Document Detail

Identification of the nuclear factor kappa-beta (NF-kB) in cortical of mice Wistar using Technovit 7200 VCR®.
MedLine Citation:
PMID:  20526271     Owner:  NLM     Status:  MEDLINE    
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.
Marcos B Salles; Bruno König; Sergio Allegrini; Marcelo Yoshimoto; Marília T Martins; Paulo G Coelho
Publication Detail:
Type:  Journal Article     Date:  2011-01-01
Journal Detail:
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:
Created Date:  2010-12-30     Completed Date:  2011-05-03     Revised Date:  2011-08-25    
Medline Journal Info:
Nlm Unique ID:  101231694     Medline TA:  Med Oral Patol Oral Cir Bucal     Country:  Spain    
Other Details:
Languages:  eng     Pagination:  e124-31     Citation Subset:  D; IM    
Department of Anatomy, Institute of Biomedical Sciences, Universidade de São Paulo, Brazil.
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MeSH Terms
Acrylic Resins*
Bone and Bones / anatomy & histology*
Immunohistochemistry / methods*
NF-kappa B*
Rats, Wistar
Reg. No./Substance:
0/Acrylic Resins; 0/NF-kappa B; 88507-29-3/Technovit 7100

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