title: Influence of different decalcifying agents on EGF and EGFR immunostaining.
abstract: This study was designed to verify the influence of three demineralizing agents on EGF and EGFR immunostaining as well as on tissue morphology. We chose submandibular glands that are a source of EGF and its receptor and which could be analyzed using a control in which the decalcification step was not carried out. After sacrifice of adult male Wistar rats by perfusion fixation, the submandibular glands and mandibles were excised and placed together in each of the following solutions: (a) 5% nitric acid in 4% formaldehyde; (b) 4.13% EDTA pH 7.4; (c) 5% trichloroacetic acid. Mandibles served as a parameter for decalcification time in each demineralizing solution. A control group was performed with submandibular glands that were not placed in any demineralizing solution. After mandibles were completely decalcified, glands were processed by embedding in Paraplast® and immunohistochemical staining was made to detect EGF and EGFR. It was observed that decalcification did not produce noticeable differences in terms of EGF and EGFR immunoreactivity, but had an effect on the quality of the morphology and staining. Our results indicate there is no problem performing immunostaining of EGF and EGFR in tissues that require decalcification. 4.13% EDTA (pH 7.4) is the best choice for decalcification in cases that are not urgent.
authors.affiliation: Department of Morphology, Piracicaba Dental School, State University of Campinas - UNICAMP, Piracicaba, SP, Brazil.
authors.forename: Juliana Dos Santos
authors.initials: Jdos S
authors.lastname: Neves
authors.forename: Nádia Fayez
authors.initials: NF
authors.lastname: Omar
authors.forename: Eliene Aparecida Orsini
authors.initials: EA
authors.lastname: Narvaes
authors.forename: José Rosa
authors.initials: JR
authors.lastname: Gomes
authors.forename: Pedro Duarte
authors.initials: PD
authors.lastname: Novaes
mesh_terms: D000818:Animals; D003648:Decalcification Technique; D004492:Edetic Acid; D004815:Epidermal Growth Factor / Q000032:analysis* / Q000378:metabolism; D066246:ErbB Receptors / Q000032:analysis* / Q000378:metabolism; D005557:Formaldehyde; D007150:Immunohistochemistry / Q000379:methods; D008297:Male; D008334:Mandible / Q000378:metabolism* / Q000473:pathology; D017942:Nitric Acid; D051381:Rats; D017208:Rats, Wistar; D012680:Sensitivity and Specificity; D013194:Staining and Labeling; D013363:Submandibular Gland / Q000378:metabolism* / Q000473:pathology; D016707:Tissue Fixation; D014238:Trichloroacetic Acid
chemical_list: D005557:Formaldehyde; D017942:Nitric Acid; D014238:Trichloroacetic Acid; D004815:Epidermal Growth Factor; D004492:Edetic Acid; C483803:Egfr protein, rat; D066246:ErbB Receptors