Flavokawain B induces apoptosis of human oral adenoid cystic cancer ACC-2 cells via up-regulation of Bim and down-regulation of Bcl-2 expression.

Novel effective drugs are still urgently needed in the prevention and treatment of oral adenoid cystic carcinoma (ACC). In this study, we have assessed the antitumor potential and molecular mechanisms of flavokawain B (FKB) as a kava chalcone on the ACC-2 cell line in vitro. The results demonstrated that FKB could significantly inhibit the cell proliferation of ACC-2 in a dose-dependent manner that was associated with induced apoptosis and cell cycle G2-M arrest, and the half maximal inhibitory concentration (IC50) of flavokawain-B treatment for 48 h was estimated to be 4.69 ± 0.43 µmol/L. Mechanistically, FKB could induce the release of cytochrome c from mitochondria into the cytosol, and activate the cleavage of caspase-3 and, eventually, the poly(ADP-ribose) polymerase (PARP), in a dose-dependent manner, leading to marked apoptotic effect of ACC-2 cells. The apoptotic action of FKB was associated with the increased expression of proapoptotic proteins: Bim, Bax, Bak and a decreased expression of antiapoptotic Bcl-2. Among them, Bim expression was significantly induced by FKB, and knockdown of Bim expression by short-hairpin RNAs attenuated the inhibitory effect induced by FKB on ACC-2 cells. These results suggest Bim may be one of the potential transcriptional targets, and suggests the potential usefulness of FKB for the prevention and treatment of ACC.

Magnetic fields of 10mT and 120mT change cell shape and structure of F-actins of periodontal ligament cells.

Dental magnetic attachments, usually applied locally to oral cavities, produce stray fields (flux leakage) spreading in adjacent tissues. It has been found that human periodontal ligament (PDL) cells change their geometry and the structure of their cytoskeleton F-actins when the cell cultures are exposed to B-field strengths of B = 10mT and 120mT, respectively, which are similar to those generated by dental magnetic attachments. Analytically, after long-time exposures to B-fields for 12h, 36 h and 60 h, respectively, cytoskeleton F-actins are labeled with a fluorescent dye and observed under a laser scanning confocal microscope. The geometrical cell parameters of cell length and cell width and the fluorescence emission of labeled F-actins, respectively, were determined and subjected to an automatic image analysis using a special software. The results on cell shrinkage and filament reorganizations were statistically analyzed by the program ANOVA (P < 0.05). It was found that only long-time (hours) exposure to high fields in the order of 0.1T may produce tissue irritations during long-time medical treatments using open- and closed-field dental magnetic attachments.

[Determination and Weibull analysis of the strength of dental veneering porcelains].

OBJECTIVE: To determine the strength of the veneering porcelain stored in artificial saliva. METHODS: Standard beams with a size of 3 mm x 2 mm x 20 mm made from the body, incisal, and translucent porcelains of SHOFU Vintage were stored in the artificial saliva for 30 days. Twenty samples from each material were tested for the strength by the 3-point flexural test at a crosshead speed of 0.5 mm. The data were analyzed by the two-parameter Weibull method. RESULTS: The strength, Weibull modulus (m) and characterization strength (male0) were (75.5 +/- 5.6) MPa, 17.3, and 77.6 MPa for Body porcelain, respectively; (73.6 +/- 5.0) MPa, 16.5, and 76.0 MPa for incisal porcelain, respectively; and (72.9 +/- 4.5) MPa, 16.1, and 75. 5 MPa for translucent porcelain, respectively. CONCLUSION: There are no differences in strength between the three dental porcelains. Weibull analysis describes the strength of the porcelains better than the average and standard deviation.

Effects of static magnetic fields on the voltage-gated potassium channel currents in trigeminal root ganglion neurons.

To evaluated the effects of moderate-intensity static magnetic fields (SMF) on two types of voltage-gated potassium channel (VGPC) currents: I(K,A) and I(K,V), whole-cell patch-clamp experiments were conducted on acute dissociated rat trigeminal root ganglion (TRG) neurons. The results demonstrated that 125 mT SMF could influence the inactivation kinetics of these two VGPC currents by altering the inactivation rate and velocity. No significant change was observed in the activation properties. These findings supported the hypothesis that biological membrane would be deformed in moderate-intensity SMF and the physiological characteristics of ion channels on the membrane would be influenced. The mechanism underlying the different effects of SMF on the I(K,A) and I(K,V) inactivation was also discussed.

[Effects of static magnetic field on the activity of superoxide dismutase of human gingival fibroblasts].

OBJECTIVE: To investigate the effects of static magnetic fields (SMF) generated by the open-circuit and closed-circuit dental magnetic attachments on the enzymology of human gingival fibroblasts. METHODS: In this study, a cellular SMF exposure system developed by our department of prosthetic dentistry was used to impose the static magnetic force on the cultured human gingival fibroblasts in vitro. The activity of superoxide dismutase in those fibroblasts of the test group and that of the control group were measured and compared. RESULTS: No significant difference (P > 0.05) in superoxide dismutase activity was found between the groups when they were loaded with different intensity of magnetic fields or for different period of time. CONCLUSION: Under the conditions in this experiment, the static magnetic fields of the open-circuit magnetic attachments, and those of the closed-circuit ones as well, have no biological effects on the superoxide dismutase of human gingival fibroblasts.

[The research for magnetic attachments producing static magnetic fields to influence on cytoskeletons of human periodontal ligament cells].

OBJECTIVE: The present research aimed to investigate that the static magnetic field (SMF) generated by dental magnetic attachments influenced on the cytoskeletons of human periodontal ligament cells (HPDLCs). METHODS: In vitro cultured HPDLCs were exposed to 10 mT and 120 mT SMF which simulated those of from the closed-field and open-field magnetic attachments respectively for 12-60 hours in a cellular SMF exposure system. The control group cells were cultured outside the SMF exposure system and were only exposed to geomagnetic field of about 0.03-0.07 mT. After finishing the SMF exposure, the cytoskeletons were observed under a laser scanning confocal microscope (LSCM). The image analysis software was utilized to measure and analyze the cell area, length/width ratios and the contents of F-actin of cells. RESULTS: There were certain changes observed in the cytoskeletons after HPDLCs exposed to 10 mT and 120 mT SMF for 12-60 hours. With the SMF strength increasing and the loading time extending, the cell microfilaments got ashorened change and a disordered arrangement inside cells, and the length/width ratios of cells decreased (P < 0.05). The areas of cells decreased after SMF loading for 60 hours (P < 0.05). The cell F-actin contents decreased after SMF loading for 12 hours (P < 0.05), but in the meantime there was no significant difference happening between the 36 or 60 hours SMF loading group and the control group (P > 0.05). CONCLUSION: In this study the SMF of magnetic attachments can lead the certain changes to the cytoskeletons in HPDLCs.

[Effects of hypotonic solution on the proliferation, alkaline phosphatase activity and [Ca2+]i concentration of osteoblast-like cells].

OBJECTIVE: To investigate the effects of static-stretch from the hypotonic solution on the proliferation, alkaline phosphatase activity and [Ca2+]i in the osteoblast-like cells. METHODS: Mechanical loading was introduced by swelling in the hypotonic solution. In vitro cultured MG63 were incubated under continuous swelling of 277, 240 and 163mOsm for 2h, 4h, 8h, 12h, 24h and 48h. The cell proliferation was detected by MTT assay. ALPase and [Ca2+]i were determined by modified enzyme dynamic method and OCPC respectively. RESULTS: The cell proliferation, ALP activity and [Ca2+]i increased slowly under continuous static-stretch of 277 and 240mOsm. The cell proliferation was inhibited under 163mOsm, with a sharp increase of [Ca2+]i at 8h (11.383 +/- 0.111) and an increase of ALPase activity (0.326 +/- 0.002). CONCLUSION: The static-stretch induced from the hypotonic solution has an impact on the proliferation, differentiation, ALPase and Ca2+-ATPase of the MG63. The [Ca2+]i is correlated with the ALPase.

[Compatibility between high-strength dental ceramic (type A) and vintage AL veneering porcelain].

OBJECTIVE: To investigate the interface bond strength and compatibility between High-Strength Dental Ceramic (type A) and Vintage AL veneering porcelain. METHODS: Twenty bar-shape specimens (ten Vintage AL and ten Vitadur alpha) were fabricated, and shear test was conducted to determine the bond strength. A bilayered composite (1 mm core ceramic and 0.8 mm Vintage AL) was prepared and then fractured for scanning electron microscopy (SEM) and energy dispersive spectroscopy (EDS) analysis. Ten all-ceramic anterior crowns were fabricated and the temperatures of thermal shock resistance were tested. RESULTS: The mean values of the bond strength measured were (55.52 +/- 14.64) MPa and (59.37 +/- 13.93) MPa for Vintage AL and Vitadur alpha respectively (P>0.05). SEM showed tight connection between the High-Strength Dental Ceramic (type A) and the veneering porcelain. Element diffusion was also confirmed by energy dispersive spectroscopy (EDS) analysis. The temperature of thermal shock resistance of this system was (179 +/- 15) degrees C. CONCLUSION: Vintage AL veneering porcelain has good thermal and chemical compatibility with High-Strength Dental Ceramic (type A).

[Study on the interface of Ti-porcelain fused under different surface treatment].

OBJECTIVE: This study was made to observe the topography and ionic diffusion of the elements titanium, silicon, et cetera in the Ti-porcelain interface area which was pretreated with different methods. METHODS: Ti-bond porcelain was fused on commercial pure titanium (CPT) with 8 different pretreatment methods; according to the temperature of the preoxidation, there were 300 degrees C, 400 degrees C, 500 degrees C and 600 degrees C groups, and based on the oxidizing-time in the air, there were 5 min, 15 min, 2 h and 24 h groups. The topography and ionic diffusion of elements in the Ti-porcelain interface area were observed with scanning electron microscopy (SEM) and electron probe microanalysis (EPMA) respectively. RESULTS: There were clefts and exfoliations in the 500 degrees C pre-oxidation group and 600 degrees C pre-oxidation group in SEM images. The distribution of titanium in the interlayer in EPMA images decreased from the titanium surface to porcelain, which was opposite to the distribution of silicon. CONCLUSION: The clefts and exfoliations suggested the position of the fracture and brittleness of alpha-Ti[o] layer. The widths of ionic diffusion in the eight groups were different, but it was difficult to measure and compare them exactly.

Laboratory evaluation of a new denture adhesive.

OBJECTIVE: The ideal denture adhesive demonstrates a neutral or slightly basic pH, satisfactory bonding load, and minimal toxicity to the oral mucosa. The purpose of this study was to evaluate the properties of a new denture adhesive (Comfort-DA, developed by the authors). METHODS: Comfort-DA was evaluated in the laboratory to determine the clinical effect of pH, bonding strength, and cytotoxicity. Firstly, the material was diluted at 1.0, 2.0, 3.3, 5.0, and 10% concentrations. Then the pH values were measured at 1-, 2-, 3-, 4-, 5-, 6-, and 8-hour intervals. Secondly, bonding load test was performed between acrylic resin samples on a universal testing machine. An existing product, Fittydent, was used as a control. Finally, the cytotoxicity of several dilutions of Comfort-DA was recorded with a MTT assay on human oral fibroblast cells. Cell viability was recorded by optical density after culturing in an atmosphere of 5% CO2 and 95% air at 37 degrees C for 2, 3 and 4 days, respectively. Wells containing fresh medium alone served as controls. All the results were compared with ANOVA or Tukey intervals (alpha = 0.05). RESULTS: Comfort-DA remained above the critical pH for hydroxyapatite (pH 6.5) in all dilutions throughout the study. Results of bonding strength test indicated that Comfort-DA was significantly stronger than Fittydent (p < 0.05). The statistical analysis of results for cytotoxicity showed that Comfort-DA only displayed slight cytotoxicity. SIGNIFICANCE: The generally favorable laboratory performance of Comfort-DA formulations indicates that this product may be an efficacious denture adhesive.

[Measurements of the flux densities of static magnetic fields generated by two types of dental magnetic attachments and their retentive forces].

OBJECTIVE: To measure and analyze the flux densities of static magnetic fields generated by two types of commonly used dental magnetic attachments and their retentive forces, and to provide guidance for the clinical application of magnetic attachments. METHODS: A digital Gaussmeter was used to measure the flux densities of static magnetic fields generated by two types of magnetic attachments, under four circumstances: open-field circuit; closed-field circuit; keeper and magnet slid laterally for a certain distance; and existence of air gap between keeper and magnet. The retentive forces of the magnetic attachments in standard closed-field circuit, with the keeper and magnet sliding laterally for a certain distance or with a certain air gap between keeper and magnet were measured by a tensile testing machine. RESULTS: There were flux leakages under both the open-field circuit and closed-field circuit of the two types of magnetic attachments. The flux densities on the surfaces of MAGNEDISC 800 (MD800) and MAGFIT EX600W (EX600) magnetic attachments under open-field circuit were 275.0 mT and 147.0 mT respectively. The flux leakages under closed-field circuit were smaller than those under open-field circuit. The respective flux densities on the surfaces of MD800 and EX600 magnetic attachments decreased to 11.4 mT and 4.5 mT under closed-field circuit. The flux density around the magnetic attachment decreased as the distance from the surface of the attachment increased. When keeper and magnet slid laterally for a certain distance or when air gap existed between keeper and magnet, the flux leakage increased in comparison with that under closed-field circuit. Under the standard closed-field circuit, the two types of magnetic attachments achieved the largest retentive forces. The retentive forces of MD800 and EX600 magnetic attachments under the standard closed-field circuit were 6.20 N and 4.80 N respectively. The retentive forces decreased with the sliding distance or with the increase of air gap between keeper and magnet. CONCLUSION: The magnetic attachments have flux leakages. When they are used in patients' oral cavities, if keeper and magnet are not attached accurately, the flux leakage will increase, and at the same time the retentive force will decrease. Therefore the keeper and magnet should be attached accurately in clinical application.

[Experimental study on the reconstruction of mandibular defects with sinboneHT bone replacement in rabbits].

OBJECTIVE: After sinboneHT bone replacement (SBR) was implanted in animals, to evaluate the biocompatibility of SBR and compounded in autogenetic bone in the proportion of one to one in order to prepare for the clinical applications in the future. METHODS: Bone defects of 10 mm x l0 mm x 2 mm was made at the mandibular of rabbits, then SBR with different granule diameter and autogenetic bone was compounded in the proportion of being applied in the left defects, while autogenetic bone was implanted in the right defects and nothing was used in the right reformed defects. Animals were sacrificed at 2, 4 and 8 weeks respectively. The biologic capacity was evaluated with anatomy, X-rays studies and histology. RESULTS: SBR has better biocompatibility, which can effectively accelerate the reconstruction of bone defects and help the new bone by being compounded with autogenetic bone. It provides the appropriate scaffold or template which would allow cellular infiltration, attachment and multiplication. CONCLUSION: SBR is a kind of bone substitute material with good biocompatibility. SBR compounded with self-bone has a better regeneration function.

[Bi-axial flexure strength and fracture mode of alumina glass-infiltrated core/veneer ceramic composites].

PURPOSE: To determine the bi-axial flexural strength and fracture mode of bilayered alumina glass-infiltrated core and the veneering porcelain. METHODS: Forty disk specimens were made from alumina glass-infiltrated core (HSDC-A) and veneer porcelain (Vintage AL), and equally divided into four groups as follows: monolithic specimens of veneer(MV),monolithic specimens of core material(MC),bilayered specimens with the veneer on top (BC) and bilayered specimens with core material on top(BV). Mean flexure strength, standard deviation and associated Weibull modulus were determined using bi-axial flexure (ball-on-ring) for each group. The results were analyzed with one-way ANOVA and the Weibull distribution with SPSS 13.0 software package. Both optical and scanning electron microscopy were employed for identification of the fracture mode and origin. RESULTS: The strength in the group MC and BC were significantly stronger than that in the group MV and BV. The frequency of specimen delamination, Hertzian cone formation and sub-critical radial cracking in the bilayered discs were dependent on the surface loaded in tension. CONCLUSION: Material which lies on the bottom surface dictates the strength and fracture mode of the specimens. Supported by Shenzhen Municipal Technological Project (Grant No.200903082).

[Influence of multiple sintering on wear behavior of Cercon veneering ceramic].

OBJECTIVE: To investigate the influence of multiple sintering on wear behavior of Cercon veneering ceramic. METHODS: Samples were fabricated according to the manufacture's requirement for different sintering times (1, 3, 5, 7 times). The wear test was operated with a modified MM-200 friction and wear machine in vitro. The wear scars were characterized by scanning electron microscope (SEM) and atomic force microscopy (AFM). RESULTS: With the sintering times increasing, the wear scar width became larger. The correlation was significant at the 0.01 level. Significant difference was observed in wear scar width among different samples (P < 0.05). SEM and AFM results showed that veneering ceramic wear facets demonstrated grooves characteristic of abrasive wear. CONCLUSION: Multiple sintering can decrease the wear ability of Cercon veneer, and the wear pattern has the tendency to severe wear.

[Effect of different dynamic mechanical strain on the proliferation and synthetic function of human periodontal ligament fibroblasts in vitro].

OBJECTIVE: To study the changes in the proliferation and synthetic function of human periodontal ligament fibroblasts (HPDLF) in response to dynamic mechanical strains of different modes, magnitudes and durations. METHODS: Using a 4-point bending system, the effect of dynamic mechanical strains of different modes, magnitudes and durations on the proliferation of HPDLF was investigated by analyzing the cell cycle changes with flow cytometry (FCM), and the total protein level and the activity of alkaline phosphatase (ALP) in HPDLF were assayed by quantitative analysis. RESULTS: The percentage of G(0)/G(1) cell decreased, S phase cells increased, and the proliferation index (PI), total protein level and activity of ALP were augmented significantly in response to dynamic mechanical micro-strains. These changes showed close correlations to the magnitude and duration of the strain. The mode of strain caused significant changes in G(0)/G(1), S, and G(2)/M phase cell percentages as well as the PI, total protein level and ALP activity of the cells. In the gradient strain group, the cell proliferation activity, total protein level and ALP activity were obviously higher than those in 1000 and 4000 microstrain groups. CONCLUSION: The changes in the proliferation and synthetic function of HPDLF are closely correlated to the mode, magnitude and duration of the strains.

[Osteoblastic early attachment onto the surface of bioadhesive peptide modified pure titanium].

OBJECTIVE: To investigate the long-term integrity and the biological function of interface between the bioadhesive peptide modified implant surface and peri-implant tissue. METHODS: A short bioadhesive peptide containing Glycine-Tyrosine-Arginine-Glycine-Asparticacid-Serine (GYRGDS) sequence was immobilized onto the titanium implant surface by means of sol-gel coating technique and self-assembled monolayers (SAM). The chemical composition and organic functional groups on the titanium surfaces were characterized using XPS (X-ray photoelectron spectroscopy) and FTIR (Fourier transform infrared spectrometer). The adhesive strength and stability of osteoblasts on various implant surfaces were compared under flow condition. RESULTS: The results showed that alkali/hot water aging treatment could apparently improve the content of -OH functional groups of titanium surface. The chemical reactive Ti-O-Ti bonding at the surface of titanium played a vital role in inducing the formation of organosilane SAM. GYRGDS peptide can be covalently grafted onto the surface of titanium by SAM technique. The resistance of freshly adherent osteoblasts to detachment by flow was shear time dependent. When the four groups were compared under the same flow stress condition (2.05 Pa) at three specific time spans (30 min, 1 h, 2 h), the cells retention rates in GYRGDS-grafted groups were 93.0%, 54.4%, 34.4% respectively and were much higher than those in non-coated groups. CONCLUSIONS: It was suggested that GYRGDS might have positive effects on maintaining stability and adherence of cells onto the substrates under flow condition.

[Regulation of integrin beta1 mRNA expression by mechanical stress in human periodontal ligament fibroblasts].

OBJECTIVE: To investigate the effect of different kinds of mechanical stress on the mRNA expression of integrin beta1 subunit in cultured human periodontal ligament fibroblasts (hPDLF). METHODS: To scalp and remove the periodontal ligament attached to the mid-third part of the fresh root of young premolars extracted for the cause of orthodontics. Cultured hPDLF by the method of digesting by I-type collagenase combining with tissue adhering. Then hPDLF was isolated and purified by cells passage. The sixth passage's cells were selected to be loaded. A new cyclic strain loading apparatus. Forcel four point bending device was used for mechanically loading. Cells were loaded by three levels (1000, 2000, 4000 microstrain) of tensional and compressive forces and collected at different times (0, 0.5, 1, 4, 8, 12 h) course after strain loading. The quantity of integrin beta1 mRNA in every group was analyzed by means of quantitative real-time PCR with the special primers of up- and down-regulated genes. RESULTS: Dynamic mechanical forces down-regulated the expression of integrin beta1 subunit mRNA in hPDLF and the difference in groups by different magnitude, different kinds, and different time of mechanical forces loading were statistically significant. The stronger stimulated forces, the more down-regulated expression. Compression down-regulated the expression of integrin beta1 subunit mRNA more than tension did. CONCLUSION: Dynamic mechanical forces could regulate the expression of integrin beta1 subunit mRNA. The difference among all the groups by different magnitudes, different kinds, and different time of mechanical forces loading were statistically significant.

[Regulation of collagen type I and fibronectin mRNA expression by mechanical stress in cultured human periodontal ligament fibroblasts].

OBJECTIVE: To investigate the effect of different dynamic tensional and compressive stress on the mRNA expression of collagen type I and fibronectin in cultured human periodontal ligament fibroblasts (hPDLF), and explore the regularity of functional change in hPDLF. METHODS: A new cyclic strain loading apparatus was used for mechanically loading. Cells cultured in vitro were loaded with three levels (1000 microstrain, 2000 microstrain, 4000 microstrain) of tensional and compressive forces and collected at different time (0 h, 0.5 h, 1 h, 4 h, 8 h,12 h) course after strain loading. The quantity of collagen type I and fibronectin mRNA was analyzed by means of quantitative real-time PCR with special primers of up- and down-regulated genes. Data were analyzed using SPSS version 10.0 software. RESULTS: Different magnitude and different kinds of mechanical forces as well as the force application time significantly changed the expression of collagen type I and fibronectin mRNA in hPDLF. CONCLUSIONS: Dynamic mechanical forces could regulate the expression of collagen type I and fibronectin mRNA in hPDLF. Collagen type I and fibronectin participated in the mechanical signal transduction in human periodontal ligament fibroblasts.

[Effect of simulating static magnetic field of magnetic attachment on osteoblastic morphology and surface ultrastructure].

OBJECTIVE: To investigate effects of the static magnetic field (SMF) generated by dental magnetic attachment on osteoblastic morphology and surface ultrastructure. METHODS: The in vitro cultured rat osteoblasts were exposed continuously to 12.5 mT, 125 mT, and 250 mT static magnetic fields for 1, 3, 5, and 7 days. After exposed in SMF, osteoblasts were observed under a phase contrast microscope, and then HE stained and observed under a light microscope. In addition, the cells were observed under a scanning electron microscope (SEM). RESULTS: By continuous exposure, the different intensities of SMF exposure did not change the vital osteoblast growth pattern or distribution. The SEM photos showed that there were certain changes in cellular microstructures for osteoblasts after exposed to 12.5 mT for 5 to 7 days, as well as 125 mT and 250 mT for 3 to 7 days. The more exposure time increased, the more microvesicles on the surfaces of cells were observed. CONCLUSIONS: Continuous SMF-stimulation could not affect the shape, distribution, and growth pattern of osteoblasts. The SMF of magnetic attachments could lead to certain changes in surface ultrastructures of osteoblasts in this study.

[Clinical application of implant-supported restoration with all-ceramic crown fused to small-size titanium abutment].

OBJECTIVE: To get aesthetic implant-supported restorations by means of the all-ceramic crown fused to small-size titanium abutment and evaluate the quality of the restorations. METHODS: A small-size titanium abutment that had extremely thin circumferential collar and axial wall was fabricated, while a Cercon all-ceramic crown made with computer aided manufacture (CAM). The crown was fused to the abutment by firing the opaque porcelain. A total of 6 restorations in 5 patients were installed and assessed according to the California Dental Association (CDA) quality evaluation system. RESULTS: All the restorations were fabricated well and ranked in clinic evaluation excellent for surface, anatomical form, marginal quality, and color at baseline and one year after insertion, respectively. CONCLUSIONS: The restoration of all-ceramic crown fused to titanium small-size abutment is a new aesthetic alternative for the implant-supported restoration.