Design of experiment and research methods

Experimental work on creation of model of caries of a dentine is executed on 28 rats of line Wistar in mass of 200-220 and 3 not purebred dogs in mass of 15,0 kg, 10,0 kg and 12,0 kg. The experimental model was formed taking into account literary data in own updating (Gizzatullina L.

L from co-workers., 2008; Erofeeva L.M. from co-workers., 2013). Conditions of the maintenance and feeding of experimental animals were carried out according to to Sanitary rules on the device, the equipment and the maintenance of experimentally-biological clinics, confirmed Order MZSSSR 1179 from 10.10.83. Researches were carried out according to the European convention on protection of the vertebrate animals used for experiments or in other scientific purposes (ETS 123) (Strasbourg, on March, 18th 1986.

All animals contained in standard conditions of a vivarium on a high-grade ration and were one age. All interventions on rats were spent under a chloroformic narcosis, on dogs under a narcosis with use of preparations "Zoletil" 0,5 ml and "Ksila" 1 ml, with observance of rules of an asepsis and antiseptics.

Preliminary, time of action and a dose of a chloroformic narcosis for rats have been by practical consideration defined. The narcotic dream of rats, in of 5 minutes, was reached by use of 8 drops of Chloroformium on an animal in mass of 200-210 gramme. Then an animal placed on the fixing device with clamps for extremities.

to Dogs the premedication of 0,7 ml r-ra kardiamina and 1 ml of Dimedrol was spent. For a narcosis intramusculary entered 0,5 ml "Zoletil" and 1 ml "Ksila", the narcotic dream for 30-60 minutes was thus reached. For deducing from a narcosis to dogs entered 5 ml of Pyracetamum. Rats cleared up natural by.

For this time interval at rats dissected the first molar tooths of the top jaw in limits okolopulparnogo a dentine on II class according to classification Black (1889), at dogs dissected on 4 premolar tooths on the top jaw and 4 premolar tooths on a mandible in limits okolopulparnogo a dentine on I class according to classification Black. The stomatologic portable equipment "Vladmiva" was used. Dissected cavities konusnymi diamond borons of the smallest size at rats, at dogs used diamond round burs 2. The generated cavities corresponded to width and height of a head of boron. A teeth with the generated cavities processed the sterile distilled water.

Experimental animals were sectioned on groups on control and skilled. In skilled group animals were sectioned on terms and quantity of a fence of an investigated material (Table 2.1, 2.2).

Table 2.1 - Distribution of experimental rats on terms and kol-vu teeth



Quantity of a teeth Quantity of preparations
the Control 4 8 23
Experience 24 48 134
1 series

1 days

8 16 48
2 series

2 days

8 16 46
3 series

4 days

8 16 40
Total 28 56 157

Table 2.2 - Distribution by quantity of a teeth and preparations at dogs

Quantity of a teeth Quantity of preparations
the Control 4 18
Experience 24 100

the Teeth animal (rats and dogs) was sectioned on two groups; I group (control) - the usual method of treatment (cavity preparing, entering adgeziva OptiBond (Kerr), applying of seal Point 4 (Kerr) was spent. In the first group treatment 8 teeth at rats and 4 at dogs is spent.

In the second group - treatment similar to I group, but before applying of a constant seal was spent deep fluorination of a dentine dentin-germetizirujushchej by a liquid with influence on it nizkointensivnym laser radiation. In the second group treatment of 48 teeth at rats and 24 at dogs is spent.

For morphological research firm tissues of a teeth were used. Research is spent at level of light microscopy (ZEIZZ AXIO Imager A2c) with use of digital chamber AXIO Cam ER c 5s. The set conditions of experiment (mikroskopirovanie) were supported automatically by program Axio Vision 4.8.2 (Carl Zeizz Micro Imaging Gmbh, Germany). Tissues ekstragirovannyh teeth were fixed in a solution of 10 % of neutral formalin. A decalcification spent a preparation "Biodeck" (BioVitrum). Dehydrated in alcohols and filled in in parafintselloidinovuju an admixture. Prepared sections in the thickness 5 6 microns and painted them on Van Gizon, a hematoxylin and eosine. Immunotsitohimichesky research is spent immunoperoksidaznym by a method with use of monoclonal antibodies of manufacture of firms NOVOCASTRA, DAKO and Lab Vision to a marker of an endothelium of vessels, antigen CD31 (allows to visualise vessels in tissues). For the purpose of visualisation of result of reaction of linkage of an antigen with an antibody used system detektsii UltraVision LP Value HRP Polymer (the goat antibodies to a rabbit and the mouse), Lab Vision the USA.

M research carried out on preparations with use of program Axio Vision and G.G.Avtandilova's (1990) standard grid, and also screw ocular-micrometer -1-154,2.

In connection with high rate reparativnyh properties of firm tissues of a teeth of rats, the fence of a material for morphological research was spent to control terms for 4 days after an intervention. As the fence of an intact teeth for the control was spent. At dogs the material fence was spent in
control terms for 10 days, for the control the fence of an intact teeth was spent. The ex-traction of a teeth was spent at rats after Chloroformium lulling to slip in pairs. To dogs entered intramusculary 3 ml of a solution "Adelin" then made an ex-traction of a teeth.

For statistical processing of the received data commercial program Staistica 12 has been used. Estimated parametrical t-criterion Stjudenta with amendment Bonferroni.


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Scientific source KOVALEVA MARIA SERGEEVNA. Clinico-morphological research of a condition of tissues of a tooth at treatment of caries of a dentine with application of deep fluorination and nizkointensivnogo laser radiation. The dissertation on competition of a scientific degree of the candidate of medical sciences. Great Novgorod - & 2014. 2014

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