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Microwave-induced fast decalcification of rat bone for electron microscopic analysis: an ultrastructural and cytochemical study
Pitol, Dimitrius Leonardo; Caetano, Flavio Henrique; Lunardi, Laurelúcia Orive.
  • Pitol, Dimitrius Leonardo; University of São Paulo. School of Dentistry of Ribeirão Preto. Department of Morphology, Stomatology and Physiology. Ribeirão Preto. BR
  • Caetano, Flavio Henrique; São Paulo State University. Institute of Biosciences. Department of Biology. Rio Claro. BR
  • Lunardi, Laurelúcia Orive; São Paulo State University. Institute of Biosciences. Department of Biology. Rio Claro. BR
Braz. dent. j ; 18(2): 153-157, 2007. ilus
Article in English | LILACS | ID: lil-466510
ABSTRACT
Bone decalcification is a time-consuming process. It takes weeks and preservation of the tissue structure depends on the quality and velocity of the demineralization process. In the present study, a decalcification methodology was adapted using microwaving to accelerate the decalcification of rat bone for electron microscopic analysis. The ultrastructure of the bone decalcified by microwave energy was observed. Wistar rats were perfused with paraformaldehyde and maxillary segments were removed and fixed in glutaraldehyde. Half of specimens were decalcified by conventional treatment with immersion in Warshawsky solution at 4ºC during 45 days, and the other half of specimens were placed into the beaker with 20 mL of the Warshawsky solution in ice bath and thereafter submitted to irradiation in a domestic microwave oven (700 maximum power) during 20 s/350 W/±37ºC. In the first day, the specimens were irradiated 9 times and stored at 40ºC overnight. In the second day, the specimens were irradiated 20 times changing the solution and the ice after each bath. After decalcification, some specimens were postfixed in osmium tetroxide and others in osmium tetroxide and potassium pyroantimonate. The specimens were observed under transmission electron microscopy. The results showed an increase in the decalcification rate in the specimens activated by microwaving and a reduction of total experiment time from 45 days in the conventional method to 48 hours in the microwave-aided method.
RESUMO
A preservação da estrutura de ossos é dependente da qualidade e da velocidade em que ocorre o processo de desmineralização. Neste estudo foi observada a ultraestrutura de maxila de rato descalcificada utilizando microondas. Ratos Wistar sofreram perfusão com paraformaldeído e o segmento de maxila retirado e fixado em glutaraldeído. Após esta etapa algumas amostras foram descalcificadas por imersão em solução de Warshawsky durante 45 dias a 4(0)C. Outras amostras foram submetidas a irradiação por microondas (forno de microondas doméstico 700 Watts de potência), durante 20 s/350 W/ ± 37ºC. No primeiro dia foram realizadas um total de 9 irradiações e os espécimes foram deixadas posteriormente a 4ºC por 12 h na solução descalcificadora sem agitação. No segundo dia, os fragmentos foram submetidos à nova irradiação totalizando 20 banhos, trocando-se a solução e o gelo a cada banho. A seguir algumas amostras foram pós-fixadas com tetróxido de ósmio e outras com tetróxido de ósmio e piroantimonato de potássio. As amostras foram observadas em microscópio eletrônico de transmissão. Os resultados mostraram que o processo de descalcificação ativado por microondas reduziu para 48 h o período de descalcificação, o qual pelo método tradicional ocorre em 45 dias.
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Full text: Available Index: LILACS (Americas) Main subject: Bone and Bones / Decalcification Technique / Microwaves Limits: Animals Language: English Journal: Braz. dent. j Journal subject: Dentistry Year: 2007 Type: Article Affiliation country: Brazil Institution/Affiliation country: São Paulo State University/BR / University of São Paulo/BR

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Full text: Available Index: LILACS (Americas) Main subject: Bone and Bones / Decalcification Technique / Microwaves Limits: Animals Language: English Journal: Braz. dent. j Journal subject: Dentistry Year: 2007 Type: Article Affiliation country: Brazil Institution/Affiliation country: São Paulo State University/BR / University of São Paulo/BR