The effect of glutathione on 2-hydroxyethylmethacrylate cytotoxicity and on resin-dentine bond strength.

AIM: To evaluate the influence of reduced glutathione (GSH) application on 2-hydroxyethylmethacrylate (HEMA) cytotoxicity on rat pulpal cells and evaluate the effect of etched-dentine treatment with GSH on the immediate microtensile bond strength (µTBS) of etch-and-rinse adhesive. METHODOLOGY: The cytotoxicity of 10 mmol L(-1) HEMA, 10 mmol L(-1) HEMA + 1 mmol L(-1) GSH, 10 mmol L(-1) HEMA + 5 mmol L(-1) GSH and 10 mmol L(-1) HEMA + 10 mmol L(-1) GSH was compared (6 h and 24 h). Cells viability was measured by means of 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay, followed by morphological observation of cells. Etched-dentine surfaces were rinsed and treated with one of the following solutions: 2% GSH, 5% GSH or 10% GSH, bonded with Adper Single Bond Plus (3M, ESPE, St. Paul, MN, USA) and restored with resin composite. The control group received no GSH treatment. After 1 day of water-storage at 37 °C, the specimens were subjected to µTBS testing. Cytotoxicity and µTBS data were analysed by one-way anova and Tukey post hoc tests (P < 0.05). RESULTS: There were significant differences between the groups. HEMA elicited a remarkable toxic effect. 10 mmol L(-1) GSH prevented HEMA-induced damage at both exposure times. Whilst 5 mmol L(-1) GSH lost its protective effect at 24-h exposure time and 1 mmol L(-1) GSH showed no protective effect at both exposure times, GSH had no significant effect on the immediate µTBS; however, 5% GSH had higher bond strength value when compared to 10% GSH (P = 0.003). CONCLUSION: Controlled concentrations of GSH had a protective effect against HEMA cytotoxicity. GSH had neither positive nor negative influence on µTBS.

Mechanism of the immunosuppressive effect in vivo of novel immunosuppressive drug beta-SQAG9, which inhibits the response of the CD62L+ T-cell subset.

INTRODUCTION: We synthesized sulfo-glycolipid, beta-SQAG9 (designate square beta-SQAG9 liposome, because it efficiently forms a liposome structure) that possessed immunosuppressive effects such as inhibition of T-cell responses in human allogeneic MLR and skin allograft survival in rats, and bound to CD62L (L-selectin) in vitro. In this study, we further investigated the immunosuppressive mechanism in vivo by beta-SQAG9 liposome in a skin-allografted rat model. METHODS: ACI rats (RT1(a)) were grafted skin of LEW rats (RT1(1)) treated with PBS or beta-SQAG9 liposome IV once a day for 7 days. Subsequently, we investigated the population of T cells and CD62L(+) T-cell subset in the spleen, axillary lymph nodes (ALNs), and peripheral blood of skin-allografted rats by two-color flow cytometry. RESULTS: Five of 11 (45.5%) rats that were treated with 50 mg/kg beta-SQAG9 liposome showed graft survival and another showed moderate rejection in graft. The CD62L(+) T-cell subset population in ALNs of beta-SQAG9 liposome-treated rats decreased in a dose-dependent manner. No significant difference in the T-cell population was observed between the beta-SQAG9 and control groups. These data suggest that beta-SQAG9 could bind to the CD62L(+) T-cell subset in vivo as well as in vitro and affect T-cell migration, which might lead to T-cell tolerance in vivo.

Hypersensitivity to thromboxane A2 in cholesterol-rich human platelets.

The effect of changes in platelet membrane cholesterol content on thromboxane A2 (TXA2)-induced platelet activation was studied. Concentrations of 9,11-epithio-11,12-methano-TXA2 (STA2), a stable analogue of TXA2 which can cause half-maximal aggregation and release of [14C]serotonin in cholesterol-rich platelets were significantly lower than those in cholesterol-normal platelets. STA2-induced increase in cytosolic calcium concentration and [32P]phosphatidic acid formation in cholesterol-rich platelets were significantly greater than those in cholesterol-normal platelets. The maximal concentration of binding site (Bmax) for SQ29548 was significantly increased in cholesterol-rich platelets compared with cholesterol-normal platelets, while the equilibrium dissociation rate constant (Kd) for SQ29548 did not differ between cholesterol-rich and cholesterol-normal platelets. The present study suggested that sensitivity to TXA2 was increased by the incorporation of cholesterol into platelet membrane and that the cause of hypersensitivity to TXA2 in cholesterol-rich platelets may be partly explained by an increase in binding capacity for TXA2.

Immunohistochemical localization of metallothionein in the developing teeth of cadmium-injected rats.

The ability of cadmium (Cd) to induce the synthesis of metallothionein (MT) in the developing teeth of the rat was investigated. Rats were given daily intraperitoneal injections of cadmium chloride (1.5 mg Cd/kg) for 7 days. The induction of MT synthesis in incisor teeth after Cd treatment was investigated immunohistochemically using a polyclonal antibody to MT. Immunoreactivity to MT was observed in the papillary layer of epithelial cells of the secretory zone, in one layer of epithelial cells of the presecretory zone and within ameloblasts of the postsecretory zone. Normal control rats did not exhibit MT staining. These results indicate that Cd induces MT synthesis within specific epithelial cells of the enamel organ of the rat. It is proposed that these findings demonstrate an adaptive cellular mechanism that protects these cells from cadmium toxicity.

Gene expression and immunolocalization of amelogenin in enamel hypoplasia induced by successive injections of bisphosphonate in rat incisors.

Successive injections of 1-hydroxyethylidene-1, 1-bisphosphonate (HEBP) in rats induce enamel hypoplasia. To elucidate the pathogenesis of this hypoplasia, male Wistar rats were daily injected with HEBP or physiological saline for 7 days. After the last injection, they were killed under anaesthesia and their maxillary incisors were examined using an in situ hybridization technique and immunohistochemical staining to detect the gene expression and localization of amelogenin protein, respectively. In the HEBP-injected rats, several islets of partially mineralized enamel were present along crown-analogous surface of the incisor in the secretory stage of amelogenesis and enamel-free zones existed between these islets. In situ hybridization demonstrated amelogenin gene expression over the ameloblasts facing the islets of the matrix enamel as well as over those of the enamel-free zones. Immunohistochemical studies using rabbit antiamelogenin antibody revealed positive reaction both in the enamel matrix of the control group and in the islets of enamel matrix of the HEBP-injected group. Some small granules immunoreactive to amelogenin antibody were found in the distal portions of the ameloblasts in the HEBP-injected rats. The results indicate that HEBP does not alter amelogenin gene expression over ameloblasts, or the protein's existence in enamel matrix. There appeared to be some accumulation of amelogenin in the HEBP-treated ameloblasts. It is therefore suggested that the enamel hypoplasia in this experiment may not be due to a disturbance in amelogenin synthesis but to a disturbance in a later process, presumably of protein secretion.

The immunohistochemical localization of Bax and Bcl-2 and their relation to apoptosis during amelogenesis in developing rat molars.

Bax and Bcl-2 are members of a family of intracellular, membrane-associated proteins that regulate programmed cell death. It has been suggested that, when Bax predominates, programmed cell death is accelerated and the apoptosis inhibitory activity of Bcl-2 is suppressed. The present study was undertaken to immunohistochemically (IHC) localize Bax and Bcl-2 in the cells of the enamel organ during amelogenesis in rat molars. Also, apoptotic cells were detected by TUNEL staining. The IHC intense localization of Bcl-2 and light staining for Bax in the pre-ameloblasts suggest that apoptosis is inhibited in the proliferating pre-ameloblasts. This is consistent with an absence of TUNEL staining for apoptosis in these cells. However, in the late secretory and transition ameloblasts, and adjacent stratum intermedium, evidence of apoptosis of the ameloblasts was observed. Bax and Bcl-2 were co-localized in the proximal ends of late secretory, transition and early maturation-stage ameloblasts, but immunoreactivity for Bax markedly increased in the proximal ends of late secretory and transition ameloblasts, while the Bcl-2 staining appeared to be lighter. This suggests that Bax antagonized Bcl-2 function, limiting the ability of Bcl-2 to prolong cell survival. In the early maturation stage, Bax staining faded while the immunoreactivity for Bcl-2 increased. Evidence of distinct apoptosis was reduced in the early maturation stage ameloblasts. When related to the occurrence of apoptosis during amelogenesis, the relative intensity of expression of Bax and Bcl-2 changed in a pattern consistent with that observed in other cell lines. This indicates that these proteins play essential roles in the process of amelogenesis, as predicted by their proposed mechanisms of action in the control of apoptosis.

Network structures and thermal properties of polyurethane films prepared from liquefied wood.

Polyurethane (PU) films were prepared by solution-casting after co-polymerization of liquefied woods (LWs) and polymeric methylene diphenylene diisocyanate (PMDI). The resulting PU films had various [NCO]/[OH] ratios ranging from 0.6 to 1.4 and contained 5.0-16.8% dissolved woody components at the [NCO]/[OH] ratio of 1.0. The crosslink densities of the films with [NCO]/[OH] ratios of 0.6-1.4 increased remarkably with increasing [NCO]/[OH] ratio. Similarly, there were large increases in glass transition temperatures (Tg). These characteristics could be attributed to effective incorporation of PMDI into the LW. The crosslink densities and Tg of the PU films prepared at the [NCO]/[OH] ratio of 1.0 increased because the amounts of dissolved woody components in the films increased. It is concluded that the dissolved woody components acted as crosslinking points in PU network formations. The thermal degradation of the PU films at an [NCO]/[OH] ratio of more than 0.8 or with more than 10.6% dissolved wood started above 262 degrees C under an N2 atmosphere. The thermostability was lost at low crosslink density or with large amount of co-polymerized glycerol structures in the PU networks.

[An adult case of purulent meningitis secondary to retropharyngeal and deep neck abscess after treatment of odontogenic infection].

Retropharyngeal and deep neck abscess, which may follow odontogenic infection, is uncommon in adults, but can be fatal. Furthermore, bacterial meningitis secondary to this disorder is extremely rare. A 67-year-old man was brought to our hospital because he had developed neck pain, trismus, and disturbance of consciousness over several days. A few days prior to the appearance of neck pain, he had the periodontitis treated. Based on CSF, cervical X-ray and CT findings, he was diagnosed as having bacterial meningitis secondary to deep neck abscess. Culture of the CSF yielded gram-positive cocci, later identified as Gemella species, that is a rare organism for bacterial meningitis. Although the administration of antibiotics and drainage of the abscess resulted in gradual improvement of the infectious process, neurologically he remained with apallic syndrome. We would like to stress the importance of odontogenic and pharyngolaryngogenic sources as potential foci of purulent meningitis.

[Limitation of myocardial infarct size by polyethylene glycol-conjugated superoxide dismutase in a canine model of 90 min coronary occlusion followed by 4 days of reperfusion].

The role of oxygen free radicals in the genesis of myocardial reperfusion injury is supported by the studies with superoxide dismutase (SOD) which provides protection against the extention of myocardial injury. However, the efficacy of SOD may be of limited value due to its short plasma half-life of 5-6 min. Conjugation of SOD to polyethylene glycol (PEG-SOD) increases the half-life to greater than 30 hours. Forty-two male, mongrel anesthetized dogs were subjected to occlusion of left circumflex coronary artery for 90 min followed by 4 days of reperfusion. Dogs were randomized to receive either PEG-SOD (1,000 U/kg) or PEG-Albumin via left atrium, starting 15 min before reperfusion and ending simultaneously with reperfusion, and were sacrificed 4 days later. The hearts were stained by ex vivo dual perfusion technique for the determination of myocardial infarct size. Infarct size expressed as a percent of area-at-risk in PEG-SOD (n = 13) and PEG-Albumin (n = 13) treated dogs differed significantly between groups: 29.2 +/- 1.6% vs 44.2 +/- 2.6%, respectively (p less than 0.01) with no observed difference in the size of area-at-risk: 46.0 +/- 1.6% vs 44.4 +/- 1.4% (n.s.). Hemodynamic parameters between groups did not differ during coronary artery occlusion and could not account for the effect of PEG-SOD on infarct size. Collateral blood flows to the inner 2/3 of the ischemic myocardium for both groups did not differ. Plasma SOD activity in PEG-SOD group initially exceeded 20 U/kg and sustained significantly for 4 days.(ABSTRACT TRUNCATED AT 250 WORDS)

Development of the needle insertion robot for percutaneous vertebroplasty.

Percutaneous Vertebroplasty (PVP) is an effective and less invasive medical treatment for vertebral osteoporotic compression fractures. However, this operative procedure is quite difficult because an arcus vertebra, which is narrow, is needled with accuracy, and an operator's hand is exposed to X-ray continuously. We have developed a needle insertion robot for Percutaneous Vertebroplasty. Its experimental evaluation on the basic performance of the system and needle insertion accuracy are presented. A needle insertion robot is developed for PVP. This robot can puncture with accuracy and an operator does not need to be exposed to X-ray. The mechanism of the robot is compact in size (350 mm x D 400 mm x H270 mm, weight: 15 kg) so that the robot system can be inserted in the space between C-arm and the patient on the operating table. The robot system is controlled by the surgical navigation system where the appropriate needle trajectory is planned based on pre-operative three-dimensional CT images. The needle holding part of the robot is X-ray lucent so that the needle insertion process can be monitored by fluoroscopy. The position of the needle during insertion process can be continuously monitored. In vitro evaluation of the system showed that average position and orientation errors were less than 1.0 mm and 1.0 degree respectively. Experimental results showed that the safety mechanism called mechanical fuse released the needle holding disk properly when excessive force was applied to the needle. These experimental results demonstrated that the developed system has the satisfactory basic performance as needle insertion robot for PVP.

Cadmium induces expression of metallothionein in rat dental pulp.

Metallothionein (MT) is a metalloprotein with high affinity for certain heavy metals. In the present study, the ability of cadmium (Cd) to induce the synthesis of MT in dental pulp was investigated. Rats were injected subcutaneously with CdCl2 (1.5 mg Cd/kg) daily for 7 days. Expression of the MT gene was demonstrated by reverse transcription polymerase chain-reaction techniques. The results indicate that Cd induced MT in dental pulp of rat incisor and this may be involved in the cellular defense mechanism against Cd toxicity.

Cytological and cytogenetic effects of the fluoroquinolone ofloxacin on human periodontal ligament fibroblasts.

The fluoroquinolone ofloxacin (OFLX) is one of the candidates of antibacterial agents to be topically used against periodontitis. To estimate the maximum concentration of OFLX which exerts little or no adverse effect on the periodontal ligament, cytological and cytogenetic effects of OFLX on human periodontal ligament fibroblasts (Pel cells) were examined. Treatment of Pel cells with < or =0.3 mM OFLX for 24 or 48 h had little inhibitory effect on cellular growth and survival. DNA, RNA and protein syntheses in Pel cells did not decrease in response to treatment with < or =0.3 mM OFLX. The constitutive level of alkaline phosphatase mRNA was retained in cells treated with < or = 0.03 mM OFLX for 24 or 48 h. The level of type I procollagen mRNA was not affected by treatment with < or = 0.003 mM OFLX for 24 or 48 h. Cytogenetic effects of OFLX were evaluated by the ability of OFLX to induce chromosome aberrations in Pel cells. Treatment with OFLX at 0.3-3.0 mM for 6, 24, or 48 h failed to induce chromosome aberrations in Pel cells. The failure of OFLX to induce chromosome aberrations was seen even in the presence of exogenous metabolic activation using a 5% rat liver post-mitochondrial supernatant mixture. These results indicate that treatment of Pel cells with < or =0.003 mM OFLX has few or no adverse effects on the cytological and cytogenetic endpoints examined, suggesting that there would be little adverse effect on growth and differentiation of the periodontal ligament, as well as little cytogenetic activity, if OFLX were to be topically administered to the gingival crevice at the minimal inhibitory concentration (MIC90) against periodontopathic bacteria (< or = 0.0027 mM). It is important to note, however, that extrapolation of these findings to in vivo conditions has yet to be undertaken.

Influence of the muscle tension on the masseteric silent period in children and adults.

Three subject groups who have shown different biting forces were examined in order to investigate whether the physiological phenomenon that the duration of the silent period (SPD) of the masseter muscle would be influenced by background activities of the muscles. They were classified into child (CN, n = 10) and adult groups; the adult group was then further subdivided according to the strength of maximum biting force into a normal group (AN, n = 10) and a low biting force group with some symptoms (AS, n = 5). SPD was observed in the masseter muscles by applying chin tap stimulations during both clenching teeth (10%, 50% and maximum) and biting on a transducer (50 N, 100 N and maximum). With increase of muscle activities, SPD was significantly decreased in the AN and CN groups, whereas no significant changes were found in the AS group. When comparing SPDs among the three groups at the same voluntary effort, the AN group exhibited the shortest SPD followed by the CN and AS groups. The results suggest that SPD was positively influenced by the strength of the background activities of the muscles and that immaturity of the masseters in terms of muscle activity could affect the SPD.

Co-ordination of tongue movements and peri-oral muscle activities during nutritive sucking.

A feeding bottle equipped with micro-video-camera and pressure sensor was devised to show the inside of the mouth and record sucking pressure. Activities of the temporal (TM), masseter muscle (MM), orbicular muscle of the mouth (OM) and suprahyoid muscles (SM) of 25 healthy infants were examined. Tongue and jaw movements, EMGs and sucking waves were scanned simultaneously. The tongue movements included elevation of the medial part of the tongue in a backward-moving peristaltic wave; significant correlations were found between jaw motion, tongue movement and sucking pressure. The TM, MM and OM were most active when the sucking pressure became positive and the jaw was closing, the SM showing highest activity in the negative-pressure phase. These findings show that each suckling cycle is biphasic, with sucking pressure, peri-oral muscle activities and jaw motion all closely correlated.

Development of perioral muscle activity during suckling in infants: a cross-sectional and follow-up study.

The activity of the perioral muscles during breastfeeding in infants was investigated using EMGs. Fifty-six infants aged from 1 to 5 months were classified into five groups according to month of age in the cross-sectional study. Follow-up was carried out on 18 infants whose mean age was 2.5 months at the initial examination, and 4.8 months at the second. During suckling, EMGs were recorded unilaterally from the temporalis (TM), the masseter (MM), the orbicularis oris (OM), and the suprahyoid (SM) muscle groups. The activity of the SM increased significantly with age, while there was no appreciable increase in the activity of the TM, MM, and OM in either the cross-sectional study or the follow-up. However, total muscle activity was shown to increase significantly in both parts of the study. These findings suggest that the active tongue- and jaw-lowering movement may play a primary role in increasing sucking strength during the suckle-feeding period in infants.

[Study on occlusal sounds in children. (1). Influences of the velocity of tooth tapping on the occlusal sound].

The purpose of this study was to evaluate the influence of the maximum velocity of the mouth closing phase on the occlusal sound. Six subjects examined were divided into two groups (children and young adults). Occlusal sounds generated by the tooth tapping were obtained with in microelectronic condenser microphone placed on infra-orbital and external auditory canal. Three parameters were used for the measuring points such as 1) maximum velocity of the closing phase, 2) maximum amplitude of the occlusal sound, 3) duration of the occlusal sound. The results obtained in the present study were as follows: 1) The waveform of occlusal sounds obtained from infra-orbital showed more clearly than that obtained from the external auditory canal. 2) With increasing velocity, both the values of maximum amplitude and duration showed a tendency to increase in both groups. 3) The prolonging of the duration was apparent in the children. The results obtained in the study suggest that the analysis of the occlusal sounds generated by tooth tapping can apply to aid in the functional diagnosis of occlusion in children.

[Study on occlusal sounds in children. 2. Fast Fourier Transform (FFT) analysis on the occlusal sounds].

The purpose of this study was to evaluate the Fast Fourie Transform (FFT) analysis of the occlusal sounds generated by tooth tapping which may apply in assisting in the functional diagnosis of occlusion in children. The fifteen subjects examined were divided into three groups: the first group with deciduous dentition, the second with mixed dentition and the third with permanent dentition. Occlusal sounds generated by tooth tapping at the rate of 76 times per minute were obtained with a micro-electronic condenser microphone placed on the infra-orbital region. For the time wave, the wave pattern of the occlusal sound was classified into two groups: 1) Impact sound, 2) Slide sound, and the ratio of appearance was examined. For the spectrum, three parameters were chosen for the measuring points such as 1) frequency range, 2) peak frequency, 3) strength of the peak frequency. The results obtained in the present study were summarized as follows: 1) Occlusal sounds in the deciduous dentition, the appearance of sliding sound was noticed in over 50% of the cases while on the permanent dentition, the sliding sound was dominant. 2) The power spectrum of the occlusal sound showed generally two or three peaks. 3) The range of frequency was recognized to be extended on the sliding sound when compared with the impact sound.

[Analysis of occlusal contacts of children by the T-Scan system. (1) The reproducibility of the sensor].

The present study was designed to discuss the reproducibility and the clinical usage of the T-Scan system in children. A plastic model of a normal permanent dentition was used for the study. The model was pressed on the sensor of the T-Scan system in maximum intercuspation under various assigned pressures (10, 20, 30, 40 and 50 kg) which were controlled by a press machine (Auto Graph AG-5000C). The pressings were made under following conditions, such as 1) the sensor handle was fixed, 2) the sensor handle was free and 3) the sensor handle was free but the maxilla arch form was marked on the sensor by the silicone impressional material. The total number of tooth contacts were counted and sagittal and the vertical distance from midline to the tooth contacts were measured. The results obtained in the present study are summarized as follows: 1) The number of tooth contacts became stable, when the pressing sensor was exercised several times. 2) The more pressure raised on the sensor, the more the number of tooth contacts increased, and the more the coefficient of variation tended to decline. 3) The arch model displayed on the T-Scan system was not exactly represented in the actual dental arch. 4) Due to the marking by silicone impressional material on the sensor, the reproducibility of the T-Scan system was further improved.

Reduction of dental plaque deposition in humans by oolong tea extract.

The inhibitory effect of oolong tea extract (OTE) containing polymerized polyphenols on plaque deposition was examined in 35 human volunteers. Thirty-five human volunteers, aged 18-29 years, who received extensive oral prophylactic procedures were requested to refrain from all oral hygiene procedures for 4 days, and to rinse their mouth with 0.5 mg/ml OTE solution in 0.2% ethanol before and after every intake of food and before sleeping at night. No restriction regarding meals was given during the test period, except to refrain from teas or coffee. Plaque deposition was evaluated after disclosing the teeth with Erythrocin at the termination of this experiment. The study was repeated 1 week after the first trial, but only 0.2% ethanol without OTE was used for mouthrinsing in the second trial. OTE was found to significantly inhibit plaque deposition in volunteers, although mouthrinsing with OTE solution had no significant effect on the number of mutans streptococci in unstimulated whole saliva.