The MMP-7 and MMP-9 antibody-staining status were identified as either negative or positive. Immunohistochemical positive staining was defined by the presence of brown chromogen detection on the edge of the hematoxylin-stained cell nucleus, distributed within the cytoplasm or in the membrane via evaluation by light microscopy as previously described.28 (link) Positive controls consisted of tissue specimens with known antigenic positivity. Sections treated with PBS without the primary antibodies served as negative controls. Seven fields of about 600.000 μm2, randomly selected from each section, were considered for morphometric and densitometric analysis. The percentage areas (morphometric analysis) stained with MMP-7 and MMP-9 antibodies were expressed as % positive, dark brown pixels of the analyzed fields. While, the levels (high/low) of staining intensity of positive areas (densitometric analysis) were expressed as densitometric count (pixel2) of positive, dark brown pixels of the analyzed fields. These parameters were calculated using software for image acquisition (AxioVision Release 4.8.2 - SP2 Software, Carl Zeiss Microscopy GmbH, Jena, Germany). Data were expressed as mean ± standard deviation (SD). Digital micrographs were taken and fitted as previously described.