Petra E. Lockwood

Effects of HEMA on water evaporation from water-HEMA mixtures.

OBJECTIVES The aims of this research were: (1) to determine the relative rates of evaporation of water and HEMA, and (2) to determine the effects of increasing concentrations of HEMA on the rate of evaporation of water from water and HEMA mixtures. METHODS Ten microliters of each solution (100% H2O, 75% H2O-25% HEMA, 50% H2O-50% HEMA, 25% HEMA, 100% HEMA) were placed on the pan of a thermogravimetric analysis instrument held at 37 degrees C. The rate of spontaneous weight loss was measured as a function of time and relative humidity (RH) and compared statistically using ANOVA and Scheffé F test. RESULTS The rate of evaporation of pure water was 32-fold higher than that of 100% HEMA. Addition of HEMA to water lowered the rate of evaporation of water from the water-HEMA mixtures in a manner that was proportional to its effect on lowering the vapor pressure of water (p < 0.05 comparing 50% HEMA with 75% HEMA). The rate of evaporation of water from water-HEMA mixtures was higher (p < 0.05) when the ambient gas was at 0% RH than when it was at 51% RH. SIGNIFICANCE The results indicate that as water evaporates from water-HEMA mixtures, the concentration of HEMA rises because it is relatively non-volatile. This rise in HEMA concentration lowers the vapor pressure of water making it more difficult to remove the last amounts of water. This residual water may interfere with polymerization of adhesive monomers, thereby lowering the quality of the hybrid layer.

Fatigue of Dental Ceramics in a Simulated Oral Environment

Fatigue in ceramics refers to the subcritical growth of cracks, aided by the combined influence of water and stress. The dynamic fatigue (constant stressing rate) method was used to obtain subcritical crack growth parameters for three dental ceramics: a feldspathic porcelain, an aluminous porcelain, and a fine-grain, polycrystalline core material. The constant stressing rate experiments were carried out at 37°C for all three ceramics in distilled water, and, for the feldspathic porcelain, in artificial saliva as well. Considerable differences were found in the value of the crack growth exponent (n) among the three ceramics. The feldspathic porcelain was lowest in n-value, while the fine-grain ceramic had the highest n-value. No differences were found for the feldspathic porcelain with respect to n measured in water and in the artificial saliva. Lifetime prediction curves in 37°C water, constructed from the n-values and inert strengths, showed that fatigue failure within five years is a good possibility for feldspathic porcelain specimens at stress levels which can reasonably be anticipated to occur in the oral environment. Little likelihood of failure was perceived for the fine-grain ceramic. The aluminous porcelain was intermediate between these two materials with respect to failure probability.

Ability of Ni-containing biomedical alloys to activate monocytes and endothelial cells in vitro

Nickel-containing alloys commonly are used in medical and dental applications that place them into long-term contact with soft tissues. The release of Ni ions from these alloys is disturbing because of the toxic, immunologic, and carcinogenic effects that have been documented for some Ni compounds. In particular, Ni ions in solution recently have been shown to cause expression of inflammatory mediators, such as interleukin-1beta (IL-1beta), tumor necrosis factor-alpha (TNF-alpha), and intercellular adhesion molecules (ICAMs) from keratinocytes, monocytes, and endothelial cells. However, the ability of the solid alloys themselves to induce these inflammatory effects has not been demonstrated. An in vitro system was used to determine if Ni-containing biomedical alloys could cause secretion of either IL-1beta or TNF-alpha from monocytes or expression of ICAMs on endothelial cells. Pure nickel, titanium, and three biomedical alloys-18-8 stainless steel, NiTi, and Rexillium III-were evaluated. First, it was determined whether or not the alloys or pure metals could cause cytotoxicity to THP-1 human monocytes or human microvascular endothelial cells (HMVECs) by measuring the succinic dehydrogenase (SDH) activity of the cells. Then, using identical conditions of exposure, the secretion of IL-1beta or TNF-alpha from monocytes or ICAM-1 expression on the HMVECs was determined. Only pure nickel suppressed (by 48% compared to Teflon controls) the SDH activity of the HMVECs or THP-1 monocytes. No alloy or metal caused the HMVECs to express ICAM-1, but the NiTi alloy caused a significant (ANOVA/Tukey) secretion of IL-1beta from the THP-1 monocytes. Secretion of TNF-alpha induced by NiTi was detectable but not statistically significant. The levels of IL-1beta secretion from monocytes were sufficient to induce ICAM-1 expression on HMVECs. The release of Ni from the NiTi was a logical suspect in causing the IL-1beta secretion by monocytes, but its role was not confirmed since other alloys, such as Rexillium III, released the same quantities of Ni yet did not activate the THP-1 monocytes. Within the limitations of in vitro conditions, our results indicate that NiTi alloys pose a risk of promoting an inflammatory response in soft tissues by activating monocytes. Further study is needed to substantiate this finding in vivo.

Fracture toughness of commercial dental porcelains

Abstract Ceramic materials suffer from a number of limitations which restrict their use in dental restorations. Chief among these shortcomings is their extreme brittleness which is manifested as a low ability to absorb elastic strain energy during structural failure. This property is embodied in the material parameter, fracture toughness or K IC . The fracture toughness values of two types of dental porcelains, the feldspathic and aluminous porcelains, were determined by the indentation technique. The aluminous porcelains were significantly tougher than the feldspathic porcelains. These differences in K IC were attributed to differences in the nature of crack-microstructure interaction occurring in the two types of porcelain.

In vitro cytotoxicity of resin-containing restorative materials after aging in artificial saliva

Abstract Studies have reported that dental resin-based materials release substances which have biological liabilities. However, some current methods for detecting these substances may not be adequate to detect biologically relevant concentrations. In the current study, we hypothesized that resin-based materials exhibit cytotoxic effects and alter cellular function in vitro when high-pressure liquid chromatography (HPLC-UV detection) cannot detect any release of substances. We further hypothesized that this release continues even after aging the samples in artificial saliva. Five types of composite or compomer materials (Z-100, Tetric Ceram, Dyract AP, Solitaire, and Clearfil AP-X) and one organically modified ceramic material (Definite) were tested after aging in artificial saliva for 0, 7, or 14 days. Cytotoxicity was assessed using direct contact with fibroblasts and measurement of succinic dehydrogenase activity after 48 h of exposure post aging. Release of substances from the materials was assessed using HPLC with UV detection. Altered cellular function was estimated by measuring proliferation of MCF-7 cells with sulforhodamine staining. HPLC showed that whereas initial release of substances was higher without aging, this release dropped significantly after 7 or 14 days of aging, and was equivalent to the Teflon controls after 14 days for four of the materials (Tetric Ceram, Definite, Solitaire, and Clearfil AP-X). Without aging in saliva, all materials had cytotoxicities >50% of the Teflon negative controls. After 14 days of aging, all materials except the Definite continued to show severe cytotoxicity. Only the Definite could be tested for its ability to alter cellular function because of the continuing toxicity of the other materials. This modified ceramic material caused a significant proliferative effect on the MCF-7 cells indicating that sufficient substances were released to alter cellular function. We concluded that all of these commercially available resin-based dental materials continue to release sufficient components to cause lethal effects or alter cellular function in vitro even after 2 weeks of aging in artificial saliva.

Components of Dentinal Adhesives Modulate Heat Shock Protein 72 Expression in Heat-stressed THP-1 Human Monocytes at Sublethal Concentrations

Few studies have investigated the ability of dental resins to induce cellular stress at sublethal concentrations. Cellular stress, especially in immune cells such as monocytes, may modulate the biological response to materials or the hosts ability to respond to bacterially mediated inflammation. The current study examined the ability of sublethal concentrations of 2-hydroxylethylmethacrylate (HEMA) and triethyleneglycol dimethacrylate (TEGDMA) to induce heat shock protein 72 (HSP72) in human monocytes. HEMA and TEGDMA significantly suppressed heat-induced HSP72 expression, even at sublethal levels, but did not induce HSP72 by themselves. The results of the current study suggest that components released from dental resin could modulate the HSP stress response without altering cellular metabolic activity.

Release of elements from dental casting alloys into cell-culture medium over 10 months

OBJECTIVE The release of elements from eight types of commonly used dental casting alloys into cell-culture medium was measured over a 10-month period. The release of elements was determined to provide information about the long-term biological risk these alloys may pose to the oral tissues. The current work extends previous studies of shorter time periods, and is more relevant to the in vivo situation, where dental alloys are present intraorally for years. METHOD The alloys were Au-, Ag-, Pd-, and Ni-based with nobilities ranging from 0 to 95 at.%. Alloy samples (n = 12) were exposed to cell-culture medium. The medium was changed every 30 days for 10 months. Elemental release into the medium was measured by means of atomic absorption spectrophotometry. Differences in mass release were determined using ANOVA and Tukey multiple comparison intervals (alpha = 0.05). RESULTS The release of elements continued through 10 months, and it appeared that the release was constant throughout most of the experiment. Higher initial rates were suspected but not verified. Most alloys reached a constant rate after < 100 days of exposure to the medium. Long-term element release was not generally intuitive based on the amount of an element in an alloy or the overall nobility of the alloy. Total mass lost over the 10-month period ranged from < 2 micrograms/cm2 for the Au-Pd alloy to 55 micrograms/cm2 for the Au-Ag-Cu alloy (Tukey interval at alpha = 0.05 was 0.8 microgram/cm2). By comparison, pure Cu lost 4500 micrograms/cm2 during this period. SIGNIFICANCE Tests which assess biological risk from elemental release must consider longer-term release because elemental release continues for extended periods. Longer-term mass loss from a given alloy is often not intuitive based on its overall composition or noble metal content.

Effect of silver, copper, mercury, and nickel ions on cellular proliferation during extended, low‐dose exposures

Previous studies have demonstrated and quantified the cytotoxicity of metal ions in vitro, but the data from these reports have been limited to short-term exposures of metal ions to cells (24-72 h). Yet, the longer-term, low-dose effects of metal ions are most relevant to the clinical use of dental and other biomedical alloys. Thus, the purpose of the current study was to assess longer-term effects of ions of silver, copper, mercury, and nickel - four metal ions known to be released from dental alloys - on monocytes. THP-1 human monocytes were exposed to the metal ions for up to 4 weeks. Concentrations of the metal ions were 1-10% of those known to cause cytotoxicity with 24-h exposures. Cellular proliferation and cellular viability were measured weekly. Ag(1+) and Hg(2+) did not alter the percentage of nonviable cells, but Cu(2+) and Ni(2+) increased the nonviable component as a function of metal concentration. These effects were cumulative over the 4 weeks only for Ni(2+). All metal ions caused a significant reduction in cellular proliferation, but the pattern of the effect was unique to each metal ion, and the effects were often not evident until 3 or 4 weeks of exposure. The results of the current study indicate that metal ions released from metallic biomaterials may have adverse biological effects at concentrations lower than previously reported.

The effect of glaze on porcelain strength

The self-glazing technique provides an esthetic and hygienic surface for crowns and fixed partial dentures that use porcelain veneers. A study of the biaxial flexure strengths of polished vs. glazed specimens is needed to verify that current laboratory methods are appropriate for planned fatigue studies. Four groups of 50 porcelain disk specimens each were subjected to the following polishing and firing procedures: group one was fired, glazed-no hold, and polished; group two was fired, polished, and glazed-no hold; group three was fired, polished and glazed-1 min. hold; group four was fired, polished, and not glazed. The piston-on-three-ball method was used for testing biaxial flexure strengths. Significantly lower differences in biaxial flexure strengths were noted when group two values were compared with values from groups one, three and four. The results show that the Weibull distribution is an appropriate model for our studies. Differences in glaze thickness among the groups were noted in SEM examination; however, bulk (interior) microcrack density differences were absent. The specimens that were fired, polished to a 1 micron surface finish, and not glazed (group four) were significantly higher in flexure strength than groups one and three at the p less than 0.001 level. The hypothesis that glazing of porcelain surfaces improves the biaxial flexure strength of test specimens was rejected.

Elemental release from dental casting alloys into biological media with and without protein.

OBJECTIVE The objective of this study was to determine the role of proteins in affecting elemental release from a variety of clinically available dental casting alloys. An important role for proteins was suspected based on previous reports about the corrosion of stainless steel and the cytotoxicity of alloys after exposure to a saline-protein solution. METHODS Clinically available alloys with compositions ranging from 0 to 94at.% noble elements were exposed for 1 week to either saline, saline with 3% bovine serum albumin (BSA), or complete cell-culture medium with 3% serum. Atomic absorption spectrophotometry was used to measure the release of elements from the alloys. Elemental release was normalized for the exposed surface area of the alloys. RESULTS In general, more elemental release occurred into the saline-BSA solution compared to saline alone for all released elements (Ag, Cu, Pd, and Zn) except for Ni. Ni release from the NiCr alloy was lower in the presence of BSA. Each element responded somewhat differently with Pd being the least predictable in its behavior. Elemental release was less in the cell-culture medium than in the saline-BSA solution for most elements. For alloys which released multiple elements, all elements responded similarly but not identically to the presence of protein. A high elemental release during exposure to the saline-BSA solution correlated with a low alloy cytotoxicity post-exposure to the saline-BSA. SIGNIFICANCE This study demonstrates the importance of defining exactly the composition of biological solutions used to assess in vitro corrosion and biocompatibility of dental casting alloys. Other molecules in addition to proteins appeared to be critical to the corrosion of these alloys in vitro.