Figure 6.

Inhibition of osteoclast bone resorptive activity by PIIBNP in vivo. The calvarial bones were harvested from mice treated with PBS, LPS, LPS + GST, and LPS + PIIBNP. (a) Upper panels showed that the histological sections stained for TRAP and counterstained with hematoxylin. Red cells are TRAP-stained osteoclasts. Lower panels showed that the histlogical staining for in situ TUNEL staining and counterstained with methyl green. Black arrow indicates TUNEL positive cells. The histological sections were fixed with 4% neutral buffered formalin for 10 min and apoptotic cells were determined using TUNEL assay kit (Wako) according to the manufacturer's protocol. Bar =100 μm. (b) Quantitation of osteoclast numbers. Columns represent mean ± S.D. of the osteoclasts number/ mm².

(c) Percent of eroded surface occupied by osteoclasts. Columns represent mean ± S.D. of the percentage of eroded bone surface length covered by osteoclasts. For each animal, the sections with the largest diameter of bone marrow were used for calculations. (d) Quantitation of serum type I collagen fragment (C-telopeptide, Ctx) at the time of sacrifice. Columns represent mean ± S.D. of the Ctx concentration in serum. Each assay was performed in triplicate wells and three independent experiments. *indicates p<0.05 versus samples treated with LPS + GST.