CN110204703B - Diatomite-based composite material and preparation method thereof - Google Patents
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Abstract
The invention belongs to the field of drug loading and tissue engineering biomedical science, and discloses a diatomite-based composite material and a preparation method thereof. The diatomite-based composite material is prepared by dissolving acidified diatomite and aniline oligomer into an organic solvent under the protection of inert gas, uniformly mixing to obtain a mixed solution A, adding a degradable high-molecular solution B into the mixed solution A, uniformly stirring, adding a catalyst at 30-80 ℃ for reaction, filtering, washing and drying. The diatomite-based composite material can promote cell adhesion, proliferation and differentiation in the environment of an external electromagnetic field, and has the characteristics of biodegradability, good biocompatibility, simple preparation method, simple synthesis, few byproducts and the like.
Description
Technical Field
The invention belongs to the field of drug loading and tissue engineering biomedical science, and particularly relates to a diatomite-based composite material and a preparation method thereof.
Background
The diatomite is made of SiO2The unique and regular three-dimensional porous microstructure of the main biomineral material enables the biomineral material to have high porosity, hydrophilicity, high strength, high specific surface area and good adsorbability. Polyaniline (PANI) is a conductive polymer material with a P-electron conjugated structure in a molecular chain, has good biocompatibility and certain electric propertyThe magnetic field can promote cell differentiation, proliferation and growth. The degradable high polymer material has excellent biocompatibility, can be gradually decomposed into absorbable or dischargeable micromolecules under the environment of body fluid to attract attention, and is further developed in the field of medical materials.
Polyaniline is mostly blended with degradable polymer through a physical bonding mode, or the polyaniline conductive polymer material is wrapped after surface treatment of the degradable polymer material after electrostatic spinning, the two molding modes can be used as the conductive degradable polymer material in the fields of tissue engineering supports, biosensors and the like to a certain extent, but the bonding force of polyaniline and the degradable polymer material is weaker, the degradable polymer is degraded in vivo, the macromolecular conductive polymer of the surface wrapping is left in vivo, is not easy to degrade and discharge, can produce toxicity to surrounding tissues, and causes local inflammation. Thus, the search for a readily degradable, electrically conductive polymeric material is an important direction and goal in the field of tissue engineering.
Disclosure of Invention
In order to solve the above-mentioned disadvantages and drawbacks of the prior art, the present invention aims to provide a diatomite-based composite material.
The invention also aims to provide a preparation method of the diatomite-based composite material.
The invention further aims to provide application of the diatomite-based composite material.
The purpose of the invention is realized by the following technical scheme:
under the protection of inert gas, the diatomite-based composite material is prepared by dissolving acidified diatomite and aniline oligomer in an organic solvent and then uniformly mixing to obtain a mixed solution A, adding a degradable high-molecular solution B into the mixed solution A, uniformly stirring, adding a catalyst at 30-80 ℃ for reaction, filtering, washing and drying.
Preferably, the particle size of the diatomite is 1-500 mu m.
Preferably, the acidizing fluid is one or more of phosphoric acid, hydrochloric acid, sulfuric acid, dodecylbenzene sulfonic acid, naphthalene sulfonic acid or camphor sulfonic acid, and the concentration of the acidizing fluid is 0.1-2.5 mol/L.
Preferably, the aniline oligomer is aniline trimer, aniline tetramer, aniline pentamer, aniline hexamer, aniline heptamer or aniline octamer.
Preferably, the organic solvent is one or more of ethanol, diethyl ether, acetone, dichloromethane, chloroform, carbon disulfide, toluene, tetrahydrofuran, N-dimethylformamide, benzoic acid, or N-methylpyrrolidone.
Preferably, the degradable polymer is polycaprolactone, polylactic acid, polyglycolic acid, polylactic acid-glycolic acid copolymer, polyethylene glycol or derivatives of the above materials.
Preferably, the mass ratio of the aniline oligomer to the acidified diatomite is 1: (1-9); the molar ratio of the aniline oligomer to the degradable polymer is (1-9): 1; the mass ratio of the organic solvent to the aniline oligomer is (10-1000): 1.
preferably, the catalyst is lipase or stannous octoate; the catalyst is 0.1-10 wt% of aniline oligomer.
Preferably, the reaction time is 2-300 h; the reaction stirring speed is 100-800 r/min; the inert gas is N2Or Ar.
The preparation method of the diatomite-based composite material comprises the following specific steps:
s1, adding acidified diatomite and aniline oligomers into an organic solvent under inert gas, and uniformly stirring;
s2, adding degradable macromolecules dissolved in the same organic solution into the mixed solution, stirring and mixing uniformly to obtain a mixed solution A, and adding a solution B of the degradable macromolecules into the solution A to obtain a mixed solution C;
and S3, adding a catalyst into the mixed solution C at the temperature of 30-80 ℃, stirring for reaction, and filtering, washing and drying after the reaction is finished to obtain the diatomite-based composite material.
Compared with the prior art, the invention has the following beneficial effects:
1. the diatomite-based composite material has the characteristics of certain porosity, conductivity in an electromagnetic field environment, capability of promoting cell adhesion, proliferation and differentiation, biodegradability and good biocompatibility.
2. The invention uses enzyme catalysis to form the synthesis of new copolymer of aniline oligomer and degradable macromolecule, because aniline oligomer exists in the form of bond on the surface of diatomite, but aniline oligomer and degradable macromolecule form new chemical covalent bond.
3. The method has the advantages of simple synthesis, less by-products and high reaction efficiency.
Drawings
Fig. 1 is a schematic structural view of a diatomite-based composite material according to the present invention.
Fig. 2 is an SEM photograph of the diatomite-based composite material obtained in example 1.
Detailed Description
The following examples are presented to further illustrate the present invention and should not be construed as limiting the invention.
Example 1
Adding 1.2.0g of diatomite acidified by 2.5mol/L hydrochloric acid and 1.2g of Aniline Trimer (AT) into 40ml of toluene, uniformly mixing, magnetically stirring AT normal temperature for 30min, and performing ultrasonic treatment AT 37 ℃ for 10min to obtain a mixed solution I; dissolving 0.3g of Polycaprolactone (PCL) in 20ml of toluene, heating and dissolving at 45 ℃ to obtain a polycaprolactone solution (II);
2.0.1g of lipase (lipase) is dissolved in 10ml of ultrapure water and is magnetically stirred for 30min to obtain a lipase aqueous solution (③);
3. adding the solution II into the solution I, magnetically stirring the solution I at 40 ℃ for 30min at 400r/min to obtain a mixed solution IV; adding the solution (c) into the mixed solution (c) and magnetically stirring for 30min at a speed of 400r/min to obtain a mixed solution (c);
4. placing the mixed solution in an argon atmosphere, performing oil bath AT 40 ℃, and performing magnetic stirring AT 120r/min for 30 hours to synthesize an AT-PCL mixed solution; carrying out vacuum filtration on the AT-PCL mixed solution, and cleaning with Tetrahydrofuran (THF) to obtain a precipitate; the precipitate was dried in a vacuum oven at 60 ℃ for 24h to give 0.93g of finished powder, 62% yield and 7.5% porosity.
The obtained diatomite-based composite material is dissolved in DMF, and is made into a scaffold material through electrostatic spinning for culturing osteoblasts, under the action of a proper electromagnetic field, the growth state of the osteoblasts is normal, the cell density is increased by 25% compared with that of a blank group, and after the scaffold material is cultured for 7 days, the mass loss of the scaffold material is 40%.
Fig. 1 is a schematic structural view of a diatomite-based composite material according to the present invention. Wherein, 1 represents diatomite, 2 represents aniline trimer, 3 represents biodegradable polymer material, and any one of Polycaprolactone (PCL), polylactic acid (PLA), polyglycolic acid (PGA), polylactic-co-glycolic acid (PLGA) and polyethylene glycol (PEG) and mixture or derivative thereof. As can be seen from FIG. 1, the aniline trimer is supported on a diatomaceous earth structure, and forms a copolymer with a degradable polymer under the action of lipase. Fig. 2 is an SEM photograph of the diatomite-based composite material obtained in example 1. As can be seen from FIG. 2, aniline trimer was successfully supported on the surface of diatomaceous earth, and PCL was present on the surface of aniline trimer.
Example 2
1.2.5g of diatomite acidized by 2mol/L dodecylbenzene sulfonic acid and 2.5g of Aniline Tetramer (ATe) are added into 100ml of N' -N-Dimethylformamide (DMF) to be uniformly mixed, magnetically stirred at normal temperature for 30min and ultrasonically treated at 37 ℃ for 10min to obtain a mixed solution I; 2.5g of Polycaprolactone (PCL) is dissolved in 100ml of DMF and heated at 45 ℃ to obtain polycaprolactone solution (II);
2.0.5g of lipase (lipase) is dissolved in 10ml of ultrapure water and is magnetically stirred for 30min to obtain a lipase aqueous solution (③);
3. adding the solution II into the solution I, magnetically stirring the solution I at 40 ℃ for 30min at 400r/min to obtain a mixed solution IV; adding the solution (c) into the mixed solution (c) and magnetically stirring for 30min at a speed of 400r/min to obtain a mixed solution (c);
4. placing the mixed solution in nitrogen atmosphere, performing oil bath at 40 ℃, and performing magnetic stirring at 120r/min for 100 hours to synthesize ATe-PCL mixed solution; vacuum filtering ATe-PCL mixed solution, and cleaning with ethanol to obtain precipitate; the precipitate was dried in a vacuum oven at 60 ℃ for 24h to give 2.80g of finished powder with a yield of 56% and a porosity of 5.8%.
The obtained diatomite-based composite material is dissolved in toluene, the bracket material is prepared by electrostatic spinning and used for culturing osteoblasts, the growth state of the osteoblasts is normal under the action of a proper electromagnetic field, the cell density is increased by 32% compared with that of a blank group, and the mass loss of the bracket material is 60% after the bracket material is cultured for 14 days.
Example 3
1.1.0g of diatomite and diatomite which are acidified by 1mol/L naphthalenesulfonic acid and 2.0g of aniline trimer are added into 50ml of toluene to be uniformly mixed, and then the mixture is magnetically stirred for 30min at normal temperature and is subjected to ultrasonic treatment for 10min at 37 ℃ to obtain a mixed solution I; dissolving 4.0g of Polycaprolactone (PCL) in 100ml of toluene, heating and dissolving at 45 ℃ to obtain polycaprolactone solution (II);
2.0.7g of stannous isooctanoate is dissolved in 10ml of ultrapure water and is magnetically stirred for 30min to obtain a stannous isooctanoate aqueous solution (c);
3. adding the solution II into the solution I, magnetically stirring the solution I at 40 ℃ for 30min at 400r/min to obtain a mixed solution IV; adding the solution (c) into the mixed solution (c) and magnetically stirring for 30min at a speed of 400r/min to obtain a mixed solution (c); placing the mixed solution in an argon atmosphere, performing oil bath AT 40 ℃, and performing magnetic stirring AT 120r/min for 180 hours to synthesize an AT-PCL mixed solution;
4. carrying out vacuum filtration on the AT-PCL mixed solution, and cleaning with Tetrahydrofuran (THF) to obtain a precipitate; the precipitate was dried in a vacuum oven at 70 ℃ for 24h to give 3.79g of finished powder with a yield of 63.2% and a porosity of 4.6%.
The obtained diatomite-based composite material is dissolved in dichloromethane, the bracket material is prepared by electrostatic spinning and used for culturing osteoblasts, the growth state of the osteoblasts is normal under the action of a proper electromagnetic field, the cell density is increased by 30% compared with that of a blank group, and the mass loss of the bracket material is 65% after the bracket material is cultured for 28 days.
Example 4
Adding 1.4g of diatomite acidified by 0.5mol/L camphorsulfonic acid and 1.8g of Aniline Pentamer (AP) into 50ml of dichloromethane, uniformly mixing, magnetically stirring at normal temperature for 30min, and performing ultrasonic treatment at 37 ℃ for 10min to obtain a mixed solution I; 2.0g of Polycaprolactone (PCL) is dissolved in 20ml of dichloromethane and heated and dissolved at 45 ℃ to obtain polycaprolactone solution (II);
2.0.5g of lipase (lipase) is dissolved in 10ml of ultrapure water and is magnetically stirred for 30min to obtain a lipase aqueous solution (③);
3. adding the solution II into the solution I, magnetically stirring the solution I at 40 ℃ for 30min at 400r/min to obtain a mixed solution IV; adding the solution (c) into the mixed solution (c) and magnetically stirring for 30min at a speed of 400r/min to obtain a mixed solution (c);
4. placing the mixed solution in an argon atmosphere, performing oil bath at 40 ℃, and performing magnetic stirring at 120r/min for 100 hours to synthesize the AP-PCL mixed solution; carrying out vacuum filtration on the AP-PCL mixed solution, and cleaning with Tetrahydrofuran (THF) to obtain a precipitate; the precipitate was dried in a vacuum oven at 60 ℃ for 24h to give 1.8g of finished powder with a yield of 47.4% and a porosity of 6.9%.
The obtained diatomite-based composite material is dissolved in chloroform, a scaffold material is prepared by electrostatic spinning and used for culturing osteoblasts, the osteoblasts are in a normal growth state under the action of a proper electromagnetic field, the cell density is increased by 32% compared with that of a blank group, and the mass loss of the scaffold material is 25% after 3 days of culture.
The above embodiments are preferred embodiments of the present invention, but the present invention is not limited to the above embodiments, and any other changes, modifications, substitutions, combinations and simplifications which do not depart from the spirit and principle of the present invention should be construed as equivalents thereof, and all such changes, modifications, substitutions, combinations and simplifications are intended to be included in the scope of the present invention.
Claims (6)
1. The diatomite-based composite material is characterized in that under the protection of inert gas, acidified diatomite and aniline oligomers are dissolved in an organic solvent and then are uniformly mixed to obtain a mixed solution A, a degradable high-molecular solution B is added into the mixed solution A, the mixture is uniformly stirred, a catalyst is added at the temperature of 30-80 ℃ for reaction, and the diatomite-based composite material is prepared through filtering, washing and drying; the aniline oligomer is aniline tripolymer, aniline tetramer, aniline pentamer, aniline hexamer, aniline heptamer or aniline octamer; the degradable polymer is polycaprolactone, polylactic acid, polyglycolic acid, polylactic acid-glycolic acid copolymer or polyethylene glycol; the mass ratio of the aniline oligomer to the acidified diatomite is 1: (1-9); the molar ratio of the aniline oligomer to the degradable polymer is (1-9): 1; the mass ratio of the organic solvent to the aniline oligomer is (10-1000): 1; the catalyst is lipase or stannous octoate; the catalyst is 0.1-10 wt% of aniline oligomer.
2. The diatomite-based composite material according to claim 1, wherein the diatomite has a particle size of 1-500 μm.
3. The diatomite-based composite material according to claim 1, wherein the acidizing fluid is one or more of phosphoric acid, hydrochloric acid, sulfuric acid, dodecylbenzene sulfonic acid, naphthalene sulfonic acid and camphor sulfonic acid, and the concentration of the acidizing fluid is 0.1-2.5 mol/L.
4. The diatomite-based composite material according to claim 1, wherein the organic solvent is one or more of ethanol, diethyl ether, acetone, dichloromethane, chloroform, carbon disulfide, toluene, tetrahydrofuran, N-dimethylformamide, benzoic acid, or N-methylpyrrolidone.
5. The diatomite-based composite material according to claim 1, wherein the reaction time is 2-300 h; the reaction stirring speed is 100-800 r/min; the inert gas is N2Or Ar.
6. The process for the preparation of a diatomite-based composite material according to any one of claims 1 to 5, characterized in that it comprises the following specific steps:
s1, under the inert gas, adding the acidified diatomite and aniline oligomer into the organic solvent and uniformly stirring to obtain a mixed solution A;
and S2, adding the degradable high molecular solution B into the mixed solution A, uniformly stirring, adding a catalyst at 30-80 ℃ for reaction, and filtering, washing and drying to obtain the diatomite-based composite material.
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| CN1651514A (en) * | 2005-01-07 | 2005-08-10 | 华东理工大学 | Fibrous polyaniline / diatomite nano-conductive composite material |
| WO2016172461A1 (en) * | 2015-04-23 | 2016-10-27 | The University Of Connecticut | Stretchable organic metals, composition, and use |
| CN107413305A (en) * | 2017-05-05 | 2017-12-01 | 兰州理工大学 | Polyaniline diatomite/Fe3 O4The preparation method of Chitosan Composites |
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|---|---|---|---|---|
| CN1651514A (en) * | 2005-01-07 | 2005-08-10 | 华东理工大学 | Fibrous polyaniline / diatomite nano-conductive composite material |
| WO2016172461A1 (en) * | 2015-04-23 | 2016-10-27 | The University Of Connecticut | Stretchable organic metals, composition, and use |
| CN107413305A (en) * | 2017-05-05 | 2017-12-01 | 兰州理工大学 | Polyaniline diatomite/Fe3 O4The preparation method of Chitosan Composites |
Non-Patent Citations (2)
| Title |
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| Large-area fibrous network of polyaniline formed on the surface of diatomite;Xingwei Li et al.;《Applied Surface Science》;20090616;第255卷;8276-8280 * |
| Universal Two-Step Approach to Degradable and Electroactive;Baolin Guo et al.;《Macromolecules》;20110615;第44卷;5227-5236 * |
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