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Biomedical subjects

W R Hume

Publications and source records attributed to W R Hume.

At least 19 recordsLinked to original sources

A proposal for a new classification of lesions of exposed tooth surfaces.

In the presence of improved methods of identification and treatment of lesions on the exposed surfaces of teeth, it should now be acknowledged that the GV Black "classification of carious cavities" is out of date. This paper describes a new system, proposed in 1997, discussed broadly throughout the profession, and eventually modified. The system has been adopted in several regions around the world as being a useful corollary to the current developing concept of minimal intervention dentistry. It is now desirable to adopt a new approach to the identification and recording of the lesions caused by both caries and non-carious tooth loss. A major advantage arising from its adoption would be that it would encourage the profession to minimise the amount of normal healthy tooth structure that is often sacrificed in pursuit of the cavity designs as suggested by Black. The authors are members of a Project Group of the FDI Science Committee, and this paper explains the concept and offers justification for the adoption of the system.

Dental Caries↗

Resin monomer 2-hydroxyethyl methacrylate (HEMA) is a potent inducer of apoptotic cell death in human and mouse cells.

Mechanisms by which the resin monomer 2-hydroxyethyl methacrylate (HEMA) induces hypersensitivity reactions in humans are not well-established, nor have the direct effects of HEMA on cell death been fully characterized. The objective of this study was to establish whether HEMA is capable of inducing apoptotic cell death, and whether differences exist in the levels of apoptotic death induced by HEMA in cells obtained from healthy individuals and from patients with established HEMA hypersensitivity. HEMA induced apoptotic death in Peripheral Blood Mononuclear Cells (PBMCs) obtained from both healthy and HEMA-sensitized patients and in the murine RAW cells in a dose-dependent manner. However, induction of cell death by HEMA was lower in PBMCs obtained from patients in comparison with healthy individuals. Studies reported in this paper demonstrate that HEMA induces apoptotic death, and that decreased susceptibility of lymphocytes to HEMA-mediated death might be an important mechanism for the generation and persistence of hypersensitivity reactions in patients.

Animals↗

Biological clearance of HEMA in guinea pigs.

No toxicokinetic data are available about the dental composite component 2-hydroxyethylmethacrylate (HEMA) in vivo in the literature. Therefore, the excretion of HEMA in feces and urine in vivo and, using the pendular perfusion technique with segments of jejunum and colon, in the biliary and enteric excretion in situ were investigated in anesthetized guinea pigs. In the in situ experiments, guinea pigs (n = 4) received HEMA (0.02 mmol/kgbw labelled with a tracer dose 14C-HEMA 0.3 kBq/gbw) injected into the jugular vein. In the in vivo experiments, guinea pigs (n = 4) received HEMA (+ 14C-HEMA, same dose as above) via gastric tube. Urine and feces were collected for 24h. In the in situ experiments, organs from guinea pigs were removed 60 min after the beginning of the experiment, and then the 14C-radioactivity was measured. During the 60 min perfusion period the calculated amount of 14C-activity excreted into the total jejunum and colon was 6.0 +/- 1.0% and 2.7 +/- 0.7% of the dose administered, respectively (mean +/- sem). Of the 14C-HEMA dose, 5.3 +/- 0.3% was found in the bile. Significantly (p < 0.05) higher bile/blood concentration ratios were found at 10-40 min after the injection of HEMA, as compared to the ratio at 60 min. The total 14C-recovery in all organs tested was 20.0 +/- 2.6%. During 24h the amounts of 14C-activity excreted in the feces and urine were 1.1 +/- 0.1% or 17.1 +/- 1.50% of the dose administered, respectively (mean +/- sem). The total 14C-recovery in all organs tested was 11.6 +/- 0.6%. In a second series of in vivo experiments, exhaled air from the animals was captured during the 24h experimental period. 14C was exhaled to 63.6 +/- 2.11% of the administered 14C-HEMA dose (mean +/- sem; n = 4) as 14C-carbondioxide. The results indicate a rapid clearance of 14C-HEMA and/or 14C-HEMA metabolite(s) from the organism, exhalation being the major route of elimination.

Animals↗

Biological clearance of TEGDMA in guinea pigs.

The excretion of the dental composite component triethylene glycol dimethacrylate (TEGDMA) in feces and urine in vivo and, using the pendular perfusion technique with segments of jejunum and colon, the biliary and enteric excretion in situ were investigated in anesthetized guinea pigs. In the in situ experiments guinea pigs (n = 4) received TEGDMA (0.02 mmol/kg body weight labelled with a tracer dose 14C-TEGDMA 0.7 kBq/g body weight) injected into the jugular vein. In the in vivo experiments guinea pigs (n = 4) received TEGDMA (+14C-TEGDMA; same dose as above) via a gastric tube. Urine and feces were collected for 24 h. In the in situ experiments organs were removed from the guinea pigs 60 min after the beginning of the experiment, and the 14C radioactivity measured. During the 60-min perfusion period the calculated amount of 14C radioactivity excreted into the total jejunum and colon was 0.9 +/- 0.2% and 1.9 +/- 0.1% of the dose administered, respectively (means +/- SEM). Of the 14C-TEGDMA dose, 3.7 +/- 0.2% was found in the bile. A significantly (P < 0.05) higher bile/blood concentration ratio was found 10 min after injection of TEGDMA as compared with the ratios at 20 to 60 min. The following 14C activities (percent of the dose) per total organ were found in guinea pigs (in situ experiment; means +/- SEM): 6.9 +/- 1.7 (muscle), 3.9 +/- 0.5 (kidney), 3.3 +/- 0.1 (skin), 1.4 +/- 0.1 (blood), and 1.2 +/- 0.1 (liver). The 14C activity in all other organs was < 0.4%. The total 14C recovery in all organs tested was 17.5 +/- 1.8%. Over 24 h the amounts of 14C activity excreted in the feces and urine were 0.5 +/- 0.1% and 14.7 +/- 1.8% of the dose administered, respectively (means +/- SEM). The following 14C activities (percent of the dose) per total organ or contents of organs were found (means +/- SEM): 1.4 +/- 0.3 (liver), 0.8 +/- 0.3 (muscle), 0.5 +/- 0.1 (skin), and 0.5 +/- 0.1 (contents of cecum). The 14C activity in all other organs was < 0.2%. The total 14C recovery in all organs tested was 3.9 +/- 0.9%. In a second series of in vivo experiments exhaled air from the animals was captured during the 24-h experimental period. Of the administered dose, 61.9 +/- 4.6% of the 14C (means +/- SEM; n = 4) was exhaled as 14C-carbon dioxide. The results indicate a rapid clearance of 14C-TEGDMA and/or 14C-TEGDMA metabolite(s) from the organism and exhalation is the major route of elimination.

Animals↗

Cytotoxicity of dental composite components and mercury compounds in lung cells.

OBJECTIVE: The effect of dental composite components triethyleneglycoldimethacrylate (TEGDMA) and hydroxyethylmethacrylate (HEMA), as well as mercuric chloride (HgCl2) and methylmercury chloride (MeHgCl) was investigated on the release of lactatedehydrogenase (LDH) from alveolar epithelial lung cell lines in vitro. METHODS: The confluent cell layers from the A549 (human, malignant) and the L2 cells (rat) were incubated with various concentrations of HEMA, TEGDMA, MeHgCl and HgCl2 at 37 degrees C in 2% (v/v) CO2 atmosphere for 8h. In further experiments the L2 cells were incubated with the same compounds for 6-48 h. LDH release was measured and the values were expressed as percentage of the LDH content. The values were plotted on a concentration log-scale and the substance concentration at the maximum slope was assessed as effective concentration (EC50). RESULTS: A significant (p<0.05) increase in the LDH release was found in the L2 cells after 8-h incubation with HEMA (4 mmol/l), TEGDMA (2 mmol/l), MeHgCl (0.01 mmol/l) and HgCl2 (0.015 mmol/l), and in A549 cells with HEMA (14 mmol/l), TEGDMA (15 mmol/l), MeHgCl (0.15 mmol/l) and HgCl2 (0.05 mmol/l), compared to controls. The EC50 values from compounds in the L2 cells are shown in the following table (mean; sem in parentheses; n=3-6; #n=1): [see text]. SIGNIFICANCE: The toxic effect of HgCl2 and MeHgCl from the L2 cells was about 100-700-fold higher than of the dental composite components. A significant (p<0.05) time dependent increase of toxicity was observed with TEGDMA, HEMA and MeHgCl.

Animals↗

Surface hardness change of restorative filling materials stored in saliva.

OBJECTIVES: This study was to investigate the effect of saliva used as storage liquid and the length of storage effect on surface hardnesses of Fuji IX (GP) (FIX), Dyract (DR), Z-100 and Estio LC (ELC). METHODS: The materials were mixed according to the manufacturers' instructions and immersed in distilled water or human parotid saliva. Vickers hardness number (HVN) was measured 1, 7, 20 and 40 days after the materials were mixed. HVN was calculated from the indentation diameter after 100 or 300g loading on their surface for 15s. The two methods of characterization used in this work were X-ray photoelectron spectroscopy (XPS) for surface chemical composition and electron probe microanalysis (EPMA) for depth profile analysis. RESULTS: Only in FIX, did HVN increase with time at both storage conditions, distilled water and saliva. The increase rate of the value was higher when stored in saliva than distilled water. After 40 days storage in saliva, the HVN value of FIX increased by 39%. The increase for storage in saliva for DR was 22%, ELC 16%, and Z100 3%, compared to 1 day storage in distilled water. Ca and P peaks caused by saliva were detected by XPS and EPMA analysis, but these peaks did not exist in either composite resin or polyacid-modified composite resin by EPMA analysis. SIGNIFICANCE: Saliva has the remarkable effect of increasing surface hardness of Fuji IX (GP).

Compomers↗

Distribution and excretion of TEGDMA in guinea pigs and mice.

The monomer triethyleneglycoldimethacrylate (TEGDMA) is used as a diluent in many resin-based dental materials. It was previously shown in vitro that TEGDMA was released into the adjacent biophase from such materials during the first days after placement. In this study, the uptake, distribution, and excretion of 14C-TEGDMA applied via gastric, intradermal, and intravenous administration at dose levels well above those encountered in dental care were examined in vivo in guinea pigs and mice as a test of the hypothesis that TEGDMA reaches cytotoxic levels in mammalian tissues. 14C-TEGDMA was taken up rapidly from the stomach and small intestine after gastric administration in both species and was widely distributed in the body following administration by each route. Most 14C was excreted within one day as 14CO2. The peak equivalent TEGDMA levels in all mouse and guinea pig tissues examined were at least 1000-fold less than known toxic levels. The study therefore did not support the hypothesis.

Animals↗

Induction of apoptotic cell death in peripheral blood mononuclear and polymorphonuclear cells by an oral bacterium, Fusobacterium nucleatum.

It is largely unknown why a variety of bacteria present in the oral cavity are capable of establishing themselves in the periodontal pockets of nonimmunocompromised individuals in the presence of competent immune effector cells. In this paper we present evidence for the immunosuppressive role of Fusobacterium nucleatum, a gram-negative oral bacterium which plays an important role in the generation of periodontal disease. Our studies indicate that the immunosuppressive role of F. nucleatum is largely due to the ability of this organism to induce apoptotic cell death in peripheral blood mononuclear cells (PBMCs) and in polymorphonuclear cells (PMNs). F. nucleatum treatment induced apoptosis of PBMCs and PMNs as assessed by an increase in subdiploid DNA content determined by DNA fragmentation and terminal deoxynucleotidyltransferase-mediated dUTP-biotin nick end-labeling assays. The ability of F. nucleatum to induce apoptosis was abolished by either heat treatment or proteinase digestion but was retained after formaldehyde treatment, suggesting that a heat-labile surface protein component is responsible for bacterium-mediated cell apoptosis. The data also indicated that F. nucleatum-induced cell apoptosis requires activation of caspases and is protected by NF-kappaB. Possible mechanisms of F. nucleatum's role in the pathogenesis of periodontal disease are discussed.

Apoptosis↗

Effect of dental materials on gluconeogenesis in rat kidney tubules.

The effect of dental composite components triethyleneglycoldimethacrylate (TEGDMA) and hydroxyethylmethacrylate (HEMA) as well as mercuric chloride (HgCl(2)) and methylmercury chloride (MeHgCl) on gluconeogenesis was investigated in isolated rat kidney tubules. From starved rats kidney tubules were prepared and isolated by digestion with collagenase. Every 10 min up to 60 min 1-ml samples were drawn from the cell suspension for quantitating the glucose content. Glucose formation in controls was 3.3 +/- 0.2 nmol/mg. per min (mean +/- SEM, n=21). Relative rates of glucose formation were obtained by expressing individual rates as a percentage of the corresponding control. X-Y concentration curves (effective concentration, EC) of the substances were calculated by fitting a four-parametric sigmoid function to the relative rates of glucose formation at various test concentrations. At the end of the incubation period cell viability was assessed by trypan blue exclusion. Cell viability decreased within the 60 min interval from 90 to approx. 80% (controls), <25 (HEMA), <20 (TEGDMA), <10 (MeHgCl), and <10% (HgCl(2)). Values of 50% effective concentration (EC(50)) were calculated from fitted curves. EC(50) values were (mmol; mean +/- SEM; n=4): HEMA, 17.7 +/- 2.9; TEGDMA, 1.8 +/- 0.2; MeHgCl, 0.018 +/- 0.0005; and HgCl(2), 0. 0016 +/- 0.0005. The toxic effect of HgCl(2) was approximately 1000 or 10 000 higher than that of the dental composite components TEGDMA or HEMA, respectively.

Animals↗

Component release from light-activated glass ionomer and compomer cements.

The purpose of this study was to identify and quantify any component released from seven commercially available light-cured or resin-modified glass ionomer and compomer cements. Twenty-one separate cylindrical stainless steel moulds 6 mm in diameter and 1.0 mm deep were filled with one of seven glass ionomer or compomer cements, light activated and then immediately immersed in separate containers of distilled water. Water samples were retrieved over a time period of up to 30 days and retained for analysis. An occlusal cavity 6 mm in diameter was prepared in extracted human third molar teeth with a remaining dentin thickness of 1.6-2.0 mm. A polypropylene chamber was attached to the cemento-enamel junction of each tooth to contain 1 mL of distilled water. Ten teeth were each filled with one of three cements and light activated. Water samples (eluates) were retrieved over a period of time. All samples were analysed by high performance liquid chromatography. Only one component, hydroxyethyl methacrylate (HEMA), was detected in the eluates from both tooth and mould samples. Analysis of diffusion of the HEMA through dentin showed a relatively sustained movement into the pulp space during the first day, with exponential decline thereafter. Our data show that HEMA was released from all of the light activated glass ionomer cements studied and from the compomer, both directly into water and through dentin. This release may be relevant both to the risk of adverse pulpal responses in patients and to the risk of allergy in patients and dental personnel.

Chromatography, High Pressure Liquid↗

Rapid and quantitative detection of Streptococcus mutans with species-specific monoclonal antibodies.

Streptococcus mutans is known to be a prime etiologic agent for the initiation and progression of human dental caries. Rapid, accurate, and quantitative detection of S. mutans will help us better understand the pathogenic mechanisms of dental caries and will help to develop methods for caries diagnosis and risk assessment. This study describes the development of three highly species-specific monoclonal IgG antibodies against S. mutans. The antibodies were used to develop a number of methods that quantitatively detect S. mutans in less <1 min and are sensitive enough to detect a single bacterial cell. These methods could be widely used in basic and clinical studies related to S. mutans and in the clinical diagnosis and treatment of caries in humans.

Antibodies, Bacterial↗

A new cavity classification.

With the development of adhesive restorative materials and a far better understanding of the action of the fluoride ion it is suggested that the time has arrived for a reassessment of the traditional cavity classification as set out by G.V. Black over one hundred years ago. When preventive measures and remineralization fail and a carious lesion has progressed through the enamel into the dentine there is a need to remove the infected dentine, and possibly some of the affected dentine as well, to eliminate cavitation and avoid further accumulation of plaque. In most situations this will involve removal of enamel to achieve access to the infected dentine but, in the presence of fluoride, both enamel and dentine are capable of being remineralized and therefore conserved, at least to a degree. The principle of minimal extension must be encouraged to allow maximum preservation of natural tooth structure. A new cavity classification is proposed which is designed to make the most of the potential for healing which is inherent in both enamel and dentine. However, it must be accepted that a considerable proportion of restorative dentistry is carried out to replace failed restorations and, in this case, cavity design will be complicated by existing loss of tooth structure.

Adhesives↗

A study of component release from resin pit and fissure sealants in vitro.

OBJECTIVE: A recent study reported that an estrogenic chemical, bisphenol-A, was released from a fissure sealant. The aim of this study was to identify and quantify the major (or detectable) components released from any of seven commercially-available, light-cured pit and fissure sealants in vitro. METHODS: The fissure systems of ten extracted, third molar teeth were filled with sealant, light-activated and immersed in separate containers of distilled water. Separate, cylindrical stainless steel molds were filled with sealant which was then light-activated and immersed. Each mold or tooth with sealant was moved to a new container of water at defined times and each remaining water sample (eluate) then analyzed by high performance liquid chromatography (HPLC). RESULTS: Triethylene glycol dimethacrylate (TEGDMA) was present in all eluates from each of the sealants tested. 2,2-bis[4'-(2'-hydroxy-3'-methacryloyloxy)phenyl]propane (BisGMA) was detected at much lower levels (about one thousand-fold less) in eluates from one sealant only. Bisphenol-A was not detected in any eluates. The rates of TEGDMA and BisGMA release were highest on first immersion and decreased thereafter. The total amount of TEGDMA released was on the order of 0.25 mg per tooth. Most release occurred during the first day. SIGNIFICANCE: Because bisphenol-A release could not be detected from any of the seven sealants tested, these results call into question earlier concerns expressed about possible adverse effects of bisphenol-A released from resin sealants.

Air Pollutants, Occupational↗

The effect of dentine thickness on diffusion of resin monomers in vitro.

Forty extracted human third molar teeth were divided into four groups, each of 10 teeth, to test the hypothesis that dentine thickness variation influences diffusion of the monomers 2-hydroxyethylmethacrylate (HEMA) and triethylene glycol dimethacrylate (TEGDMA) from light-cured bonding resin-composite resin restorations to the pulp space. An occlusal cavity 6 mm in diameter was prepared in each tooth of four groups with remaining dentine thickness of 3.4-3.6, 2.4-2.6, 1.4-1.6 and 0.4-0.6 mm, respectively. A polypropylene chamber was attached to the cemento-enamel junction of each tooth to contain 1 mL of distilled water. Each cavity was treated with Scotchbond Multipurpose (3 M, U.S.A.) then restored with Z100 (3 M) and light activated for 30 s. Water samples were retrieved over a time course up to 30 days and analysed by high performance liquid chromatography. Both HEMA and TEGDMA were detected in the pulp samples for all teeth. Decreasing dentine thickness substantially increased pulpward diffusion rate of both HEMA and TEGDMA during the first day after placement, as well as the total release of both components from a bonding resin-composite combination in vitro.

Chromatography, High Pressure Liquid↗

Diffusion of resin monomers through human carious dentin in vitro.

The diffusion of 2-hydroxyethylmethacrylate (HEMA) and triethylene glycol dimethacrylate (TEGDMA) from light cured bonding resin-composite resin restorations through human carious dentin was investigated. Extracted human molar teeth with different degrees of caries were obtained from consenting donors. Teeth were classified into three groups according to caries severity (mild, moderate and severe) using subjective criteria. The outer carious lesions were then removed guided by a proprietary caries detector dye. Teeth with exposure of the pulp space after caries removal were excluded from the study. A polypropylene chamber was attached to the cemento-enamel junction of each tooth to contain 1 ml distilled water. Each cavity was restored with a HEMA containing bonding resin then a TEGDMA-containing resin composite. Water samples were retrieved over a time course and analyzed by high performance liquid chromatography. There was great variation between teeth in HEMA and TEGDMA permeability. The cumulative amounts released were of similar magnitude to those observed in non-carious teeth for the mild and moderately-severe groups. However, the cumulative amounts released were markedly greater in severely carious teeth than in those with moderate or mild caries.

Adult↗