Alexander Franz

Metal ion-induced toxic histamine release from human basophils and mast cells

Recent data suggest that distinct metal ions can be released from dental alloys or other biomaterials, and may cause toxic effects on various cells. In this study, the effects of 14 metal ions on histamine release from human blood basophils (n = 4), isolated tissue mast cells (lung n = 8, uterus n = 2, skin n = 1, gingiva n = 1), the basophil cell line KU-812, and the mast cell line HMC-1 were analyzed. Of the 14 metal ions, Ag+ (0.33 mM) and Hg2+ (0.33 mM) were found to induce release of histamine in blood basophils, KU-812, mast cells, and HMC-1. The effects of Ag+ and Hg2+ were dose dependent and were observed within 60 min of incubation. In primary mast cells and basophils, AU3+ (0.33 mM) also induced histamine release, whereas no effects of Au3+ on HMC-1 or KU-812 cells were seen. The other metal ions showed no effects on primary or immortal cells within 60 min. However, Pt4+ (0.33 mM) induced histamine liberation in HMC-1 and lung mast cells after 12 h. The Ag+- and Hg2+-induced rapid release of histamine from HMC-1 was associated with ultrastructural signs of necrosis, but not apoptosis. In contrast, prolonged exposure to Pt4+ (0.33 mM, 14 h) induced apoptotic cell death in HMC-1 cells, as assessed by electron microscopy and DNA analysis. Together, certain metal ions induce distinct cytopathogenic effects in mast cells and basophils. Whereas Ag+, Hg2+, and Au3+ cause direct toxicity, Pt4 causes cell death through induction of apoptosis. Whether such effects contribute to local adverse reactions to metal-containing biomaterials in vivo remains to be determined.

Cytotoxicity of a calcium aluminate cement in comparison with other dental cements and resin-based materials

The objective of this study was to compare the cytotoxic effects of a calcium aluminate cement with several currently used direct restorative materials. Specimens of three composites (QuiXfil, Tetric Ceram, Filtek Supreme), one zinc phosphate cement (Harvard Cement), one glass ionomer cement (Ketac Molar), and one calcium aluminate cement (DoxaDent), were used fresh or after 7-days’ preincubation in cell culture medium at 37°C, pH 7.2. PVC strips for ISO 10993-5 cytotoxicity test were used as positive control and glass specimens as negative control. L-929 fibroblasts (5-ml aliquots, containing 3×104 cells/ml), cultivated in DMEM with 10% FCS, 1% glutamine, and 1% penicillin/streptomycin at 37°C/5% CO2 and trypsinized, were exposed to the specimens for 72 h. The cells were harvested, centrifuged, and resuspended in 500 µl DMEM and then counted in 500 µl DMEM for 30 s with a flow cytometer at 488 nm. The analysis of variance comparing the six materials showed different influences on L-929 fibroblast cytotoxicity (p<0.0001). The cytotoxicity of all specimens diminished with increasing preincubation time (p<0.0001). Fresh DoxaDent exhibited the lowest cytotoxicity, followed by QuiXfil. Ketac Molar showed the highest cytotoxicity. After 7 days of preincubation, Harvard Cement and Filtek Supreme demonstrated more cytotoxicity than the other materials (p<0.005).

Cytotoxicity and shear bond strength of four orthodontic adhesive systems.

The objective of this study was to compare the cytotoxicity of four orthodontic bonding systems, Light Bond, Enlight, Concise, and Transbond, and to evaluate their shear bond strength (SBS). These orthodontic bonding materials were applied to metal brackets (Mini Diamond). Glass specimens were used as controls in all experiments. Only Concise was a chemically cured system, the other systems were light cured. The specimens were added to L-929 fibroblast cultures immediately after fabrication or after pre-incubation for 7 days. The incubation time was 72 hours and the cells were counted by flow cytometry. One hundred and fifty-seven freshly extracted human third molars were used for testing the SBS in a universal testing machine. Statistical significance was determined using analysis of variance followed by post hoc comparisons for multiple-level alpha control. Pairwise comparisons showed a significant difference only between Light Bond and Concise (P = 0.0126). The highest SBS was obtained with Light Bond (23.23 +/- 1.53 MPa) followed by Transbond (20.39 +/- 1.18 MPa) and Enlight, (20.32 +/- 1.06 MPa). Concise (17.87 +/- 1.04 MPa) showed the lowest SBS. The cytotoxicity of all light-cured systems for fresh specimens was comparable, whereas the chemically cured system, Concise, was significantly more cytotoxic. After 7 days of pre-incubation, all systems were significantly less cyotoxic than fresh specimens (P < 0.001). Brackets alone were not cytotoxic. All bonding systems showed a clinically satisfactory bond strength higher than 10 MPa, with the chemically cured system showing the lowest SBS.

The influence of internote-interval variation of the advertisement call on the phonotactic behaviour in male Allobates femoralis (Dendrobatidae)

Acoustic conspecific recognition depends on spectral and temporal call properties, but not all of these are necessary for species identification. To test the importance of the advertisement calls internote interval of the dendrobatid frog Allobates (=Epipedobates) femoralis, we conducted field playback experiments with modified conspecific calls and investigated phonotactic responses. Allobates femoralis males showed positive phonotaxis when calls with internote intervals varying up to ±60% of the populations mean were presented. No phonotactic approach was observed in response to modified advertisement calls containing internote intervals of the syntopic heterospecific leptodactylid frog Adenomera hylaedactyla. However, 26.7% of the A. femoralis males tested showed positive phototoxic when altered advertisement calls with internote intervals of the syntopic heterospecific dendrobatid frog Epipedobates cf. hahneli were presented. Our results show that only a combination of different acoustic call parameters provides correct species-specific recognition.

Effects of dental amalgam and heavy metal cations on cytokine production by peripheral blood mononuclear cells in vitro

The effects of dental amalgam on cytokine production by human peripheral blood mononuclear cells (PBMC) from healthy donors were analyzed. To induce cytokine production, PBMC were stimulated with lipopolysaccharide, phytohemagglutinin, or staphylococcal enterotoxin A and cultured for 48 h in the presence of either freshly prepared amalgam, aged amalgam, or amalgam-conditioned culture medium (ACCM). The concentrations of several cytokines were measured in PBMC supernatants by enzyme-amplified sensitivity immunoassays (EASIAs). Freshly prepared amalgam as well as ACCM induced a decrease in the production of interferon-gamma (IFN-gamma) and interleukin-10 (IL-10), and an increase in the concentrations of tumor necrosis factor-alpha (TNF-alpha). Both fresh amalgam and ACCM showed no effects on IL-2, IL-6, or granulocyte-macrophage colony-stimulating factor levels. Amalgam aged for 6 weeks did not affect the concentration of any of the above cytokines. To investigate which heavy metal cations released from amalgam caused the observed immunomodulatory effects, Cu2+, Hg2+, and Sn2+, which were detected in amalgam supernatants by inductively coupled plasma atomic spectrophotometry, were added as salts to the cultures. Cu2+ and Hg2+ induced a decrease in IFN-gamma and IL-10 levels, and Hg2+ an increase in TNF-alpha concentrations. Cytokine production was not significantly modulated by Sn2+. Under these experimental conditions, release of Ag+ into culture medium was not detectable. However, Ag+ markedly suppressed the production of IFN-gamma, IL-10, and TNF-alpha. In summary, our results show that fresh amalgam, but not amalgam aged for 6 weeks, causes changes in the cytokine pattern of PBMC in vitro, and that these effects are due to the release of Cu2+ and Hg2+.

Influence of dental amalgam and heavy metal cations on in vitro interleukin-1β production by human peripheral blood mononuclear cells

The influence of dental amalgam and heavy metal cations on interleukin-1beta (IL-1beta) expression by peripheral blood mononuclear cells from healthy donors was studied. A marked decrease in the production of IL-1beta was caused by freshly prepared amalgam or amalgam-conditioned culture medium, but not by amalgam aged for 6 weeks. When metal cations were added as salts, Cu(2+), Hg(2+), and Ag(+) at high concentrations (33.3 and 333.3 microM) were highly inhibitory. Among other heavy metal cations, Au(3+), Pt(4+), Ni(2+), Pd(2+), but not Ga(3+) or Sn(2+), inhibited IL-1beta production in a concentration-dependent manner. Flow cytometry studies indicated that Hg(2+) and Ag(+) strongly reduced the percentage of CD14(+) cells containing IL-1beta intracellularly. As shown by Northern blot analysis, Hg(2+) inhibited the level of IL-1beta-specific mRNA by 28% at 3.3 microM and completely at 33.3 microM. Only slight inhibitory effects were induced by Cu(2+) at 33.3 microM. Interestingly, Ag(+) at a concentration of 3.3 microM increased twofold the amount of IL-1beta-specific mRNA. Our data show that IL-1beta production is altered at protein and mRNA levels by components released from fresh amalgam and by other heavy metal cations, suggesting a role of these cations in changes in the cell phenotype and IL-1-mediated cell functions.

Reprocessing of dental instruments in washer-disinfectors: does a representative test soil exist in dentistry?

Background: Reprocessing of medical devices, being classified as semi-critical B is recommended to be performed in a washer-disinfector. In order to estimate, whether the expected contaminants of the various medical disciplines can be effectively removed by this washer-disinfector, different so called “test soils” have been proposed to be tested as a marker of cleaning efficacy of the disinfector. Todays described test soils are optimised for the testing of contaminations occurring in surgical procedures, but not for dental procedures. Methods: In this study the test soils being proposed in the EN 15883-5 (e.g. KMNE soil, recipe by Koller and coagulated sheep’s blood) were compared with 8 reference substances used in the conservative-prosthetic dental practice. The success of the cleaning efficacy in the washer-disinfector was checked visually and by determining the residual protein concentration on the contaminated instruments after the cleaning procedure. Results: It could be shown that in contrast to the proposed test soils of the EN 15883-5, the used reference substances of the dental practice could not be removed by the washer-disinfector. Removal of these reference substances was only possible after manual or ultrasonic cleaning. Conclusions: Since blood plays a subordinate role as a contaminant of instruments during conservative-prosthetic dental treatments, testing of the cleaning efficacy of the washer-disinfector with test soils according to the proposals of the EN 15883-5 is not representative in this discipline of dentistry. Most of the materials used in dental practice can only be removed manually or with the help of the ultrasound bath.

Evaluation of the Morphological Characteristics of Laser-Irradiated Dentin

OBJECTIVE The aim of this study was to investigate the effect of different energy settings of Er:YAG laser irradiation on dentin surface morphology with respect to the number of opened dentinal tubules. BACKGROUND DATA An ideally prepared dentin surface with opened dentinal tubules is a prerequisite for adhesive fixation. No study, however, has yet compared the numbers of opened dentinal tubules with regard to statistical differences. METHODS Conventional preparations using a bur with or without additional acid etching acted as control groups. Dentin specimens were prepared from human third molars and randomly divided into eight groups according to the energy settings of the laser (1, 1.5, 4, 6, 7.5, and 8 W) and two controls (bur and bur plus acid etching). After surface preparation, dentin surfaces were analyzed with a scanning electron microscope, and the number of opened dentinal tubules in a defined area was counted. RESULTS The control groups showed smooth surfaces with (bur plus acid etching) and without opened dentinal tubules (bur), whereas all laser-irradiated surfaces showed rough surfaces. Using the energy setting of 4 W resulted in significantly more opened dentinal tubules than the conventional preparation technique using the bur with additional acid etching. In contrast, the energy setting of 8 W showed significantly fewer opened dentinal tubules, and also exhibited signs of thermal damage. CONCLUSIONS The Er:YAG laser with an energy setting of 4 W generates a dentin surface with opened dentinal tubules, a prerequisite for adhesive fixation.

The Effects of CO2 Laser with or without Nanohydroxyapatite Paste in the Occlusion of Dentinal Tubules

The aim of this study was to evaluate a new treatment modality for the occlusion of dentinal tubules (DTs) via the combination of 10.6 µm carbon dioxide (CO2) laser and nanoparticle hydroxyapatite paste (n-HAp). Forty-six sound human molars were used in the current experiment. Ten of the molars were used to assess the temperature elevation during lasing. Thirty were evaluated for dentinal permeability test, subdivided into 3 groups: the control group (C), laser only (L−), and laser plus n-HAp (L+). Six samples, two per group, were used for surface and cross section morphology, evaluated through scanning electron microscope (SEM). The temperature measurement results showed that the maximum temperature increase was 3.2°C. Morphologically groups (L−) and (L+) presented narrower DTs, and almost a complete occlusion of the dentinal tubules for group (L+) was found. The Kruskal-Wallis nonparametric test for permeability test data showed statistical differences between the groups (P < 0.05). For intergroup comparison all groups were statistically different from each other, with group (L+) showing significant less dye penetration than the control group. We concluded that CO2 laser in moderate power density combined with n-HAp seems to be a good treatment modality for reducing the permeability of dentin.

Cytotoxicity of post and core composites as a function of environmental conditions

OBJECTIVES In the revised version of ISO 7405 there are so far no detailed recommendations concerning temperature and humidity during specimen production for light curing and chemically setting dental materials. The main objective of the present study was to observe if different environmental conditions during specimen production influence cytotoxicity and degree of conversion of four post and core composite materials and to investigate if cytotoxicity of post and core materials is influenced by their corresponding bonding substances. METHODS Specimens of four different post and core composite materials (LuxaCore - Dual, Core X-Flow, Flow White and MultiCore Flow) were produced in a climate test chamber at 23°C/50% relative humidity or 37°C/95% relative humidity and were dual-cured or self-cured, with or without their corresponding bonding substances. Specimens were added to cell cultures immediately after production or after preincubation for 7 days. Specimens were incubated with L-929 fibroblasts for 72h and cell numbers determined by a flow cytometer. FTIR spectroscopic measurements of post and core materials were performed at the same temperature conditions as for the cytotoxicity assay (23°C or 37°C). RESULTS Dual-cured specimens of all post and core composites exhibited less cytotoxicity under both environmental conditions than self-cured specimens. All self-cured specimens manufactured at 37°C/95% showed less cytotoxicity than specimens produced at 23°C/50%. All dual-cured specimens showed similar cytotoxicity at both environmental conditions. After 7 days of preincubation most dual-cured specimens produced at 23°C/50% showed less cytotoxicity than self-cured specimens (with the exception of Flow White). Compared to fresh specimens, 7-day aged specimens of most materials showed reduced cytotoxicity. Materials already showing low cytotoxicity as fresh specimens did not further reduce their cytotoxicity after 7 days of preincubation. For dual-cured materials the degree of conversion was higher compared to self-cured materials. SIGNIFICANCE Different temperatures during specimen production have an impact on cytotoxicity and degree of conversion of dual-curing composite materials. Detailed recommendations for standardization concerning environmental conditions during specimen production are required.