CN105412934A - Multifunctional drug carrier material based on nanometer rare earth doped with hydroxyapatite - Google Patents

Abstract

The invention relates to a nano-rare earth-doped hydroxyapatite-based multifunctional drug carrier material, which is characterized in that the rare earth-doped hydroxyapatite fluorescent material is used as the basis to construct fluorescence, mesoporous, magnetic, nano-core-shell morphology, etc. Functional nano-core-shell structure drug carrier particles; its components from the inside to the outside are iron ferric oxide as the magnetic core of the core-shell structure, a non-porous silica layer, a rare earth-doped hydroxyapatite fluorescent layer, and a mesoporous Silicon oxide layer; core-shell structure diameter is 80-100nm; its expression is: Fe ₃ O ₄ nonporous-SiO ₂ HAP-EuMesoporous-SiO ₂ . The material has good application prospects in biomarkers, cell imaging, DNA detection, lesion detection, and biosensing.

Description

A nano-rare earth-doped hydroxyapatite-based multifunctional drug carrier material

technical field

The invention relates to a nano-rare earth-doped hydroxyapatite-based multifunctional drug carrier material, which belongs to the field of biomedical material science.

Background technique

Biomedical materials are materials used to diagnose, treat, repair or replace damaged tissues and organs of living organisms or enhance their functions, also known as biomaterials. It mainly includes general biomedical materials, tissue engineering biomedical materials, modern diagnostic systems and advanced controlled release systems. Biomaterials that can undertake multiple functions have promising application prospects in biomarkers, cell imaging, DNA detection, lesion detection, and biosensing. Nano-scale drug carrier is a sub-particle drug carrier delivery system belonging to the category of nano-scale microcosm. Encapsulating drugs in sub-particles can adjust the release rate, increase the permeability of biofilms, change the distribution in the body, and improve bioavailability, etc. Screen and combine nanoparticle carrier materials to obtain a suitable drug release rate; use surface chemical methods to modify the surface of nanoparticles to improve targeting ability and change the targeting site; optimize the preparation process to increase drug loading and clinical application To make it suitable for industrial production; to explore the dynamics of in vivo engineering, so as to correctly describe the changes of drugs in the blood and target organs. The drug loading capacity is greater than 30%, the encapsulation rate is higher than 80%, it has the characteristics of suitable preparation and purification methods, biodegradability, low or no toxicity, appropriate particle size and particle shape, and long circulation time in the body. It is an essential condition for an ideal drug carrier.

Nano-drug carriers with mesoporous structures have many advantages in clinical applications. Mesoporous materials refer to a class of porous materials with pore sizes ranging from 2 to 50 nm. General biomacromolecules such as proteins, enzymes, nucleic acids, and viruses have a molecular weight below 30 nanometers, making mesoporous materials very suitable for the immobilization and separation of proteins, enzymes, etc. For example, synthetic mesoporous materials such as maltose can immobilize enzymes on the basis of retaining enzyme activity. Continuous membrane materials can be generated on substrates of different mesoporous materials for the separation of cells and DNA; mesoporous materials have a large specific surface area and specific pore volume, and can be loaded with porphyrin, pyridine, or Immobilize and embed biological drugs such as proteins, and improve the durability of drug effects by modifying functional groups to control the release of drugs. Utilizing the bioguiding effect, it can effectively and accurately hit targets such as cancer cells and lesion sites, and give full play to the curative effect of drugs. Superparamagnetic iron oxide nanoparticles have emerged as a new class of probes for in vitro and in vivo cellular and molecular imaging. The use of superparamagnetic contrast agents in NMR has the advantage of producing stronger proton relaxation than paramagnetic contrast agents. Thus, less contrast agent doses need to be injected into the body. Putting superparamagnetic iron oxide in an AC electromagnetic field can make the magnetic direction change randomly between parallel and antiparallel, so that the magnetic energy can be transferred to the particles in the form of heat, which can be used to destroy diseased cells in organisms. Tumor cells are more sensitive to temperature than healthy cells. Magnetic nanoparticles with external magnetic fields and/or magnetizable implants can deliver the particles to the target area, and fix the particles at the local site when the drug is released, so that the drug can be released locally to achieve targeted drug delivery.

Contents of the invention

Aiming at the deficiencies of the prior art, the problem to be solved in the present invention is to provide a nano-rare earth-doped hydroxyapatite-based multifunctional drug carrier material, which is characterized in that the rare-earth-doped hydroxyapatite fluorescent material is used as the basis to construct a collection of fluorescent materials. Nano-core-shell drug carrier particles with functions such as mesoporous, magnetic, and nano-morphology; its components are from the inside to the outside, ferric oxide as the magnetic core of the core-shell structure, non-porous silica layer, rare earth doped Hydroxyapatite fluorescent layer, mesoporous silica layer; core-shell structure diameter is 80-100 nm; its expression is: Fe ₃ O ₄ @nonporous-SiO ₂ @HAP:Eu@Mesoporous-SiO ₂ .

Among them, Fe ₃ O ₄ represents the ferroferric oxide magnetic core; nonporous-SiO ₂ represents the nonporous silica layer; HAP:Eu represents the rare earth-doped hydroxyapatite fluorescent nanolayer; Mesoporous-SiO ₂ represents the mesoporous silica layer.

As the core of the nano-core-shell structure, ferroferric oxide is characterized by: superparamagnetic, spherical, with a diameter of 5-30 nm; the thickness of the non-porous silicon dioxide layer is 10-15 nm, which plays a role in the magnetic and fluorescent functional areas. Form an isolation barrier between them to prevent the magnetic and fluorescent functions from interfering with each other and weaken their functions; rare earth doped hydroxyapatite HAP:Eu fluorescent layer thickness is 10-30nm, hydroxyapatite has the same as human bone and animal bone Similar structure of inorganic components, biocompatibility, biostability and non-toxicity, rare earth doped hydroxyapatite fluorescent material has excitation light in the infrared region, large tissue penetration depth, can achieve zero background detection, and has fluorescent properties Hydroxyapatite has incomparable characteristics of organic dyes and quantum dots; the thickness of the mesoporous silicon oxide layer is 1-20 nm, and the mesopore size is continuously adjustable. It acts as a carrier to anchor drugs or proteins, enzymes, etc. Anchored object delivery and detachment functions.

The invention relates to a hydroxyapatite-based multifunctional functional core-shell structure biomedical carrier material with good microscopic appearance, high purity, good dispersibility and high biocompatibility. The invention also provides a nanoscale high dispersibility A method for preparing a functional nano-core-shell biomedical carrier material with application capabilities such as magnetic targeting, high-capacity drug anchoring, fluorescent lesion detection, and biosensing.

A nano-rare earth-doped hydroxyapatite-based multifunctional drug carrier material according to the present invention is realized by the following steps:

(1) Preparation of magnetic nano ferric oxide particles, 1.988 g FeCl ₂ 4H ₂ O and 3.244 g FeCl ₃ 6H ₂ O were dissolved in 100 mL deionized water, transferred to a 250 mL three-necked bottle, and placed in a 25°C water bath In the three-necked bottle, argon was introduced into the bottle for protection, 4.599 g of KOH was dissolved in 100 mL of deionized water, and added into the three-necked bottle under stirring at 400 rpm. When the solution turned black, the stirring was continued for 5 h. To obtain Fe ₃ O ₄ nanoparticles, the synthesized Fe ₃ O ₄ nanoparticles were washed alternately with deionized water and absolute ethanol three times, and dried in vacuum at 80°C for later use;

(2) Preparation of non-porous silica layer, weigh 0.1 g of Fe ₃ O ₄ prepared in step (1) and add it to 80 mL of absolute ethanol, sonicate for 20-40 min, then add 2 mL of 28% ammonia water and 2 mL of deionized, Fe ₃ O ₄ were uniformly dispersed in the solvent, and 3 mL of TEOS absolute ethanol solution with a volume concentration of 25% ethyl orthosilicate was added to it, and the ultrasound was continued for 3-6 h, and it was allowed to stand at room temperature for 12 h. The product is separated by a strong magnetic field, washed with absolute ethanol and deionized water for 3-6 times, and the cleaned nanoparticles are dispersed into 50 mL of deionized water for later use;

(3) Preparation of rare earth-doped hydroxyapatite layer, transfer the solution obtained in step (2) into a 250 mL three-neck flask, and then add 0.1584 g of (NH ₄ ) ₂ HPO ₄ and 0.1 g of hexadecyl Trimethylammonium bromide CTAB, 0.01-0.06 g Eu(N0 ₃ ) ₃ 6H ₂ O Use ammonia water to adjust the pH value to 10-12, weigh the corresponding amount of CaCl ₂ according to Ca/P=1.67 and dissolve in 50 mL In deionization, the three-necked bottle is placed in a water bath, the temperature of the water bath is 80-90 °C, stirred at 400 rpm, and argon is introduced, and then the CaCl ₂ solution is added dropwise to the three-necked bottle, and kept After stirring for 6 h, the product solution was transferred into a hydrothermal reactor with a polytetrafluoroethylene liner. After hydrothermal treatment at 160 °C for 5 h, a HAP:Eu package with a certain microscopic morphology, high crystallinity, and a single component was obtained. Coating, the product is separated by a strong magnetic field, washed with absolute ethanol and deionized water for 3-6 times, and then dissolved in 100 mL of deionized water for later use;

(4) Preparation of mesoporous silica layer, take step (3) solution and add 0.01-0.05 g CTAB and 0.01-0.05 g NaOH under electromagnetic stirring, then slowly add 0.03-0.1 mL TEOS, continue stirring for 5 h , aged for 12 h, separated the solution in a strong magnetic field, washed 3-5 times with deionized water and ethanol, and dried in vacuum at 80°C, and put the dried product and 100 mL of acetone into a 250 mL three-neck flask and refluxed at 70°C 24 h, then separated by a strong magnetic field, washed twice with acetone, dried in a drying oven at 80 °C for 10 h in vacuum, and then ground to obtain a nano-rare earth-doped hydroxyapatite-based multifunctional drug carrier material with a diameter of 80-100 nm , expressed as: Fe ₃ O ₄ @nonporous-SiO ₂ @HAP:Eu@Mesoporous-SiO ₂ .

detailed description

The present invention will be further described below in conjunction with the examples, but the protection content of the present invention is not limited to the examples.

Example 1:

(1) Preparation of magnetic nano ferric oxide particles, 1.988 g FeCl ₂ 4H ₂ O and 3.244 g FeCl ₃ 6H ₂ O were dissolved in 100 mL deionized water, transferred to a 250 mL three-necked bottle, and placed in a 25°C water bath In the three-necked bottle, argon was introduced into the three-necked bottle for protection, 4.599 g KOH was dissolved in 100 mL deionized water, and added to the three-necked bottle under stirring at 400 rpm, and when the solution turned black, continued to stir for 5 h to obtain For Fe ₃ O ₄ nanoparticles, the synthesized Fe ₃ O ₄ nanoparticles were washed alternately with deionized water and absolute ethanol three times, and dried in vacuum at 80 °C for later use;

(2) Preparation of non-porous silica layer. Weigh 0.1 g of Fe ₃ O ₄ prepared in step (1) and add it to 80 mL of absolute ethanol, sonicate for 30 min, then add 2 mL of 28% ammonia water and 2 After Fe ₃ O ₄ was uniformly dispersed into the solvent, 3 mL of TEOS absolute ethanol solution with a volume concentration of 25% tetraethyl orthosilicate was added to it, and ultrasonication was continued for 4 h, and the product was left to stand at room temperature for 12 h. Magnetic field for separation, washed with absolute ethanol and deionized water for 4 times, and the cleaned nanoparticles were dispersed into 50 mL of deionized water for later use;

(3) Preparation of rare earth-doped hydroxyapatite layer, transfer the solution obtained in step (2) into a 250 mL three-neck flask, and then add 0.1584 g of (NH ₄ ) ₂ HPO ₄ and 0.1 g of hexadecyl Trimethylammonium bromide CTAB, 0.03 g Eu(N0 ₃ ) ₃ 6H ₂ O Use ammonia water to adjust the pH value to 10-12, weigh the corresponding amount of CaCl ₂ according to Ca/P=1.67 and dissolve in 50 mL deionized , the three-necked flask was placed in a water bath, the temperature of the water bath was 80 °C, stirred at 400 rpm, argon was passed through, and then the CaCl ₂ solution was added dropwise to the three-necked flask, and kept stirring for 6 h after the addition was completed. The product solution was transferred into a hydrothermal reaction kettle lined with polytetrafluoroethylene, and after hydrothermal treatment at 160 °C for 5 h, a HAP:Eu coating layer with a certain microscopic shape, high crystallinity, and single component was obtained. The product was separated by a strong magnetic field, washed four times with absolute ethanol and deionized water, and then dissolved in 100 mL of deionized water for later use;

(4) Preparation of the mesoporous silica layer, take the solution in step (3) and add 0.03 g CTAB and 0.03 g NaOH under electromagnetic stirring, then slowly add 0.03 mL TEOS, continue to stir for 5 h, and age for 12 h. The solution was separated by a strong magnetic field, washed five times with deionized water and ethanol, and dried in vacuum at 80 °C. The dried product and 100 mL of acetone were placed in a 250 mL three-necked bottle at 70 °C for 24 h, and then separated by a strong magnetic field. After that, it was washed twice with acetone, vacuum-dried in a drying oven at 80 °C for 10 h, and ground after grinding to obtain a nano-rare earth-doped hydroxyapatite-based multifunctional drug carrier material with a diameter _of 80-100 nm, expressed as: Fe3 O ₄ @nonporous-SiO ₂ @HAP-Eu@Mesoporous-SiO ₂ .

Example 2:

(1) Preparation of magnetic nano ferric oxide particles, 1.988 g FeCl ₂ 4H ₂ O and 3.244 g FeCl ₃ 6H ₂ O were dissolved in 100ml deionized water, transferred to a 250 mL three-necked bottle, and placed in a 25°C water bath , put argon into the three-necked bottle for protection, take 4.599 g KOH and dissolve it in 100 mL deionized water, add it into the three-necked bottle under stirring at 400 rpm, when the solution turns black, continue to stir for 5 h to get For Fe ₃ O ₄ nanoparticles, the synthesized Fe ₃ O ₄ nanoparticles were washed alternately with deionized water and absolute ethanol three times, and dried in vacuum at 80 °C for later use;

(2) Preparation of non-porous silica layer. Weigh 0.1 g of Fe ₃ O ₄ prepared in step (1) and add it to 80 mL of absolute ethanol, sonicate for 40 min, then add 2 mL of 28% ammonia water and 2 After Fe ₃ O ₄ was uniformly dispersed in the solvent, 3 mL of TEOS absolute ethanol solution with a volume concentration of 25% tetraethyl orthosilicate was added to it, and ultrasonication was continued for 6 h, and the product was left at room temperature for 12 h. Magnetic field for separation, washed with absolute ethanol and deionized water for 5 times, and the cleaned nanoparticles were dispersed into 50 mL of deionized water for later use;

(3) Preparation of rare earth-doped hydroxyapatite layer, transfer the solution obtained in step (2) into a 250 mL three-neck flask, and then add 0.1584 g of (NH ₄ ) ₂ HPO ₄ and 0.1 g of hexadecyl Trimethylammonium bromide CTAB, 0.06 g Eu(N0 ₃ ) ₃ 6H ₂ O Use ammonia water to adjust the pH value to 12, weigh the corresponding amount of CaCl ₂ according to Ca/P=1.67 and dissolve it in 50 mL deionized water, The three-necked bottle was placed in a water bath, the temperature of the water bath was 90 °C, stirred at 400 rpm, and argon gas was introduced, and then the CaCl solution was added dropwise to the three _- necked bottle, and the product was stirred for 6 h after the addition was completed. The solution was transferred into a hydrothermal reactor with a polytetrafluoroethylene liner, and after hydrothermal treatment at 160 °C for 5 h, a HAP:Eu coating layer with a certain microscopic shape, high crystallinity, and single component was obtained, and the product was used separated by a strong magnetic field, washed with absolute ethanol and deionized water for 6 times, and then dissolved in 100 mL of deionized water for later use;

(4) Preparation of the mesoporous silica layer, take the solution in step (3) and add 0.05 g CTAB and 0.05 g NaOH under electromagnetic stirring, then slowly add 0.06 mL TEOS, continue to stir for 5 h, and age for 12 h. The solution was separated by a strong magnetic field, washed 3-5 times with deionized water and ethanol, and dried in vacuum at 80 °C. The dried product and 100 mL of acetone were placed in a 250 mL three-necked bottle at 70 °C for 24 h, and then used a strong After magnetic field separation, wash with acetone twice, vacuum-dry in a drying oven at 80 °C for 10 h, and then grind to obtain a nano-rare earth-doped hydroxyapatite-based multifunctional drug carrier material with a diameter _of 80-100 nm, expressed as: Fe3 O ₄ @nonporous-SiO ₂ @HAP:Eu@Mesoporous-SiO ₂ .

Claims (1)

1. A nano-hydroxyapatite-based multifunctional drug carrier material, characterized in that: based on rare-earth-doped hydroxyapatite fluorescent materials, it has built functions such as fluorescence, mesoporous, magnetic properties, and nano-core-shell morphology. Nano-core-shell structure drug carrier particles; its components from the inside to the outside are ferroferric oxide as the magnetic core of the core-shell structure, non-porous silica layer, rare earth doped hydroxyapatite fluorescent layer, mesoporous silica layer; the diameter of the core-shell structure is 80-100 nm; its expression is: Fe ₃ O ₄ @nonporous-SiO ₂ @HAP:Eu@Mesoporous-SiO ₂ ; as the core of the nano-core-shell structure, ferric oxide is characterized by : Superparamagnetic, spherical, with a diameter of 5-30 nm; the thickness of the non-porous silica layer is 10-15 nm, and its function is to form an isolation barrier between the magnetic and fluorescent functional areas to prevent the magnetic and fluorescent functional areas from Mutual interference weakens their respective functions; rare earth-doped hydroxyapatite HAP:Eu fluorescent layer has a thickness of 10-30nm, has excitation light in the infrared region, has a large tissue penetration depth, can achieve zero background detection, and has excellent biological Compatibility; the thickness of the mesoporous silica layer is 1-20 nm, and the mesoporous size is continuously adjustable. It acts as a carrier to anchor drugs or proteins, enzymes, etc., and undertakes the function of delivering and separating anchored objects; nano The rare earth-doped hydroxyapatite-based multifunctional drug carrier material is realized by the following steps: (1) Preparation of magnetic nano-ferric oxide particles, 1.988 g FeCl ₂ ∙4H ₂ O and 3.244 g FeCl ₃ ∙6H ₂ O were dissolved in 100 mL deionized water, transferred to a 250 mL three-necked bottle, and placed at 25 ℃ water bath, put argon into the three-necked bottle for protection, take 4.599 g KOH and dissolve it in 100 mL deionized water, add it into the three-necked bottle under stirring at 400 rpm, when the solution turns black, continue to stir for 5 h, to obtain Fe ₃ O ₄ nanoparticles, wash the synthesized Fe ₃ O ₄ nanoparticles with deionized water and absolute ethanol three times alternately, and dry them in vacuum at 80°C for later use; (2) Preparation of non-porous silica layer, weigh 0.1 g of Fe ₃ O ₄ prepared in step (1) and add it to 80 mL of absolute ethanol, sonicate for 20-40 min, then add 2 mL of 28% ammonia water and 2 mL of deionized, Fe ₃ O ₄ were uniformly dispersed in the solvent, and 3 mL of TEOS absolute ethanol solution with a volume concentration of 25% ethyl orthosilicate was added to it, and the ultrasound was continued for 3-6 h, and it was allowed to stand at room temperature for 12 h. The product is separated by a strong magnetic field, washed with absolute ethanol and deionized water for 3-6 times, and the cleaned nanoparticles are dispersed into 50 mL of deionized water for later use; (3) Preparation of rare earth-doped hydroxyapatite layer, transfer the solution obtained in step (2) into a 250 mL three-neck flask, and then add 0.1584 g of (NH ₄ ) ₂ HPO ₄ and 0.1 g of hexadecyl Trimethylammonium bromide CTAB, 0.01-0.06 g Eu(N0 ₃ ) ₃ 6H ₂ O Use ammonia water to adjust the pH value to 10-12, weigh the corresponding amount of CaCl ₂ according to Ca/P=1.67 and dissolve in 50 mL In deionization, the three-necked bottle is placed in a water bath, the temperature of the water bath is 80-90 °C, stirred at 400 rpm, and argon is introduced, and then the CaCl ₂ solution is added dropwise to the three-necked bottle, and kept After stirring for 6 h, the product solution was transferred into a hydrothermal reactor with a polytetrafluoroethylene liner. After hydrothermal treatment at 160 °C for 5 h, the HAP:Eu coating layer was obtained. Wash with water, ethanol and deionized water for 3-6 times, then dissolve in 100 mL of deionized water for later use; (4) For the preparation of the mesoporous silica layer, take the solution in step (3) and add 0.01-0.05 g CTAB and 0.01-0.05 g NaOH, then slowly add 0.03-0.1 mL TEOS, continued to stir for 5 h, aged for 12 h, separated the solution in a strong magnetic field, washed 3-5 times with deionized water and ethanol, and dried in vacuum at 80 °C, and put the dried product and 100 mL of acetone into 250 mL The three-neck bottle was refluxed at 70 °C for 24 h, then separated by a strong magnetic field, washed twice with acetone, dried in a drying oven at 80 °C for 10 h in vacuum, and then ground to obtain nano-rare earth-doped hydroxyapatite-based multifunctional drug carrier materials. Its diameter is 80-100 nm.