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.

[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.

[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).

[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.

[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.

[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.

[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.

[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.

[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.

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.

[Wear behavior of enamel and veneering ceramics].

OBJECTIVE: To compare the wear between the enamel and two types of dental decoration porcelains for all-ceramic restorations (Vita-alpha, Vintage AL). METHODS: Friction coefficients, wear scar width, element concentrations and wear surface evolution were considered relatively to the tribology of that in vivo situation. The wear scars of the samples were characterized by means of dynamic atomic force microscopy (DFM). The different element concentrations of the surface before/after the wear test were determined with energy dispersion spectrometry (EDS). RESULTS: The friction coefficient varied from time in each kind of material. The statistical differences between materials were observed in wear scar width and properties of materials (P<0.05). DFM results showed wear surface of natural tooth full of abrasive particles and denaturation of dental texture. Wear surface of veneering ceramics consisted mainly of abrasive particles, plough and microcracking. EDS results showed that the element concentration of Fe was obviously found on the samples after wear. CONCLUSION: The main underlying mechanisms of natural teeth wear are abrasive, and denaturation of dental texture. Abrasive wear, adhesion and fatigue of veneering ceramics characterize the wear patterns which plays different role in Vita-alpha and Vintage AL. The wear patterns of veneering ceramics can be described as mild wear.

[Effects of simulating static magnetic fields of magnetic attachments on osteoblasts proliferation, cell cycle distribution and apoptosis ratio].

OBJECTIVE: To investigate the effects of the static magnetic field (SMF) generated by dental magnetic attachments on osteoblastic proliferations, cell cycle distribution, and apoptosis ratio. METHODS: By simulating those of the closed-field, the closure process and the open-field Magnedisc 800 magnetic attachments respectively, the in vitro cultured rat osteoblasts were exposed continuously to 12.5, 125, 250 mT SMF. The effects of the SMF on the proliferation of the cells were examined. MTT colorimetry test was performed to detect the effect of the SMF on the vitalities of cells. Flow cytometry was utilized to analyze the cell cycles and cell apoptosis rates. RESULTS: The SMF exposure didn't change the vital osteoblasts number, the cell cycle distribution and proliferation activities of osteoblasts. The cell apoptosis situation were not observed statistical differences. CONCLUSION: No matter the closed-field, the closure process and the open-field magnetic attachments respectively, continuous simulating SMF-stimulation of magnetic attachments couldn't change osteoblasts proliferation activity, cell cycle distribution, and apoptosis ratio.

[Study on color matching of equal volume in dental ceramic prothesis materials].

OBJECTIVE: To study the color changes on brightness and hue in dental ceramic prothesis materials. METHODS: The different color mark of VINTAGE series was sintered on 15 pieces Ni-Cr alloy denture materials. The sample group and control group was given the sintering of color blending with equivalent volume. The order of color mark was of A1 and A2, A2 and A3, A3 and A4, B1 and B2, B2 and B3, B3 and B4 in sample group, and A1, A2, A3, A3.5, A4, B1, B2, B3 and B4 orderly in control group. The chromatic analysis on samples of VITA had been taken through applying PR 650 spectrum detector and adopted the statistic software of Photo Research and SPSS 12.0 for analysis. RESULTS: The statistical difference (P<0.05) of L*a*b* value was found in the different color mark of the same color serials. A non-linearity relation had been found in the chromatic parameters a*b*. It means the change of color was visible. CONCLUSION: Set A has a bigger grading change of brightness than Set B. The change of red-green is obviously in Set A, as well as yellow-blue in Set B.

[Effect of vacuum deposition technology on the metal-porcelain bond strength of a new type of CO-CR ceramic and framework dental alloy].

OBJECTIVE: To investigate the effect of a new engineering technique of vacuum deposition-plasma magnetron reactive sputter deposition technique on the metal-porcelain bond strength of a new type of Co-Cr ceramic and framework dental alloy. METHODS: Before porcelain painted on the specimens, the standardized metal strips made from DA9-4 dental alloy were coated with a thin Al2O3 ceramic film by plasma magnetron reactive sputter deposition technique. The conformation, structure and thickness of the ceramic film were analyzed. The specimens for three-point bending test made from DA9-4 alloy and VMK95 porcelain were used for metal-porcelain bond strength measurement, in the same time the interface of metal-porcelain and element distribution were also observed. RESULTS: The flexural bonding strength of metal-porcelain of sputtering group and control group were (180.55+/-16.45) MPa and (143.80+/-24.49) MPa. The flexural bonding strength of metal-porcelain of sputtering group was higher than control group significantly through statistical analysis (P<0.01). CONCLUSION: The plasma magnetron reactive sputter deposition technique has a positive effect in improving the bonding strength of DA9-4 dental alloy and ceramic.

[Study of biologic effects of simulating static magnetic field of magnetic attachment on human periodontal ligament fibroblasts].

OBJECTIVE: To evaluate the biologic effects of simulating static magnetic field of magnetic attachment on human periodontal ligament fibroblasts (HPDLF). METHODS: HPDLF were cultured in vitro and exposed to simulating static magnetic fields of magnetic attachments for 4 days with magnetic flux density of 12.5, 125 and 250 mT respectively, while the control group free of additional static magnetic field was also set up. The activities of cellular alkaline phosphatase (ALP) and superoxide dismutase (SOD) were measured with a spectrophotometer, and cell cycle was analyzed by flow cytometry. RESULTS: An increase were detected, induced by 250 mT static magnetic field exposure for 4 days in terms of activity of ALP in HPDLF, as compared with the control group (P < 0.05). However, no corresponding effects were found in activity measurements of SOD (P > 0.05). Also there were no statistical differences observed on cell cycle distribution (P > 0.05) or cell proliferation index (P > 0.05) under such static magnetic fields. CONCLUSION: The static magnetic fields of magnetic attachment used in this study have improved the activity of ALP, and do little inference to the activity of SOD and the cell cycle distribution.

[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.

[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.

[Study on the surface roughness of three dental alloys soaked in artificial saliva].

PURPOSE: The aim of this study was to investigate the corrosion behavior of three metal materials for dental use (Co-Cr alloy, Ni-Cr alloy, titanium) by measurement of their surface roughness after soaked in artificial saliva with two different pH values. METHODS: Standard samples were casted respectively from 3 different alloys and grided into the same roughness .Then the samples were exposed to artificial saliva with two different pH values. After three months,the surface roughness of the samples was tested, and the microstructure of surfaces was studied by metalloscope. The data were analyzed with SPSS11.0 software package for one-way ANOVA and t test. RESULTS: There was significant difference in surface roughness of three alloys after exposed to artificial saliva with pH5.6. The surface roughness values of three alloys were listed as follows: Ni-Cr alloy>Co-Cr alloy >titanium(P<0.01). When exposed to artificial saliva with pH7.0, Ni-Cr alloy was found rougher than Co-Cr alloy and titanium(P<0.01), while there was no significant difference in roughness between Co-Cr alloy and titanium(P>0.05).Ni-Cr alloy and Co-Cr alloy soaked in artificial saliva with pH5.6 were found rougher than the same material in artificial saliva with pH7.0(P<0.01).The microstructure of the samples' surface verified the results. CONCLUSION: Titanium was stable not only in neutral environment but also in acid environment. The corrosion resistance of Co-Cr alloy and Ni-Cr alloy was superior in neutral environment than in acid environment. The corrosion resistance of the three alloys was: Titanium >Co-Cr alloy >Ni-Cr alloy.

[The fabrication and chroma study of a kind of custom shade guide with metal substrate].

PURPOSE: This study was to explore the chroma of a custom shade guide with metal substrate. METHODS: A custom shade guide sample was fabricated using the central maxillary incisor as a model,which was corresponding to the colour of VITAPAN shade guide.The colour of each shade tab was measured with PR-650 spectrophotometer when every porcelain was added. RESULTS: From the opaque to glaze, the lightness was decreased and the chroma was increased distinctively; When the body porcelain were baked over their corresponding opaque, the lightness was decreased and the chroma was increased; When the enamel porcelain and the translucent porcelain were added on, the lightness and chroma were decreased slightly; the glaze could increase the lightness. CONCLUSION: The "add on" porcelain can affect the final color of the restoration, and the effects of each porcelain layer were different from each other.