CN102053148A - Method for detecting exposure toxicity of nanoparticles and toxicant exposure cabinet used thereby - Google Patents
Method for detecting exposure toxicity of nanoparticles and toxicant exposure cabinet used thereby Download PDFInfo
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- CN102053148A CN102053148A CN2009101978970A CN200910197897A CN102053148A CN 102053148 A CN102053148 A CN 102053148A CN 2009101978970 A CN2009101978970 A CN 2009101978970A CN 200910197897 A CN200910197897 A CN 200910197897A CN 102053148 A CN102053148 A CN 102053148A
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Abstract
The invention discloses a method for detecting the exposure toxicity of nanoparticles and a toxicant exposure cabinet used thereby. In the toxicant exposure cabinet, the bottom is provided with a fan, and the middle-upper part is provided with a metal net on which test animals can be placed; and meanwhile, the middle part of the cabinet wall of the toxicant exposure cabinet is provided with an opening which is a sample adding hole, a seal plug is arranged in the opening, and the top of the cabinet is provided with a cabinet cover which can be sealed and fixed. The method comprises the following steps: (1) fixing a test animal by a fixator, wherein the outside surface of the fixator is coated with smooth materials, and only the head of the test animal is exposed outside; (2) putting the fixed test animal on the metal net in the toxicant exposure cabinet, and sealing and fixing the toxicant exposure cabinet; and (3) starting the fan, scattering nanoparticles into the toxicant exposure cabinet from the sample adding hole, adjusting the air speed, and controlling the nanoparticles to be blown away, thereby realizing static toxicant exposure. Through simulating the approaches by which people touch the dust in an actual environment, the method and toxicant exposure cabinet provided by the invention are strong in feasibility, easy for operation and convenient to use; and meanwhile, the toxicant exposure cabinet is small in volume, small in sample use amount and low in cost, and the detection simulation conditions of the exposure toxicology can be preferably achieved.
Description
Technical field
The invention belongs to the animal experiment method field, particularly a kind of nanoparticle that detects exposes the method for toxicity and used toxicant exposure cabinet thereof.
Background technology
Along with developing rapidly of nanosecond science and technology, nanoparticle is widely used in different fields as new material, and because grain diameter reaches nanoscale, the essence variation has also taken place in their physicochemical property, will show small-size effect, surface effect, effects such as quantum size, therefore compare with traditional material, huge variation has taken place in the character of nanoparticle, the corresponding variation that matter also takes place of its biological safety.In daily life or because the influence of working environment, many workers can touch nano particle, are exactly SiO and contact maximum
2Particle often causes damage because of sucking nano particle to health, and cause injury of lungs such as people because of the contact nanometer particle, even cause the case of silicosis more and more to come into one's own, as Mine Dust, the fine particle that ceramic tile processing produces, cement mill chimney dust etc.But so far but and imperfection for the safety evaluatio of nanoparticle.In fact, present many national researchers have realized that this problem, and increasing worker begins to pay close attention to the biological effect of the material of nanoscale, to environment and health affected.
In recent years, scientists has been carried out the research of nanoparticle exposure toxicity in succession, mainly concentrate on the molecular cell level, directly investigate the influence of nanoparticle to red blood cell and rat pulmonary alveolus macrophage, also there is the researcher to pass through the tracheae injection method and directly injects nanoparticle in the rat tracheae, investigate the damage of nanoparticle tissues such as rat lungs.These methods have higher requirement to experiment operator usually, complicated operation, consider that in addition the central people of actual life contact the approach and the mode of superfine microparticle, these methods lack simulating reality environment and investigate the environment of nanoparticles to people, the influence of health.
Summary of the invention
Therefore, the technical problem to be solved in the present invention is exactly that toxicity assessment at existing nanoparticle only stops molecular cell or organizes level, also there is not to contact the deficiency of the method for evaluating safety of superfine microparticle as people in the middle of the actual life, provide a kind of nanoparticle that detects to expose the method for toxicity and used device thereof, the method is contaminated in the mode that people during simulating reality is lived contact dust as far as possible, selected nanoparticle, carry out the static state contamination, simple, thus realize nanoparticle is carried out safety evaluatio.
The present invention solves the problems of the technologies described above one of technical scheme of being adopted: a kind of toxicant exposure cabinet, in this toxicant exposure cabinet, the bottom is a blower fan, the middle and upper part is the wire netting that can place experimental animal, there is an opening at cabinet wall middle part at toxicant exposure cabinet, be well, have sealing-plug can clog this opening in this opening, the cabinet top is salable fixing cabinet lid.
According to the present invention, preferable, use circular arc material chamfering to adhere to for interior four jiaos of toxicant exposure cabinet to remove the dead angle.The inwall of toxicant exposure cabinet is 4 circular arc wall hooks, uses for the support metal net.Wire netting is stainless (steel) wire preferably.Cabinet has covered O-ring seal, so that cabinet is covered sealing and fixing.The preferable of the material of toxicant exposure cabinet cabinet wall is organic glass.
According to the present invention, preferable, between blower fan and the toxicant exposure cabinet bottom cylinder that is fixed at the bottom of the cabinet, to support blower fan, make itself and toxicant exposure cabinet bottom keep certain space.Preferable, toxicant exposure cabinet cabinet wall is being opened a pore with the horizontal fan place, and the blower fan electric wire passes through this pore outside toxicant exposure cabinet.The preferably adjustable wind speed blower fan of blower fan, more preferably circular adjustable wind speed blower fan.
According to the present invention, preferable, also comprise an experimental animal fixator, the smooth outer surface densification of this fixator.Described fixator can be that conventional animal is used fixator, and the health of experimental animal has only a nose to expose all in fixator inside.The preferably cylindrical fixator of this fixator.What the fixator material was preferable is plastics.
The present invention solves the problems of the technologies described above two of the technical scheme that adopted: a kind ofly detect the method that nanoparticle exposes toxicity, may further comprise the steps:
(1) be covered with the fixator fixation test animal of lubricious material with outside surface, the head of a retention test animal is exposed in the surrounding environment;
(2) experimental animal that fixes is placed on the wire netting in the aforesaid toxicant exposure cabinet the good toxicant exposure cabinet of sealing and fixing;
(3) open blower fan, be sprinkled into nanoparticle by well, regulate wind speed, the control nanoparticle is blown afloat, static contamination.
In the step (1), preferable, before the fixation test animal, adhere to water-absorbing sponge, with excretas such as absorption urines at the experimental animal afterbody.What fixator was preferable is the cylindrical fixator of class.What the fixator material was preferable is stainless steel.The lubricious material that the fixator surface is covered is stainless steel substrates preferably, preservative film etc.
In the step (2), the mean grain size of described nanoparticle can be any, preferred 10nm～10 μ m, more preferably 20nm～800nm.
Step (3) is in order experimental animal to be exposed in the nanoparticle dust, to carry out the static state contamination.Wherein preferable, regulate wind speed by speed regulator, the control dust can blow afloat and get final product.It is any time that static contamination time can continue, and carries out many times, and preferably static contamination time is 2 hours, and each 2 rats are not taken food during the experimental animal contamination and do not drink water, the other times feed of freely drinking water.After contamination finishes, require experimental animal is observed, detect xicity related index, the exposure toxicity of assessment nanoparticle according to experimental program.
Raw material that the present invention is used or reagent except that specifying, all commercially available getting.
Than prior art, beneficial effect of the present invention is as follows: people contact the approach of dust in the middle of the simulating reality environment of the present invention, and feasibility is strong, and easy operating is easy to use.And should static state toxicant exposure cabinet volume little, sample dosage is little, and be with low cost.Toxicologic detection simulated conditions have been realized exposing preferably.The present invention detects the method that nanoparticle exposes toxicity, can be used to investigate nanoparticle and expose toxicology and the exposure security of nanoparticle is assessed etc., has realistic meaning and experimental feasibility preferably.
Description of drawings
Below in conjunction with description of drawings feature of the present invention and beneficial effect.
Fig. 1 is the toxicant exposure cabinet overall diagram.1, toxicant exposure cabinet cabinet wall; 2, well; 3, wire netting; 4, the wall hook of support metal net; 5, the cabinet arc interior angle of toxicant exposure cabinet; 6, the cabinet lid; 7, blower fan; 8, the blower fan stilt.
Fig. 2 is an experimental animal fixer structure synoptic diagram.9, animal holder; 10, a nose outlet; 11, lid to be opened/closed; 12, the stationary installation of absorbent materials such as sponge; 13, the afterbody outlet; 14, fixed angles.
Fig. 3～9th, the pathological section figure of rat lung.Wherein, Fig. 3 is the blank group; Fig. 4 is the 20nm experimental group; Fig. 5 is the 50nm experimental group; Fig. 6 is the 80nm experimental group; Fig. 7 is the 140nm experimental group; Fig. 8 is the 280nm experimental group; Fig. 9 is the 800nm experimental group.
Embodiment
Further specify the present invention with embodiment below, but the present invention is not limited.The experimental technique of unreceipted actual conditions in the following example, usually according to normal condition, or the condition of advising according to manufacturer.
The preparation of embodiment 1 static toxicant exposure cabinet
Static toxicant exposure cabinet, material is an organic glass, thickness is 8mm, gluing attached with special-purpose organic glass, length * wide * height=42cm * 42cm * 60cm, cumulative volume are 100L, use circular arc organic glass chamfering to adhere to remove the dead angle for four jiaos in the toxicant exposure cabinet, well is dug out at cabinet wall middle part, inlay 4 circular arc wall apart from the inwall of 40cm at the bottom of the cabinet and collude, diameter is 5mm, uses for supporting entanglement, O-ring seal is added on the toxicant exposure cabinet top, carries out sealing and fixing with plexiglass cover again.Device is as Fig. 1.
Circular adjustable wind speed axial flow blower is settled in the toxicant exposure cabinet bottom, model: 150FZY2-D, length * wide=16cm * 16cm apart from toxicant exposure cabinet bottom 15cm, uses the organic glass cylindrical body supports, and be adhered fixed, open a pore on the toxicant exposure cabinet wall, height is consistent with the blower fan setting height(from bottom), after blower fan control electric wire is wrapped up with the cylindrical plastic pipe, cause outside the toxicant exposure cabinet, and load onto speed regulator.
The fixing exposed device of embodiment 2 rats
Rat is fixed with cylindrical fixator, and the fixator material is plastics, is fixedly adhering to water-absorbing sponge at rat tails before the rat, with excretas such as absorption urines.The fixator surface only allows rat head to be exposed in the surrounding environment coated with the lubricious material preservative film.Fixator such as Fig. 2.
The static contamination of embodiment 3 rats
The rat that fixes is put into toxicant exposure cabinet, behind the good toxicant exposure cabinet of sealing and fixing, open blower fan, be sprinkled into 20nm respectively by well, 50nm, 140nm, 280nm, the SiO of 800nm particle diameter
2Particle 40mg, every group of 15 rats are regulated wind speed by speed regulator, and the control dust can blow afloat and get final product.Static contamination time is 2 hours (h), and each 2 rats are not taken food during the rat contamination and do not drink water, the other times feed of freely drinking water.
After contamination finishes, observe the physiology living condition of rat, death condition.In contamination back 24h, 48h, 72h puts to death each experimental group rat, opens the thoracic cavity immediately, and the whole right lung of ligation extracts right lung and is used for histopathologic slide's observation.After isolating pars cervicalis tracheae, open one in the tracheae lower end " T " the shape osculum, adopt external diameter be the ductus venosus of 1.8mm as trachea cannula, treat that intubate arrives the trachea carina position after, deflection enters left principal bronchus and gets final product left.With stroke-physiological saline solution left lung is carried out alveolar wass, totally 6 times, be followed successively by 3ml, 2ml, 2ml, 2ml, 2ml, 2ml, 2ml.Collect all bronchoalveolar lavage fluids, adopt Allegra X-15R centrifuge (U.S. BECKMAN COULTER) hydro-extractor, 4 ℃ of temperature controls, refrigerated centrifuge 2000r/min, refrigerated centrifuge 15min.Draw supernatant and carry out total protein (TPr), lactic dehydrogenase (LDH) determination of activity, pipe end sedimentation cell is made cell suspension with the dilution of 5ml stroke-physiological saline solution, dilutes 5 times again, carries out total cellular score (total cellular score, TCS) counting.
The result shows, dye dirt after, rat TPr, LDH content all have the control group that is higher than in various degree, and the visible significantly bag of TCS counting cytosis, macrophage increases.See Table 1,2,3.
Pathological section shows that all experimental group rats all have lung inflammation damage in various degree, mainly shows alveolar septum thickening in various degree, with inflammatory cell infiltration, mainly is macrophage, lymphocyte, or neutrophil leucocyte.See Fig. 3～9.
It is feasible to illustrate that the relevant nanoparticle of the present invention exposes toxicologic test method, more than describing is explanation of the invention, is not that institute of the present invention restricted portion is referring to claims to the restriction of invention, under the situation of spirit of the present invention, the present invention can make any type of modification.
Table 1. different-grain diameter SiO
2Dye behind the dirt total cellular score in the different time bronchoalveolar lavage fluid
Table 2. different-grain diameter SiO
2Dye behind the dirt total protein content in the different time bronchoalveolar lavage fluid
Table 3. different-grain diameter SiO
2Dye behind the dirt LDH content in the different time bronchoalveolar lavage fluid
Claims (10)
1. a toxicant exposure cabinet is characterized in that, in this toxicant exposure cabinet, the bottom is a blower fan, and the middle and upper part is the wire netting that can place experimental animal, at the cabinet wall middle part of toxicant exposure cabinet one opening is arranged, be well, have sealing-plug can clog this opening in this opening, the cabinet top is salable fixing cabinet lid.
2. toxicant exposure cabinet as claimed in claim 1 is characterized in that, uses circular arc material chamfering to adhere to remove the dead angle for interior four jiaos of toxicant exposure cabinet; The inwall of toxicant exposure cabinet is 4 circular arc wall hooks, uses for the support metal net; Cabinet has covered O-ring seal, so that cabinet is covered sealing and fixing.
3. toxicant exposure cabinet as claimed in claim 1 is characterized in that, is the cylinder that is fixed at the bottom of the cabinet between blower fan and the toxicant exposure cabinet bottom; Toxicant exposure cabinet cabinet wall is being opened a pore with the horizontal fan place, and the blower fan electric wire passes through this pore outside toxicant exposure cabinet.
4. toxicant exposure cabinet as claimed in claim 1 is characterized in that, blower fan is circular adjustable wind speed blower fan.
5. toxicant exposure cabinet as claimed in claim 1 is characterized in that, also comprises an experimental animal fixator, the smooth outer surface densification of this fixator.
6. one kind is detected the method that nanoparticle exposes toxicity, it is characterized in that, may further comprise the steps:
(1) be covered with the fixator fixation test animal of lubricious material with outside surface, the head of a retention test animal is exposed in the surrounding environment;
(2) experimental animal that fixes is placed on as on the wire netting in each described toxicant exposure cabinet of claim 1～5 the good toxicant exposure cabinet of sealing and fixing;
(3) open blower fan, be sprinkled into nanoparticle by well, regulate wind speed, the control nanoparticle is blown afloat, static contamination.
7. method as claimed in claim 6 is characterized in that, in the step (2), the mean grain size of described nanoparticle is 10nm～10 μ m.
8. method as claimed in claim 7 is characterized in that the mean grain size of described nanoparticle is 20nm～800nm.
9. method as claimed in claim 6 is characterized in that, in the step (1), adheres to water-absorbing sponge at the experimental animal afterbody before the fixation test animal, with excretas such as absorption urines; The static contamination time of step (3) is 2 hours, after contamination finishes, requires experimental animal is observed according to experimental program, detects toxicity index, the exposure toxicity of assessment nanoparticle.
10. method as claimed in claim 6 is characterized in that described experimental animal is a rat.
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| CN2009101978970A CN102053148A (en) | 2009-10-29 | 2009-10-29 | Method for detecting exposure toxicity of nanoparticles and toxicant exposure cabinet used thereby |
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Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104873296A (en) * | 2014-08-06 | 2015-09-02 | 南通大学 | Operating method for up-and-down type carbon monoxide poisoning device through filtration |
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| CN2838538Y (en) * | 2005-06-29 | 2006-11-22 | 武汉大学 | Static inhalation contamination cabinet |
| CN201171714Y (en) * | 2007-12-04 | 2008-12-31 | 中国科学院高能物理研究所 | Self-inhalation type nano-granule contaminated box |
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2009
- 2009-10-29 CN CN2009101978970A patent/CN102053148A/en active Pending
Patent Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN2838538Y (en) * | 2005-06-29 | 2006-11-22 | 武汉大学 | Static inhalation contamination cabinet |
| CN201171714Y (en) * | 2007-12-04 | 2008-12-31 | 中国科学院高能物理研究所 | Self-inhalation type nano-granule contaminated box |
Non-Patent Citations (1)
| Title |
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| 孙亮等: "静式吸入染毒柜制作初试", 《医学动物防制》 * |
Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104873296A (en) * | 2014-08-06 | 2015-09-02 | 南通大学 | Operating method for up-and-down type carbon monoxide poisoning device through filtration |
| CN104873295A (en) * | 2014-08-06 | 2015-09-02 | 南通大学 | Vertical type filtered carbon monoxide poisoning device capable of significantly improving working efficiency |
| CN104873295B (en) * | 2014-08-06 | 2016-08-17 | 南通大学 | Filtered vertical type carbon monoxide poisoning device |
| CN104873296B (en) * | 2014-08-06 | 2016-09-28 | 南通大学 | Filtered vertical type carbon monoxide poisoning device |
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Application publication date: 20110511 |