Biomedical subjects

John C Wataha

Publications and source records attributed to John C Wataha.

27 records · Page 2Linked to original sources

PubMed2003

Effect of subtoxic concentrations of metal ions on NFkappaB activation in THP-1 human monocytes.

THP-1 human monocytes and human peripheral blood monocytes have altered inflammatory cytokine secretion profiles after exposure to a variety of metal ions known to be released from biomaterials. Transcriptional regulation of these cytokines often involves activation of the transcription factor NFkappaB. The present study was designed to determine whether metal ion treatment of monocytes results in changes in levels of activated NFkappaB. THP-1 cells were grown in suspension in the presence of sublethal concentrations of ions of Ag(+), Co(2+), Cu(2+), Hg(2+), Ni(2+), and Pd(2+). After 24 h of exposure to metal ions, the cells were harvested, counted, and the nuclear proteins extracted. Electrophoretic mobility shift assays were performed using a (32)P-ATP end-labeled oligonucleotide consensus sequence for the NFkappaB transcription factor. DNA/protein complexes were quantified by phosphorimage analysis and compared by ANOVA (Tukey, alpha = 0.05). Exposure of THP-1 cells to 100 microM of Pd(2+) caused a significant increase in activated NFkappaB (p < 0.05) whereas treatment with 5 microM of Ag(+) resulted in significantly decreased levels of nuclear NFkappaB (p < 0.05). No other metal ions tested caused a significant change in basal levels of nuclear NFkappaB (Co(2+), Hg(2+), Ni(2+), and Cu(2+)). However, exposure to 50 microM of Cu(2+) resulted in a reproducible, though not significant, increase in nuclear NFkappaB levels. These results indicate that inflammatory responses to some metal ions may be influenced by NFkappaB-mediated transcriptional regulation.

Autoradiography↗

PubMed2003

In vitro cytotoxicity of traditional versus contemporary dental ceramics.

STATEMENT OF PROBLEM: The biocompatibility of new dental ceramics has not been assessed with the same scrutiny as has been applied to alloys and composites. Yet, the biocompatibility of ceramics is critical to the long-term success of dental prostheses because ceramics are in close contact with oral tissues for extended periods. MATERIAL AND METHODS: Five dental ceramics (2 traditional feldspathic veneer porcelains [Vita Omega and Duceragold], 2 lithium disilicate pressable materials [Stylepress and Empress-2], and a pressable leucite-based material [Empress-1]) were tested for their ability to alter cellular mitochondrial dehydrogenase activity after fabrication using a tetrazolium assay, after aging for 2 weeks in a biologic solution and after post-aging polishing with either a fine diamond or diamond polishing paste. Cellular responses were compared with polytetrafluoroethylene controls (analysis of variance, Tukey pairwise post-hoc comparison, alpha=.05). RESULTS: The feldspathic porcelains caused only mild (<25% of controls) mitochondrial suppression regardless of aging or polishing. The pressable leucite-based material initially caused a 5% stimulation (not significant) of mitochondrial activity, which decreased significantly (P<.05) by 30% with aging to levels comparable to the feldspathic porcelains, and did not change with polishing. Both lithium disilicate materials caused an initial suppression of mitochondrial activity that decreased significantly with aging, but Empress-2 was severely cytotoxic initially (<20% of controls, P<.01), and became more cytotoxic again after polishing. Stylepress was less cytotoxic initially (85% of controls, not significant) and did not become cytotoxic again after polishing. CONCLUSIONS: Dental ceramics are not equivalent in their in vitro biologic effects, even within the same class of material, and biologic safety should not be assumed. Most ceramics caused only mild in vitro suppression of cell function to levels that would be acceptable on the basis of standards used to evaluate alloys and composites. However, 1 Li-disilicate material (Empress-2) exhibited cytotoxicity that would not be deemed biologically acceptable on the basis of prevailing empirical standards for dental alloys and composites.

Aluminum Silicates↗

PubMed2003

In vitro biological response to core and flowable dental restorative materials.

OBJECTIVES: In vitro cytotoxicities of commercially available core and flowable dental restorative materials were assessed and compared to traditional resin composites. Our hypothesis was that the increased resin diluents added to achieve higher flow in flowables would increase cytotoxicities, whereas the higher filler content of core materials would decrease cytotoxicities relative to traditional resin composites. METHODS: Specimens were made under aseptic conditions, then extracted into an artificial saliva solution for 0-4 weeks, to assess the effect of aging on cytotoxicity. After extraction, specimens were tested for cytotoxicity in vitro using Balb/c fibroblasts in direct contact format. Cells were exposed to the materials for 48h, after which the mitochondrial activity of the cells was measured (MTT method). Cellular activity was normalized to Teflon negative controls. RESULTS: Core materials were uniformly and severely (<50% of Teflon cellular activity) cytotoxic initially, but several materials (Corepaste, Definite core) improved somewhat with aging in artificial saliva. Flowable materials were uniformly and severely cytotoxic with no trend toward improvement with aging. The Definite-flow was the least cytotoxic of the flowable materials, but it too was severely cytotoxic. SIGNIFICANCE: Commercially available core and flowable restorative materials showed severe in vitro cytotoxicities that are worse than some traditional composites and most dental casting alloys and amalgams used today. Of particular note was the persistent cytotoxicity of these materials after 4 weeks of extraction with artificial saliva. These cytotoxicities indicate a continuing release of mass from these materials at levels that have biological relevance in vitro. In vivo relevance of these cytotoxicities is less clear, but these results indicate a higher biological risk for these materials compared to traditional materials that exhibit less initial toxicity and improve with aging time.

3T3 Cells↗

PubMed2003

Sublethal, 2-week exposures of dental material components alter TNF-alpha secretion of THP-1 monocytes.

OBJECTIVES: The aim of this study was to investigate the hypothesis that dental material components alter cytokine secretion from monocytes if applied for several weeks at sublethal doses. The current study significantly extended exposure times of monocytes to the components over times published in previous studies. These exposure times approached the estimated average life span of monocytes in the bloodstream. METHODS: Human THP-1 monocytes were exposed to 2-hydroxyethylmethacrylate (HEMA, 0-1.2mmol/l), triethyleneglycoldimethacrylate (TEGDMA, 0-0.75mmol/l), Hg(2+) (0-2 micromol/l), or Ni(2+) (0-20 micromol/l) for 2 weeks. The cells were then collected and additionally incubated for 24h, with or without bacterial lipopolysaccharide (LPS), a common component of dental plaque. TNF-alpha secretion from THP-1 was determined using by enzyme-linked immunosorbent assay. RESULTS: None of the dental material components induced TNF-alpha from THP-1 by themselves, but LPS alone strongly induced TNF-alpha secretion as expected. HEMA and TEGDMA significantly suppressed (40-70%) TNF-alpha secretion from cells stimulated with LPS. Hg(2+) at 2.0 micromol/l doubled TNF-alpha secretion from THP-1s stimulated with LPS over LPS alone. Ni(2+) did not significantly affect TNF-alpha secretion, with or without LPS exposure. Significance. The results in this study suggest that sublethal, 2-week exposures of some dental material components may alter TNF-alpha secretion from THP-1 monocytes when the cells are challenged. These alterations may influence the biological response of tissues to materials in an inflammatory intraoral environment.

Analysis of Variance↗

PubMed2003

Microtensile bond strength between adhesive cements and root canal dentin.

OBJECTIVES: The hypotheses tested were that the bond strength of adhesive cements to root canal dentin (1) would be reduced as a function of configuration factor, polymerization process and type of luting material and (2) would be lowered near the apex of the tooth. METHODS: Human canines and premolars were prepared for post cementation using Single Bond/Rely X ARC, ED Primer/Panavia F, C and B Metabond, and Fuji Plus. The specimens were divided into two groups. For intact roots, the posts were luted using standard clinical procedures. For flat roots, the posts were applied directly into flat ground canals. All roots were sectioned into 0.6 mm thick slices, trimmed mesio-distally and stressed to failure at 1 mm/min. The muTBS of each slab was calculated as the force at failure divided by the bonded cross-sectional surface area. The results were compared using a one-way ANOVA and Tukey multiple comparison intervals (alpha=0.05). Least squares linear regression analysis was used to assess the effect of dentin location on bond strength. RESULTS: All cements showed significantly (p</=0.05) lower bond strengths in intact vs. flat roots. The muTBS of posts to intact roots were not significantly different for Single Bond/Rely X ARC and Panavia F, but both were significantly lower (p</=0.05) than the bonds produced by C and B Metabond and Fuji Plus cements. For Single Bond/Rely X ARC and Fuji Plus a significant decrease in bond strength was observed in dentin closer to the apex of the root. SIGNIFICANCE: Stresses from polymerization shrinkage and problems with adequate access to the root canal complicate the formation of high-strength bonds when cementing endodontic posts with resin cements.

Analysis of Variance↗

PubMed2003

Effect of hydrolyzed surface layer on the cytotoxicity and chemical resistance of a low fusing porcelain.

OBJECTIVE: The objectives of this study were to verify the formation of a hydrolyzed surface layer on Duceram LFC and to determine its effect on the cytotoxicity of the porcelain as measured by cellular activity and concentrations of leached ionic species. METHODS: Specimens were fabricated from dentin porcelain by a vibration blotting technique. Half of the specimens underwent accelerated aging by subjecting them to the standard test for hydrolytic resistance (ISO 6872). Fibroblast cell cultures were placed in direct contact with specimens. Cell viability was assessed using succinic dehydrogenase activity. Chemical resistance was determined by leaching specimens with a continuous flow of deionized water and measuring the concentrations of soluble ions in the leachates. Porcelain surface topography was examined using atomic force microscopy, and Fourier transform infrared spectroscopy was used to detect the composition of the surface layer. RESULTS: Hydrolyzation treatment created a smooth texture on the porcelain surfaces but did not result in a hydrolyzed surface layer with increased hydroxyl content. There was a decreased alkali ion content in the surface layer of hydrolyzed specimens. Mean cellular SDH activities for non-hydrolyzed and hydrolyzed porcelain were 75+/-7 and 80+/-5% of Teflon controls, respectively. Only sodium ions were present in significant concentrations in collected leachates. The sodium concentration decreased over the initial 4.5h of leaching. SIGNIFICANCE: The changes in surface layer texture and composition as the result of the aging treatment had little effect on the cytotoxicity and chemical resistance of Duceram LFC. The results suggest that Duceram LFC would pose no biocompatibility risk even after extended exposure to the oral environment. These data also provide baseline material properties to be used in future studies of the effects of porcelain additives.

3T3 Cells↗

PubMed2003

Viscoelastic properties of demineralized dentin matrix.

OBJECTIVES: To evaluate the viscoelastic properties of demineralized dentin matrix. Stress-relaxation studies were done on matrices in tension and strain elongation or creep studies were done in both tension and compression. METHODS: Mid-coronal dentin disks were prepared from extracted unerupted human third molars. Disks were 0.5 mm thick for stress-relaxation or tensile creep experiments and 0.2-0.3 mm thick for compressive creep studies. 'I' beam specimens were prepared from dentin disks and the middle region was demineralized in 0.5 M EDTA (pH 7) for 4 days. The specimens were held in miniature friction grips in water and pulled at 100 micro m s(-1) to strains of 5, 10, 15 or 20% and then held for 10 min to follow the decay of stress over time. Creep was determined on demineralized dentin immersed in water in tension and in compression. Compressive creep was measured using an LVDT contact probe with loads of 0.02-0.5 N. Strain data were converted to compliance-time curves (strain/stress) and expressed as total compliance (J(t)), instantaneous elastic compliance (J(o)), retarded elastic compliance (J(R)) and viscous response (t/eta) or creep. RESULTS: The dentin matrix exhibits both stress-relaxation and creep behavior. Stress-relaxation and tensile creep were independent of strain but compressive creep rates were inversely related to compressive strain. Creep values were about 10% at low compressive strains, but fell progressively to 1% at high strains. Compliance-time curves fell with stress and came closer together. However, tensile creep was about 3% regardless of the strain. SIGNIFICANCE: The dentin matrix exhibits viscoelastic properties, but is not linearly viscoelastic. The relatively high creep rates of the matrix under low compressive loads may cause viscous deformations in poorly infiltrated hybrid layers in resin-bonded teeth under function.

Analysis of Variance↗

PubMed2002

Effect of toothbrushing on the toxicity of casting alloys.

STATEMENT OF PROBLEM: The biological properties of casting alloys have been assessed largely under passive conditions. The effect of common intraoral stresses such as brushing, toothpastes, and low pH on alloy toxicity are not known. PURPOSE: This study assessed the toxicity of 5 types of casting alloys commonly used in prosthodontics after toothbrushing, brushing in an acidic environment, or brushing with toothpaste. These toxicities were compared with those observed without any brushing. MATERIAL AND METHODS: Au-Pt, Au-Pd, Pd-Cu-Ga, Ni-Cr-Be, and Ni-Cr (no Be) alloys were brushed for 48 hours in a toothbrushing machine at 90 strokes/minute and 200g force. Alloys were brushed with either saline at pH 7, saline at pH 4 (acidified with sodium lactate), or saline with 1:7 (wt/wt) toothpaste. After the brushing regimen, the cytotoxicity of the alloys was assessed in a standard in vitro test. Cytotoxicities of the alloys after different brushing treatments were compared with unbrushed (control) specimens. Analysis of variance (ANOVA) and Tukey multiple comparison intervals (alpha=.05) were used to identify significant differences among brushing conditions. RESULTS: Brushing at pH 7 significantly increased the toxicity of the Pd-Cu-Ga alloy (15% to 20% over unbrushed specimens). Brushing at pH 4 increased the toxicity of the Au-Pt and Au-Pd alloys by 30% and the Pd-Cu-Ga alloy by >40%. The Ni-based alloys were not affected by acid. After being brushed with toothpaste, both Ni-based alloys were significantly more toxic, but Ni-Cr-Be was the worst, increasing more than 60% in toxicity over the controls. The toxicity of the Au-Pd alloy also increased significantly (15%). CONCLUSION: Brushing dental casting alloys may increase their cytotoxicity in vitro, but the increase depends heavily on the alloy type and brushing condition.

3T3 Cells↗

PubMed2002

Alloys for prosthodontic restorations.

The numbers and types of alloys for prosthodontic restorations have increased dramatically over the past 25 years, making selection of an alloy for a given clinical situation difficult. Factors such as cost, the need for better strength, and worries about alloy corrosion have pressured the alloy market to change significantly. A number of properties--including yield strength, hardness, elastic modulus, microstructural phases, grain size, corrosion performance, coefficient of thermal expansion, oxide color, and melting range--are relevant to the proper selection of an alloy for a given clinical problem. In this article, a brief historical look at prosthodontic alloys and the nomenclature for alloys is followed by a discussion of the most important physical properties of alloys for clinical practice. A summary of the types of alloys available today and their classification is then presented. Finally, speculations about future trends for alloys are made, and simple guidelines are suggested to help dentists choose appropriate alloys for their practices. This review excludes implant alloys, dental amalgam, and alloys for orthodontic and endodontic applications.

Biocompatible Materials↗