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Originally published as JHC exPRESS on February 6, 2006.
doi:10.1369/jhc.5A6853.2006
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Journal of Histochemistry and Cytochemistry
Volume 54 (6): 713-725, 2006
Copyright ©The Histochemical Society, Inc.

Immunocytochemical Study of Amelogenin Deposition during the Early Odontogenesis of Molars in Alendronate-treated Newborn Rats

Luciana F. Massa, Vivian Bradaschia-Correa and Victor E. Arana-Chavez

Laboratory of Mineralized Tissue Biology, Department of Cell and Developmental Biology, Institute of Biomedical Sciences, University of São Paulo, São Paulo, Brazil

Correspondence to: Dr. Victor E. Arana-Chavez, Laboratory of Mineralized Tissue Biology, Department of Cell and Developmental Biology, Institute of Biomedical Sciences, University of São Paulo, 05508-900 São Paulo, SP, Brazil. E-mail: vearana{at}usp.br

Newborn rats were treated with sodium alendronate to study how enamel is formed and the effect of alendronate during early odontogenesis. Ultrastructural analysis combined with high-resolution immunocytochemistry for amelogenin was carried out. Twelve rats were subjected to daily SC injections of sodium alendronate (2.5 mg/kg/day) for 3 days on their dorsal region, whereas three rats were daily injected with saline solution as a control. Molar tooth germs from 3-day-old rats were fixed under microwave irradiation in 0.1% glutaraldehyde + 4% formaldehyde buffered at pH 7.2 with 0.1 M sodium cacodylate. The specimens were left undecalcified, postfixed with osmium tetroxide, dehydrated, and embedded in LR White resin. Ultrathin sections were incubated with a chicken anti-24-kDa rat amelogenin antibody, a secondary antibody, and finally with a protein A–gold complex. Large patches of amelogenin were present over the unmineralized mantle dentin and at early secretory ameloblasts. At more advanced stages, they were also detected at the enamel matrix, as well as in the mineralized dentin, at the periodontoblastic space of the dentinal tubules, and at the predentin. It is likely that the main effect of alendronate at early stages of odontogenesis is the increase of synthesis/secretion of amelogenin, promoting its deposition within the forming dentin and enamel. (J Histochem Cytochem 54:713–725, 2006)

Key Words: amelogenin • alendronate • odontogenesis • ameloblasts • odontoblasts • bisphosphonates • immunocytochemistry


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