Influence of different decalcifying agents on EGF and EGFR immunostaining.
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.
Neves
Juliana Dos Santos
Jdos S
Department of Morphology, Piracicaba Dental School, State University of Campinas - UNICAMP, Piracicaba, SP, Brazil.
Omar
Nádia Fayez
NF
Narvaes
Eliene Aparecida Orsini
EA
Gomes
José Rosa
JR
Novaes
Pedro Duarte
PD
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
D005557:Formaldehyde; D017942:Nitric Acid; D014238:Trichloroacetic Acid; D004815:Epidermal Growth Factor; D004492:Edetic Acid; C483803:Egfr protein, rat; D066246:ErbB Receptors