Identification of the nuclear factor kappa-beta (NF-kB) in cortical of miceWistar using Technovit 7200 VCR®

ABSTRACT

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 adultmale Wistar rats.Study Method: A 1.8 mm diameter defect was performed 0.5mm from the femur proximal joint by means of around bur. Experimental groups were divided according to fixing solution prior to histologic processing Group1- 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 nonsurgical 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 4oC for 7days, and were dehydrated in a series of alcohols at -20oC. The polymer embedding procedure was performed atethanol/polymer ratios of 70%-30%, 50%-50%, 30%-70%, 100%, and 100% for 72 hours at -20oC. Polymerization followed the manufacturer’s recommendation. The samples were grounded and polished to 10-15ìm thickness,and were deacrylated. The sections were rehydrated and were submitted to the primary polyclonal antibody anti-NF-kB on a 175 dilution for 12 hours at room temperature.Results: Microscopy showed that the Group 2 presented positive reaction to NF-kB, diffuse reactions for NSPGand SPBE, and no reaction for the NC group.Conclusion: The results obtained support the feasibility of the developed immunohistochemistry technique (AU)