CN104528676A - Method for quickly preparing polyethylene glycol regulated and controlled nano hydroxyapatite - Google Patents
Method for quickly preparing polyethylene glycol regulated and controlled nano hydroxyapatite Download PDFInfo
- Publication number
- CN104528676A CN104528676A CN201410785658.8A CN201410785658A CN104528676A CN 104528676 A CN104528676 A CN 104528676A CN 201410785658 A CN201410785658 A CN 201410785658A CN 104528676 A CN104528676 A CN 104528676A
- Authority
- CN
- China
- Prior art keywords
- nanometer hydroxyapatite
- calcium
- polyoxyethylene glycol
- phosphate
- hydroxyapatite
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Pending
Links
- 229910052588 hydroxylapatite Inorganic materials 0.000 title claims abstract description 61
- XYJRXVWERLGGKC-UHFFFAOYSA-D pentacalcium;hydroxide;triphosphate Chemical compound [OH-].[Ca+2].[Ca+2].[Ca+2].[Ca+2].[Ca+2].[O-]P([O-])([O-])=O.[O-]P([O-])([O-])=O.[O-]P([O-])([O-])=O XYJRXVWERLGGKC-UHFFFAOYSA-D 0.000 title claims abstract description 61
- 239000002202 Polyethylene glycol Substances 0.000 title claims abstract description 5
- 229920001223 polyethylene glycol Polymers 0.000 title claims abstract description 5
- 238000000034 method Methods 0.000 title abstract description 13
- 230000001105 regulatory effect Effects 0.000 title abstract 2
- 238000002360 preparation method Methods 0.000 claims abstract description 24
- 238000006243 chemical reaction Methods 0.000 claims abstract description 18
- 238000003756 stirring Methods 0.000 claims abstract description 16
- 159000000007 calcium salts Chemical class 0.000 claims abstract description 10
- 238000005406 washing Methods 0.000 claims abstract description 7
- 229910019142 PO4 Inorganic materials 0.000 claims abstract description 6
- 238000010438 heat treatment Methods 0.000 claims abstract description 6
- NBIIXXVUZAFLBC-UHFFFAOYSA-K phosphate Chemical compound [O-]P([O-])([O-])=O NBIIXXVUZAFLBC-UHFFFAOYSA-K 0.000 claims abstract description 6
- 239000010452 phosphate Substances 0.000 claims abstract description 6
- LYCAIKOWRPUZTN-UHFFFAOYSA-N Ethylene glycol Chemical compound OCCO LYCAIKOWRPUZTN-UHFFFAOYSA-N 0.000 claims description 43
- -1 polyoxyethylene Polymers 0.000 claims description 25
- 229920003171 Poly (ethylene oxide) Polymers 0.000 claims description 22
- WGCNASOHLSPBMP-UHFFFAOYSA-N hydroxyacetaldehyde Natural products OCC=O WGCNASOHLSPBMP-UHFFFAOYSA-N 0.000 claims description 21
- HEMHJVSKTPXQMS-UHFFFAOYSA-M Sodium hydroxide Chemical compound [OH-].[Na+] HEMHJVSKTPXQMS-UHFFFAOYSA-M 0.000 claims description 15
- 238000001035 drying Methods 0.000 claims description 8
- 239000008367 deionised water Substances 0.000 claims description 6
- 229910021641 deionized water Inorganic materials 0.000 claims description 6
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Chemical compound O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims description 6
- 238000013019 agitation Methods 0.000 claims description 5
- 238000005119 centrifugation Methods 0.000 claims description 5
- 238000001816 cooling Methods 0.000 claims description 5
- 239000008187 granular material Substances 0.000 claims description 5
- 238000010792 warming Methods 0.000 claims description 5
- ZCCIPPOKBCJFDN-UHFFFAOYSA-N calcium nitrate Chemical compound [Ca+2].[O-][N+]([O-])=O.[O-][N+]([O-])=O ZCCIPPOKBCJFDN-UHFFFAOYSA-N 0.000 claims description 4
- BNIILDVGGAEEIG-UHFFFAOYSA-L disodium hydrogen phosphate Chemical compound [Na+].[Na+].OP([O-])([O-])=O BNIILDVGGAEEIG-UHFFFAOYSA-L 0.000 claims description 4
- 229910000397 disodium phosphate Inorganic materials 0.000 claims description 4
- 235000019800 disodium phosphate Nutrition 0.000 claims description 4
- 239000001506 calcium phosphate Substances 0.000 claims description 3
- WHQOKFZWSDOTQP-UHFFFAOYSA-N 2,3-dihydroxypropyl 4-aminobenzoate Chemical compound NC1=CC=C(C(=O)OCC(O)CO)C=C1 WHQOKFZWSDOTQP-UHFFFAOYSA-N 0.000 claims description 2
- SIWNEELMSUHJGO-UHFFFAOYSA-N 2-(4-bromophenyl)-4,5,6,7-tetrahydro-[1,3]oxazolo[4,5-c]pyridine Chemical compound C1=CC(Br)=CC=C1C(O1)=NC2=C1CCNC2 SIWNEELMSUHJGO-UHFFFAOYSA-N 0.000 claims description 2
- UNMYWSMUMWPJLR-UHFFFAOYSA-L Calcium iodide Chemical compound [Ca+2].[I-].[I-] UNMYWSMUMWPJLR-UHFFFAOYSA-L 0.000 claims description 2
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 claims description 2
- NKWPZUCBCARRDP-UHFFFAOYSA-L calcium bicarbonate Chemical compound [Ca+2].OC([O-])=O.OC([O-])=O NKWPZUCBCARRDP-UHFFFAOYSA-L 0.000 claims description 2
- 229910000020 calcium bicarbonate Inorganic materials 0.000 claims description 2
- YYRMJZQKEFZXMX-UHFFFAOYSA-L calcium bis(dihydrogenphosphate) Chemical compound [Ca+2].OP(O)([O-])=O.OP(O)([O-])=O YYRMJZQKEFZXMX-UHFFFAOYSA-L 0.000 claims description 2
- LVGQIQHJMRUCRM-UHFFFAOYSA-L calcium bisulfite Chemical compound [Ca+2].OS([O-])=O.OS([O-])=O LVGQIQHJMRUCRM-UHFFFAOYSA-L 0.000 claims description 2
- 229910001622 calcium bromide Inorganic materials 0.000 claims description 2
- VTYYLEPIZMXCLO-UHFFFAOYSA-L calcium carbonate Substances [Ca+2].[O-]C([O-])=O VTYYLEPIZMXCLO-UHFFFAOYSA-L 0.000 claims description 2
- 235000010216 calcium carbonate Nutrition 0.000 claims description 2
- YALMXYPQBUJUME-UHFFFAOYSA-L calcium chlorate Chemical compound [Ca+2].[O-]Cl(=O)=O.[O-]Cl(=O)=O YALMXYPQBUJUME-UHFFFAOYSA-L 0.000 claims description 2
- WGEFECGEFUFIQW-UHFFFAOYSA-L calcium dibromide Chemical compound [Ca+2].[Br-].[Br-] WGEFECGEFUFIQW-UHFFFAOYSA-L 0.000 claims description 2
- 235000010260 calcium hydrogen sulphite Nutrition 0.000 claims description 2
- 229940046413 calcium iodide Drugs 0.000 claims description 2
- 229910001640 calcium iodide Inorganic materials 0.000 claims description 2
- JXRVKYBCWUJJBP-UHFFFAOYSA-L calcium;hydrogen sulfate Chemical compound [Ca+2].OS([O-])(=O)=O.OS([O-])(=O)=O JXRVKYBCWUJJBP-UHFFFAOYSA-L 0.000 claims description 2
- 229960000935 dehydrated alcohol Drugs 0.000 claims description 2
- MNNHAPBLZZVQHP-UHFFFAOYSA-N diammonium hydrogen phosphate Chemical compound [NH4+].[NH4+].OP([O-])([O-])=O MNNHAPBLZZVQHP-UHFFFAOYSA-N 0.000 claims description 2
- ZPWVASYFFYYZEW-UHFFFAOYSA-L dipotassium hydrogen phosphate Chemical compound [K+].[K+].OP([O-])([O-])=O ZPWVASYFFYYZEW-UHFFFAOYSA-L 0.000 claims description 2
- 235000019691 monocalcium phosphate Nutrition 0.000 claims description 2
- 229910000150 monocalcium phosphate Inorganic materials 0.000 claims description 2
- 150000003016 phosphoric acids Chemical class 0.000 claims description 2
- 238000012545 processing Methods 0.000 claims description 2
- 239000003814 drug Substances 0.000 abstract description 6
- 206010028980 Neoplasm Diseases 0.000 abstract description 3
- 201000011510 cancer Diseases 0.000 abstract description 3
- 239000000463 material Substances 0.000 abstract description 3
- 238000011160 research Methods 0.000 abstract description 3
- 238000002156 mixing Methods 0.000 abstract description 2
- 239000000843 powder Substances 0.000 abstract description 2
- 231100000331 toxic Toxicity 0.000 abstract description 2
- 230000002588 toxic effect Effects 0.000 abstract description 2
- 239000012266 salt solution Substances 0.000 abstract 2
- 239000000243 solution Substances 0.000 abstract 2
- 239000013598 vector Substances 0.000 abstract 2
- VHUUQVKOLVNVRT-UHFFFAOYSA-N Ammonium hydroxide Chemical compound [NH4+].[OH-] VHUUQVKOLVNVRT-UHFFFAOYSA-N 0.000 abstract 1
- 235000011114 ammonium hydroxide Nutrition 0.000 abstract 1
- 239000012620 biological material Substances 0.000 abstract 1
- 238000013270 controlled release Methods 0.000 abstract 1
- 229940079593 drug Drugs 0.000 abstract 1
- 238000001415 gene therapy Methods 0.000 abstract 1
- 238000009776 industrial production Methods 0.000 abstract 1
- 238000004519 manufacturing process Methods 0.000 abstract 1
- 210000004027 cell Anatomy 0.000 description 16
- 239000002245 particle Substances 0.000 description 10
- 230000000694 effects Effects 0.000 description 7
- UXVMQQNJUSDDNG-UHFFFAOYSA-L Calcium chloride Chemical compound [Cl-].[Cl-].[Ca+2] UXVMQQNJUSDDNG-UHFFFAOYSA-L 0.000 description 6
- 230000002349 favourable effect Effects 0.000 description 5
- 239000002105 nanoparticle Substances 0.000 description 5
- 230000005540 biological transmission Effects 0.000 description 4
- 230000008569 process Effects 0.000 description 4
- VWDWKYIASSYTQR-YTBWXGASSA-N sodium;dioxido(oxo)azanium Chemical compound [Na+].[O-][15N+]([O-])=O VWDWKYIASSYTQR-YTBWXGASSA-N 0.000 description 4
- 229910002651 NO3 Inorganic materials 0.000 description 3
- NHNBFGGVMKEFGY-UHFFFAOYSA-N Nitrate Chemical compound [O-][N+]([O-])=O NHNBFGGVMKEFGY-UHFFFAOYSA-N 0.000 description 3
- 238000002474 experimental method Methods 0.000 description 3
- 230000004044 response Effects 0.000 description 3
- 108010087230 Sincalide Proteins 0.000 description 2
- 238000010609 cell counting kit-8 assay Methods 0.000 description 2
- 210000005229 liver cell Anatomy 0.000 description 2
- 231100000252 nontoxic Toxicity 0.000 description 2
- 230000003000 nontoxic effect Effects 0.000 description 2
- 239000000047 product Substances 0.000 description 2
- 108090000623 proteins and genes Proteins 0.000 description 2
- IZTQOLKUZKXIRV-YRVFCXMDSA-N sincalide Chemical compound C([C@@H](C(=O)N[C@@H](CCSC)C(=O)NCC(=O)N[C@@H](CC=1C2=CC=CC=C2NC=1)C(=O)N[C@@H](CCSC)C(=O)N[C@@H](CC(O)=O)C(=O)N[C@@H](CC=1C=CC=CC=1)C(N)=O)NC(=O)[C@@H](N)CC(O)=O)C1=CC=C(OS(O)(=O)=O)C=C1 IZTQOLKUZKXIRV-YRVFCXMDSA-N 0.000 description 2
- 239000001488 sodium phosphate Substances 0.000 description 2
- QORWJWZARLRLPR-UHFFFAOYSA-H tricalcium bis(phosphate) Chemical compound [Ca+2].[Ca+2].[Ca+2].[O-]P([O-])([O-])=O.[O-]P([O-])([O-])=O QORWJWZARLRLPR-UHFFFAOYSA-H 0.000 description 2
- 208000030090 Acute Disease Diseases 0.000 description 1
- 206010008342 Cervix carcinoma Diseases 0.000 description 1
- 241000700112 Chinchilla Species 0.000 description 1
- 208000017667 Chronic Disease Diseases 0.000 description 1
- 206010061902 Pancreatic neoplasm Diseases 0.000 description 1
- 239000004698 Polyethylene Substances 0.000 description 1
- 229920001030 Polyethylene Glycol 4000 Polymers 0.000 description 1
- 108020004459 Small interfering RNA Proteins 0.000 description 1
- 208000006105 Uterine Cervical Neoplasms Diseases 0.000 description 1
- 230000001154 acute effect Effects 0.000 description 1
- 230000004520 agglutination Effects 0.000 description 1
- 239000007900 aqueous suspension Substances 0.000 description 1
- 230000009286 beneficial effect Effects 0.000 description 1
- 230000015572 biosynthetic process Effects 0.000 description 1
- 229910000389 calcium phosphate Inorganic materials 0.000 description 1
- 235000011010 calcium phosphates Nutrition 0.000 description 1
- 239000000969 carrier Substances 0.000 description 1
- 125000002091 cationic group Chemical group 0.000 description 1
- 210000000170 cell membrane Anatomy 0.000 description 1
- 230000003833 cell viability Effects 0.000 description 1
- 201000010881 cervical cancer Diseases 0.000 description 1
- 239000011248 coating agent Substances 0.000 description 1
- 238000000576 coating method Methods 0.000 description 1
- 238000013461 design Methods 0.000 description 1
- 238000001514 detection method Methods 0.000 description 1
- 238000011161 development Methods 0.000 description 1
- 239000006185 dispersion Substances 0.000 description 1
- 210000003027 ear inner Anatomy 0.000 description 1
- 238000005265 energy consumption Methods 0.000 description 1
- 238000005516 engineering process Methods 0.000 description 1
- 238000004108 freeze drying Methods 0.000 description 1
- 238000001027 hydrothermal synthesis Methods 0.000 description 1
- 229910010272 inorganic material Inorganic materials 0.000 description 1
- 239000011147 inorganic material Substances 0.000 description 1
- 230000031700 light absorption Effects 0.000 description 1
- 201000007270 liver cancer Diseases 0.000 description 1
- 208000014018 liver neoplasm Diseases 0.000 description 1
- 208000015486 malignant pancreatic neoplasm Diseases 0.000 description 1
- 239000012528 membrane Substances 0.000 description 1
- 238000012986 modification Methods 0.000 description 1
- 230000004048 modification Effects 0.000 description 1
- 239000002539 nanocarrier Substances 0.000 description 1
- 239000002073 nanorod Substances 0.000 description 1
- 231100000957 no side effect Toxicity 0.000 description 1
- 230000001473 noxious effect Effects 0.000 description 1
- 230000006911 nucleation Effects 0.000 description 1
- 238000010899 nucleation Methods 0.000 description 1
- 108020004707 nucleic acids Proteins 0.000 description 1
- 150000007523 nucleic acids Chemical class 0.000 description 1
- 102000039446 nucleic acids Human genes 0.000 description 1
- 235000015097 nutrients Nutrition 0.000 description 1
- 230000008520 organization Effects 0.000 description 1
- 201000002528 pancreatic cancer Diseases 0.000 description 1
- 208000008443 pancreatic carcinoma Diseases 0.000 description 1
- 230000001717 pathogenic effect Effects 0.000 description 1
- 239000012466 permeate Substances 0.000 description 1
- 229920000573 polyethylene Polymers 0.000 description 1
- 239000002244 precipitate Substances 0.000 description 1
- 239000002904 solvent Substances 0.000 description 1
- 239000000725 suspension Substances 0.000 description 1
- 238000003786 synthesis reaction Methods 0.000 description 1
- 238000012360 testing method Methods 0.000 description 1
- 231100000419 toxicity Toxicity 0.000 description 1
- 230000001988 toxicity Effects 0.000 description 1
- 238000001890 transfection Methods 0.000 description 1
- 238000005199 ultracentrifugation Methods 0.000 description 1
Landscapes
- Medicinal Preparation (AREA)
Abstract
The invention discloses a method for quickly preparing polyethylene glycol regulated and controlled nano hydroxyapatite and belongs to the field of preparation of biomedical materials. The method comprises the following process steps: preparing a calcium salt solution and a phosphate solution respectively, and mixing the calcium salt solution and the phosphate solution with equal volumes; adjusting the pH value of a reaction system by using ammonia water while stirring; adding polyethylene glycol, and heating the reaction system; and performing centrifuging and washing after reaction, and collecting granular deposits to obtain nano hydroxyapatite granular powder. The method disclosed by the invention is simple in preparation process, strong in laboratory operability, low in production cost and high in product yield and is suitable for laboratory research application and industrial production; and moreover, the prepared rod-shaped nano hydroxyapatite has good dispersity, does not have toxic reaction to normal cells and cancer cells, and has a huge application potential in the fields of biomedicine and biological materials such as drug controlled-release vectors, gene therapy vectors and the like.
Description
Technical field
The invention belongs to technical field of biomedical materials, be specifically related to the fast preparation method of a kind of polyoxyethylene glycol regulation and control nanometer hydroxyapatite.
Background technology
Hydroxyapatite nanoparticle has extensive and important purposes in fields such as coating, medical science, pharmacy, and can be used as a kind of nano inorganic material having good organization's consistency, have no side effect, for transmission and the slow-released carrier of gene or medicine.But hydroxyapatite is in preparation process, size can be expanded owing to constantly growing, cannot effective transmitter loss be carried out.Therefore, how simple and easyly nano-scale is prepared and the hydroxyapatite with good dispersion becomes the focus of research at present.In existing preparation method, have been reported the nanometer hydroxyapatite can producing different structure and pattern, as [Klesing J such as Klesing J, Chernousova S and Epple M. Freeze-dried cationic calcium phosphate nanorods as versatile carriers of nucleic acids (DNA, siRNA). Journal of Materials Chemistry, 2012, 22 (1): 199-204] polymine is utilized to carry out surface modification to hydroxyapatite, unconjugated polymine is removed with ultracentrifugation, carry out resuspended again, the nanometer club hydroxyapatite of 100-200nm is obtained by freeze-drying, this hydroxyapatite has good aqueous suspension, but particle is oversize, and agglutination phenomenon is serious, [the Wu X such as Wu X, Ding D, Jiang H, et al. Transfection using hydroxyapatite nanoparticles in the inner ear via an intact round window membrane in chinchilla. Journal of Nanoparticle Research, 2012, 14 (1): 1-13] hydroapatite particles is prepared with hydrothermal synthesis method, finishing is carried out again with polymine, obtain the hydroxyapatite nanoparticle of a kind of 50-100 nm, but this particle need at high temperature carry out, operational safety degree is low, preparation cycle is long, and obtained granule-morphology heterogeneity, purity is low.In recent years, the development of inorganic nano carrier technique brings new opportunity to develop to the preparation of hydroxy apatite powder, and has made new requirement to the characteristic of hydroxyapatite.Carry safety of medicine for making carrier and arrive pathogenic site, carrier must have that purity is high, size is little and the performance such as favorable dispersity.And various experimental technique all cannot prepare that a kind of size is little, the hydroxyapatite nanoparticle of favorable dispersity now, thus in the urgent need to develop a kind of preparation condition simple, consuming time short, energy consumption is low, particle size is little, the nanometer hydroxyapatite preparation method of favorable dispersity.
Summary of the invention
For prior art Problems existing, the present invention is with the synthesis of polyoxyethylene glycol regulation and control nanometer hydroxyapatite, in reaction process, with the nanometer hydroxyapatite of polyoxyethylene glycol parcel nucleation, it is suppressed to increase further, and properties modifying to hydroxyapatite surface by polyoxyethylene glycol, object be to design a kind ofly can to prepare in laboratory, technique is simple, preparation time is short, specific surface area is comparatively large, the nanometer hydroxyapatite of favorable dispersity.
The fast preparation method of described a kind of polyoxyethylene glycol regulation and control nanometer hydroxyapatite, is characterized in that by following processing step:
1) calcium salt soln of 15-50mM is configured respectively, 9-30mM phosphate solution;
2) calcium salt soln getting 50-200mL in 0 DEG C of cooling, by same volume phosphate solution with 10-60 drip/min speed joins in calcium salt soln, and stirs with 500-1000rpm/min gradient;
3) step 2 is maintained with sodium hydroxide solution) pH of reaction system is 9-14, until be added dropwise to complete;
4) in step 3) reaction system, slowly add 0.5-10mg polyoxyethylene glycol, and stop stirring;
5) after 1h by step 4) reaction system with 500-1000rpm/min rapid stirring, and be warming up to 70-100 DEG C, Keep agitation 1h;
6) stop heating, after step 5) system is cooled to room temperature, resuspended washing granule centrifugal under the cold condition of 2-8 DEG C with deionized water 4-6 time, obtains throw out;
7) throw out step 6) obtained, with filter paper capping, carries out dustless drying treatment, the nanometer hydroxyapatite of obtained polyoxyethylene glycol regulation and control;
Described nanometer hydroxyapatite is club shaped structure, major diameter is 20-80nm, minor axis is 10-15nm, length-to-diameter ratio is (1-3): 1, organic content 1-5% in nanometer hydroxyapatite, nanometer hydroxyapatite content is 85-95%, and nanometer hydroxyapatite Z current potential is (-0.02)-(-0.1) mV, and described polyethylene glycol relative molecular weight is 200-2000.
The fast preparation method of described a kind of polyoxyethylene glycol regulation and control nanometer hydroxyapatite, is characterized in that calcium salt described in step 1) is any one in monocalcium phosphate, nitrocalcite, Calcium hydrogen carbonate, calcium bisulfate, calcium bisulfite, Losantin, Calcium Bromide, calcium iodide, calcium chlorate, calcium perchlorate or calcium permanganate.
The fast preparation method of described a kind of polyoxyethylene glycol regulation and control nanometer hydroxyapatite, is characterized in that the phosphoric acid salt described in step 1) is any one in Secondary ammonium phosphate, Sodium phosphate dibasic, dipotassium hydrogen phosphate.
The fast preparation method of described a kind of polyoxyethylene glycol regulation and control nanometer hydroxyapatite, it is characterized in that the centrifugal speed in step 6) is 4000-16000rpm/min, centrifugation time is 2-30min.
The fast preparation method of described a kind of polyoxyethylene glycol regulation and control nanometer hydroxyapatite, it is characterized in that in step 7), throw out suspends with dehydrated alcohol when drying, drying temperature is 40-60 DEG C, and time of drying is 12-24h.
Compared with prior art, the present invention has following beneficial effect:
1) the present invention can in the indoor operation of routine experimentation, preparation method is extremely simple, be easy to control, gained nanometer hydroxyapatite major diameter is 20-80nm, and minor axis is 10-15nm, length-to-diameter ratio is (1-3): 1, organic content in nanometer hydroxyapatite is 1-5%, and nanometer hydroxyapatite content is 85-95%, and nanometer hydroxyapatite Z current potential is (-0.02)-(-0.1) mV, favorable dispersity, specific surface area is larger;
2) the present invention uses polyoxyethylene glycol to regulate and control to prepare nanometer hydroxyapatite, and polyoxyethylene glycol is washed away in washing of precipitate, and the hydroapatite particles biocompatibility obtained is good; The ownership system does not relate to noxious solvent for medicament, and preparation condition gentleness is controlled;
3) the nanometer hydroxyapatite size prepared of the present invention is little, there is the more excellent length-to-diameter ratio of medicine carrying carrier, can permeates cell membranes easily, and the nanometer hydroxyapatite of preparation has good biocompatibility, to normal cell and cancer cells all non-toxic reactions, can be used as the transmission carrier of gene or medicine, various acute and chronic diseases is treated.
Accompanying drawing explanation
Fig. 1 is the field emission scanning electron microscope figure of product in embodiment 1;
Fig. 2 is the transmission electron microscope figure of product in embodiment 1;
Fig. 3 be in embodiment 4 nanometer hydroxyapatite to the effect of vigor of L-02 cell;
Fig. 4 be in embodiment 4 nanometer hydroxyapatite to the effect of vigor of Hep-3B cell;
Fig. 5 be in embodiment 4 nanometer hydroxyapatite to the effect of vigor of PANC-1 cell;
Fig. 6 be in embodiment 4 nanometer hydroxyapatite to the effect of vigor of HeLa cell.
Embodiment
In order to make the present invention easier to understand, 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, NM specific experiment method in the following example, experimental technique carries out routinely usually.
Embodiment 1
Configure 15mM ca nitrate soln, 9mM ammonium dibasic phosphate solution respectively.First, 50mL ca nitrate soln is placed in 0 DEG C of cooling, then 50mL ammonium dibasic phosphate solution is joined in ca nitrate soln with 10/min by constant flow pump.In dropping process, progressively raise stirring velocity to 500rpm/min with the speed of 10rpm/min, and in reaction process with 1M sodium hydroxide solution maintain reaction system pH be 9.After being added dropwise to complete, dropwise adding polyoxyethylene glycol-200 0.5mg, after being added dropwise to complete, stopping stirring, leave standstill 1h.With 500rpm/min rapid stirring system, and be warming up to 70 DEG C, Keep agitation 1h.Stop heating, after question response system is cooled to room temperature, with deionized water 4 DEG C of low-temperature centrifugations, resuspended, washing granule 4 times.Gained throw out is placed on 12h in 40 DEG C of constant temperature ovens to dry, collects nano-hydroapatite particles.
The scanning electron microscopic picture of this nano-hydroapatite particles powder is shown in Fig. 1; Fig. 2 is shown in by transmission picture.
Embodiment 2
Configure 50mM calcium chloride solution, 30mM disodium phosphate soln respectively.First, 50mL calcium chloride solution is placed in 0 DEG C of cooling, then 200mL disodium phosphate soln is joined in calcium chloride solution with 60/min by constant flow pump.In dropping process, progressively raise stirring velocity to 1000rpm/min with the speed of 20rpm/min, and in reaction process with 1M sodium hydroxide solution maintain reaction system pH be 14.After being added dropwise to complete, dropwise adding Polyethylene glycol-2000 10mg, after being added dropwise to complete, stopping stirring, leave standstill 1h.With 1000rpm/min rapid stirring system, and be warming up to 100 DEG C, Keep agitation 1h.Stop heating, after question response system is cooled to room temperature, with deionized water 4 DEG C of low-temperature centrifugations, resuspended, washing granule 6 times.Gained throw out is placed on 24h in 60 DEG C of constant temperature ovens to dry, collects nano-hydroapatite particles.
Embodiment 3
Configure 40mM calcium chloride solution, 24mM ammonium dibasic phosphate solution respectively.First, 40mL calcium chloride solution is placed in 0 DEG C of cooling, then 160mL ammonium dibasic phosphate solution is joined in calcium chloride solution with 40/min by constant flow pump.In dropping process, progressively raise stirring velocity to 800rpm/min with the speed of 15rpm/min, and in reaction process with 1M sodium hydroxide solution maintain reaction system pH be 10.After being added dropwise to complete, dropwise adding PEG-4000 6mg, after being added dropwise to complete, stopping stirring, leave standstill 1h.With 800rpm/min rapid stirring system, and be warming up to 90 DEG C, Keep agitation 1h.Stop heating, after question response system is cooled to room temperature, with deionized water 4 DEG C of low-temperature centrifugations, resuspended, washing granule 6 times.Gained throw out is placed on 18h in 50 DEG C of constant temperature ovens to dry, collects nano-hydroapatite particles.
Embodiment 4
The nanometer hydroxyapatite 5mg that Example 1 obtains, is dispersed in 50mL deionized water, ultrasonic mixing, is mixed with the suspension of 0.1mg/mL, and carry out aseptically process.Nanometer hydroxyapatite is detected to the toxicity of various cell by CCK-8 cytoactive detection kit.Spread normal liver cell L-02, liver cancer cell Hep-3B, pancreatic cancer cell PANC-1, cervical cancer cell HeLa 96 orifice plate respectively, cell plantation density is 8000/hole, and volume is 100 μ L/ holes.In 96 orifice plate corresponding apertures, add the nanometer hydroxyapatite of 10 μ L different concns after 24h, continue to cultivate at 37 DEG C, 5%CO
248h is cultivated in constant incubator.In the 96 every holes of orifice plate, add 10 μ L CCK-8 solution, continue cultivation 1.5 hours, under microplate reader, test the light absorption value of each hole at 450nm place, and obtain the impact of different concns hydroxyapatite on cell viability.The final concentration of each nanometer hydroxyapatite in nutrient solution is respectively: 0,0.2,0.4,0.8,1.6,3.2,5.6,8,10mg/L.
The effect of vigor of this nanometer hydroxyapatite to L-02 cell is shown in Fig. 3, sees Fig. 4 to the effect of vigor of Hep-3B cell, sees Fig. 5 to the effect of vigor of PANC-1 cell, sees Fig. 6 to the toxic effect of HeLa cell.Can find out from Fig. 3-6, this nanometer hydroxyapatite does not have non-toxic reaction to normal liver cell and cancer cells.
The nano-hydroapatite particles that embodiment 2 and 3 obtains all has the technical parameter identical with the nanometer hydroxyapatite that embodiment 1 obtains.
Claims (5)
1. a fast preparation method for polyoxyethylene glycol regulation and control nanometer hydroxyapatite, is characterized in that by following processing step:
1) calcium salt soln of 15-50mM is configured respectively, 9-30mM phosphate solution;
2) calcium salt soln getting 50-200mL in 0 DEG C of cooling, by same volume phosphate solution with 10-60 drip/min speed joins in calcium salt soln, and stirs with 500-1000rpm/min gradient;
3) step 2 is maintained with sodium hydroxide solution) pH of reaction system is 9-14, until be added dropwise to complete;
4) in step 3) reaction system, slowly add 0.5-10mg polyoxyethylene glycol, and stop stirring;
5) after 1h by step 4) reaction system with 500-1000rpm/min rapid stirring, and be warming up to 70-100 DEG C, Keep agitation 1h;
6) stop heating, after step 5) system is cooled to room temperature, resuspended washing granule centrifugal under the cold condition of 2-8 DEG C with deionized water 4-6 time, obtains throw out;
7) throw out step 6) obtained, with filter paper capping, carries out dustless drying treatment, the nanometer hydroxyapatite of obtained polyoxyethylene glycol regulation and control;
Described nanometer hydroxyapatite is club shaped structure, major diameter is 20-80nm, minor axis is 10-15nm, length-to-diameter ratio is (1-3): 1, organic content 1-5% in nanometer hydroxyapatite, nanometer hydroxyapatite content is 85-95%, and nanometer hydroxyapatite Z current potential is (-0.02)-(-0.1) mV, and described polyethylene glycol relative molecular weight is 200-2000.
2. the fast preparation method of a kind of polyoxyethylene glycol regulation and control nanometer hydroxyapatite according to claim 1, is characterized in that calcium salt described in step 1) is any one in monocalcium phosphate, nitrocalcite, Calcium hydrogen carbonate, calcium bisulfate, calcium bisulfite, Losantin, Calcium Bromide, calcium iodide, calcium chlorate, calcium perchlorate or calcium permanganate.
3. the fast preparation method of a kind of polyoxyethylene glycol regulation and control nanometer hydroxyapatite according to claim 1, is characterized in that the phosphoric acid salt described in step 1) is any one in Secondary ammonium phosphate, Sodium phosphate dibasic, dipotassium hydrogen phosphate.
4. a kind of polyoxyethylene glycol regulates and controls the fast preparation method of nanometer hydroxyapatite according to claim 1, and it is characterized in that the centrifugal speed in step 6) is 4000-16000rpm/min, centrifugation time is 2-30min.
5. a kind of polyoxyethylene glycol regulates and controls the fast preparation method of nanometer hydroxyapatite according to claim 1, and it is characterized in that in step 7), throw out suspends with dehydrated alcohol when drying, drying temperature is 40-60 DEG C, and time of drying is 12-24h.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201410785658.8A CN104528676A (en) | 2014-12-18 | 2014-12-18 | Method for quickly preparing polyethylene glycol regulated and controlled nano hydroxyapatite |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201410785658.8A CN104528676A (en) | 2014-12-18 | 2014-12-18 | Method for quickly preparing polyethylene glycol regulated and controlled nano hydroxyapatite |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN104528676A true CN104528676A (en) | 2015-04-22 |
Family
ID=52844372
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201410785658.8A Pending CN104528676A (en) | 2014-12-18 | 2014-12-18 | Method for quickly preparing polyethylene glycol regulated and controlled nano hydroxyapatite |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN104528676A (en) |
Cited By (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105327364A (en) * | 2015-12-15 | 2016-02-17 | 浙江理工大学 | Nanometer-hydroxyapatite-siRNA compound and preparing method thereof |
| CN106927442A (en) * | 2017-03-29 | 2017-07-07 | 河南师范大学 | A kind of preparation method of cellular porous hydroxyapatite |
| CN108046227A (en) * | 2017-12-21 | 2018-05-18 | 衢州学院 | A kind of method of the needle-shaped hydroxy calcium apatite of low temperature preparation micron order |
| CN109928374A (en) * | 2019-02-26 | 2019-06-25 | 大连理工大学 | A kind of preparation method for the nano hydroxyapatite material that draw ratio is controllable |
| CN110775953A (en) * | 2019-11-27 | 2020-02-11 | 中山市科信生物技术有限公司 | Method for synthesizing thermodynamically stable hydroxyapatite with microscopic kinetic reaction limitation |
| CN114560454A (en) * | 2022-03-16 | 2022-05-31 | 浙江理工大学 | Chrysanthemum-shaped calcium phosphate nanocluster based on polydopamine regulation and control as well as preparation method and application thereof |
Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1308016A (en) * | 2000-11-10 | 2001-08-15 | 中国科学院上海硅酸盐研究所 | Preparation of low temperature sinterable hydroxyapatite powder |
| CN101385856A (en) * | 2008-10-17 | 2009-03-18 | 同济大学 | Controllable slow-releasing nano hydroxyapatite with efficient absorption for antalzyme and preparation method thereof |
| CN103553012A (en) * | 2013-11-07 | 2014-02-05 | 中山大学 | Method for preparing nano hydroxyapatite |
-
2014
- 2014-12-18 CN CN201410785658.8A patent/CN104528676A/en active Pending
Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1308016A (en) * | 2000-11-10 | 2001-08-15 | 中国科学院上海硅酸盐研究所 | Preparation of low temperature sinterable hydroxyapatite powder |
| CN101385856A (en) * | 2008-10-17 | 2009-03-18 | 同济大学 | Controllable slow-releasing nano hydroxyapatite with efficient absorption for antalzyme and preparation method thereof |
| CN103553012A (en) * | 2013-11-07 | 2014-02-05 | 中山大学 | Method for preparing nano hydroxyapatite |
Cited By (9)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105327364A (en) * | 2015-12-15 | 2016-02-17 | 浙江理工大学 | Nanometer-hydroxyapatite-siRNA compound and preparing method thereof |
| CN105327364B (en) * | 2015-12-15 | 2019-03-12 | 浙江理工大学 | A kind of nanometer hydroxyapatite-siRNA compound and preparation method thereof |
| CN106927442A (en) * | 2017-03-29 | 2017-07-07 | 河南师范大学 | A kind of preparation method of cellular porous hydroxyapatite |
| CN106927442B (en) * | 2017-03-29 | 2018-10-12 | 河南师范大学 | A kind of preparation method of cellular porous hydroxyapatite |
| CN108046227A (en) * | 2017-12-21 | 2018-05-18 | 衢州学院 | A kind of method of the needle-shaped hydroxy calcium apatite of low temperature preparation micron order |
| CN109928374A (en) * | 2019-02-26 | 2019-06-25 | 大连理工大学 | A kind of preparation method for the nano hydroxyapatite material that draw ratio is controllable |
| WO2020173265A1 (en) * | 2019-02-26 | 2020-09-03 | 大连理工大学 | Preparation method for nano hydroxyapatite material with controllable length-to-diameter ratio |
| CN110775953A (en) * | 2019-11-27 | 2020-02-11 | 中山市科信生物技术有限公司 | Method for synthesizing thermodynamically stable hydroxyapatite with microscopic kinetic reaction limitation |
| CN114560454A (en) * | 2022-03-16 | 2022-05-31 | 浙江理工大学 | Chrysanthemum-shaped calcium phosphate nanocluster based on polydopamine regulation and control as well as preparation method and application thereof |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN104528676A (en) | Method for quickly preparing polyethylene glycol regulated and controlled nano hydroxyapatite | |
| Xiang et al. | Fabrication of a pH-responsively controlled-release pesticide using an attapulgite-based hydrogel | |
| Lai et al. | Hydrothermal fabrication of porous hollow hydroxyapatite microspheres for a drug delivery system | |
| Lvov et al. | Clay nanotube encapsulation for functional biocomposites | |
| Barick et al. | Nanoscale assembly of mesoporous ZnO: A potential drug carrier | |
| Qi et al. | Hydroxyapatite hierarchically nanostructured porous hollow microspheres: rapid, sustainable microwave‐hydrothermal synthesis by using creatine phosphate as an organic phosphorus source and application in drug delivery and protein adsorption | |
| Karaman et al. | Shape engineering vs organic modification of inorganic nanoparticles as a tool for enhancing cellular internalization | |
| CN104491871B (en) | A kind of nano microgel of the pH based on polyglutamic acid and cystamine and reduction-sensitive | |
| Simovic et al. | Silica materials in drug delivery applications | |
| CN103803565B (en) | A kind of preparation method of monodispersity hollow mesoporous silicon dioxide nano particle | |
| Kang et al. | Enzyme-responsive polymeric supra-amphiphiles formed by the complexation of chitosan and ATP | |
| CN104961115A (en) | Hollow hydroxyapatite microsphere and preparation method thereof | |
| Dzido et al. | Rapid continuous microwave-assisted synthesis of silver nanoparticles to achieve very high productivity and full yield: from mechanistic study to optimal fabrication strategy | |
| Li et al. | Chitosan stabilized Prussian blue nanoparticles for photothermally enhanced gene delivery | |
| CN105329929A (en) | Spherical calcium carbonate particles and preparation method thereof | |
| Wang et al. | Shape matters: morphologically biomimetic particles for improved drug delivery | |
| Jiang et al. | Strontium-substituted, luminescent and mesoporous hydroxyapatite microspheres for sustained drug release | |
| CN101608020A (en) | Magnetic Fe prepared by hydrothermal method3O4Polymer submicron sphere and application | |
| CN104525966B (en) | A kind of xanthan gum-argentum nano composite material and preparation method thereof | |
| CN109111575B (en) | Preparation method and application of metal-organic framework nano-particles | |
| CN105327364B (en) | A kind of nanometer hydroxyapatite-siRNA compound and preparation method thereof | |
| Xuan et al. | Synthesis of dodecylamine-functionalized graphene quantum dots and their application as stabilizers in an emulsion polymerization of styrene | |
| CN105348548A (en) | Hydrogel microspheres based on glucan and preparation method thereof | |
| CN107931628A (en) | A kind of load type floriform hierarchy nano-noble metal material and preparation method thereof | |
| Qian et al. | Facile synthetic Photoluminescent Graphene Quantum dots encapsulated β-cyclodextrin drug carrier system for the management of macular degeneration: Detailed analytical and biological investigations |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| PB01 | Publication | ||
| C10 | Entry into substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| RJ01 | Rejection of invention patent application after publication |
Application publication date: 20150422 |
|
| RJ01 | Rejection of invention patent application after publication |