Sc 9068

After the periods of 7, 15, 30, and 60 days, the animals were euthanized by an anesthetic overdose. The polyethylene tubes were removed with the surrounding tissue and fixed in 10% buffered formalin at pH 7.0. The specimens were embedded in paraffin, serially cut into 5 μm sections, and stained with hematoxylin-eosin or submitted to immunohistochemistry by using an indirect immunoperoxidase technique for FN (primary antibody rabbit, SC-9068, Santa Cruz Biotechnology, CA, USA) and TN (primary antibody rabbit, SC-20932, Santa Cruz Biotechnology, CA, USA). The specimens were also submitted to the procedures suppressing the use of primary antibodies to the negative control.
The inflammatory infiltrate at the open end of the tubes was scored according to previous studies[4 (link)9 ] as follows: 0, few inflammatory cells or no reaction; 1, <25 cells and mild reaction; 2, between 25 and 125 inflammatory cells and moderate reaction; and 3, 125 or more inflammatory cells and severe reaction (×400 magnification). Fibrous capsules were considered thin when <150 μm and thick when ≥150 μm. Immunolabeling for FN and TN was defined as the presence of brownish color in the extracellular matrix. The criteria for establishing the adopted scores were 0, absence of immunolabeling; 1, low immunolabeling standard; 2, moderate immunolabeling standard; and 3, high immunolabeling standard (×1000 ).