CN105193843A - Method for killing cancer cells by use of TiO2-based nanometer material - Google Patents
Method for killing cancer cells by use of TiO2-based nanometer material Download PDFInfo
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- CN105193843A CN105193843A CN201510664732.5A CN201510664732A CN105193843A CN 105193843 A CN105193843 A CN 105193843A CN 201510664732 A CN201510664732 A CN 201510664732A CN 105193843 A CN105193843 A CN 105193843A
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- nano material
- cancerous cell
- titanium dioxide
- dioxide base
- base nano
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Abstract
The invention discloses a method for killing cancer cells by use of a TiO2-based nanometer material, and aims to explore a new way for cancer treatment. According to the method, the TiO2-based nanometer material is developed, and an experimental method for killing cancer cells and a method for evaluating the cancer cell killing efficiency are constructed. The method provided by the invention has the characteristics that the TiO2-based nanometer material is high in stability and dispersity, and has a function of killing cancer cells at relatively low temperature (4-39 DEG C), especially at normal body temperature (37 DEG C); the experimental method is simple, easy to operate, low in implementation condition requirement, and free from other requirements (such as illumination); the method for evaluating the cancer cell killing efficiency is accurate and feasible. Therefore, the method provided by the invention opens up a new direction for cancer treatment.
Description
[technical field]
The present invention establishes a kind of method utilizing titanium dioxide base nano material to kill cancerous cell.Utilize the titanium dioxide base nano material newly developed at a lower temperature (4 ~ 39 DEG C), particularly carry out the experiment of killing cancerous cell normal human body temperature's (37 DEG C), and cancerous cell efficiency is killed to it carry out testing evaluation.This new method belongs to field of cancer.
[background technology]
Cancer is the world today's directly one of disease threatening human life, and global cancer morbidity is in rising trend in recent years.At present clinically mainly contain the methods such as operative treatment, chemotherapy, radiotherapy; Surgical therapy is only limited to topical therapeutic, and chemotherapy and radiotherapy are all along with larger side effect, and therefore exploratory development is little to human body toxicity, and the new method of effectively killing cancerous cell is the primary study direction in Current cancer treatment field.
The invention discloses a kind of method utilizing titanium dioxide base nano material to kill cancerous cell.The method be utilize development titanium dioxide base nano material at a lower temperature (4 ~ 39 DEG C), particularly effectively kill cancerous cell normal human body temperature's (37 DEG C), and method is simple to operation, implementation condition requires low, and the efficiency rating of killing cancerous cell is accurate.The method utilizing nano material to kill cancerous cell normal human body temperature's (37 DEG C) at present has no report, means that the present invention has potential application prospect in Therapeutic cancer field.
[summary of the invention]
The present invention establishes a kind of method utilizing titanium dioxide base nano material to kill cancerous cell, its summary of the invention comprises following aspect: 1. with dopant ion salt, titanate esters or titanate for presoma, with carbon-based material (as Graphene etc.) for composite, the methods such as collosol and gel are adopted to prepare titanium dioxide base nano material; 2. establish and utilize this nano material (4 ~ 39 DEG C) at a lower temperature, particularly effectively kill the experimental technique of cancerous cell normal human body temperature's (37 DEG C) and kill the evaluation methodology of cancerous cell efficiency.3. utilize titanium dioxide base nano material to carry out tumoricidal experiment.Result shows, titanium dioxide base nano material is (4 ~ 39 DEG C) at a lower temperature, particularly under normal human body temperature's (37 DEG C) condition, there is the stronger ability of killing cancerous cell, and experimental technique simple, be easy to operation and implementation condition requires low, the evaluation methodology of killing cancerous cell efficiency is accurately feasible.
In order to realize the object of the invention, the invention provides a kind of method utilizing titanium dioxide base nano material to kill cancerous cell, it comprises: development titanium dioxide base nano material, establishes the experimental technique killing cancerous cell and the evaluation methodology of killing cancerous cell efficiency; The feature of described titanium dioxide base nano material is with dopant ion salt, titanate esters or titanate for presoma, adopts sol-gal process to prepare metal or nonmetal doping TiO
2nano material or carbon-based material composition metal or nonmetal doping TiO
2nano material; Good dispersion, the stability of this nano material are strong, and the efficiency of killing cancerous cell at the lower temperature of 4 ~ 39 DEG C is high; The experimental technique killing cancerous cell and the evaluation methodology of killing cancerous cell efficiency are characterised in that cancerous cell cultivates in containing 1640 culture medium of 10% hyclone, 100U/mL streptomycin and 100 μ g/mL penicillins, are placed in 37 DEG C, containing 5%CO
2saturated humidity incubator in; First be in the cancerous cell of exponential phase with trypsinization, make during concentration is every milliliter and have 1 × 10
5the cancer cell suspension of individual cell, injects each hole 200 μ L in 96 orifice plates, ensures that each hole has 2 × 10
4individual cell; Then cultivate 24 hours in incubator, shift out original culture medium, and 1640 culture medium injected containing variable concentrations titanium dioxide base nano material, cultivation is continued 16 hours in the incubator of saturated humidity, guarantee that nano material and cancerous cell interact, finally adopt the MTT method of standard to detect the survival rate of cancerous cell, evaluate tumoricidal efficiency.
Further technical scheme is, described lower temperature is normal human body temperature 37 DEG C.
Further technical scheme is, the described method preparing titanium dioxide base nano material is as follows: (1) doped Ti O
2prepared by nanoparticle: at normal temperatures, configure certain density dopant ion saline solution, add diluent again, acidulant and titanate esters or titanate is added under Keep agitation condition, obtained mixed sols, under room temperature aging 24 hours, obtains mixed gel, through 90 DEG C ~ 150 DEG C oven dry, in air, 450 DEG C ~ 800 DEG C calcining 2.5 ~ 6h obtain doped Ti O
2nanoparticle; (2) the composite mixed TiO of Graphene
2preparation method of nano material is as follows: in beaker, add appropriate concentrated sulphuric acid under ice-water bath condition; Under continuous stirring, crystalline flake graphite and Chile saltpeter is added; Control reaction temperature less than 20 DEG C again gradation adds potassium permanganate, obtained bottle green liquid; Reaction temperature is risen to 35 DEG C, continue to stir 1h, slowly add a certain amount of deionized water again, liquid acutely seethes with excitement and maintains the temperature at 98 DEG C is 15 ~ 30min, continue to add appropriate distilled water and hydrogen peroxide, leave standstill after 24h and obtain red-brown precipitation, finally with 5%HCl solution and deionized water wash until in filtrate sulfate radical-free be detected; Gained solid obtains graphene oxide 60 DEG C of dryings; 9: 1 ~ 3: 7 by TiO by a certain percentage
2base nanoparticle and graphene oxide are placed in 250ml beaker, add diluent, ultrasonic 80 ~ 120min; The pressed powder obtained after centrifugal is 40 ~ 80 DEG C of oven dry, and under inert gas shielding, 300 ~ 450 DEG C of calcining 2 ~ 5h, obtain the composite mixed TiO of Graphene
2nano material.
Further technical scheme is, described cancerous cell is any one in human lung cancer cell A549, human colon cancer cell, human cervical carcinoma cell or synovial cell.
Further technical scheme is, described titanium dioxide base nano material is metal or nonmetallic ion-doped titanium dioxide or graphene carbon sill composition metal or nonmetallic ion-doped titanium dioxide.
Further technical scheme is, described dopant ion is palladium, any one in indium, platinum, stannum or nitrogen metal or nonmetallic ion singly mix or any two kinds two mix.
Further technical scheme is, described acidulant is the mixing of any one or more in hydrochloric acid, nitric acid, phosphoric acid, glacial acetic acid, perchloric acid.
Further technical scheme is, described titanate esters or titanate are selected from following a kind of or several mixing: Ti (OC
4h
9)
4, Ti (OC
2h
5)
4, Ti (iso-OC
3h
7)
4, Ti (SO
4)
2or TiCl
4.
Further technical scheme is, described diluent is the mixing of any one or more in high purity water, dehydrated alcohol, absolute methanol, glacial acetic acid or formic acid.
Further technical scheme is, described noble gas is any one in nitrogen, argon or helium.
[accompanying drawing explanation]
The Raman spectrum of Fig. 1 embodiment sample is (with pure TiO
2relatively);
Fig. 2 embodiment sample kills the efficiency chart of lung cell A549 (with pure TiO
2relatively);
Fig. 3 embodiment sample kills the efficiency chart of carcinoid synovial cell (with pure TiO
2relatively);
Below in conjunction with specific embodiments and the drawings, the present invention is described further, the preparation illustrating titanium dioxide base nano material of the present invention, the experimental technique killing cancerous cell and kill the evaluation methodology of cancerous cell efficiency.
[detailed description of the invention]
The preparation method of Pd doped titanium dioxide nano material is as follows: at normal temperatures, first prepares 5mLPdCl
2solution (0.1036mol/L), then add 40mL dehydrated alcohol.After Keep agitation half an hour, add the butyl titanate of 1mL concentrated hydrochloric acid and 12mL, then at aging 24h, obtain mixed gel.Through 100 DEG C of oven dry, grind into powder, obtains Pd-TiO at 450 DEG C of calcining 2.5h
2nanoparticle.
Graphene composite titanium dioxide based nano-material preparation method is as follows: the beaker assembling 1L in ice-water bath, adds the concentrated sulphuric acid of 115mL.Add the solid mixture of 5g crystalline flake graphite and 2.5g Chile saltpeter under continuous stirring, then gradation adds 15g potassium permanganate, control reaction temperature and be no more than 20 DEG C.Then liquid is warmed up to 35 DEG C, continues to stir 60min.Slowly add the deionized water of 230mL again, liquid acutely seethes with excitement and maintains the temperature at about 98 DEG C 15min.And add 0.7L distilled water and 12.5mL hydrogen peroxide, finally with the 5%HCl solution of 1L and deionized water wash until in filtrate sulfate radical-free be detected.Last fully dry in the vacuum drying oven of 60 DEG C, obtain graphene oxide.
6: 4 by Pd-TiO in mass ratio
2be placed in 250ml beaker with graphene oxide, add 150ml dehydrated alcohol, ultrasonic 100min.Dry in the baking oven of 50 DEG C after centrifugal.Finally under 300 DEG C of argon shields, calcine 2h, i.e. obtained graphene/Pd-TiO
2nano material.Pure graphene adopts identical preparation method, and difference is not add Pd-TiO in preparation process
2.
Fig. 1 is and pure TiO
2relatively, the Raman spectrum of above-described embodiment sample.As can be seen from the figure, at 144cm
-1(E
g), 194cm
-1(E
g), 396cm
-1(B
1g), 516cm
-1(A
1gandB
1g) and 638cm
-1(E
g) obvious diffraction maximum appears in place, shows that the titanium dioxide prepared is typically anatase structured.In addition at 1314cm
-1and 1588cm
-1also occurred intensity peak, this distinguishes D peak and the G peak of correspondence graphene.Therefore show successfully to prepare TiO
2, Pd-TiO
2, graphene and graphene/Pd-TiO
2nano material.
The result of Fig. 2 and Fig. 3 shows TiO
2almost do not have killing cancer cell effect, cell survival rate still remains on about 85%; Sample is killing cancer cell A549 activity (see Fig. 2) under normal human body temperature, when nano material concentration is 400 μ g/mL, and graphene/Pd-TiO
2the A549 cancer cell survival rate of sample is only 7.2%; And pure TiO
2the A549 cancer cell survival rate of sample is 87.5%, Pd-TiO
2the survival rate of A549 cancerous cell be 45.8%; Sample kills synovial cell's activity (see Fig. 3) under normal human body temperature, after nano material concentration is 400 μ g/mL, reaction 16h, and graphene/Pd-TiO
2a549 synovial cell's survival rate of sample is 21.4%, and pure TiO
2synovial cell's survival rate of sample is 81%, Pd-TiO
2the survival rate of synovial membrane cancerous cell be 46.5%.
As can be seen from embodiment, graphene/Pd-TiO
2sample and pure TiO
2compare, the efficiency of killing lung carcinoma cell improves about 7.4 times, and the efficiency of killing quasi-cancer cell synovial cell improves about 4.1 times.
Claims (10)
1. utilize titanium dioxide base nano material to kill a method for cancerous cell, it comprises: development titanium dioxide base nano material, establishes the experimental technique killing cancerous cell and the evaluation methodology of killing cancerous cell efficiency; The feature of described titanium dioxide base nano material is with dopant ion salt, titanate esters or titanate for presoma, adopts sol-gal process to prepare metal or nonmetal doping TiO
2nano material or Graphene composition metal or nonmetal doping TiO
2nano material; Good dispersion, the stability of this nano material are strong, and the efficiency of killing cancerous cell at the lower temperature of 4 ~ 39 DEG C is high; The experimental technique killing cancerous cell and the evaluation methodology of killing cancerous cell efficiency are characterised in that cancerous cell cultivates in containing 1640 culture medium of 10% hyclone, 100U/mL streptomycin and 100 μ g/mL penicillins, are placed in 37 DEG C, containing 5%CO
2saturated humidity incubator in; First be in the cancerous cell of exponential phase with trypsinization, make during concentration is every milliliter and have 1 × 10
5the cancer cell suspension of individual cell, injects each hole 200 μ L in 96 orifice plates, ensures that each hole has 2 × 10
4individual cell; Then cultivate 24 hours in incubator, shift out original culture medium, and 1640 culture medium injected containing variable concentrations titanium dioxide base nano material, cultivation is continued 16 hours in the incubator of saturated humidity, guarantee that nano material and cancerous cell interact, finally adopt the MTT method of standard to detect the survival rate of cancerous cell, evaluate tumoricidal efficiency.
2. the method utilizing titanium dioxide base nano material to kill cancerous cell according to claim 1, is characterized in that, described lower temperature is normal human body temperature 37 DEG C.
3. the method utilizing titanium dioxide base nano material to kill cancerous cell according to claim 1, is characterized in that, the described method preparing titanium dioxide base nano material is as follows: (1) doped Ti O
2prepared by nanoparticle: at normal temperatures, configure certain density dopant ion saline solution, add diluent again, acidulant and titanate esters or titanate is added under Keep agitation condition, obtained mixed sols, under room temperature aging 24 hours, obtains mixed gel, through 90 DEG C ~ 150 DEG C oven dry, in air, 450 DEG C ~ 800 DEG C calcining 2.5 ~ 6h obtain doped Ti O
2nanoparticle; (2) the composite mixed TiO of Graphene
2preparation method of nano material is as follows: in beaker, add appropriate concentrated sulphuric acid under ice-water bath condition; Under continuous stirring, crystalline flake graphite and Chile saltpeter is added; Control reaction temperature less than 20 DEG C again gradation adds potassium permanganate, obtained bottle green liquid; Reaction temperature is risen to 35 DEG C, continue to stir 1h, slowly add a certain amount of deionized water again, liquid acutely seethes with excitement and maintains the temperature at 98 DEG C is 15 ~ 30min, continue to add appropriate distilled water and hydrogen peroxide, leave standstill after 24h and obtain red-brown precipitation, finally with 5%HCl solution and deionized water wash until in filtrate sulfate radical-free be detected; Gained solid obtains graphene oxide 60 DEG C of dryings; 9: 1 ~ 3: 7 by doped Ti O by a certain percentage
2nanoparticle and graphene oxide are placed in 250ml beaker, add diluent, ultrasonic 80 ~ 120min; The pressed powder obtained after centrifugal is 40 ~ 80 DEG C of oven dry, and under inert gas shielding, 300 ~ 450 DEG C of calcining 2 ~ 5h, obtain the composite mixed TiO of Graphene
2nano material.
4. the titanium dioxide base nano material that utilizes according to any one of claims 1 to 3 kills the method for cancerous cell, it is characterized in that: described cancerous cell is any one in human lung cancer cell A549, human colon cancer cell, human cervical carcinoma cell or synovial cell.
5. the titanium dioxide base nano material that utilizes according to any one of claims 1 to 3 kills the method for cancerous cell, it is characterized in that: described titanium dioxide base nano material is metal or carbon-based material composition metal or the nonmetallic ion-doped titanium dioxide such as nonmetallic ion-doped titanium dioxide or Graphene.
6. the titanium dioxide base nano material that utilizes according to any one of claims 1 to 3 kills the method for cancerous cell, it is characterized in that: any one in the metals such as described dopant ion is palladium, indium, platinum, stannum or nitrogen or nonmetallic ion singly mix or any two kinds two mix.
7. the method utilizing titanium dioxide base nano material to kill cancerous cell according to claim 3, is characterized in that: described acidulant is the mixing of any one or more in hydrochloric acid, nitric acid, phosphoric acid, glacial acetic acid, perchloric acid.
8. the titanium dioxide base nano material that utilizes according to any one of claims 1 to 3 kills the method for cancerous cell, it is characterized in that: described titanate esters or titanate are selected from following a kind of or several mixing: Ti (OC
4h
9)
4, Ti (OC
2h
5)
4, Ti (iso-OC
3h
7)
4, Ti (SO
4)
2or TiCl
4deng.
9. the method utilizing titanium dioxide base nano material to kill cancerous cell according to claim 3, is characterized in that: described diluent is the mixing of any one or more in high purity water, dehydrated alcohol, absolute methanol, glacial acetic acid or formic acid.
10. the method utilizing titanium dioxide base nano material to kill cancerous cell according to claim 3, is characterized in that: described noble gas is any one in nitrogen, argon or helium.
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|---|---|---|---|
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Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US20060210798A1 (en) * | 2005-03-16 | 2006-09-21 | Clemens Burda | Doped metal oxide nanoparticles and methods for making and using same |
| CN101301619A (en) * | 2008-07-03 | 2008-11-12 | 南开大学 | Method for preparing high efficiency metallic, non-metallic ion co-doped nano-TiO2 visible-light responsive photocatalyst |
| CN101444724A (en) * | 2008-12-24 | 2009-06-03 | 南开大学 | Method for preparing high activity non-metallic ion co-doped titanium dioxide photochemical catalyst |
| CN101890344A (en) * | 2010-07-27 | 2010-11-24 | 华东理工大学 | Preparation method of graphene/titanium dioxide composite photocatalyst |
-
2015
- 2015-10-15 CN CN201510664732.5A patent/CN105193843B/en active Active
Patent Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US20060210798A1 (en) * | 2005-03-16 | 2006-09-21 | Clemens Burda | Doped metal oxide nanoparticles and methods for making and using same |
| CN101301619A (en) * | 2008-07-03 | 2008-11-12 | 南开大学 | Method for preparing high efficiency metallic, non-metallic ion co-doped nano-TiO2 visible-light responsive photocatalyst |
| CN101444724A (en) * | 2008-12-24 | 2009-06-03 | 南开大学 | Method for preparing high activity non-metallic ion co-doped titanium dioxide photochemical catalyst |
| CN101890344A (en) * | 2010-07-27 | 2010-11-24 | 华东理工大学 | Preparation method of graphene/titanium dioxide composite photocatalyst |
Non-Patent Citations (1)
| Title |
|---|
| LI,ET AL: "Study on the visible-light-induced photokilling effect of nitrogen-doped TiO2 nanoparticles on cancer cells", 《NANOSCALE RESEARCH LETTERS》 * |
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