Cytotoxicity of bisphenol A glycidyl methacrylate on cytochrome P450-producing cells

Of the cytochrome P450 (CYP) family of carcinogen-activating enzymes, CYP3A is the major form found in human livers. The purpose of this study was to investigate the cytotoxic effects of dental resin monomers after being metabolized by CYP3A4 and CYP3A7, using a colony formation assay and a neutral red assay. Specimen wells were plated with transfected cells derived from the Chinese hamster lung at 100 cells well(-1). The experimental group consisted of CYP-producing 3A4-10 and 3A7-40 cells, while the control group consisted of non-CYP-producing CR-119 cells. Bisphenol A (BPA) and bisphenol A glycidyl methacrylate (Bis-GMA) and a positive control (Aflatoxine Bl) were added separately to each well and cultured for 7 days. After cultivation, the number of the colonies was counted and IC50 values were determined. The data were statistically analysed by a Student's t-test. The resultant of IC50 values indicated that the monomers were not metabolically activated by CYP3A4 or CYP3A7 as compared with the control (P < 0.05). We also confirmed that these monomers act neither as activators nor as inhibitors of CYP3A4 and CYP3A7.     

IL-17A stimulates the expression of inflammatory cytokines via celecoxib-blocked prostaglandin in MC3T3-E1 cells

Objective

The prostaglandins (PGs) released from osteoblasts can alter the process of bone remodelling. Recently, we showed that compressive force induced the expression of pro-inflammatory cytokine interleukin (IL)-17s and their receptors in osteoblastic MC3T3-E1 cells and that IL-17A was expressed most highly. Consequently, in the current study we examined the effect of IL-17A and/or celecoxib on PGE₂ production and the expression of cyclooxygenases (COXs) and inflammatory cytokines in MC3T3-E1 cells. We also examined the effects of PGE₂ and cyclohexamide on the expression of inflammatory cytokines.

Methods

Cells were cultured with or without IL-17A (0.1, 1.0, or 10 ng/ml) in the presence or absence of 10 μM celecoxib, a specific inhibitor of COX-2, for up to 72 h. Cells were pretreated with or without 10 μg/ml cycloheximide, protein synthesis inhibitor, for 30 min, and then cultured with 10 ng/ml IL-17A for 24 h. Cells were also cultured with or without 1.5 ng/ml PGE₂ for 24 h. PGE₂ production was determined by ELISA. The expression of COX-1, COX-2, IL-1α, IL-6, IL-8, IL-11, and TNF-α mRNAs and proteins was determined by real-time PCR and ELISA, respectively.

Results

The expression of COX-2, IL-1α, IL-6, IL-8, IL-11, and TNF-α, as well as PGE₂ production increased in the presence of IL-17A, whereas COX-1 expression did not change. Celecoxib blocked the stimulatory effect of IL-17A on the expression of COX-2, IL-1α, IL-6, IL-8, and IL-11 as well as PGE₂ production, whereas it did not block TNF-α expression. Cycloheximide pretreatment suppressed the expression of IL-17-induced inflammatory cytokines. The expression of IL-1α, IL-6, IL-8, and IL-11 increased by the addition of PGE₂, whereas TNF-α expression was not affected.

Conclusion

These results suggest that IL-17A stimulates the expression of bone resorption-related inflammatory cytokines through an autocrine mechanism involving celecoxib-blocked PGs, mainly PGE₂, in osteoblasts.     

Degree of immobilization of synthetic RGDS(PO(3)H(2))PA peptides on titanium surfaces

The purpose of this study was to characterize the chemical interaction between titanium surfaces and the peptide RGDS(PO(3)H(2))PA (P-RGD) synthesized from RGD peptide (RGD) and o-phospho-L-serine (P-Ser), and to determine the degree of peptide immobilization on the titanium surface. X-ray photoelectron spectroscopy showed that the adsorption amount of RGD was significantly smaller than those of P-Ser and P-RGD (p<0.05). Furthermore, although it appeared that P-RGD bonded to the surface, ultrasonic rinsing with water caused it to dissociate, releasing RGD and leaving only S(PO(3)H(2))PA bonded to the surface. These findings show that although it remains difficult to obtain a stable P-RGD layer, the phosphate functional group greatly improves immobilization of the molecule on titanium surfaces.     

Gamma-interferon enhances expression of CD14/MyD88 and subsequent responsiveness to lipopolysaccharide from Actinobacillus actinomycetemcomitans in human gingival fibroblasts

OBJECTIVES: CD14, toll-like receptor 4 (TLR4) and MyD88 have been shown to mediate responsiveness in host cells to lipopolysaccharide. We investigated here the regulatory effects of inflammatory cytokines on the expression of membrane CD14 (mCD14), TLR4 and MyD88, and on subsequent responsiveness to lipopolysaccharide from Actinobacillus actinomycetemcomitans in human gingival fibroblasts. MATERIALS AND METHODS: Following treatment with either interleukin-1beta, tumor necrosis factor-alpha (TNF-alpha) or gamma-interferon (IFN-gamma), expression of mCD14/TLR4 and MyD88 was determined by flow cytometry and western blotting, respectively. After pretreatment with IFN-gamma, cells were pre-incubated with either anti-CD14 antibody MY4 or anti-TLR4 antibody HTA125 and subsequently treated with A. actinomycetemcomitans lipopolysaccharide. Then, phosphorylation of mitogen-activated protein (MAP) kinases and IkappaBalpha was examined by western blotting, and production of interleukin-6 and interleukin-8 was measured by their respective enzyme-linked immunosorbent assay (ELISA) kits. RESULTS: IFN-gamma stimulated expression of mCD14, whereas -1beta and TNF-alpha did not. Expression of MyD88 but not TLR4 was also enhanced by IFN-gamma. The lipopolysaccharide activated MAP kinases, such as extracellular signal-regulated kinase (ERK), c-Jun N-terminal kinase (JNK) and p38, and IkappaBalpha and stimulated production of interleukin-6 and interleukin-8. The lipopolysaccharide-stimulated interleukin-6 and interleukin-8 production was markedly inhibited by MY4 or HTA125. Pretreatment with IFN-gamma augmented the following activation of MAP kinases and IkappaBalpha and production of interleukin-6 and interleukin-8 in response to the lipopolysaccharide. CONCLUSIONS: These results suggest that the augmentation by IFN-gamma of the responsiveness to A. actinomycetemcomitans lipopolysaccharide, such as activation of MAP kinases and IkappaBalpha and terminal cytokine production in human gingival fibroblasts, may be partially mediated by up-regulation of CD14 and MyD88 expression.     

Occlusal force distribution on the dental arch during various levels of clenching

The purpose of this study was to explore the occlusal force distribution on the dental arch in the intercuspal position and to evaluate the relationship between the clenching strength and the occlusal force distribution. These variables were recorded using the Dental Prescale System in 16 healthy young adults. The number of tooth contacts, occlusal force and occlusal contact area increased linearly as clenching strength increased. The distribution of the occlusal force was greatest at the molar region followed by the premolar and anterior teeth region. The proportion of occlusal force (occlusal force at each region/total occlusal force) on molar regions increased as clenching strength increased. On the contrary, the proportion of occlusal force on the premolar and anterior teeth regions decreased as clenching strength increased. These findings suggest that control of occlusal force is important in diagnosis of the nature of occlusal contacts.     

In situ electrochemical scanning tunneling microscopy of Co(0001) single crystal electrodes in acidic solution

The surface of a Co(0001) single crystal electrode was examined by in situ electrochemical scanning tunneling microscopy (STM) in 0.05 M Na₂SO₄ at pH 3 under potential control. Cyclic voltammetry indicated that the anodic dissolution of Co proceeds without passivation, and the anodic current was almost proportional to applied potential. The Co(0001)-(1 × 1) structure was observed on atomically flat terraces in the potential range between the hydrogen evolution and the onset of the anodic dissolution. At the foot of the rising anodic current-potential curve, a hexagonal surface lattice with (5 × 5) symmetry corresponding closely to that of CoO(111) or Co(OH)₂(0001) was also observed. In the early stages of anodic dissolution, the process proceeded in the layer-by-layer mode, forming atomically flat terraces extending over large areas. By reapplying a negative potential after the anodic dissolution, well-defined terrace-step structures were restored, suggesting that the electrodeposition also took place preferentially at step edges.     

Importance of the changes in joint effusion shown by magnetic resonance imaging before and after arthroscopic lysis and lavage of the temporomandibular joint

We investigated the changes in the amount of joint effusion estimated from T2-weighted magnetic resonance imaging (MRI) before and after arthroscopic lysis and lavage of the temporomandibular joint (TMJ). We studied 29 consecutive patients, each with internal derangement and osteoarthritis in one TMJ. Before operation, the MRI showed joint effusions in 22 of the patients (76%). After operation, the amount of the effusion decreased in 16 and increased in 2 patients. Effusions developed postoperatively in four of the seven patients who had no effusion before operation. In four of the six patients in whom the effusion increased, the symptoms had almost resolved by the time the MRI was taken. There was no significant correlation between changes in the amount of joint effusion and the clinical condition of the patients before and after the operation. In conclusion, changes in the amount of joint effusion in the TMJ are not related to the patient's clinical condition.     

The fibrous structure of the cemento-dentinal junction in human molars shown by scanning electron microscopy combined with NaOH-maceration

The cemento dentinal junction was studied in acellular and cellular cementum of human mandibular third molars by scanning electron microscopy combined with NaOH-maceration. Scanning electron microscopy with NaOH-maceration was applied to observe the fibrous structure in detail through long sections of the cemento-dentinal junction. In macerated specimens, the cemento dentinal junction was a fibril-poor groove. Some cemental fibrils or fibril bundles penetrated the groove and appeared to intermingle with dentinal fibrils. Prolonged maceration caused detachment of the cemento-dentinal junction irrespective of fibril intermingling allowing observation of the inner cementum surface facing the root dentin. Observations suggested that the fibril intermingling was point-like and present only in places at the cemento-dentinal junction. It was established that NaOH-maceration removes interfibrillar substances effectively in connective tissues and does no damage to the collagen fibril structure and architecture. This study showed the 3-dimensional fibrous structure of the cemento-dentinal junction in human mandibular third molars, and suggested that interfibrillar adhesive substances are more important than the fibril intermingling for the cemento-dentinal attachment.     

Flexural properties of eight flowable light-cured restorative materials, in immediate vs 24-hour water storage

This study evaluated the flexural strength, flexural modulus, modulus of resilience and water sorption of eight flowable light-cured restorative materials compared with two conventional restoratives (as control). Forty specimens of each material were made. Twenty specimens were immediately flexural tested, while the remaining 20 were weight-measured and immersed in distilled water in a 37 degrees C incubator. After 24 hours, the samples were weight-measured again to identify water sorption and they were flexural tested. The findings were statistically analyzed using t-test, one-way ANOVA, Tukey test and Pearson's Product-Moment Correlation. The results of the flexural strength test were also analyzed using Weibull statistic. All flowable light-cured restorative materials except Palfique Estelite Low Flow exhibited immediate flexural strength values between the conventional ones. All flowable light-cured restorative materials showed 24-hour flexural strength values between the conventional ones. The Weibull modulus for immediate flexural strength of the materials varied from 6.37 to 15.23, while for 24-hour flexural strength, the strength varied from 8.10 to 14.30. In both conditions, all flowable light-cured resin composites showed lower flexural moduli but higher modulus of resilience than the conventional ones. The water sorption of all resin composites was lower than the flowable light-cured compomer. There was a distinct relation (r=-0.84, p<0.01) between the increasing ratio in modulus of resilience and the amount of water sorption.