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Biomedical subjects

J C Wataha

Publications and source records attributed to J C Wataha.

At least 73 records · Page 4Linked to original sources

Biological effects of palladium and risk of using palladium in dental casting alloys.

In dentistry, palladium is a very common component of dental casting alloys of all types, and its use has increased over the past several decades in response to the increased cost of gold. However, there have been recent controversies, particularly in Germany, over possible adverse biological effects of using palladium in dental alloys. Therefore, the purpose of this paper is to review the known biological effects of palladium and the likelihood that these effects can be caused by dental alloys which contain palladium. In an ionic form and at sufficiently high concentrations, palladium has toxic and allergic effects on biological systems. Palladium allergy almost always occurs in individuals who are sensitive to nickel. The carcinogenic potential of the palladium ion is still unclear, although there is some evidence that it is capable of acting as a mutagen. However, there are no well documented cases of adverse biological reactions to palladium in the metallic state. Furthermore, in spite of the potential adverse biological effects of palladium ions, the risk of using palladium in dental casting alloys appears to be extremely low because of the low dissolution rate of palladium ions from these alloys.

Animals↗

Materials for endosseous dental implants.

The goal of placement of endosseous dental implants is to achieve osseointegration or biointegration of the bone with the implant. A wide variety of materials has been used for these implants, but only a few promote osseointegration and biointegration. Titanium and titanium alloy (Ti6A14V) have been the most widely used of these materials. The surface oxide of titanium appears to be central to the ability of this material to osseointegrate. The oxide limits dissolution of elements and promotes the deposition of biological molecules which allow bone to exist as close as 30 A to the surface of the implant. The details of the ultrastructure of the gap between the implant and bone remain undefined, and the consequences of elements which are released on the interface over time are not known. These areas of investigation are particularly important in defining the differences between commercially pure titanium implants and those made of titanium, aluminium and vanadium. The epithelial interface between the gingiva and titanium appears to contain many of the structural characteristics of the native tooth-gingiva interface, but details are still vague. The connective tissue interface with the titanium appears to be one of tightly fitting tissues rather than adhesion. Ceramic coatings appear to improve the ingrowth of bone and promote chemical integration of the implant with the bone. The characteristics of these coatings are complex and affect the bony response, but the mechanisms remain obscure. The degradation of the coatings is an issue of particular controversy. Progress in dental implantology is likely to continue as the interface between the material and bone is more clearly understood, and biological molecules and artificial tissues are developed.

Alloys↗

Biocompatibility of posterior restorative materials.

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.

Acrylic Resins↗

In vitro reaction of macrophages to metal ions from dental biomaterials.

OBJECTIVES: This study was conducted to 1) measure the sensitivity of human and mouse macrophages to metal ions which are released from dental biomaterials, 2) compare these sensitivities with those of other cell types in the oral cavity, and 3) determine if metal ions alter the metabolism and synthetic processes of these cells at lower concentrations than are required to lyse the cells. This information will help define the biological risks associated with the release of metal ions into the oral cavity. METHODS: Macrophages were exposed to a range of concentrations of Ag1+, Au3+, Cu2+, Hg2+, Ni2+, Pd2+, Pt4+, and Zn2+ for 24 h in cell culture. The concentrations which caused a 50% decrease in succinic dehydrogenase (SDH) activity, protein production, and lactate dehydrogenase (LDH) release were measured and compared with these values for fibroblasts and osteoblasts. RESULTS: Most metal ions caused alteration in SDH activity and protein production at lower concentrations than were required to induce LDH release. There were exceptions to this trend, and the differences were not always statistically significant. Furthermore, although the macrophages sometimes had statistically different sensitivities to metal ions than fibroblasts or osteoblasts, these differences were less than one order of magnitude. Macrophage response to the metal ions was highly dependent on the metal ion and the species of macrophage. SIGNIFICANCE: Macrophages react adversely to metal ions at similar concentrations as other cell types found in the oral cavity. Furthermore, the concentrations which affect cell metabolism and protein production are generally lower than those which lyse the cells. Thus, non-lethal concentrations of metal ions may alter the secretion of protein inflammatory mediators such as cytokines which direct the inflammatory response in tissues.

Analysis of Variance↗

Cytotoxic interactive effects of dentin bonding components on mouse fibroblasts.

Previous studies have shown a wide range of pulpal reactions to dentin bonding systems and a poor correlation between in vitro and in vivo toxicity of dentin bonding agents. Because dentin bonding agents are composed of multiple components which may diffuse through dentin, we hypothesized that these components may cause cytotoxicity through interactive (synergistic) effects. We investigated the cytotoxicities of four dentin bonding components--HEMA, Bis-GMA, TEGDMA, and UDMA--and interactive effects for three binary combinations of the dentin bonding components--HEMA and Bis-GMA, Bis-GMA and TEGDMA, and TEGDMA and UDMA. Cytotoxicities to Balb/c 3T3 mouse fibroblasts were measured by the MTT assay. Concentrations which caused 50% toxicity compared with controls (TC50 values) were compared, and the interactive effects were determined by evaluation of the differences between observed and expected MTT activities of the cells. The ranks of toxicity of the dentin bonding components in terms of TC50 values were as follows: Bis-GMA > UDMA > TEGDMA >>> HEMA (least toxic) after 24- and 72-hour exposures. As binary combinations, the three combinations of dentin bonding components interacted in three ways--synergism, additivism, and antagonism--which were influenced by the concentrations of both components. The longer period of exposure resulted in a significant increase in the cytotoxicity of the dentin bonding components and combinations. The findings indicate that both exposure time and the interactions between the dentin bonding components may be important parameters in determining the cytotoxicity of dentin bonding agents in vivo.

3T3 Cells↗

In vitro effects of metal ions on cellular metabolism and the correlation between these effects and the uptake of the ions.

The effects of Ag+1, Au+3, Cd+2, Cu+2, Ga+3, In+3, Ni+2, Pd+2, and Zn+2 on DNA synthesis, protein synthesis, succinic dehydrogenase activity, and total cellular protein of mammalian fibroblasts were measured for exposures less than 12 h. The rates at which these cellular functions responded to metal ion exposure were compared and related to the uptake rate of the ions into the cells. These rates of response were significantly different: DNA synthesis decreased the fastest, followed by protein synthesis, succinic dehydrogenase activity, and total protein. This order of response was similar for most metal ions. At 4 h, the rate of uptake of the metal ions correlated most closely with depression of succinic dehydrogenase activity, whereas at 8 h, the uptake correlated most closely with depression of protein synthesis. The similar response of cells to all metal ions may imply that these ions act on cells by similar mechanisms. The rates of uptake of Ag+1, Cu+2, and Zn+2 were sufficiently fast that in vivo exposures of tissues to these metals for periods less than 12 h would be capable of disrupting cellular metabolism.

Animals↗

Effect of cell line on in vitro metal ion cytotoxicity.

OBJECTIVES: The choice of cell line for in vitro biological tests which assess the cytotoxicity of dental materials remains controversial, yet this issue is important because these tests are widely used to rate the biocompatibility of new and existing materials, and many different cell lines are commonly used. The purpose of the current study was to quantify the responses of four cell lines (Balb/c 3T3, L929, ROS 17/2.8 and WI-38) to 14 metal ions which are released from dental materials, and relate these responses to the metabolic activity and population doubling times of these cells. METHODS: Succinic dehydrogenase (SDH) activity was used to monitor metabolic activity and cytotoxic response. RESULTS: The cell lines responded differently to most metal ions. In general, the Balb/c 3T3 line was the most sensitive, and the WI-38 line was the least sensitive. However, there were many exceptions depending on the metal ion. The passage number of the cells also affected the cytotoxic response. It was concluded that the cytotoxicity of materials which release metal ions will be significantly different depending on which cell line is selected and its passage number. SIGNIFICANCE: Based on the findings that cell lines ranked the toxicities of the metal ions similarly, it seems reasonable to use these types of in vitro tests to rank the cytotoxicities of materials. However, if these types of tests are used to predict in vivo cytotoxicity, care should be taken to choose conditions and cells which are relevant.

3T3 Cells↗

Correlation of cytotoxicity with element release from mercury- and gallium-based dental alloys in vitro.

OBJECTIVES: An in vitro screening test was used to compare the cytotoxicity and elemental release from mercury- and gallium-based dental restorative materials. METHODS: The test employed three sequential extractions of the samples into cell-culture medium which were then used to evaluate the cytotoxicity of the samples and the release of elements from the samples. Cytotoxicity was measured by placing the extract in contact with Balb/c mouse fibroblasts for 24 h and measuring the succinic dehydrogenase activity of the cells. The release of elements was measured by means of atomic absorption spectrophotometry. RESULTS: Samples of Tytin (Kerr) showed no cytotoxicity compared to Teflon controls. Dispersalloy (Johnson and Johnson) was severely cytotoxic initially when Zn release was greatest, but was less toxic between 48 and 72 h as Zn release decreased. Gallium Alloy GF (Tokuriki Honten) was moderately cytotoxic after 8 h, and increased in cytotoxicity thereafter, which correlated with a substantial and persistent release of Ga from this material. SIGNIFICANCE: The results of the current study concurred with in vivo assessments of these materials, and the use of sequential extractions was useful in determining trends in the cytotoxicity and elemental release from these materials.

3T3 Cells↗

Cytotoxicity of components of resins and other dental restorative materials.

Cytotoxicity testing of dental restorative materials must be viewed as an assessment of hazards, that is the potential of the material to cause pulpal problems. In this context, composites, glass ionomers, amalgams, zinc-based cements and peroxide bleaching agents are all possible hazards to the pulp. The risks that these materials will cause pulpal toxicity in vivo can be partly estimated by assessing the cytotoxicity of the substances which are released from these materials in vitro and comparing these cytotoxic concentrations with those concentrations that are present in vivo. The resin components of composites, metal ions and hydrogen peroxide, all of which are released from dental restorative materials, have been shown to be cytotoxic in vitro in sufficient concentrations. The potencies of these substances are quite diverse. However, the cytotoxicity of these substances in usage tests, and therefore the risks of pulpal toxicity, depends on their ability to diffuse through the dentine and accumulate in the pulp.

Animals↗

Permeability of biological and synthetic molecules through dentine.

The diffusion through dentine by a number of biological and synthetic molecules, including resins and dyes, is reported. In vitro measurements were derived by experiments with a modified 'split-chamber device'. Diffusion was found to be indirectly proportional to dentine thickness for all molecules. Permeability of water-soluble molecules and ethanol-soluble molecules was proportional to the molecular weights, except for fluorescein, hydrogen peroxide (H2O2) and urethane dimethacrylate. The resin components tested are not soluble enough in an aqueous medium to diffuse through 0.5 mm dentine at sufficient concentrations to cause cytotoxicity to pulpal cells.

Blood Proteins↗

Uptake of metal cations by fibroblasts in vitro.

Atomic absorption spectroscopy was used to assess uptake of Ag+1, Au+4, Cd+2, Cu+2, In+3, Ni+2, Pd+2, and Zn+2 by in vitro cultures of Balb/c 3T3 fibroblasts as a function of concentration of the cations in cell-culture medium. Reversibility of this uptake was also measured. Metal cations exhibited a 400-fold difference in their tendency to accumulate in the cells; In+3 tended to accumulate the most, whereas Pd+2 accumulated the least. Uptake of the cations in the cells increased linearly with the concentration of the cation in the medium for all cations up to their 50% toxicity concentrations. Reversal of this uptake was slower than that of the initial uptake for three cations studied in more detail (Cd+2, In+3, and Ni+2). The duration of the initial exposure affected the proportion of the metal cations that were retained by the cells 5 h after the cations were removed from the medium. The proportion of retained Cd+2 did not change when the initial exposure was increased from 2 h to 6 h, whereas the proportion of retained In+3 decreased and Ni+2 increased over the same period. The tendency of the cells to accumulate these cations correlated with their cytotoxic potency (measured previously).

Animals↗

The effect of repeated stretching on the force decay and compliance of vulcanized cis-polyisoprene orthodontic elastics.

Compliance measurements, used in the past to measure the viscoelastic properties of dental impression materials, were used to assess these properties in vulcanized cis-polyisoprene orthodontic elastics, and the results were compared with traditional force decay measurements. Both methods were also used to evaluate the effect of repeated stretching on these elastics. Compliance measurements successfully characterized the viscoelastic behavior of the elastics, and the results agreed with force decay measurements. Repeated stretching significantly reduced the force and the compliance of the elastics. There was no statistical difference in the force or compliance measurements after the elastics were stretched more than 200 times. Stretching for 1000 cycles of 400% extension reduced elastic force by approximately 12%.

Analysis of Variance↗

The effect of cell monolayer density on the cytotoxicity of metal ions which are released from dental alloys.

The effect of cell density (number of cells per unit area of a monolayer culture) on the in vitro cytotoxicity of metal ions which are known to be released from dental materials was investigated. The effects of cell density (1) may explain previous discrepancies in in vitro tests, (2) may be important in wound healing where cell density changes over time, and (3) may help clarify the mechanisms of cytotoxicity of metal ions. Balb/c 3T3 fibroblasts were plated at cell densities ranging from 10,000-80,000 cells/cm2 and were exposed to 8 concentrations of 10 different metal ions. After 24 h, the succinic dehydrogenase activity and DNA synthesis were measured to quantify the cytotoxic effect. Higher cell densities markedly reduced the sensitivity of these fibroblasts to all metal ions except Al+3 and Zn+2, but the magnitude of the reduction was metal dependent. In addition, the DNA synthesis was inhibited more than the succinic dehydrogenase activity for all metal ions except Zn+2. The unique effect of cell density on each metal ion supported the hypothesis that the effect was not simply caused by a dilution of the number of metal ions per cell. Given these results, the effect of cell density should be carefully selected in in vitro cytotoxicity tests.

3T3 Cells↗

Cytotoxicity and dentin permeability of carbamide peroxide and hydrogen peroxide vital bleaching materials, in vitro.

There has been recent concern about the inadvertent exposure of dentin with patent tubules as well as gingiva to bleaching systems containing 10-15% carbamide peroxide or 2-10% hydrogen peroxide for more than a few minutes. The aims of the present study were: (1) to determine the cytotoxicity of dilutions of hydrogen peroxide in cell culture; (2) to measure hydrogen peroxide diffusion from bleaching agents through dentin in vitro; and (3) to determine the risk of hydrogen peroxide-induced cytotoxicity from exposure of dentin to these vital bleaching agents. The 50% inhibitory dose (ID50) of hydrogen peroxide to succinyl dehydrogenase activity in cultured cells was found to be 0.58 mmol/L after 1 h. All bleaching materials demonstrated diffusion of hydrogen peroxide through dentin in an "in vitro pulp chamber" device. The one- and six-hour diffusates of all bleaching agents through 0.5-mm dentin exceeded the ID50 in monolayer cultures. Inhibition of succinyl dehydrogenase activity corresponded to the amount of hydrogen peroxide that can rapidly diffuse through dentin in vitro and reach concentrations which are toxic to cultured cells in less than 1 h.

3T3 Cells↗

In vitro synergistic, antagonistic, and duration of exposure effects of metal cations on eukaryotic cells.

Effects of duration of exposure of single metal cations (Ag+1, Cd+2, Cu+2, Ga+3, Ni+2, and Zn+2) on 3H-thymidine (3H-Tdr) incorporation and 3-[4,5-dimethyl-thiazol-2yl-]-2,5-diphenyl tetrazolium bromide-formazan (MTT-f) production in cultured murine fibroblasts (Balb/c 3T3) were investigated, and the synergistic and antagonistic effects of two metal cations applied simultaneously to the fibroblasts were assessed. The effects of duration of exposure were quantified using TC50 values (concentration of an element required to cause 50% toxicity compared with controls) measured after 24, 48, or 72 h. Using MTT-f production, Cd+2, Cu+2, Ga+3, and Ni+2 showed significantly lower TC50 values with increasing time of exposure, whereas the TC50 values for Ag+1 and Zn+2 remained constant. The TC50 values using 3H-Tdr incorporation exhibited a similar pattern with time of exposure, but the effects were less pronounced. The TC50 values for 3H-Tdr and MTT-f tended to equalize at 72 h. All combinations of cations tested (Ag-Cu, Ag-Zn, Ag-Ni, Cu-Zn, Cu-Ni, Ga-Ni, and Cu-Cd) exhibited synergistic and antagonistic effects as measured by MTT-f production. Synergistic and antagonistic effects were not necessarily mutually exclusive in the same system.

3T3 Cells↗

Precision of and new methods for testing in vitro alloy cytotoxicity.

Previous studies have utilized in vitro alloy cytotoxicity tests to evaluate dental casting alloys. The purposes of this study were to: (1) evaluate the precision of the optical density and visual tests previously used, (2) evaluate a new test measuring absorbance of solubilized formazan dyes, and (3) test the correlation between these tests for cytotoxicity. Balb/c 3T3 cells were plated in 24-well culture trays at 25,000 cells/cm2 around ten types of dental casting alloys (six samples/alloy) and incubated for 72 h. Cells were histochemically stained with MTT (3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyl tetrazolium bromide)/succinate for 2 h, then fixed, washed, and dried. Toxicity was measured by optical densitometer (OD) scanning, visual assessment, and 560-nm absorbance of DMSO-solubilized dyes. Measurements of rings of inhibition were not used, because they did not provide precise data, and correlated poorly with the other methods. The results were analyzed by ANOVA, Tukey intervals, and coefficients of variation (CV's). MTT required shorter incubation times for adequate staining, allowed for solubilization of the monolayers, and was less expensive than NBT (2,2'-di-p-nitro-phenyl-5,5'-diphenyl-3,3'-dimethoxy-[3,3'-dimethoxy-4,4 '-biphenylene] ditetrazolium chloride). Results showed that all three methods ranked alloy toxicities similarly (p = 0.05). The solubilization method was most discriminating due to lower CV's. Correlation between densitometer and solubilization methods was excellent (R2 = 0.96). Between-experiment CV's were generally less than 20%, and often less than 10%. Between-sample CV's were generally less than 20%.(ABSTRACT TRUNCATED AT 250 WORDS)

Analysis of Variance↗

Delineation of cytotoxic concentrations of two dentin bonding agents in vitro.

Until adhesiveness of dentin bonding agents and other restorative materials to dental structures can be assured, microleakage into resulting "gaps" and dentin permeability will remain major concerns in cases of pulpal irritation. The objectives of the present study were to (a) delineate the kinds and levels of metabolic cytotoxicity of the GLUMA and Scotchbond 2 systems as well as glutaraldehyde and 2-hydroxyethylmethacrylate, and (b) compare the effects of these same materials after diffusion through dentin discs approximately 0.5-mm thick. In monolayer cultures, glutaraldehyde was much more cytotoxic than 2-hydroxyethylmethacrylate. However, GLUMA sealer and Scotchbond 2 adhesive exhibited similar cytotoxicity in monolayer cultures. After diffusion through dentin, glutaraldehyde and 2-hydroxyethylmethacrylate effects were diluted 14.7 and 26.7 times, respectively. The postdiffusional effects of the GLUMA and Scotchbond 2 systems were not significantly different and less than those effects in monolayer cultures. This study should help in the evaluation of possible causes of pulpal irritation following restorative procedures.

3T3 Cells↗

The effects of cleaning on the kinetics of in vitro metal release from dental casting alloys.

The kinetics of the release of elements from six dental casting alloys into cell-culture medium was assessed by means of atomic absorption spectroscopy. Alloys were evaluated in the polished and polished-cleaned conditions so that the effects of cleaning could be determined. Auger scanning microscopy was used for analysis of the surfaces of selected alloys before and after exposure to the cell-culture medium. Release patterns for each element were characterized by the shape of the dissolution vs. time curve, concentration of the element at 12 h as a percentage of the 72-hour concentration, and the relative slope of the curve from 48 to 72 h. Three patterns of release were observed for elements in these alloys. Type I patterns had logarithmic shapes with relatively large 12-hour concentrations and low 48-72-hour slopes. Type II patterns had logarithmic shapes but with moderate 12-hour concentrations and 48-72-hour slopes. Type III patterns were polynomial in shape, had relatively low 12-hour concentrations, and had large 48-72-hour slopes. Cleaning did not change the pattern of release but did generally significantly decrease the quantities of elements released (p = 0.05). The type of dissolution vs. time curve appeared to be dependent upon the element and the composition of the alloy. When cleaning reduced dissolution, surface analyses showed that the cleaning process increased the abundance of elements such as Au and Pd and reduced the abundance of Ag and Cu. Elements which were released from the alloys were more abundant on the surface than in the bulk in both polished and polished-cleaned conditions.(ABSTRACT TRUNCATED AT 250 WORDS)

Cadmium↗