Fig 2: Expression of NFATc1 in osteoblasts located in the third apical after tooth extraction on day 7 (A, B, C, D) and day 14 (E, F, G, H) using a 400x magnification light microscope is shown by the yellow arrows. Under a 400x magnification light microscope, the yellow arrow appears. A, E: The control group was given PEG gel after extraction. B, F: Extraction followed by administration of BBG gel. C, G: Extraction followed by administration of EEP gel. D, H: Extraction followed by administration of a combination of EEP-BBG gel.

Fig 3: Mean number of osteoblast cells expressing BMP7 on day 7 and day 14 and osteoclasts expressing NFATc1 on day 7 and day 14.

Fig 4: Inhibition of Rankl induced osteoclastogenesis in macrophages from Nampt+/- mice.a Bone marrow-derived macrophages from Nampt+/+ and Nampt+/- mice were treated with M-CSF (20 ng/ml) for 2 days. Non-adherent cells were washed out and cells were further cultured with M-CSF (20 ng/ml) and Rankl (100 ng/ml) for an additional 3 days. TRAP staining was performed 3 days later and cells having 3 or more nuclei were counted as osteoclasts. The frequency of osteoclasts is expressed as mean ± SD (N = 5). An average of 7 fields/well were counted per experimental sample. b Reduced osteoclast target gene expression in Nampt-deficient macrophages from Nampt+/- mice. Osteoclast target Nfatc1, Acp5, Dc-stamp, Cathepsin K mRNA expression were measured using semi-quantitative RT-PCR. ß-Actin was used as a loading control. c Representative western blot of Nampt protein in Nampt-deficient macrophages. Quantification of Nampt protein expression in Nampt-deficient macrophages. Representative images from three Nampt+/- mice with Nampt+/+ littermate controls are presented. Quantification of semi-quantitative RT-PCR of osteoclast target genes expressed as mean ± SD (N = 3). *P < 0.05; **P < 0.005

Fig 5: Overexpression of Nampt promotes osteoclast formation.Bone marrow cells were isolated from male DBA/1J Nampt+/+ and NamptOE mice and differentiated into osteoclasts. TRAP+ cells with >3 nuclei were counted as osteoclasts as described in Materials and methods. a Images of TRAP+ cells were captured under a light microscope (magnification, 10×). b Quantification of osteoclasts. c A representative image of western blot analysis of Nampt, Nfatc1, and ß-Actin protein levels in NamptOE BMM. ß-Actin was included as a loading control. d Quantification of Nampt and Nfatc1 protein levels. Data were presented as relative expression levels based on the value of Nampt+/+ wild type (WT) mice as 1. *P < 0.05 vs. Nampt+/+ mice and ***P < 0.001 vs. Nampt+/+ WT mice