Yong-Keun Lee

Lipid rafts are membrane platforms for signaling molecules that regulate various cellular functions. During osteoclastogenesis, lipid rafts also have been shown to play a crucial role for cell fusion upon RANK/RANKL interaction and further activation of osteoclasts, resulting in bone resorption. To investigate proteins that localize in lipid rafts, we conducted two-dimensional (2D) gel electrophoresis followed by MALDI-TOF mass spectrometry. We identified 12 functional proteins among 34 proteins that were detectable by silver staining of the 2D gel. Among them, a subunit of vacuolar H⁺-ATPase that is involved in resorption activity of osteoclasts was identified showing an approximate molecular weight of 56.94 kDa and pI of 5.4. The solubilization of osteoclast membrane proteins followed by sucrose density gradient fractionation confirmed that V-ATPase colocalized with flotillin-1, a marker of lipid rafts, in low-density detergent-insoluble fractions. Disruption of rafts with cholesterol-sequestering agents or introduction of a dominant-negative mutant of TRAF6 impaired V-ATPase activity. These data indicate that integrity of lipid rafts regulates the activity of V-ATPase in osteoclasts, suggesting that cholesterol-lowering agents might be useful for inhibiting osteoclast-dependent bone resorption.

Interleukin-17 (IL-17) is a cytokine secreted primarily by T_H-17 cells. Although IL-17 is primarily associated with the induction of tissue inflammation, the other biological roles of IL-17, including non-immune functions, have yet to be thoroughly explored. Here, we report that T-cell-produced IL-17 can induce proliferation of human bone marrow-derived mesenchymal stem cells (hMSCs) in a manner dependent on the generation of reactive oxygen species (ROS). Rac1 GTPase and NADPH oxidase 1 (Nox1) are activated by IL-17 to produce ROS, which in turn stimulates hMSC proliferation. The activation of the MEK-ERK pathway is also crucial for IL-17-dependent hMSC proliferation. TRAF6 and Act1 are required to activate Nox 1 and to phosphorylate MEK on IL-17 stimulation. Interestingly, IL-17 not only accelerates the proliferation of hMSCs, but also induces their migration, motility, and osteoblastic differentiation. Furthermore, IL-17 induces the expression of M-CSF and receptor activator of NF-κB ligand (RANKL) on hMSCs, thereby supporting osteoclastogenesis both in vivo and in vitro. On the basis of these results, we suggest that IL-17 can function as a signal to induce extensive bone turnover by regulating hMSC recruitment, proliferation, motility, and differentiation.

To determine the influence of opalescence and fluorescence properties on the light transmittance of resin composites as a function of wavelength (410-750 nm). Spectral distribution of seven resin composites of A2 shade was measured according to the CIELAB color scale relative to the standard illuminant D65 in the reflectance and transmittance modes. Opalescence spectrum (OPS) was calculated as the subtraction spectrum (i.e., the spectrum measured in the transmittance mode subtracted at each wavelength from the spectrum measured in the reflectance mode). UV component of the illuminant was included and excluded to calculate the fluorescence spectrum (FLR and FLT in the reflectance and transmittance mode, respectively). Contrast ratio (CR) was calculated as the ratio of reflectance over a black background and over a white background. The total transmittance spectral distribution (TSD) value was used as the parameter to indicate masking ability of the resin composites over background color. Multiple regression analyses were performed among TSD and other optical parameters at the significance level of 0.05. In all the resin composites and wavelength range, correlation between CR and TSD was very high (r = -0.99). Correlations between each parameters varied by the wavelength range of fluorescence (410-500 nm) and no-fluorescence (510-750 nm). Correlation between OPS and TSD varied by the wavelength range (r = -0.86 to -0.94, P< 0.05). Fluorescence of resin composites increased the TSD values. Based on this in vitro study, the influence of opalescence and fluorescence of resin composite varied by the wavelength.

The objective of this study was to investigate the changes in the translucency of resin composites following a series of immersion treatments in organic and chemical substances. Color of resin composites was measured according to the CIELAB color scale relative to the standard illuminant D65 over a white and a black background. Translucency parameter (TP) of resin composites was calculated at baseline, and after sequential immersion: Step 1, porcine liver esterase (a substitute for a salivary esterase); Step 2, organic substances (mucin and serum) and phosphate‐buffered saline (PBS) as a control; Step 3, chemical alteration agents [chlorhexidine (CH) and carbamide peroxide (CP)]; and Step 4, 2% methylene blue. Porcine liver esterase caused small changes in TP (ΔTP = = −0.5 to 0.2). After Step 2, ΔTP values of three groups were similar in the range of −0.7 to 1.2. After Step 3, ΔTP values of all groups were small in the range of −1.5 to 2.2. After Step 4, ΔTP values were high and differences that varied by the resin composite and the immersion protocol were clearly observed (ΔTP = −13.4 to −2.5). Changes in TP were mainly influenced by resin composite. After Step 4, mucin and serum groups showed generally small changes in TP compared to PBS group. It is a possibility that the high changes in TP after immersion in methylene blue is an indication of dye absorption which might be an indication of the degree of resin composite degradation. © 2005 Wiley Periodicals, Inc. J Biomed Mater Res Part B: Appl Biomater, 2006