machine for ultrasonic treatment for 30 seconds, stirring for 5 minutes at the rotating speed of (500-.
(2) Calcium carbonate nanoparticle drug loading
Loading of water-soluble drugs: the preparation method comprises the steps of selecting an anticancer mode drug doxorubicin hydrochloride (DOX) as a water-soluble drug, preparing the DOX into an aqueous solution with the concentration of 0.5mg/ml by using pure water, then weighing a certain amount of calcium carbonate nanoparticles, adding the calcium carbonate nanoparticles into the aqueous solution of the DOX, adsorbing the calcium carbonate nanoparticles for 24 hours, and finally washing the calcium carbonate nanoparticles for 2 times by using the pure water to obtain the DOX-loaded calcium carbonate nanoparticles.
② loading of fat-soluble medicine: lipid soluble drugs Paclitaxel (PTX) was chosen as the drug loaded. Dissolving paclitaxel in ethanol to obtain 0.5mg/ml solution, weighing a certain amount of calcium carbonate nanoparticles, adding into PTX solution, and allowing to adsorb for 24 h. And finally washing for 2 times to obtain the PTX-loaded calcium carbonate nano-particles.
(3) Preparation of polyelectrolyte multilayer film: preparing a Chitosan (CHI) solution: accurately weighing 0.2-0.5g of water-soluble chitosan dissolved in 2.5M NaCl solution to prepare 100mL of solution for later use; preparing a sodium Alginate (ALG) solution: weighing (0.2-0.5) g of sodium alginate, dissolving in 100mL of 2.5M NaCl solution, and preparing into sodium alginate solution for later use; thirdly, weighing a certain amount of dried calcium carbonate nano particles as a template, and carrying out LBL technologyAnd (3) alternately adsorbing CHI and ALG of 2mg/ml on the surfaces of the calcium carbonate porous particles, washing the calcium carbonate particles with deionized water twice before transferring the calcium carbonate particles from one solution to the other solution, and removing the excess polyelectrolyte solution which is not bonded on the surfaces to ensure the uniformity of the membrane. Repeating the above steps for 4 times to form 4 double-layer (CHI/ALG) composite membranes, and obtaining polyelectrolyte multilayer membrane adsorbed composite nanoparticles (CHI/ALG @ CaCO)3)4(ii) a And fourthly, crosslinking: will be prepared (CHI/ALG)4And (3) soaking the composite film-calcium carbonate nano particles in a carbodiimide (EDC) solution for crosslinking for 12 h.
(4) Synthesis and use of photosensitizer:
the gold nanoparticles were synthesized using a chemical synthesis method: heating 0.01% chloroauric acid solution to boiling, heating 1% sodium citrate and stirring for 15min to return to room temperature while stirring, finally, centrifuging at 12000rpm for 5min to collect granules, and washing with deionized water for 3 times. The gold nanoparticles prepared by the experiment are of a rod-shaped structure, have negative charges in a solution, and are modified in the middle layer of the polyelectrolyte multilayer film by an electrostatic adsorption method, so that the effective number of photosensitive particles in the polyelectrolyte multilayer film-calcium carbonate nano drug carrier is ensured.
Second, characterization and application data of material
Characterization of calcium carbonate nanoparticles and polyelectrolyte multilayer films:
calcium carbonate nano-particles and polyelectrolyte multilayer film chitosan/sodium alginate [ (PSS/PAH) after corrosion of template particles4)@CaCO3]Morphology of vesicles under a scanning electron microscope, CaCO visible from SEM picture 13The surface of the nano particle is loose and porous, the specific surface area is large, and the size is 400-800 nm. SEM picture 2 polyelectrolyte multilayer film can keep intact vesicle structure after removing the template support, can effectively protect the medicine loss that treatment medicine slowly diffuses in the course of body fluid circulation transportation.
Protection experiment of polyelectrolyte multilayer film on morphology of calcium carbonate nanoparticles:
the polyelectrolyte multilayer film can reduce the osmotic loss of the adsorbed drug in the transportation process in the organism by wrapping the calcium carbonate, and meanwhile, a large number of experiments prove that the porous calcium carbonate can be subjected to shape change in the solution, namely, the porous calcium carbonate is gradually changed into cubic particles with smooth surfaces from a porous circular structure, so that the drug property of the calcium carbonate is greatly reduced, and meanwhile, the adsorbed drug molecules face a large amount of loss. The polyelectrolyte multilayer film is adsorbed on the surface of the medicine-carrying calcium carbonate, so that the morphology of porous calcium carbonate particles can be effectively maintained, and the effective quantity of the medicine carried by the calcium carbonate carrier is ensured (figure 3).
The same amount of calcium carbonate and polyelectrolyte multilayer film protected calcium carbonate [ (PSS/PAH)4)@CaCO3]And dispersing the nano particles in deionized water respectively, keeping shaking for 48 hours at room temperature, centrifugally washing for 2 times, drying the sample, and preparing a scanning electron microscope sample. A scanning electron microscope SEM image 4a shows that the appearance of the calcium carbonate nanoparticles which are not protected by the polyelectrolyte multilayer film is obviously changed, the spherical structure in the sample completely disappears, and the spherical structure is completely changed into a square structure with a smooth surface; fig. 4b shows polyelectrolyte multilayer film-modified calcium carbonate particles with intact spherical structure and no significant change in particle size.
Fig. 5 and fig. 6 are respectively an ultraviolet absorption spectrum chart and a transmission electron microscope chart of the photosensitizer-gold nanoparticle. The gold nanoparticles have different sizes and different ultraviolet absorption peak positions, and fig. 5 shows that the gold nanoparticles have stronger absorption peaks in the near infrared 500-600nm range, and the photosensitive gold nanoparticles are rod-shaped structures, relatively uniform in size and 10nm in particle size, as can be seen from a transmission electron microscope picture.
Comparison of drug loading for different templates
Comparing the drug loading rates of different template particles, calcium carbonate, manganese carbonate and silicon dioxide nanoparticles with the same size, respectively weighing the 3 particles with the same mass, dispersing the particles in a 2mg/ml PTX solution with the same volume, standing and adsorbing for 36h, washing with deionized water for three times to remove a suspended drug solution, and collecting the absorption value of an ultraviolet spectrophotometer at an absorption peak of 227nm after the template particles are corroded. As shown in fig. 7, the highest drug loading of calcium carbonate, the next highest manganese carbonate, and the lowest silica drug loading were confirmed by comparing the absorption values.
Cytotoxicity test:
CaCO3、(CHI/ALG)4@CaCO3the cytotoxicity of (a) was evaluated by an MTT method.
200 μ l of MCF-7 cells were plated in triplicate in 96-well plates to a density of 5000 cells per well, then incubated at 37 ℃ in 5% CO2Is cultured in a cell culture box for 24 hours. The culture medium was removed, and 200. mu.l of CaCO3, (CHI/ALG)4@CaCO3The suspension (1. mu.g/mL to 250. mu.g/mL) was added to the wells, the supernatant removed after 24 hours, washed 2 times with PBS, and 180. mu.L of fresh medium and 5mg/mL of 20. mu.L of LMTT solution were added to each well and incubated for an additional 4 h. Finally, removing the supernatant, adding 200 mul DMSO into each hole, shaking for 10min, detecting the OD value at the wavelength of 570nm by using a microplate reader, and calculating the cell activity.
As shown in FIG. 8, the results of the experiment revealed that even in the case of CaCO3, (CHI/ALG)4@CaCO3At concentrations as high as 250. mu.g/mL, cell viability was greater than 95%, demonstrating CaCO3, (CHI/ALG)4@CaCO3Does not affect the vitality of the cells and the biological safety of the cells.
In vitro efficacy evaluation:
in vitro detection of DOX loaded CaCO by MTT method3And (CHI/ALG)4@CaCO3(1. mu.g/mL to 200. mu.g/mL) killing ability against cancer cells. Firstly, shaking two carrier particles with the same drug loading amount in a PBS (phosphate buffer solution) solution for 3h to simulate the process that the drugs enter a body to participate in body fluid circulation, removing supernate, washing for three times, and co-culturing the gradient of the two drug loading particles with cancer cells for 48h respectively. The results of the experiment demonstrate (fig. 9) that the extent of killing of cancer cells is greater with increasing particle concentration. Furthermore (CHI/ALG)4@CaCO3Relative CaCO3The nano particles have larger killing degree on cancer cells, and further prove that the polyelectrolyte multilayer film can effectively protect the drug loss caused by slow diffusion of the therapeutic drugs in the process of circulating and transporting body fluid.
The calcium carbonate drug carrier wraps the drug small molecules, and the release rate of the calcium carbonate drug carrier is increased along with the increase of the temperature. To contrast the natural permeation rate with the photothermal-induced release rate, theCaCO coated with same mass of adriamycin3And (CHI/ALG)4@CaCO3Dispersing the mixture into a pure water solvent, and periodically detecting the concentration of the adriamycin in the solvent. The adriamycin has obvious ultraviolet absorption peak at about 500nm, and the drug release rate of the carrier particles is judged by the increase of the absorbance of the adriamycin in the solvent phase. As shown in FIG. 10a, CaCO3The diffusion speed of the adriamycin of the nano particles in the pure water solvent is the fastest within 2-5h, the diffusion speed is reduced along with the time extension, and the concentration of the adriamycin of the solvent is basically unchanged after 24 h. As shown in FIG. 10b, coated with the same mass of doxorubicin (CHI/ALG)4@CaCO3Drug micromolecule exudation rate and CaCO in ultrapure water solvent3The nano particles are similar, and the seepage rate is maximum within 2-5 h. The drug release amount is increased instantly after 808nm laser irradiation, the absorbance of the adriamycin in the solvent is maximum (the maximum value is 14.5%), and the concentration of the adriamycin in the solvent is not changed after 24 h.
Claims (10)
1. The preparation method of the polyelectrolyte multilayer film-calcium carbonate nano-drug carrier is characterized by comprising the following steps:
preparing calcium carbonate nanoparticles;
calcium carbonate nano particles are used as a template core, and are assembled on a template layer by layer through layer-by-layer self-assembly by utilizing the electrostatic adsorption effect between polyanion and polycation, and the steps are repeated for a certain number of times to obtain the polyelectrolyte multilayer film;
soaking the polyelectrolyte multilayer film in a proper solution for crosslinking for a period of time;
gold nanoparticles with a photothermal function are modified on the middle layer of the polyelectrolyte multilayer film by an electrostatic adsorption method to obtain the polyelectrolyte multilayer film-calcium carbonate nano-drug carrier.
2. The method according to claim 1, wherein the calcium carbonate nanoparticles are prepared by a method comprising:
when the temperature of the system is 40 ℃, the reaction solution is prepared by mixing the following components in a volume ratio of 1: 1 of 0.01-0.1M NaCO3Adding glycerol in the volume ratio of 1: 1 of 0.01-0.1M CaCl2In the mixed solution of glycerol and water, adding the mixture of glycerol and water,and putting the mixed solution into an ultrasonic machine for ultrasonic treatment for 30 seconds, stirring for 5 minutes at the rotating speed of 500-1000rpm/min by virtue of a magnetic stirring platform to obtain a product, and carrying out fractional centrifugation and collection on the product.
3. The method of claim 1, wherein the polyelectrolyte multilayer film adsorbed composite nanoparticles are synthesized by using positively charged chitosan and negatively charged sodium alginate as polyelectrolyte multilayer film synthesis components during self-assembly.
4. The method of claim 3, wherein the solution formulation of chitosan comprises:
0.2-0.5g of water-soluble chitosan was weighed and dissolved in 2.5M NaCl solution to prepare 100mL of solution for use.
5. The method as claimed in claim 3, wherein the solution of sodium alginate is prepared by the following steps:
0.2-0.5g of sodium alginate is weighed and dissolved in 100mL of NaCl solution with the concentration of 2.5M to prepare sodium alginate solution for later use.
6. The method of claim 1, wherein the method of making the polyelectrolyte multilayer film comprises:
weighing a certain amount of dry calcium carbonate nano particles as a template, alternately adsorbing chitosan and sodium alginate on the surfaces of calcium carbonate porous particles through layer-by-layer self-assembly, washing the calcium carbonate particles with deionized water twice before transferring the calcium carbonate particles from one solution to another solution, and removing the excess polyelectrolyte solution which is not bonded on the surfaces to ensure the uniformity of the membrane; repeating for 4 times to form 4 double-layer chitosan/sodium alginate composite membranes to obtain the polyelectrolyte multilayer membrane.
7. The method of claim 1, wherein the gold nanoparticles are synthesized by a method comprising:
heating a 0.01% chloroauric acid solution to boiling, heating 1% sodium citrate and stirring for 15min to recover to room temperature while stirring, finally, centrifuging at 12000rpm for 5min to collect particles, and washing with deionized water for 3 times to obtain gold nanoparticles with a rod-like structure, wherein the gold nanoparticles have negative charges in the solution.
8. The polyelectrolyte multilayer film-calcium carbonate nano-drug carrier synthesized by any one of the methods.
9. The use of the polyelectrolyte multilayer film-calcium carbonate nano-drug carrier as claimed in claim 8 in the preparation of anticancer drugs.
10. The use of claim 9, wherein the drug is a broad spectrum anticancer drug soluble or water soluble encapsulated by the polyelectrolyte multilayer film-calcium carbonate nano-drug carrier.
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Citations (5)
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