FIGURE 3.

Local delivery of nuclear factor kappa B (NF-κB) decoy oligodeoxynucleotide (ODN) mitigated the number of osteoclast-like cells induced by cPE and increased the percentage of anti-alkaline phosphatase (ALP) staining positive area in chronic inflammation. Representative pictures of TRAP and ALP staining in all groups (a). Leukocyte tartrate resistant acid phosphatase (TRAP) staining was used to identify the osteoclast-like cells with multi-nuclei stained brown (arrows), which were located on the bone perimeter within the resorption lacunae in the upper photomicrographs. Osteoblast-like cells were stained brown (arrowheads) by ALP staining in the lower photomicrographs. (b) Quantitative assessment for cells positively stained with TRAP. Using three randomly selected views with ×200 magnification in each specimen, in a blind manner, two investigators manually counted the number of TRAP staining positive cells, which were finally normalized by the bone area measured using the ImageJ software (National Institutes of Health). (c) Quantitative assessment of the percentage of ALP staining positive area based on based on the whole area of each specimen. Using the entire image of each specimen with ×100 magnification, the percentage of brown, positively stained area (arrowheads) for ALP was also quantified based on the entire area of each section measured using the ImageJ software (National Institutes of Health). Color threshold of ALP staining positive area was determined by consensus of three of the investigators. Black bar: 50 μm. *p < 0.05, **p < 0.001, ***p < 0.0001