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The aim of this article is to present the decalcification process dynamic …


Biology Articles » Anatomy & Physiology » Anatomy, Animal » Decalcification Dynamic of Dog Mineralized Tissue by Microwaves » Material and Method

Material and Method
- Decalcification Dynamic of Dog Mineralized Tissue by Microwaves

It was used healthy teeth, provided from Central Animals House of the University of Sao Paulo, Ribeirao Preto, Sao Paulo, Brazil and according to the rules proposed by the Ethics Committee. The left molars attached to the mandible arch were removed and fixed in formaldehyde 10%, during 24 hours.

Microwave oven standardization. The standardization method used in this research was proposed by Login & Dvorak (1990), in that was used a domestic microwave oven (Panasonic /700W/2450mHZ) that had its rotative plate turned off. A Becker of glass containing lOOmL of distilled water was inserted on the left side ofthe pre-heated oven in maximal potency by 2 minutes. An Agar/Giemsa block (0.5cm), prepared with 5% of agar/agar in saline solution in that was added Giemsa stained solution (Sigma Chemical St Louis, MO), was soaked in 5mL of fixative solution or decalcification solution, in Petri plastic plate, 35mm, inserted in various positions inside ofthe oven and irradiated with 100% potency in different periods of time. The color alteration ofthe Agar/ Giemsa blocks was used to watch the better irradiation focus and the temperature change. Establishing the area with most microwave incidence into the oven, as well as the exposition time, it was performed the decalcification process.

Decalcification. Considering the individual differences on teeth mineralization, it was decided to use the same teeth in the analysis by two decalcification methods. Thus, the teeth were longitudinal sectioned in bucolingual orientation, separated into three fragments and decalcified with nitric acid or Washawsky solution (EDTA 8.5% pH 7.4). The full teeth were initially weighted and sectioned into three samples, and the measures were respectively, 1.7x1.0cm of width and 2.6cm of length. All the fragments showed the same structures, enamel, dentin, alveolar bone and soft tissues. After fixative period, it was inserted 30mL of decalcified solution and irradiated for 1 minute in ice bath microwave, in sequence the decalcified solution was despise and replaced by another for new irradiation period. Each five irradiation periods, one sample ofthe decalcified solution, was collected for calcium dosage in atomic spectrophotometer. It was performed 10 irradiations per day and the sample stayed into the decalcified solution at freezer until the next day. After fixative period, the group A (control) was placed in 30mL ofthe decalcified solution and this was replaced by another after the sample remotion for calcium dosage, each 5 days.

Calcium dosage. The calcium concentration ofthe selected samples was determined by atomic absorvance spectrophotometry (Shimadzu AA680G spectrophotometer).

Microscopic study. After decalcification period, the samples were dehydrated, included in paraffin and the histologic sections stained by Masson Trichrome. In sequence, they were analyzed and photographed usinga photomicroscope Jenaval, Carl-Zeiss, Ober Kochem, Germany.


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