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Objective: The objective of this study was to determine if phosphor plates used in predoctoral clinics are microbiologically contaminated and to identify the source of contamination., Study Design: Forty-five of 300 phosphor plates (15%) were randomly selected for examination. The plates were pressed into individual blood agar plates, were incubated using standard techniques at 37 degrees C, and were monitored for 72 hours. The number, size, distribution, and variety of resulting colonies were noted. A representative of each type of colony was selected to be Gram stained using the standard technique., Results: Of the plates, 42.2% were uncontaminated, 57.8% yielded bacterial colonies, and 15.6% of those colonies demonstrated hemolytic growth. The hemolytic growth included combined alpha and beta hemolysis and beta only hemolysis. Six colonies were gram-positive rods and 7 were gram-positive cocci., Conclusion: Meticulous infection-control techniques are inevitable and continuous reinforcement and training for staff and students are mandatory. Periodic gas sterilization of phosphor plates may be necessary.

Significance: Protection of glandular cells from autoimmune-induced damage would be of significant clinical benefit to Sjogren's syndrome (SS) patients. Epigallocatechin-3-gallate (EGCG) possesses anti-apoptotic, anti-inflammatory, and autoantigen-inhibitory properties., Aims: To investigate if EGCG protects against certain autoimmune-induced pathological changes in the salivary glands of the non-obese diabetic (NOD) mouse model for SS., Main Methods: Animals were provided with either water or water containing 0.2% EGCG. At the age of 8, 16 and 22 weeks, submandibular salivary gland tissue and serum samples were collected for pathological and serological analysis., Key Findings: Significant lymphocyte infiltration was observed in the salivary glands of the water-fed group at the age of 16 weeks, while the EGCG group showed reduced lymphocyte infiltration. By 22 weeks of age, water-fed animals demonstrated elevated levels of apoptotic activity within the lymphocytic infiltrates, and high levels of serum total anti-nuclear antibody, compared to EGCG-fed animals. Remarkably, proliferating cell nuclear antigen (PCNA) and Ki-67 levels in the salivary glands of water-fed NOD mice were significantly elevated in comparison to BALB/c control mice; in contrast, PCNA and Ki-67 levels in EGCG-fed NOD animals were similar to BALB/c mice. These results indicate that EGCG protects the NOD mouse submandibular glands from autoimmune-induced inflammation, and reduces serum autoantibody levels. Abnormal proliferation, rather than apoptosis, appears to be a characteristic of the NOD mouse gland that is normalized by EGCG. The evidence suggests that EGCG could be useful in delaying or managing SS-like autoimmune disorders.

Demineralized freeze-dried bone (DFDB) in matrix form must be rehydrated with a carrier medium which allows for easy manipulation during periodontal surgery. The purpose of this study was to evaluate how human DFDB suspended in a polyol matrix affects new bone formation in the rat calvarium critical-sized defect (CSD) model. Fifty-five adult male Harlan Sprague-Dawley rats were assigned to 1 of 5 treatment groups: polyol, 100% DFDB, 47% DFDB/polyol, 47% DFDB, or an unfilled control. They were then placed into 8-m calvarial CSDs. The bone donor source company for the DFDB and DFDB/polyol groups was the same. Calvaria were harvested 10 weeks after surgery and evaluated histomorphometrically. The diameter of bone particles from the 3 groups containing DFDB was measured by scanning electron microscopy. There was no statistically significant difference in the percentage of bone fill between any of the groups, although the 100% DFDB group exhibited the most bone fill. The 47% DFDB/polyol and 47% DFDB groups had similar amounts of bone formation. The average size of the demineralized bone particles from the 100% DFDB group was significantly smaller than that of the other 2 groups containing DFDB. Adding a polyol to DFDB produced similar osseous regeneration in the rat calvarium defect model vs DFDB alone. Yet from a clinical standpoint, the polyol enhanced graft handling and stability. Graft particle size may have an effect on bone fill.

The green tea polyphenol epigallocatechin-3-gallate (EGCG) regulates gene expression differentially in tumor and normal cells. In normal human primary epidermal keratinocytes (NHEK), one of the key mediators of EGCG action is p57/KIP2, a cyclin-dependent kinase (CDK) inhibitor. EGCG potently induces p57 in NHEK, but not in epithelial cancer cells. In humans, reduced expression of p57 often is associated with advanced tumors, and tumor cells with inactivated p57 undergo apoptosis when exposed to EGCG. The mechanism of p57 induction by EGCG is not well understood. Here, we show that in NHEK, EGCG-induces p57 via the p38 mitogen-activated protein kinase (MAPK) signaling pathway. In p57-negative tumor cells, JNK signaling mediates EGCG-induced apoptosis, and exogenous expression of p57 suppresses EGCG-induced apoptosis via inhibition of c-Jun N-terminal kinase (JNK). We also found that restoration of p57 expression in tumor cells significantly reduced tumorigenicity in athymic mice. These results suggest that p57 expression may be an useful indicator for the clinical course of cancers, and could be potentially useful as a target for cancer therapies.

Aim: To compare the sealing of root canals filled with two single-cone obturation systems and a warm vertical compaction technique., Methodology: Forty-two single-rooted teeth were decoronated to obtain 17-mm-long root segments. The root canals were cleaned and shaped to size 40, 0.06 taper and filled with: (i) warm vertical compaction with AH Plus (control); (ii) ActiV GP and (iii) GuttaFlow with single master cones. Leakage was evaluated by fluid filtration at 10 psi before root resection, and after 3, 6, 9 and 12 mm apical resections. Repeated measures anovas on ranks and Dunn's multiple comparison tests were performed to examine differences in fluid flow rates amongst different resection lengths for each filling technique. The surface and interior aspects of glass-ionomer filler-coated ActiV GP gutta-percha cones was evaluated with SEM., Results: No statistical difference amongst the filling techniques was seen at 0 and 3 mm root resections. ActiV GP and GuttaFlow exhibited more leakage than AH Plus at 6, 9 and 12 mm resections. AH Plus recorded the best overall results. A nonhomogeneous coating of glass-ionomer fillers on the surface of ActiV GP cones was detected., Conclusions: The two single-cone techniques examined are as effective in sealing the apex as AH Plus when the latter was used with warm vertical compaction. It is further hypothesized that the inferior coronal seal of these single-cone techniques may be improved with the placement of accessory cones to reduce sealer thickness or an immediate coronal adhesive restoration.

Sjogren's syndrome (SS) is a relatively common autoimmune disorder. A key feature of SS is lymphocytic infiltration of the salivary and lacrimal glands, associated with the destruction of secretory functions of these glands. Current treatment of SS targets the symptoms but is unable to reduce or prevent the damage to the glands. We reported previously that the major green tea polyphenol (GTP) epigallocatechin-3-gallate (EGCG) inhibits autoantigen expression in normal human keratinocytes and immortalized normal human salivary acinar cells (Hsu et al. 2005). However, it is not known whether GTPs have this effect in vivo, if they can reduce lymphocytic infiltration, or protect salivary acinar cells from tumor necrosis factor-alpha (TNF-alpha)-induced cytotoxicity. Here, we demonstrate that in the NOD mouse, a model for human SS, oral administration of green tea extract reduced the serum total autoantibody levels and the autoimmune-induced lymphocytic infiltration of the submandibular glands. Further, we show that EGCG protected normal human salivary acinar cells from TNF-alpha-induced cytotoxicity. This protection was associated with specific phosphorylation of p38 MAPK, and inhibitors of the p38 MAPK pathway blocked the protective effect. In conclusion, GTPs may provide a degree of protection against autoimmune-induced tissue damage in SS, mediated in part through activation of MAPK elements.

This in vitro study evaluated the sealing efficacy of three root-filling systems/techniques in preventing bacterial leakage. Instrumented single-rooted root segments were filled with (1) warm vertical compaction with gutta-percha/AH Plus; (2) single-cone technique with ActiV GP; and (3) single-cone technique with Gutta-Flow. A dual-chamber leakage model using S. mutans as a microbial marker was used for leakage evaluation. Bacterial penetration was monitored over a 100-day period. Leakage was recorded when turbidity was observed in the lower chamber. Gutta-percha warm vertical compaction exhibited the best seal with bacterial leakage observed in only 16.7% of the specimens between 59 and 100 days. All ActiV GP specimens leaked between 7 and 100 days; 50% of the Gutta-Flow specimens leaked between 22 and 100 days. The two contemporary single-cone techniques did not insure a durable apical seal against bacterial leakage. A warm vertical compaction technique using thermoplasticized gutta-percha and AH Plus sealer appears to be more effective in minimizing bacterial leakage.

Potential intrinsic tetracycline staining of intraradicular dentin has been observed when BioPure MTAD was employed as the final irrigant after initial rinsing with NaOCl. This study examined the effect of NaOCl-MTAD interaction on the antimicrobial substantivity of MTAD in dentin. Dentin cores previously irrigated with either MTAD, or in conjunction with 1.3% NaOCl as an initial irrigant were placed on blood agar plates inoculated with Escherichia faecalis at 10(5) cfu/ml. Dentin cores irrigated with 1.3% NaOCl only, and autoclaved dentin disks were used as the respective positive and negative controls. After anaerobic incubation, the mean diameter of bacterial inhibition zones formed around the MTAD group was significantly larger than the NaOCl/MTAD group, which, in turn, was not significantly different from the NaOCl positive control. Oxidation of MTAD by NaOCl resulted in the partial loss of antimicrobial substantivity in a manner similar to the peroxidation of tetracycline by reactive oxygen species.

Unlabelled: This study was a single-blind, randomized, controlled clinical trial. The researchers evaluated a powered brush/irrigating device (HydraBrush Oral Health System; OHS) for its safety and ability to deliver a solution to the bottom of 5-6 mm pockets, compared to rinsing alone with a solution following brushing with a powered toothbrush (Sonicare Elite 7800; SE). An evaluation technique to measure the quantity and quality of solution able to enter the pocket was also introduced in this project., Methods: Subjects were randomized in one of two-groups: brush plus simultaneous irrigation (OHS) versus brush plus rinsing (SE). Subjects used their devices at home for two weeks. At the measurement visit, subjects used the OHS to irrigate and brush simultaneously for 1 minute (30 seconds per each side of the mouth) with a 0.01% erythrosine disclosing solution in 10 oz of distilled water. Control subjects brushed for 2 minutes with a SE followed by a 1 minute rinse with an identical disclosing solution. A blinded evaluator collected six samples of approximately of 1 microL of sucular fluid from six 5-6 mm evaluation sites. This was accomplished by inserting a microcapillary tip with a 20 microL micropipette in the sulcus. Two-group repeated measures ANOVA was used to examine differences in two measures of the disclosing solution between OHS and SE subjects; the spectrometer reading of the disclosing solutions, and by visual inspection of the samples (positive/negative) to determine the presence or absence of solution in the samples. Subjects' diaries were collected. Bleeding and discomfort during the evaluation period was reported., Results: Visually, OHS had a significantly greater proportion of solution taken from the base of 5-6 mm sites than the SE (p=0.0001). However, there was no statistical difference between the two groups (p=.1359) in the spectrophotometer readings., Conclusion: The experimental device is more efficient in delivering a solution to the base of 5-6 mm pockets than rinsing following use of a control powered toothbrush. Both devices have demonstrated they are safe and well accepted by patients. The technique developed provides a useful method for quantitative and qualitative studies of solutions from the base of periodontal pockets.

The biological effects of camphorquinone (CQ), an initiator for light-polymerized resins, have been reported to relate to its ability to generate free radicals and cause radical-induced membrane damage via lipid peroxidation. However, the effects of CQ on lipids other than peroxidation may result in unfavorable tissue responses especially at concentrations that are not overtly toxic to cells. The purpose of the current study was to examine the effects of CQ on cell lipid metabolism at subtoxic concentrations, with or without visible light irradiation. HCP and THP-1 cells were exposed to CQ with or without light irradiation under clinically relevant conditions and lipid metabolism was analyzed using 14C-labeling and thin-layer chromatography. We found that CQ increased synthesis of neutral lipids, such as triglycerides, from 7 to nearly 15% of the total and diglycerides from 2% to about 3% of the total in HCP cells, while synthesis of phospholipids, such as sphingomyelin, was decreased by 1-1.5%. In THP-1 cells cholesterol synthesis increased more than 2-fold and cholesterol ester synthesis increased more than 5-fold. Light-activated CQ did not differ significantly in terms of its bioactivity compared to no-light conditions. We conclude that CQ significantly altered the metabolism of several important structural lipids in two cell types at sub-toxic concentrations that are clinically relevant. These changes in lipid metabolism may in turn affect membrane integrity and permeability and possibly lead to significant changes in cell responses.

Benzoyl peroxide (BP), a tumor promoter, has been shown to cause free-radical-induced lipid peroxidation and membrane damage at toxic concentrations. However, its effects on lipid metabolism at concentrations that were not overtly toxic have not been investigated. The purpose of the current study was to examine the effects of BP and its final degradation product, benzoic acid (BA), on lipid metabolism. Two cell lines, hamster cheek pouch (HCP) and human monocytes (THP-1), were used to determine the effects of BP, BA, and BP combined with FeCl2 on cell lipid metabolism. Cells were exposed to BP and 14C acetate for 24 h, or cells with prelabeled lipids were harvested, and the lipids were extracted and separated with the use of thin-layer chromatography. Lipid metabolism of some neutral lipids such as triglycerides was altered for both cell types in response to BP. Also, cholesterol content was reduced in THP-1 cells and a phospholipid, phosphatidylethanolamine (PE), was reduced in HCP cells. The final degradation product of BP, BA, failed to elicit any response in lipid metabolism. Subtoxic concentrations of BP induced changes in neutral lipids such as triglycerides and cholesterol. The metabolism of major phospholipids except PE remained unchanged. The effects were related to BP and its degradation and varied with the cell type.

Metallic medical devices undergo degradation in vivo and the degradation products affect the chemistry and biological responses of cells and tissues in the immediate vicinity. The responses vary with the metal and cell type. In the current study, we examined the effects of several metals on a human monocytic cell line. Monocytes are important effector cells capable of responding rapidly to inflammatory and immune stimuli in a variety of ways, including production of inflammatory proteins, differential expression of surface adhesion molecules, enhanced phagocytic activity, and activation and differentiation to macrophages. Cells were exposed in the presence of (14)C-acetate to titanium, nickel, chromium, copper, or cobalt or vanadium at concentrations that were subinhibitory or inhibitory based on cellular mitochondrial dehydrogenase activity. Cell lipids were then extracted, separated by thin layer chromatography, and quantitated by liquid scintillation spectrometry. Total cell protein also was measured. Titanium reduced cell protein content at concentrations that were noninhibitory to mitochondrial dehydrogenase activity, whereas neither chromium nor cobalt affected protein amounts at dehydrogenase-inhibitory concentrations. In cells exposed to vanadium, the protein- and dehydrogenase-inhibitory concentrations were similar. The major effects on cell lipids appeared to occur in the neutral lipids, although chromium, cobalt, and titanium produced changes in some major phospholipids. These results suggest that metals differentially affect various metabolic pathways in THP-1 cells, perhaps related to their abilities to enter the cells or interact with the membrane. These alterations to the cells may affect the cells' abilities to respond to various stimuli that can damage the tissues., (Copyright 2004 Wiley Periodicals, Inc. J Biomed Mater Res 70A: 347-353, 2004)

The increasing use of acrylate-based resins in dentistry has raised questions about the biocompatibility of these substances with oral tissues. The focus of the present investigation was to assess the responsiveness of blood vessels to the resin polymerization accelerating agent dimethylaminoethyl methacrylate (DMAEMA) and its degradation products dimethylethanolamine (DME) and methacrylic acid (MAA), using the rat aortic ring preparation as a tissue model. DMAEMA induced concentration-dependent relaxation of norepinephrine (NE)-contracted aortic rings with and without endothelium. N-nitro-L-arginine methyl ester (L-NAME) selectively inhibited the endothelium-dependent relaxation induced by DMAEMA, suggesting the release of nitric oxide from the endothelium by DMAEMA. Both indomethacin and glybenclamide attenuated the vasorelaxation elicited by DMAEMA in the presence as well as in the absence of endothelium, providing evidence for the role of vasorelaxant prostanoid(s) and K(ATP) channel activation in the responses observed. On the other hand, while MAA was without any apparent effect on the rat aorta, DMAEMA at high and DME at relatively low concentrations caused contraction of the tissues with and without endothelium in the absence of NE. The DME-induced contraction was inhibited by indomethacin, suggesting the involvement of contractile arachidonic acid metabolite(s) in the action of DME. This observation was supported by the findings of increased thromboxane A(2) (TXA(2)) production in aortic rings incubated with DME. Taken together, the data suggest that both DMAEMA and its degradation product, DME, are vasoactive, inducing vasorelaxation and contraction by various mechanisms that may involve the release of nitric oxide from the endothelium, the activation of smooth muscle K(ATP) channels, and the generation of vasorelaxant prostanoid(s) and TXA(2). These effects may play a role in tissue homeostasis and certain adverse conditions associated with the use of dental resin materials containing DMAEMA and/or DME., (Copyright 2003 Wiley Periodicals, Inc.)

The frequent use of resins in dentistry has raised the question of their compatibility with oral tissues. The present study was undertaken to determine the effects of the resin components methyl methacrylate (MMA), hydroxyethyl methacrylate (HEMA), and triethylene glycol dimethacrylate (TEGDMA) on the reactivity of blood vessels using the isolated rat aorta as a tissue model. MMA, HEMA, and TEGDMA caused endothelium-dependent and -independent relaxation of rat aortic rings in a concentration-related manner. The endothelium-dependent responses of the tissues to all the resins were significantly attenuated by N-nitro-L-arginine methyl ester (L-NAME), indicating the involvement of nitric oxide. The vasorelaxant effects of both MMA and TEGDMA on the intact and denuded aortae were markedly inhibited by indomethacin, providing evidence for the role of prostanoids in these responses. Glybenclamide selectively attenuated TEGDMA-induced relaxation of the tissues with and without endothelium to a similar extent, suggesting the activation of vascular smooth muscle K(ATP) channels by this resin. It is concluded that MMA, HEMA, and TEGDMA interfere with the function of blood vessels by inducing vasorelaxation via different mechanisms, which, depending upon the type of resin, may at least involve the release of nitric oxide and prostanoid(s), and the activation of smooth muscle K(ATP) channels. These phenomena may play a role in tissue homeostasis and certain pathophysiological conditions associated with the application of resin materials to the oral environment., (Copyright 2002 Wiley Periodicals, Inc.)

A human oral tumour progression model was established that consists of normal epithelial cells and three cell lines representing stages from dysplastic to metastatic cells. To investigate the impact of exogenous transforming growth factor-beta 1 on this model system, we analysed the responsiveness of those cells to transforming growth factor-beta 1 and explored the potential mechanism underlying the transforming growth factor-beta 1 activity. We found that the growth of all cell types, regardless of their stage of tumour progression, is inhibited by transforming growth factor-beta 1, although to different degrees. Transforming growth factor-beta 1 induced the expression of cyclin-dependent kinase inhibitors p15(INK4B), p21WAF1/(CIP1) and p27(KIP1). In contrast, transforming growth factor-beta 1 was found to stimulate the invasive potential of one cell type that represents the most advanced stage of tumour phenotype, suggesting that the impact of transforming growth factor-beta 1 on functional features of tumour cells other than cellular proliferation may play a significant role in the process of oral tumour progression.

Some components of resins used in restorative dentistry have been shown to alter metabolism in cultured oral epithelial cells. Here we have extended such studies to the underlying supportive tissue, composed of gingival fibroblasts (GF). Primary cultures of human GF were transferred to serum-free, defined medium and exposed to either 2-(dimethylamino)ethyl methacrylate (DMAEMA) or 4-methoxyphenol (MEHQ) for 24-72 h. At a DMAEMA concentration of 6.4 mM, which was well tolerated by epithelial cells, GF numbers, as estimated by crystal violet, and metabolic activity, as indicated by MTT, were reduced at least 60% within 24 h of exposure. Between 1.6 and 6.4 mM, there were dose-related reductions in cell numbers; however, at lower doses (0.32-0.64 mM), proliferation was stimulated. MEHQ, between 8 and 16 microM, did not stimulate cellular protein production. To examine the capacity of GF to respond to an inflammatory stimulus, interleukin-6 (IL-6) production by confluent cells was estimated without or with these compounds. DMAEMA (1.6- 6.4 mM) virtually eliminated the acute IL-6 response of these cells to an interleukin-1beta challenge; only at 0.32 mM DMAEMA was the response restored. MEHQ (1.6-16 microM) reduced the IL-6 response by >50%. In summary, both growth and the innate immune responsivity of GF were affected by DMAEMA and MEHQ in vitro; thus, these compounds deserve careful evaluation for biocompatibility., (Copyright 2002 John Wiley & Sons, Inc. J Biomed Mater Res 60: 30–35, 2002)

Green tea has been a popular beverage for many centuries. Only recently, however, has the anti-cancer power of green tea constituents been unveiled. Green tea polyphenols are found to induce apoptosis (programmed cell death) in many types of tumor cells, including oral cancer cells. However, mechanisms that enable normal cells to evade the apoptotic effect still are not understood. In this study, cell growth and invasion assays combined with apoptosis assays were used to examine the effects of green tea extracts, green tea polyphenols, and the most potent green tea polyphenol, (-)-epigallocatechin-3-gallate (EGCG), on normal human keratinocytes and oral carcinoma cells. The results showed that green tea and its constituents selectively induce apoptosis only in oral carcinoma cells, while EGCG was able to inhibit the growth and invasion of oral carcinoma cells. These differential responses to green tea and its constituents between normal and malignant cells were correlated with the induction of p57, a cell cycle regulator. These data suggest that the chemopreventive effects of green tea polyphenols may involve a p57 mediated survival pathway in normal epithelial cells, while oral carcinoma cells undergo an apoptotic pathway. Therefore, regular consumption of green tea could be beneficial in the prevention of oral cancer.

Green tea polyphenols are known to induce apoptosis in certain types of tumor cells. However, the mechanism(s) that enables normal cells to evade the apoptotic effect is still not understood. In this study, Western blot analysis combined with cycloheximide treatment was used to examine the effects of green tea polyphenols on the expression levels of p57, a cyclin-dependent kinase and apoptosis inhibitor, in normal human keratinocytes and in the oral carcinoma cell lines SCC25 and OSC2. The results showed that the most potent green tea polyphenol, (-)-epigallocatechin-3-gallate (EGCG), induced p57 in normal keratinocytes in a dosage- and time-dependent manner, while the levels of p57 protein in oral carcinoma cells were unaltered. The differential response in p57 induction was consistent with the apoptosis status detected by annexin V assay. The data suggest that the chemopreventive effects of green tea polyphenols may involve p57-mediated cell cycle regulation in normal epithelial cells.

Statement of Problem: Contamination of removable prostheses with microorganisms, particularly Candida albicans, is a common clinical problem. Microban, a broad-spectrum antimicrobial containing triclosan, recently has been proposed to inhibit microbial growth., Purpose: This study aimed to determine whether the addition of Microban to PermaSoft denture liner prevents the growth of C albicans and affects the cytotoxicity of the PermaSoft material., Material and Methods: Experimental specimen disks (5 x 1 mm each) with and without incorporated Microban were fabricated aseptically (n = 6) against polyester film to produce a smooth surface. To assess the cytotoxic effect of Microban, the MTT assay was used. To determine the effect of Microban on the growth of C albicans, disks were placed in Transwell dishes, covered with Sabouraud's broth containing an ATCC strain of C albicans, and incubated at 37 degrees C for 24 hours. Wells containing fluorocarbon resin disks or broth alone served as controls. The disks were rinsed to remove unattached C albicans and then sonicated in sterile water to remove surface organisms. Serial dilutions of the water extracts were plated on Sabouraud's agar and returned to the incubator for 24 hours. Colonies were counted with a Brunswick Colony Counter. Growth of C albicans in the internal aspects of the specimens was determined in a manner as previously described, with the exception that the specimens were sonicated to remove surface organisms, minced, and sonicated once more before making serial dilutions. The results were compared with ANOVA and Tukey intervals (alpha=.05)., Results: The number of colonies formed ranged from 17 to 31 x 10(5) (mean = 23 +/- 4 x 10(5)) and 14 to 69 x 10(5) (mean = 32 +/- 20 x 10(5)) for the PermaSoft with and without Microban groups, respectively. There was no statistically significant difference between PermaSoft with and without Microban., Conclusion: The addition of Microban did not significantly alter the cytotoxicity of the PermaSoft denture lining material or reduce the adherence of viable C albicans to the surface of PermaSoft material after 24 hours.

Magnetic Resonance Imaging (MRI) is the method of choice to search for epileptogenic lesions. We correlated MRI findings with the epileptogenic zone (EZ) depicted by clinical and electroencephalographic (EEG) data. We studied 400 clinical records of patients who had been submitted to MRI studies and we analyzed, retrospectively, their ictal semiology, EEG characteristics and response to treatment. They were classified into 3 groups: A) temporal lobe epilepsy, B) frontal lobe epilepsy and C) parieto-occipital epilepsy. We included 155 patients: Group A) 68 cases (43.9%), 28 men (41.1%), mean age 32 +/- 11 years old, abnormal IMR in 44 (64.7%), refractory to treatment 48 (70.5%). Group B) 68 cases (43.9%), 38 men (55.8%), mean age 30 +/- 15 years old, abnormal IMR in 26 (38.2%), refractory to treatment 30 (44.1%). Group C) 19 cases (12.2%), 13 men (68.4%), mean age 27 +/- 11 years old, abnormal IMR in 11 (57.8%), refractory to treatment 12 (63.1%). Results showed that there were higher possibilities of detecting lesions which correlate with EZ in temporal than in frontal or parieto-occipital lobes epilepsy. The chances to find abnormalities on the MRI were 5 times higher in refractory patients than in those who were non-refractory.

Objectives: Diacylglycerol-kinase (DAG-kinase) is an enzyme that phosphorylates diacylglycerol (DAG) to phosphatidic acid (PA), which serves as a precursor to phosphoglycerides involved in cell signaling or as cell membrane structural components. DAG-kinase can be inhibited by diacylethylene glycols (DAEG). We hypothesize that 2-hydroxyethyl methacrylate (HEMA) may alter phosphorylation of DAG to PA following intracellular formation of DAEG., Methods: Cultured rabbit kidney (RK13) epithelial cells were treated with HEMA, EG, or known inhibitors of DAG-kinase for 24 h, then exposed to [32P]O4- in the presence of a synthetic diacylglycerol for 2 h. Other cultures were radiolabeled with [3H]-oleic acid for 24 h, then exposed to HEMA for an additional 24 h. The cells were harvested and the lipids extracted. Radioactive lipids were separated by thin layer chromatography, located by autoradiography, and quantitated as cpm/ug protein. Cell cultures treated with HEMA were homogenized and the DAG-kinase activity was assayed and expressed as cpm/ug protein. Data were analyzed by one-way ANOVA and Newman-Keuls Multiple Comparison Test., Results: Cultures exposed to HEMA or known DAG-kinase inhibitors exhibited reduced incorporation of radioactivity in the PA fraction compared to control cultures. Direct assays of DAG-kinase activity from cells exposed to HEMA demonstrated decreased enzyme activity. Evaluation of cell phospholipid synthesis showed altered formation of phosphatidylethanolamine and phosphatidylcholine., Significance: Results suggest that HEMA impairs formation of PA, possibly by acylation of EG released by hydrolysis of the HEMA and resultant production of the inhibitor DAEG. The decreased availability of PA may alter PA-dependent cell structural lipid pathways and lipid-dependent signaling pathways, altering cell growth.

Increasing concerns over the effects of environmental estrogens on wildlife and humans have highlighted the need for screening systems to assess potentially estrogenic effects of test compounds. As a result, in vitro screening methods such as cell proliferation assays using the estrogen-responsive human breast cancer cell line, MCF-7, have been developed. The present study describes an alternative in vitro approach for the assessment of such xenoestrogens, based on estrogenic rescue of MCF-7 cells from antiestrogen-induced cytotoxicity. This method measures the ability of various estrogenic compounds to compete with a known estrogen-receptor-mediated antihormonal drug, 4-hydroxytamoxifen, using the 1-[4,5-dimethylthiazol-2-yl]-3,5-diphenylformazan (MTT) assay to assess mitochondrial activity. Because 4-hydroxytamoxifen treatment of cells results in a dramatic decrease in mitochondrial dehydrogenase activity which is directly related to their estrogen-receptor content, inhibition of this effect with estrogenic compounds represents an estrogen-receptor interaction, or estrogenic rescue. The estrogenic compounds tested include a weak xenoestrogen, bisphenol A (BPA), and two biological estrogens, 17alpha- and 17beta-estradiol. Competitive inhibition of 4-hydroxytamoxifen-induced cytotoxicity by BPA was compared to that of the biological estrogens. The results indicate that the biological estrogens can successfully compete with the antiestrogen in a dose-dependent manner. In addition, the assay is sensitive enough to detect estrogenic rescue by even the very weak xenoestrogen, BPA, albeit at high BPA concentrations. This simple in vitro method could be used as an alternative or second-line screen for potential xenoestrogens.

Oral microbial flora consist of numerous bacterial taxa, ranging from aerobes through fastidious anaerobes, and fungi, viruses, and protozoa. Many of these bacteria are unique to the oral cavity. The organisms exist in a complex interrelationship that is regulated and maintained by physical and metabolic microbial interactions, and by environmental factors, such as saliva and diet. Many of these organisms are relatively harmless, although others are significant pathogens, producing local and systemic diseases in healthy and compromised individuals.

Dimethylaminoethylmethacrylate (DMAEMA), a commonly-used component of visible-light polymerized dental resins, has the potential to elute and interact with tissue cells to cause cytotoxicity or sublethal metabolic changes. Short-term exposure of cultured oral epithelial cells to sublethal DMAEMA concentrations has been shown previously to affect cell neutral lipid and phospholipid metabolism, resulting in accumulation of significant quantities of dimethylphosphatidylethanolamine (DMPE). In non-treated cells, DMPE is a transient intermediate in phospholipid metabolism and is not detectable by standard methods. In the current study, the effects of prolonged exposure of cells to DMAEMA, and the mechanisms for formation of DMPE in the presence of DMAEMA were examined. Exposure of a keratinizing hamster buccal cheek pouch cell line (HCP cells) to 0.8 mM DMAEMA for 2, 3, 7, and 14 days resulted in reduced incorporation of [14C]acetate into several classes of phospholipids. DMPE was detectable at all time points in DMAEMA-exposed cultures and comprised between 12.48% and 18.33% of the total radiolabeled phospholipids. The results of short-term exchange experiments indicated that headgroup exchange was not the major reaction responsible for formation of DMPE in DMAEMA-treated cells; rather the formation appeared to occur through typical phospholipid metabolic pathways. The cells appeared able to re-establish and maintain homeostasis in the presence of this altered cell lipid composition.

The oral cavity is a common site for manifestations of systemic microbial diseases. Oral lesions may be typical of those seen elsewhere on the body, or the lesions may be modified by the local environment. The ease of examination within the oral cavity, however, and any site-specific features facilitates diagnosis of the systemic condition.

Recently, resin-based dental restorative materials have been targeted as potential sources of xenoestrogens, specifically bisphenol A (BPA) and bisphenol A dimethacrylate (BAD), which could contribute to overall estrogen load and result in deleterious side effects. The present study used high-pressure liquid chromatography (HPLC) to analyze twenty-eight different commercially available dental resins for the presence of BPA and/or BAD. In addition, sublines of the MCF-7 human breast tumor cell line were cultured in the presence of eluates from eleven of the dental resins and assessed for proliferative responses using the sulforhodamine B assay. Only one resin, Delton II, had detectable levels of BPA or BAD that could be verified by Fourier transform infrared spectrometry. Likewise, eluates from Delton II were the only samples that elicited a significant proliferative response in two of the MCF-7 sublines tested. Therefore, we conclude that dental resins in general do not represent a significant source of BPA or BAD exposure.

Although pit and fissure sealants have been utilized extensively in dentistry as a way of preventing occlusal caries, results described by Olea et al. (1996) raised concerns about the safety of sealants and other resin-based dental materials due to the reported presence of bisphenol A (BPA) and its dimethacrylate ester (BPA-DM). Although the release of these compounds from dental materials has not been substantiated by two subsequent studies, we believed it was important to confirm or refute the report that BPA and BPA-DM have estrogenic activity in vitro. We grew breast cancer cells (MCF-7, T-47D, ZR-75-1) known to proliferate under estrogenic stimulation in phenol red-free DMEM containing human serum and concentrations of BPA or BPA-DM ranging from 10(-8)M to 5 x 10(-6)M. After 1 week, plates were harvested for crystal violet or sulforhodamine-B assays, and the optical densities of groups of treated cells were compared with values from control cells. At concentrations at or above 10(-6)M, both BPA and BPA-DM significantly increased cell proliferation (p < 0.05), comparable to the increase seen with 10(-9)M of estrogen. Flow cytometric methods demonstrated that these mitogenic effects occurred within 24 h of exposure to estrogen, BPA, or BPA-DM. The increase in DNA synthesis was analogous to that seen with estrogen stimulation. Thus, we confirmed that BPA and BPA-DM cause cell proliferation at micromolar concentrations that exceed the effective concentrations of estrogen by 1 to 10,000-fold., (Copyright 1999 John Wiley & Sons, Inc.)

Endotoxin, cell envelope lipopolysaccharide produced by gram-negative bacteria can activate an immune response through a variety of pathways. In addition, it can stimulate bone resorption and reduce the periodontal tissue's healing capacity. Previous studies have documented the affinity of lipopolysaccharide for restorative materials. This study evaluated the affinity of lipopolysaccharide for commercially available orthodontic brackets. Stainless steel, ceramic, plastic, and "gold" brackets were exposed to 10 EU/mm2radiolabeled Porphyromonas gingivalis or Escherichia coli lipolpoysaccharide in water and incubated for 24 hours at 37 degrees C. Brackets were then transferred to fresh lipopolysaccharide-free water and incubated for 24 hours at 37 degrees C to evaluate elution. This elution transfer was continued up to 96 hours total incubation. Lipopolysaccharide adherence and elution levels were calculated after treatment, and elution solutions were evaluated through liquid scintillation spectrometry. Mean initial lipopolysaccharide adherence ranged from 2.42 +/- 0.26 EU/mm2(E. coli, plastic) to 6.75 +/- 0.34 EU/mm2 (P. gingivalis, stainless steel). P. gingivalis lipopolysaccharide adherence was significantly greater than E. coli lipopolysaccharide adherence for all bracket types. Moreover, for each lipopolysaccharide type, stainless steel brackets exhibited significantly greater lipopolysaccharide adherence. Regarding elution, only the P. gingivalis lipopolysaccharide-exposed ceramic and plastic brackets at 24 hours and the stainless steel and ceramic brackets at 48 hours eluted measurable lipopolysaccharide. Results from this study demonstrate that P. gingivalis and E. coli LPS exhibit a high affinity for orthodontic brackets. In vivo, this affinity could affect the concentration of LPS in the gingival sulcus, thereby contributing to inflammation in tissues adjacent to the brackets.

Components of dental resins such as dimethylaminoethyl methacrylate (DMAEMA) can alter cell lipid composition, presumably by esterase-mediated hydrolysis. The resulting dimethylethanolamine is incorporated into cell phospholipids, while the methacrylic acid may alter several metabolic pathways. We hypothesize that HEMA is cleaved in a similar manner and the released ethylene glycol is incorporated into cell lipids, yielding phosphatidylethylene glycol (PtEG), and the methacrylic acid alters other lipid pathways in a manner similar to that of methacrylic acid released from hydrolysis of DMAEMA. Cultures of hamster buccal pouch (HCP) and rabbit kidney (RK13) epithelial cells were exposed to subtoxic concentrations of HEMA in the presence of [14C]-acetate or [3H]-oleic acid. Other cultures were prelabeled with [14C]-acetate followed by exposure to various concentrations of HEMA. Cell lipids were extracted by the method of Bligh and Dyer and separated by thin layer chromatography on silica gel K-6 plates or SG-81 silica gel loaded chromatography paper. The fate of the ethylene glycol was traced using [14C]-ethylene glycol. Radioactive lipids were located using autoradiography and known standard lipids and quantitated by liquid scintillation spectrometry. In the presence of HEMA several classes of lipids were altered. Among the neutral lipids, the most notable changes involved sterol precursors, triglycerides, fatty acids, and cholesterol esters, while phosphatidylcholine was affected among the phospholipids. The results differed quantitatively between the two cell types. Results also suggest that EG, including that released by hydrolysis of HEMA, is incorporated into cell phospholipids, producing PtEG. The changes in neutral lipid labeling may occur by alteration of lipid synthetic pathways utilizing acetyl Co-A as well as inhibition of enzymes involved in synthesis of cholesterol from sterol precursors and hydrolysis of cholesterol esters. Synthesis of PtEG may take place via phospholipase D-mediated headgroup exchange. Alterations in the cellular lipids may affect cell membrane properties and associated cell functions.

Insulin-dependent diabetes mellitus (IDDM) is a risk factor for periodontitis. Depressed neutrophil chemotaxis has been demonstrated in IDDM and in early-onset periodontitis (EOP). HLA-DR antigens are associated with both IDDM and periodontitis. This investigation sought to determine an association of HLA-DR3, -DR4, and -DR53 with impaired neutrophil chemotaxis in an IDDM sample. The neutrophil chemotaxis index of 41 diabetics and 27 controls was determined by a modified Boyden chamber method, and certain class II HLA genotypes were determined by polymerase chain-reaction amplification of genomic DNA by means of sequence-specific primers (PCR-SSP). The mean chemotaxis index of the diabetics was significantly less than that of the controls (p < or = 0.02). HLA-DR3 (p < or = 0.002), -DR4 (p < 0.003), and -DR53 (p < or = 0.001) were associated with IDDM. Neutrophil chemotaxis and glucose metabolism were not significantly correlated. None of the HLA-DR alleles was associated with impaired neutrophil chemotaxis. Therefore, the neutrophil chemotaxis defect of IDDM appears to be independent of these HLA-DR-associated genes.

Physical property enhancement in light-cured resin composites from post-cure heating is attributed to free radicals created during initial photocuring, the number of which decreases following initial light-curing. Clinically, it is important to know when the number of remaining free radicals is too low to provide for additional conversion of monomer in post-cure-heated specimens. The hypothesis tested is that the potential for additional conversion in post-cure-heated resin composite restorations is dependent upon the time after initial light-curing at which the specimen is exposed to heat treatment. This research examined the effect of delay in post-cure heating after initial photo-activation on strength and monomer conversion of a commercial resin composite material. Discs (10 x 1 mm) of Herculite XRV (Kerr/Sybron, Orange, CA) were photocured at standardized conditions. One group was left unheated, and another was subjected to post-cure heating (Brilliant DI-500, Coltène AG, Altstätten, Switzerland) at the following times after being light-cured: 5 and 30 min, and 6, 24, 48, 72, 96, and 120 hrs. After the appropriate delay time, unheated and heated specimens (n = 10) were tested for biaxial flexural strength at a constant stressing rate. Recovered, fractured strength specimens (n = 10) were analyzed for cure by means of IR spectroscopy. Post-cure heating increased strength over that of the unheated specimens only for the shortest delay times: 5 or 30 min. Thereafter, strength values were statistically equivalent (p < 0.05). Delay in heating did not significantly enhance strength of post-cure-heated specimens, but delay in time did improve strength of the unheated groups. The greatest monomer conversion was obtained when post-cure heating was applied within 6 hrs following light-curing. The difference in cure between unheated and heated specimens remained significant up to 96 hrs of delay. Flexural strength of post-cure-heated specimens remained unchanged with time delay for heating specimens. Maximal monomer conversion of post-cured specimens is obtained only within 6 hrs of light-curing. The potential for additional conversion arising from post-cure heat treatment is dependent upon the time following initial curing at which heat is applied following initial light-curing. However, delay in heat application has no influence on flexural strength.

Both transforming growth factor-beta (TGF-beta) and platelet-derived growth factor (PDGF) have been shown to affect cell proliferation in vitro. The hypothesis being tested was that the effects of the 2 cytokines would be modulated by the presence of serum in the medium. Gingival fibroblasts, obtained from periodontally healthy patients, were maintained in primary culture. Dose response experiments were performed for each growth factor in serum-free medium and in medium containing natural or heat-inactivated fetal bovine serum (10% FBS). Changes in cell numbers were quantified by crystal violet staining. The optimal concentrations of the individual factors (10 ng/ml TGF-beta1, 20 ng/ ml PDGF-BB) were then used when the 2 factors were tested in various sequences. In serum-free medium or in medium with 10% natural serum, the response to PDGF-BB was dose-dependent up to 40 ng/ml; however, with 10% heat-inactivated serum, the maximal response was seen at 20 ng/ml. The largest increase in cell numbers was produced by the simultaneous exposure to the two cytokines, rather than a sequential presentation. The findings suggest that the 48-h growth response of human gingival fibroblasts to 10 ng/ml TGF-beta1 or 20 ng/ ml PDGF-BB in serum-free medium was equivalent to growth obtained in medium containing heat-inactivated 10% FBS without added growth factors.

The purpose of this study was to determine the ability of various solutions to maintain human periodontal ligament cell (PDL) viability in vitro. PDL cells were obtained from extracted third molars and premolars of healthy individuals. These cells were placed into 24-well culture plates containing milk, Save-A-Tooth, Save-A-Tooth supplemented with platelet-derived growth factor-BB (PDGF), or Gatorade at a concentration of approximately 80,000/well. Cells left dry served as negative controls, and cells placed in Eagles' Minimal Essential Medium served as positive controls. At 1, 2, 4, 8, and 12 h, cell viability was evaluated using an MTS assay and an ELISA plate reader to determine optical density. ANOVA and Student-Newman-Keuls tests indicated that milk and Save-A-Tooth with PDGF are suitable as transport medium for avulsed teeth and that the addition of PDGF to Save-A-Tooth may enhance its ability to maintain PDL cell viability. They also suggests that Gatorade would be unsuitable as a transport medium.

This article describes a pilot study in which the authors used aerobic bacterial cultures to compare the effects of 1:10 mouthwash, 1:20 mouthwash and 2 percent ethanol in reservoir systems with seven conventional water systems. The long-term, low-concentration antiseptic reduced bacteria to within acceptable limits.

Objective: This study compared gram-negative bacterial lipopolysaccharide (LPS) adherence to and elution from a Type III gold and a Ni-Cr-Be alloy using Escherichia coli LPS., Method: One-half of the specimens of each alloy were pre-treated with 500 micrograms non-radiolabeled E. coli LPS for 24 h at 37 degrees C. All disks were then incubated with 0.15, 15 or 150 micrograms radiolabeled E. coli LPS for 24 h at 37 degrees C. To evaluate radiolabeled LPS elution, specimens were transferred to LPS-free water and incubated for 24 h at 37 degrees C. The elution scheme, which consisted of 24 h incubations and subsequent transfer to new LPS-free water, continued for up to 96 h total elution. Radiolabeled LPS adherence and elution was determined through liquid scintillation spectrometry. Control disks not treated with LPS were evaluated throughout the study with an enzymatic assay to ensure that extraneous LPS contamination did not occur. A multifactor ANOVA (p = 0.05) was used to evaluate differences in adherence to alloy specimens based upon alloy type, pretreatment status and [3H]LPS concentration. A repeated measures analysis ANOVA (p = 0.05) was used to evaluate differences in elution patterns among groups over time. Least square means were compared in case of significant effects., Results: Toxin uptake at each treatment concentration was significantly different from the other treatment concentrations. In addition, significantly greater amounts of [3H]LPS eluted from the non-pretreated Ni-Cr-Be alloy following the 0.15 and 15 micrograms radiolabeled [3H]LPS treatment, whereas no difference in elution was found among experimental groups following the 150 micrograms [3H]LPS treatment., Significance: E. coli LPS, an LPS type representative of enteric bacteria common to the gingival sulcus, has differing affinities for the alloys. This affinity difference could influence periodontal inflammatory processes, thereby resulting in differing tissue responses adjacent to dental restorations fabricated from these materials. The interaction of other LPS types with these alloys could differ.

Methacrylates can affect cell functions by surfactant-like effects or by altering cell lipid composition. Dimethylaminoethyl methacrylate (DMAEMA), an activator widely used in visible-light polymerized dental resins has been shown to elute readily into aqueous environments. The current study examined the metabolism of this material by oral epithelial cells (HCP) and its subsequent effects on cell lipids. Cells were plated in culture medium, then exposed to DMAEMA in the presence of 14C-acetate, a precursor which labeled the cell lipids. Other cultures were prelabeled with radioisotope, then exposed to DMAEMA. After incubation, the cell lipids were extracted and separated by TLC. Radioactive lipids were located and quantitated. Exposure of the cells to DMAEMA resulted in decreased synthesis of cholesterol with a concomitant increase in sterol precursors. Cholesterol esters and triacylglycerides also increased. Among the polar lipids, phosphatidyl choline (PC) and phosphatidyl ethanolamine (PE) decreased in response to DMAEMA. However, dimethylphosphatidyl ethanolamine (DMPE), a precursor of PC not detectable in control cultures, accumulated to a significant extent in cells exposed to DMAEMA. Furthermore, changes in PC and DMPE levels persisted in the cells for at least 48 h after removal of the DMAEMA. The results indicate that DMAEMA produces alterations in the relative amounts of several cellular neutral and polar lipids. Such alterations, especially of the normal phospholipid composition, along with an alteration in cellular cholesterol, could result in altered membrane-associated cell functions.

Statement of Problem: Lipopolysaccharide (LPS) affinity for titanium implant biomaterials could affect crevicular LPS concentrations and thereby influence periimplant inflammation., Purpose of Study: The purpose of this study was to evaluate Porphyromonas gingivalis and Escherichia coli LPS affinity for titanium biomaterials groups that differed in surface oxide composition and surface roughness., Material and Method: Polished and abraded grade 1 commercially pure titanium and grade 5 alloyed extra low interstitial titanium specimens were treated with 10 EU/mm2 and radiolabeled LPS., Results: The resultant mean +/- SD LPS adherence values ranged from 4.17 +/- 0.29 to 4.79 +/- 0.40 EU/ mm2. No difference in adherence and elution was indicated on the basis of LPS type, surface oxide composition, or surface roughness. Moreover, P. gingivalis and F. coli LPS desorption was below detection., Conclusion: Clinically, the high affinity of both LPS types for titanium biomaterials may adversely influence the periimplant tissue response.

Biocompatibility of dental materials is an important consideration for the patient, clinician, laboratory technician and manufacturer. This paper examines biocompatibility testing methods and the biocompatibility of posterior restorative materials, including amalgam, casting alloys, resin composites, dentin bonding agents, cements, porcelains and ceramics.

This study evaluated the effect of pH on Porphyromonas gingivalis lipopolysaccharide (LPS) affinity for polymethyl methacrylate, polyethyl methacrylate, and polyethyl and polyisobutyl methacrylate resins. Specimens were exposed to 1,010 endotoxin units LPS in potassium phosphate buffer at pH 6, pH 7, or pH 8. Control specimens were incubated in LPS-free water. Sequence I evaluated LPS uptake and release from resin when exposure and elution pH were identical, whereas Sequence II evaluated LPS release from resin when elution pH differed from exposure pH. A slightly acidic pH decreased LPS affinity for all resins compared to pH 7. A slightly alkaline pH increased LPS affinity for the polyethyl methacrylate resin but decreased LPS affinity for the others compared to pH 7. The pH may affect resin-LPS affinity by altering LPS molecular charge.

The light-polymerized resins used in dentistry and their various constituents have been shown to produce significant levels of cytotoxicity, depending upon the material and the cell type exposed to it. These responses include altered cell growth and macromolecule synthesis. The current study examined the effects of several resin components on growth and lipid metabolism of oral epithelial cells. Resin discs were fabricated from triethyleneglycol dimethacrylate (TEGDMA) as received from the manufacturer and after removal of the stabilizer methyl ether hydroquinone (MEHQ). Some discs also contained the initiators benzoyl peroxide (BPO) and camphoroquinone (CQ), and/or an activator, dimethylaminoethyl methacrylate (DMAEMA). After polymerization, the ability of components to elute from the discs and alter cell growth and lipid synthesis were assayed by a colorimetric method and thin layer chromatography respectively. Purified TEGDMA had little effect on the cells' growth or lipid metabolism, while TEGDMA containing MEHQ did inhibit growth as well as total polar lipid synthesis. Eluates from discs containing DMAEMA inhibited cell growth as well as decreasing polar lipid formation. However, this same material produced increased synthesis of diglycerides and cholesterol esters. Eluates from BPO-containing discs, as well as those with CQ, with or without DMAEMA resulted in increased levels of diglycerides. These results demonstrate that even after polymerization, components used in dental resins may elute into the immediate environment and alter various cell metabolic processes.

Substances that elute from denture base resins may inhibit cell growth and disrupt various metabolic processes. This study investigated the effects on cell lipid metabolism of eluates from several denture base resins. Cultured oral epithelial cells were exposed in vitro to eluates of discs made from several denture base resins. Lipid metabolism of the cells was measured using isotopic labeling with 14C-acetate. Results demonstrated that the metabolism of several lipid classes found mainly in the cell membrane was altered by the resin eluates. Eluate from one resin caused the appearance of two previously unrecognized classes of lipids. The alterations of the cell lipids and the presence of the previously unrecognized lipids may be the basis for some clinically evident cytotoxic and allergic reactions.

The purpose of this study was to measure the time-sequence response of RNA and protein synthesis to transforming growth factor-beta 1 (TGF-beta 1) by human periodontal ligament (HPDLF) and gingival (HGF) fibroblasts in culture. HPDLF and HGF were cultured from explants of healthy gingival tissue and freshly extracted teeth. Cultures of 8 x 10(4) cells/ml were exposed to medium containing 3H-uridine and 35S-methionine with TGF-beta 1 at concentrations from 10(-9) M to 10(-21) M, or control medium, for up to 60 hours in order to assess RNA and protein synthesis. Protein concentrations of comparable cultures were also assayed colorimetrically. Results were reported as specific activity (CPM/microgram protein). The results indicate that 10(-9) M TGF-beta 1 treated cultures showed a significant increase in RNA synthesis by HPDLF and HGF over time, as compared to the control cultures. HPDLF showed a significant increase in protein synthesis over time while that by HGF was not significant as compared to the control cultures. Lower concentrations of TGF-beta 1 demonstrated no significant differences from control. Results suggest that the effects of TGF-beta 1 on HPDLF and HGF are both time and dose dependent, with 10(-9) M TGF-beta 1 providing the best response of those concentrations tested. These findings support the concept that TGF-beta 1 may play a role in periodontal regeneration due to its ability to promote fibroblast RNA and protein synthesis. The results also demonstrate that although these two cells types appear morphologically similar, they exhibit distinct biological responses to growth factors such as TGF-beta 1.

With the exception of plaque, the affinity of biologically active bacterial products for restorative materials and the influence of that affinity on periodontal health has not been detailed. This study recognized that Porphyromonas gingivalis endotoxin, which is cell envelope lipopolysaccharide (LPS) produced by a bacterium that is common to the crevicular microbial flora, has an affinity for dental casting alloys. Regardless of surface finish, no difference in LPS initial adherence or elution was recorded between a type III gold or nickel-chromium-beryllium alloy (p > 0.05), but LPS readily adhered and remained attached to both alloys. LPS affinity could contribute to periodontal inflammation in tissues that approximate restorations fabricated from either alloy.

To date there has been very little research into the possible effects of endodontic therapy on regeneration of the lost periodontal attachment. The objective of this study was to examine the effects of endodontic medicaments on fibroblast attachment to dentin surfaces free of a smear layer. Pulp chambers of extracted third molars were filled with one of the following medicaments: gutta-percha with Roth's zinc oxide and eugenol-based sealer, warm gutta-percha with sealer, warm gutta-percha without sealer, calcium hydroxide, formocresol, cotton pellet, or left empty. A predetermined dentin surface area was then inoculated with human gingival fibroblasts at a concentration of 2 x 10(4) cells per ml. The cells were allowed to adhere to the dentin surface for either 4 or 24 hours, then cell attachment was quantified using a methyl-tetrazolium assay. The data were analyzed using a Kruskal-Wallis one-way analysis of variance and Dunn's multiple comparison test. It was determined that fibroblast attachment was significantly reduced when exposed to formocresol or warm gutta-percha without sealer at both the 4 and 24 hour interval (P < or = 0.05). This suggests that the use of formocresol or warm gutta-percha without sealer in a root canal may impede periodontal wound healing and regeneration.

Changes in the lipid composition of a cell membrane due to the binding of one cell modulator may affect binding of a second modulator, whether that binding is receptor-mediated (specific) or non-receptor-mediated (nonspecific). Such altered binding interactions have been demonstrated in oral epithelial cells, wherein N-nitrosonornicotine (NNN), a nonspecific ligand, enhances phorbol ester binding. To characterize membrane changes that may be responsible for such an effect, the current study examined lipid changes in hamster oral epithelial (HCP) cells associated with NNN binding. HCP cultures at two cell densities, 5 x 10(6) cells/100 mm plate (subconfluent cultures) or 10 x 10(6) cells/100 mm plate (confluent cultures) were incubated in Keratinocyte-Serum-Free Medium and exposed to 10 microM NNN or DMSO (solvent control) for 48 h. Lipids were labeled with 14C-acetate, then extracted, separated by thin layer chromatography, and the 14C-lipids located by autoradiography and counted. Exposure of subconfluent cultures to NNN for 48 h, with 14C-acetate present during the final 24 h, resulted in altered phospholipid and fatty acid labeling. Phospholipid labeling increased slightly in the presence of NNN compared to controls, while fatty acid labeling showed a modest but significant decrease in the presence of NNN. Similar changes occurred in the confluent cultures. Prelabeling of lipids in subconfluent cultures, followed by exposure to NNN in the absence of radiolabel, resulted in significantly (P < 0.05) greater phospholipid labeling in the presence of NNN compared to control cultures. At the same time, fatty acid labeling decreased significantly.(ABSTRACT TRUNCATED AT 250 WORDS)

This in vitro study examined the effects of environmental pH on elution of potentially toxic substances from heat-, light-, and dual- (chemical plus light) polymerized denture base resins. Eluates were prepared by daily transfer of disks to fresh buffers at pH 4.0, 5.0, and 6.8 over a 5-day period. Oral epithelial cells were plated in culture dishes in medium containing the eluate. After 24 hours, cellular RNA synthesis was assessed by measuring tritiated uridine uptake. Effects of materials were compared to identical cultures that contained the appropriate buffer without the eluate. The results indicate that the cytotoxic components leach out of the denture base resins in different amounts and at different rates, and the amount of leaching can be affected by pH.