| Identification of the nuclear factor kappa-beta (NF-kB) in cortical of mice Wistar using Technovit 7200 VCR®. | |
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MedLine Citation:
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PMID: 20526271 Owner: NLM Status: MEDLINE |
Abstract/OtherAbstract:
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OBJECTIVE: this study aimed to develop a nondecalcified bone sample processing technique enabling immunohistochemical labeling of proteins by kappa-beta nuclear factor (NF-kB) utilizing the Technovit 7200 VCR® in adult male Wistar rats. STUDY METHOD: A 1.8 mm diameter defect was performed 0.5 mm from the femur proximal joint by means of a round bur. Experimental groups were divided according to fixing solution prior to histologic processing: Group 1--ethanol 70%; Group 2--10% buffered formalin; and Group 3--Glycerol diluted in 70% ethanol at a 70/30 ratio + 10% buffered formalin. The post-surgical periods ranged from 01 to 24 hours. Control groups included a non-surgical procedure group (NSPG) and surgical procedures where bone exposure was performed (SPBE) without drilling. Prostate carcinoma was the positive control (PC) and samples subjected to incomplete immunohistochemistry protocol were the negative control (NC). Following euthanization, all samples were kept at 4°C for 7 days, and were dehydrated in a series of alcohols at -20°C. The polymer embedding procedure was performed at ethanol/polymer ratios of 70%-30%, 50%-50%, 30%-70%, 100%, and 100% for 72 hours at -20°C. Polymerization followed the manufacturer's recommendation. The samples were grounded and polished to 10-15 m thickness, and were deacrylated. The sections were rehydrated and were submitted to the primary polyclonal antibody anti-NF-kB on a 1:75 dilution for 12 hours at room temperature. RESULTS: Microscopy showed that the Group 2 presented positive reaction to NF-kB, diffuse reactions for NSPG and SPBE, and no reaction for the NC group. CONCLUSION: The results obtained support the feasibility of the developed immunohistochemistry technique. |
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Authors:
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Marcos B Salles; Bruno König; Sergio Allegrini; Marcelo Yoshimoto; Marília T Martins; Paulo G Coelho |
Publication Detail:
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Type: Journal Article Date: 2011-01-01 |
Journal Detail:
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Title: Medicina oral, patología oral y cirugía bucal Volume: 16 ISSN: 1698-6946 ISO Abbreviation: Med Oral Patol Oral Cir Bucal Publication Date: 2011 Jan |
Date Detail:
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Created Date: 2010-12-30 Completed Date: 2011-05-03 Revised Date: 2011-08-25 |
Medline Journal Info:
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Nlm Unique ID: 101231694 Medline TA: Med Oral Patol Oral Cir Bucal Country: Spain |
Other Details:
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Languages: eng Pagination: e124-31 Citation Subset: D; IM |
Affiliation:
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Department of Anatomy, Institute of Biomedical Sciences, Universidade de São Paulo, Brazil. |
Export Citation:
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| MeSH Terms | |
Descriptor/Qualifier:
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Acrylic Resins* Animals Bone and Bones / anatomy & histology* Immunohistochemistry / methods* Male Mice NF-kappa B* Rats Rats, Wistar |
| Chemical | |
Reg. No./Substance:
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0/Acrylic Resins; 0/NF-kappa B; 88507-29-3/Technovit 7100 |
From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine
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