Journal: Bone Reports

Article Title: Mechanistic insight into the effects of Aryl Hydrocarbon Receptor activation on osteogenic differentiation

doi: 10.1016/j.bonr.2017.02.003

Figure Lengend Snippet: Differential effects of nicotine and dioxin. (A) ALP activity was assessed in BMSC grown in standard media (SM) or osteogenic media (OM). Dioxin- and nicotine-treated cells were cultured in OM. *p < 0.01, dioxin- vs. vehicle- and nicotine-treated cells. (B) BMSC migration capacity was assessed via wound-scratch assay. Significance is shown relative to both vehicle- and nicotine-treated cells grown in either SM (*p < 0.05) or OM (^p < 0.05). (C) Visualization and quantification of mineral deposition. Note that all dioxin-treated and nicotine-treated cells were grown in osteogenic media. Columns, means from at least three independent experiments *p < 0.01, dioxin-treated wells vs. all other groups.

Article Snippet: Moreover, dioxin was found to inhibit ALP activity in rat stem cells derived from the apical papilla (SCAPs) ( ).

Techniques: Activity Assay, Cell Culture, Migration, Wound Healing Assay

Journal: Bone Reports

Article Title: Mechanistic insight into the effects of Aryl Hydrocarbon Receptor activation on osteogenic differentiation

doi: 10.1016/j.bonr.2017.02.003

Figure Lengend Snippet: (A) ALP activity. All dioxin-treated and antagonist-treated cells were cultured in osteogenic media. *p < 0.001 significance of ALP activity in dioxin-only treated cells relative to all other groups. (B) Matrix mineralization. Calcium deposition in the matrix of cells grown under standard or osteogenic conditions was visualized by Alizarin red staining, which was quantified using a cetylpyridinium chloride de-stain procedure. Note that all dioxin- and antagonist-treated cells were grown in osteogenic media. Columns , means from at least three independent experiments *p < 0.01, dioxin alone-treated wells vs. all other groups.

Article Snippet: Moreover, dioxin was found to inhibit ALP activity in rat stem cells derived from the apical papilla (SCAPs) ( ).

Techniques: Activity Assay, Cell Culture, Staining

Journal: The American Journal of Pathology

Article Title: Mutation in Osteoactivin Decreases Bone Formation in Vivo and Osteoblast Differentiation in Vitro

doi: 10.1016/j.ajpath.2013.11.031

Figure Lengend Snippet: Differentiation of primary calvarial osteoblasts in neonatal D2J and D2J/Gpnmb+ mice. A: Western blots of OA glycoprotein in primary osteoblasts cultured for 7 days. B: ELISA of OA protein levels in osteoblast cell lysates, normalized to total protein concentration. C: Macro- and microscopic images of 14-day differentiated osteoblasts stained for ALP (asterisks). D: ALP activity in osteoblast matrix and cell lysates. E: Macro- and microscopic images of 14-day differentiated cultures stained for collagen (asterisks) with Sirius Red in D2J and D2J/Gpnmb+. F: Quantification of Sirius Red collagen staining in D2J and D2J/Gpnmb+ cultures. G: Percentage area fraction of Sirius Red–stained matrix in D2J and D2J/Gpnmb+ cultures. Data are expressed as means ± SEM. ∗P < 0.05, ∗∗P < 0.01. Scale bar = 100 μm.

Article Snippet: In parallel wells, ALP activity was determined in total cell lysates using an ALP activity kit (BioAssay Systems, Hayward, CA) according to the manufacturer’s protocol.

Techniques: Western Blot, Cell Culture, Enzyme-linked Immunosorbent Assay, Protein Concentration, Staining, Activity Assay

Journal: Materials

Article Title: Acceleration of Osteogenesis via Stimulation of Angiogenesis by Combination with Scaffold and Connective Tissue Growth Factor

doi: 10.3390/ma12132068

Figure Lengend Snippet: Comparison of osteogenic differentiation between CTGF-loaded ceramics and HAp ceramics (control). ( A ) Cells on CTGF-HAp ceramics were stained for ALP levels after culturing for 14 days. Arrowheads represent cell cluster with high ALP activity. ( B ) CTGF increased ALP activity in clustered regions (Higher magnification (square in A) of MC3T3-E1 on CTGF-HAp(100) ceramics). ( C ) Comparison of ALP activity of cells at day 7 and 14 in culture. Loading CTGF increased ALP activity. Experiments were repeated three times and run in triplicate. Graphs represent means ± SD. ( D ) Cells on CTGF-HAp ceramics were cultured for 28 days, and calcium deposition was assessed by Alizarin red staining.

Article Snippet: The supernatants were assayed for ALP activity (FUJIFILM Wako Pure Chemical, Osaka, Japan) according to the manufacturer’s instructions.

Techniques: Comparison, Control, Staining, Activity Assay, Cell Culture