Notoginsenoside R1 attenuates oxidative stress-induced osteoblast dysfunction through JNK signalling pathway.

Oxidative stress (OS)-induced mitochondrial damage and the subsequent osteoblast dysfunction contributes to the initiation and progression of osteoporosis. Notoginsenoside R1 (NGR1), isolated from Panax notoginseng, has potent antioxidant effects and has been widely used in traditional Chinese medicine. This study aimed to investigate the protective property and mechanism of NGR1 on oxidative-damaged osteoblast. Osteoblastic MC3T3-E1 cells were pretreated with NGR1 24 h before hydrogen peroxide administration simulating OS attack. Cell viability, apoptosis rate, osteogenic activity and markers of mitochondrial function were examined. The role of C-Jun N-terminal kinase (JNK) signalling pathway on oxidative injured osteoblast and mitochondrial function was also detected. Our data indicate that NGR1 (25 µM) could reduce apoptosis as well as restore osteoblast viability and osteogenic differentiation. NGR1 also reduced OS-induced mitochondrial ROS and restored mitochondrial membrane potential, adenosine triphosphate production and mitochondrial DNA copy number. NGR1 could block JNK pathway and antagonize the destructive effects of OS. JNK inhibitor (SP600125) mimicked the protective effects of NGR1while JNK agonist (Anisomycin) abolished it. These data indicated that NGR1 could significantly attenuate OS-induced mitochondrial damage and restore osteogenic differentiation of osteoblast via suppressing JNK signalling pathway activation, thus becoming a promising agent in treating osteoporosis.

Effect of application time of maleic acid on smear layer removal and mechanical properties of root canal dentin.

OBJECTIVES: To evaluate the effect of maleic acid (MA) on the cleaning efficacy and mechanical properties of root canal dentine with respect to different time exposure. MATERIALS AND METHODS: One hundred and eighty single-canal premolars were instrumented with rotary-files and then randomly assigned to test groups receiving 7% MA for 30 s, 45 s, 1 min, or 3 min or to control groups treated with 0.9% saline or 17% ethylenediaminetetraacetic acid for 45 s. The micro-hardness, nano-hardness and elastic modules were measured before and after treatment, while the amount of smear and erosion in the coronal, middle and apical thirds in root canal were evaluated by scanning electron microscopy, finally, the fracture strength was assessed by vertical root fracture testing. RESULTS: The efficacy of smear layer removal increased with increasing MA application time. The largest effect was observed at 45 s, even in the apical third, whereas the treatment for 1 min resulted in irreversible erosion of the dentine surface. The micro-hardness and nano-indentation testing confirmed that the micro- and nano-scale mechanical properties were significantly decreased after MA application for 1 min. Furthermore, the specimens treated with MA for 3 min presented the lowest fracture resistance among all groups. In contrast, the 45 s treatment appeared to increase the fracture resistance of the tooth. CONCLUSIONS: The cleaning efficacy and mechanical properties of root canal dentine varied with MA exposure time. The application of MA for 45 s was found to be the most promising for clinical use.

Changes in composition and enamel demineralization inhibition activities of gallic acid at different pH values.

BACKGROUND: Gallic acid (GA) has been shown to inhibit demineralization and enhance remineralization of enamel; however, GA solution is highly acidic. This study was to investigate the stability of GA solutions at various pH and to examine the resultant effects on enamel demineralization. METHODS: The stability of GA in H2O or in phosphate buffer at pH 5.5, pH 7.0 and pH 10.0 was evaluated qualitatively by ultraviolet absorption spectra and quantified by high performance liquid chromatography with diode array detection (HPLC-DAD). Then, bovine enamel blocks were subjected to a pH-cycling regime of 12 cycles. Each cycle included 5 min applications with one of the following treatments: 1 g/L NaF (positive control), 4 g/L GA in H2O or buffered at pH 5.5, pH 7.0 and pH 10.0 and buffers without GA at the same pH (negative control), followed by a 60 min application with pH 5.0 acidic buffers and a 5 min application with neutral buffers. The acidic buffers were analysed for dissolved calcium. RESULTS: GA was stable in pure water and acidic condition, but was unstable in neutral and alkaline conditions, in which ultraviolet spectra changed and HPLC-DAD analysis revealed that most of the GA was degraded. All the GA groups significantly inhibited demineralization (p < 0.05) and there was no significant difference of the inhibition efficacy among different GA groups (p > 0.05). CONCLUSIONS: GA could inhibit enamel demineralization and the inhibition effect is not influenced by pH. GA could be a useful source as an anti-cariogenic agent for broad practical application.

Mineral densities and elemental content in different layers of healthy human enamel with varying teeth age.

OBJECTIVES: This study aims to evaluate the mineral density, calcium (Ca), and phosphorus (P) weight percent of healthy human enamel of varying human teeth age and enamel layers. METHODS: Twenty human teeth were divided into "young" (18-24 years) and "old" (55 ≤ years) age groups. From each tooth, one enamel slab (approximately 8 mm × 2 mm × 1.5 mm) was obtained. All enamel slabs were embedded and scanned by micro-computed tomography. The scanned slices were reconstructed into a 3D image and regions of interest (ROIs) were defined. Then, the mineral density for each ROI was calculated. The Ca and P weight percent in the outer, middle, and inner enamel layers were quantified by scanning electron microscopy with energy dispersive spectroscopy. Finally, the data were analysed using one-way ANOVA and Student's t-test. RESULTS: In the two age groups, the mineral density, Ca and P weight percent decreased from outer to the inner enamel layer. The mineral density, Ca and P weight percent in the outer enamel layer in the old age group were significantly higher than those in the young age group (P<0.05); however, no age-dependent differences were observed for these properties in the middle and inner enamel layers (P>0.05). CONCLUSIONS: The mineral density, Ca and P weight percent decreased from the outer to the inner enamel layers in the two age groups. In the outer enamel layer, the differences in these properties between the two age groups were significantly different; however, no region-dependent differences were observed in the middle and inner enamel layers.

Combined effects of nano-hydroxyapatite and Galla chinensis on remineralisation of initial enamel lesion in vitro.

OBJECTIVES: The aim of the present study was to investigate the combined effects of nano-hydroxyapatite and Galla chinensis on remineralisation of initial enamel lesion. METHODS: Bovine enamel blocks with in vitro produced initial lesion were used. The lesions were subjected to a pH-cycling regime for 12 days. Each daily cycle includes 4 x 3 min application with one of five treatments: NaF (positive control), deionised water (negative control), crude aqueous extract of G. chinensis (GCE), nano-hydroxyapatite (nano-HA) and GCE with nano-HA. The samples were subsequently evaluated using a microhardness tester, polarised light microscopy (PLM), X-ray diffraction (XRD) and scanning electron microscopy (SEM). RESULTS: Surface hardness measurements and integrated mineral recovery value obtained from cross-sectional microhardness test (CSMH) revealed that all the treatment groups had significantly greater effect on enhancing remineralisation than that of the negative control group. Detailed investigation of both CSMH and PLM indicated that nano-HA would only help mineral deposition predominate in the outer layer of lesion and had limited capacity to reduce the lesion depth significantly. In GCE-nano-HA combined treatment group, more mineral deposition occurred in the lesion body and lesion depth was reduced significantly. Meanwhile, significantly greater mineral deposition in the outer portion of the lesion was also observed in comparison with GCE group. The results of XRD and SEM also showed that GCE could influence the deposition and adsorption of nano-HA. CONCLUSION: There was a significant synergistic effect of combined GCE and nano-HA treatment on promoting the remineralisation of initial enamel lesion.

Effect of Galla chinensis on enhancing remineralization of enamel crystals.

The aim of this scanning electron microscopy (SEM) study was to investigate the effect of chemical compounds of Galla chinensis (GCE, gallic acid) on the remineralization of enamel crystals in vitro. Bovine enamel blocks with an in vitro produced initial lesion were used. The lesions were subjected to a pH-cycling regime for 12 days. Each daily cycle included 4 x 1 min applications with one of six treatments: group A: 1000 ppm F aq. (as NaF, positive control); group B: deionized water (DDW, negative control); group C: 4000 ppm crude aqueous extract of GCE; group D: 4000 ppm gallic acid; group E: 4000 ppm GCE with 1000 ppm F; group F: 4000 ppm gallic acid with 1000 ppm F. The surface and vertical section of the enamel lesions were analyzed by SEM. The results indicated that the chemical compounds of G. chinensis could regulate the de-/remineralization balance through influencing the morphology and structure of enamel crystals, and the mechanisms seem to be different for GCE and gallic acid.