Systemic Dietary Hesperidin Modulation of Osteoclastogenesis, Bone Homeostasis and Periodontal Disease in Mice.

This study aimed to evaluate the effects of hesperidin (HE) on in vitro osteoclastogenesis and dietary supplementation on mouse periodontal disease and femoral bone phenotype. RAW 264.7 cells were stimulated with RANKL in the presence or absence of HE (1, 100 or 500 µM) for 5 days, and evaluated by TRAP, TUNEL and Western Blot (WB) analyses. In vivo, C57BL/6 mice were given HE via oral gavage (125, 250 and 500 mg/kg) for 4 weeks. A sterile silk ligature was placed between the first and second right maxillary molars for 10 days and microcomputed tomography (µCT), histopathological and immunohistochemical evaluation were performed. Femoral bones subjected or not to dietary HE (500 mg/kg) for 6 and 12 weeks were evaluated using µCT. In vitro, HE 500 µM reduced formation of RANKL-stimulated TRAP-positive(+) multinucleated cells (500 µM) as well as c-Fos and NFATc1 protein expression (p < 0.05), markers of osteoclasts. In vivo, dietary HE 500 mg/kg increased the alveolar bone resorption in ligated teeth (p < 0.05) and resulted in a significant increase in TRAP+ cells (p < 0.05). Gingival inflammatory infiltrate was greater in the HE 500 mg/kg group even in the absence of ligature. In femurs, HE 500 mg/kg protected trabecular and cortical bone mass at 6 weeks of treatment. In conclusion, HE impaired in vitro osteoclastogenesis, but on the contrary, oral administration of a high concentration of dietary HE increased osteoclast numbers and promoted inflammation-induced alveolar bone loss. However, HE at 500 mg/kg can promote a bone-sparing effect on skeletal bone under physiological conditions.

Fluoride release and uptake by various dental materials after fluoride application.

PURPOSE: To measure the amounts of fluoride released from fluoride-containing materials before and after daily topical fluoride applications. METHODS: A conventional glass-ionomer: Fuji Ionomer Type II (F2); a resin-modified glass-ionomer: Fuji Ionomer Type II LC (LC); two "giomer" materials: Reactmer Paste (RP), and Beautifil (BT); a fluoride-containing resin composite: Unifil F (UF); and a non-fluoride resin composite: AP-X (AP) were used in this study. Each material was filled into a plastic mold, with inner diameter of 9 mm wide x 3 mm high. The specimens were stored in vials filled with 8 ml distilled deionized water for 24 hours at 37 degrees C. The specimens were then removed from the vials and the amount of fluoride released into the water, over the 24-hour period, was measured. The amount of fluoride released was measured by using specific fluoride electrode and an ion-analyzer. These procedures were repeated at Days 2, 3, 7, 14, and 21. After 21 days, all specimens were exposed to 1000 ppm F NaF solution for 5 minutes once a day. This procedure and measurement of fluoride release were continued for 14 days. After 14 days, the specimens were placed in water for 7 days and fluoride release was measured. The results were statistically analyzed using Kruskal-Wallis and Mann-Whitney U-test (P< 0.05). RESULTS: At the 22nd day (1 day after starting fluoride exposure), there was no difference between the F2 and RP, though there were significant differences between the two GICs and the groups BT and UF. After that day, there were significant differences between GIC and the group RP, BT and UF. All materials showed a decrease in fluoride release 7 days after end of the fluoride immersion period. F2, LC, and UF showed no significant difference of fluoride release between Day 21 and 1 day after the end of the fluoride immersion period (P= 0.310: F2 and UF, 0.548: LC). On the other hand, RP and BT revealed lower fluoride release 1 day after the end of the fluoride immersion period as compared to Day 21 (P= 0.075: RP, 1.000: BT). For AP, fluoride release was not detected after the fluoride immersion period.