Silver nanoparticles induce apoptosis through the toll-like receptor 2 pathway.

OBJECTIVE: The objective of this study was to evaluate the effects of silver nanoparticles (AgNPs) on the Toll-like receptor 2 (TLR-2) pathway in cultured cells. STUDY DESIGN: Human chondrocytes, periodontal ligament (PDL) cells, and SCC-9 cells (squamous cell carcinoma from the tongue) were cultured and subjected to cytotoxicity assays. To evaluate the effects of AgNPs on the TLR-2 pathway, TLR-2 small interfering (si) RNA or TLR-2 antibodies were applied to the chondrocytes, followed by the application of AgNPs. RESULTS: AgNPs induced dose-dependent effects on the examined cell types in terms of both cytotoxicity and TLR-2 expression levels. AgNP-mediated apoptosis was reduced after treatment with TLR-2 siRNA in both PDL cells and chondrocytes. Furthermore, functional blocking of TLR-2 with anti-TLR2 antibodies inhibited AgNP-mediated cytotoxicity. AgNPs increased c-Jun phosphorylation, an effect that was reversed after treatment with TLR-2 siRNA. CONCLUSIONS: The results indicate that AgNP-mediated apoptosis most likely occurs via the TLR-2 pathway.

4-hexylresorcinol stimulates the differentiation of SCC-9 cells through the suppression of E2F2, E2F3 and Sp3 expression and the promotion of Sp1 expression.

The dormancy-inducing factors of bacteria inhibit tumor cell growth. In the present study, we evaluated the antitumor effects of the dormancy-inducing factor 4-hexylresorcinol (4-HR) using real-time cell electronic sensing (RT-CES) in SCC-9 cells (tongue squamous cell carcinoma cells). Treatment with 4-HR suppressed the growth of SCC-9 cells in a dose-dependent manner. We used a DNA microarray to identify genes that showed a significant change in expression upon 4-HR administration in SCC-9 cells. Among the differentially expressed genes, the protein expression of several cell proliferation related factors, including E2F1, E2F2, E2F3, E2F4, E2F5, E2F6, Sp1 and Sp3, were determined by western blot analyses. Treatment with 4-HR strongly suppressed E2F2 and slightly suppressed E2F3 but did not change the expression of E2F1, E2F4, E2F5 and E2F6 relative to no treatment. Furthermore, 4-HR increased Sp1 expression in a dose-dependent manner and decreased Sp3 expression. Therefore, the ratio of Sp1 to Sp3, an important driving force of epithelial cell differentiation, was drastically increased. Consistent with this observation, 4-HR increased the expression of the epithelial cell differentiation markers involucrin and keratin 10. Together, our results indicate that 4-HR induces the differentiation of SCC-9 via the modulation of the E2F-mediated signaling pathway.

4-Hexylresorcinol inhibits transglutaminase-2 activity and has synergistic effects along with cisplatin in KB cells.

Resistance to chemotherapy is very important in the prognosis of tumors. Transglutaminase-2 (TG-2) mediated chemotherapy resistance has been widely reported. The objective of this study was to demonstrate the effect of 4-hexylresorcinol (4-HR) on TG-2 activity in nasopharyngeal squamous cell carcinoma cells (KB cells). Treatment with a mixture of 4-HR and cisplatin significantly decreased KB cell viability compared to treatment by cisplatin alone at 10 µg/ml (p<0.001). 4-HR inhibited TG-2 activity compared to cisplatin alone at 5, 10 and 20 µg/ml (p = 0.001, 0.001 and 0.003, respectively). Nuclear translocation of TG-2 was also inhibited by 4-HR treatment. 4-HR treatment also increased the fluorescence life-time of DAPI significantly compared to the untreated control or the cisplatin treated group (p<0.001). In conclusion, 4-HR inhibited TG-2 activity and showed a synergistic effect on tumor cell growth inhibition with cisplatin.

Low molecular weight silk fibroin increases alkaline phosphatase and type I collagen expression in MG63 cells.

Silk fibroin, produced by the silkworm Bombyx mori, has been widely studied as a scaffold in tissue engineering. Although it has been shown to be slowly biodegradable, cellular responses to degraded silk fibroin fragments are largely unknown. In this study, silk fibroin was added to MG-63 cell cultures, and changes in gene expression in the MG-63 cells were screened by DNA microarray analysis. Genes showing a significant (2-fold) change were selected and their expression changes confirmed by quantitative RT-PCR and western blotting. DNA microarray results showed that alkaline phosphatase (ALP), collagen type-I alpha-1, fibronectin, and transforming growth factor-beta1 expressions significantly increased. The effect of degraded silk fibroin on osteoblastogenic gene expression was confirmed by observing up-regulation of ALP activity in MG-63 cells. The finding that small fragments of silk fibroin are able to increase the expression of osteoblastogenic genes suggests that controlled degradation of silk fibroin might accelerate new bone formation. [BMB reports 2010; 43(1): 52-56].

Electromyographic activity of the masseter muscle after radiofrequency therapy in an animal model.

PURPOSE: The purpose of this study was to examine changes in the electromyographic (EMG) activity of the masseter muscle after radiofrequency therapy (RF). METHODS: Twelve rabbits were used in this study: four in each group according to the number of RF applications. Preoperative EMG in the masseter muscle was used as the control. EMG was recorded at 1, 2, 3, and 4 weeks after RF in each rabbit. The recorded data were analyzed in terms of voltage and frequency, and changes in recorded variables were compared among the groups. The relative activity in peak voltage, root mean square of the action potential, area of voltage, and area of frequency were investigated. RESULTS: When compared to preoperative values, the variables at 3 or 4 weeks after RF application were significantly different in the single and quadruple therapy groups (P<0.05). There was no significant difference in the other groups (P>0.05). When the samples were regrouped as two groups like small number of application group (one or two point) and large number of application group (three or four points), the area of voltage and the area of frequency were significantly different between the groups at 4 weeks (P<0.05). CONCLUSIONS: Masseter muscle activity after RF was significantly decreased compared to its preoperative state. The decreased activity was related to the number of applications and time elapsed after RF.

The change in dimension of the masseter muscle in rabbits after radiofrequency therapy.

PURPOSE: The objective of this study was to evaluate the change in muscle dimension after administering radiofrequency (RF) therapy. MATERIALS AND METHODS: This study used 6 male New Zealand rabbits. Groups were divided by number of applications of RF (eg, 1 to 4 points). The dimension of the masseter muscle was measured using a computerized tomogram scan before operation, and at 1, 2, 3, and 4 weeks after RF therapy was administered under the same conditions. Two horizontal cuts were selected for measurement. RESULTS: The size of the measured areas for each group at 1 week after RF therapy was significantly increased compared with the preoperative value (P< .05). When the measurements of each group at 3 and 4 weeks after RF therapy were compared with the preoperative value, they were significantly decreased (P< .05). The dimensional change was significantly different among groups at 1 and 3 weeks post RF therapy (P< .05). The swelling at 1 week after RF therapy was increased in terms of the number of RF applications. The ratio of dimension was decreased at 3 weeks after RF therapy in terms of the number of RF applications. CONCLUSION: There was an increase in muscle dimension because of swelling in the early stages of RF therapy. However, this dimension decreased at 3 weeks post-RF administration compared with the preoperative value. Therefore, it can be concluded that the change in the masseter muscle dimension was dependent on the number of RF applications.