Basic Potential of Carbon Nanotubes in Tissue Engineering Applications
Figure 3
MWCNTs incorporated into bone-marrow-derived macrophages (BMMφ) inhibited the differentiation of BMMφ into osteoclasts. (a) BMMφ were cultured for 3 days with receptor activator of nuclear factor-κB ligand (RANKL, 100 ng/mL) and macrophage colony stimulating factor (M-CSF, 25 ng/mL) in the presence or absence of 80n-MWCNTs, 150n-MWCNTs, or carbon black (CB). Cells were fixed and stained for TRAP. . (b) BMMφ were treated with or without 80n-MWCNTs, 150n-MWCNTs, or CB for 24 or 1 h. BMMφ were further cultured for 3 days in the presence of RANKL (100 ng/mL) and M-CSF (25 ng/mL), fixed, and stained for TRAP. . #; versus control. (c) Phase-contrast microscopy of BMMφ treated with 80n-MWCNTs or CB. BMMφ were incubated for 24 h with 80n-MWCNTs or CB, fixed, and observed under a phase-contrast microscope. (d, e) Neither 80n-MWCNTs nor CB attenuated RANKL-induced signals in BMMφ. BMMφ were cultured with or without 80n-MWCNTs or CB under the same conditions as in the experiments in (b). BMMφ were then treated for 20 min with RANKL (100 ng/mL) to activate NF-κB and p38 MAPK (d). BMMφ were treated with RANKL (100 ng/mL) for 24 h to induce NFATc1 (e). Cell lysates were subjected to SDS-PAGE, followed by immunoblotting with specific antibodies [37].
