Regina L. W. Messer

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.

Biological effects of blue light from dental curing units

OBJECTIVES This study assessed the effects of three common dental photo-curing light sources (quartz-tungsten-halogen (QTH), plasma-arc (PAC), and laser) on the cellular function of fibroblasts in vitro. METHODS Mouse fibroblasts were exposed to light from dental photo-curing units for clinically relevant durations, with total energy exposures ranging from 1.3 to 60 J/cm(2). The temperature rise of the cell-culture medium was measured to assess any possible effect from temperature increases, and cellular function was assessed by succinic dehydrogenase (SDH) activity of mitochondria. To directly compare the three light sources, additional experiments were done using equivalent total energy exposures from each source by adjusting the exposure durations for each unit. RESULTS In experiments that used clinically relevant exposure durations for each light, exposures ranging from 5 J/cm(2) (laser) to 15 J/cm(2) (PAC, QTH) irreversibly suppressed SDH activity nearly 100% when compared to no-light controls up to 72 h post-exposure. For the PAC and QTH sources, exposures as low as 3.5 J/cm(2) also irreversibly suppressed SDH activity. When equivalent energies were used from each light source, exposures of 1 J/cm(2) did not suppress SDH activity for the QTH and laser sources, but significantly (50%) suppressed SDH for the PAC source, indicating a difference in the biological effects of the outputs of the different curing units. Equivalent energy exposure experiments also indicated a definite dependence of SDH activity on the total light energy of exposure. Temperature rises ranged from 2 to 9 degrees C, and elevated temperatures lasted for 60-300 s above the base temperature of 37 degrees C, but peak temperature and the duration of temperature elevation were not always related and depended on the light source used. SIGNIFICANCE Results from the current study indicate that these photo-curing sources pose some risk of disrupting cellular function in vivo. Further study is necessary in other cell types and under more clinically relevant conditions to estimate the in vivo risk of photo-curing to oral tissues.

Quaternary ammonium silane-functionalized, methacrylate resin composition with antimicrobial activities and self-repair potential.

The design of antimicrobial polymers to address healthcare issues and minimize environmental problems is an important endeavor with both fundamental and practical implications. Quaternary ammonium silane-functionalized methacrylate (QAMS) represents an example of antimicrobial macromonomers synthesized by a sol-gel chemical route; these compounds possess flexible Si-O-Si bonds. In present work, a partially hydrolyzed QAMS co-polymerized with 2,2-[4(2-hydroxy 3-methacryloxypropoxy)-phenyl]propane is introduced. This methacrylate resin was shown to possess desirable mechanical properties with both a high degree of conversion and minimal polymerization shrinkage. The kill-on-contact microbiocidal activities of this resin were demonstrated using single-species biofilms of Streptococcus mutans (ATCC 36558), Actinomyces naeslundii (ATCC 12104) and Candida albicans (ATCC 90028). Improved mechanical properties after hydration provided the proof-of-concept that QAMS-incorporated resin exhibits self-repair potential via water-induced condensation of organic modified silicate (ormosil) phases within the polymerized resin matrix.

Effects of an Experimental Calcium Aluminosilicate Cement on the Viability of Murine Odontoblast-like Cells

INTRODUCTION Quick-setting calcium aluminosilicate cement with improved washout resistance is a potential substitute for calcium silicate cements in endodontics. This study examined the effect of an experimental calcium aluminosilicate cement (Quick-Set; Primus Consulting, Bradenton, FL) on the viability of odontoblast-like cells. METHODS The biocompatibility of Quick-Set and white ProRoot MTA (WMTA; Dentsply Tulsa Dental Specialties, Tulsa, OK) cements and their eluents was evaluated using a murine dental papilla-derived odontoblast-like cell line (MDPC-23); 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay was used to examine the effects of the 2 hydraulic cements on mitochondrial metabolic activity. Flow cytometry and confocal laser scanning microscopy were used to identify the effects of the 2 cements on cell death-induced plasma membrane permeability to fluorescent dyes and DNA stains. RESULTS After the first week of immersion in culture medium, Quick-Set and WMTA were more cytotoxic than the Teflon-negative control (P < .05), and the cells exhibited more apoptosis/necrosis than Teflon (P < .05). After the second week of immersion, the 2 cements were as biocompatible as Teflon (P > .05), with cells exhibiting minimal apoptosis/necrosis. Eluents from the set cements at 1:1 dilution were significantly more cytotoxic that eluents at 1:10 or 1:100 dilution (P < .05). CONCLUSIONS Quick-Set and WMTA exhibited similar cytotoxicity profiles. They possess negligible in vitro toxicologic risks after time-dependent elution of toxic components.

Cytotoxicity of endodontic materials over 6-weeks ex vivo.

AIM To test the hypothesis that extending the time of a traditional ex vivo cytotoxicity test helps to identify trends in the behaviour of root core materials and sealers, which could ultimately aid in predicting their clinical safety and performance. METHODOLOGY Endodontic sealers and core specimens were initially tested in direct contact with L929 fibroblasts for 72 h. Cell response was estimated by measuring cellular succinate dehydrogenase activity relative to Teflon controls. Cytotoxicity (% of more active cells) was reassessed after 1, 3, 4 and 6 weeks, with the specimens stored in a physiologically balanced salt-solution between tests. RESULTS Distinct trends in cytotoxicity among both core materials and sealers were observed over the 6-week test. Four of the six sealers and two of the three core materials showed cell viabilities of <30% of Teflon after 6 weeks (>70% cytotoxicity). CONCLUSIONS The current results suggest that some endodontic materials have an elevated biological risk for extended intervals.

Periodontal ligament fibroblasts sustain destructive immune modulators of chronic periodontitis.

BACKGROUND In healthy periodontal tissue, innate immune responses effectively confine and suppress a bacterial insult. However, a disruption of the host-bacterial equilibrium may produce an overexpression of cytokines and lead to permanent, host-mediated tissue damage. Although such periodontal destruction primarily results from activated immune mechanisms, the site-specific damage suggests that local tissues participate in these pathologic changes. Periodontal ligament fibroblasts (PDLFs) are prominent in the periodontium and are critical in homeostasis and regeneration because they have the ability to produce multiple cytokines in response to a bacterial insult. These cells could play a role in the local pathogenesis of periodontal disease. METHODS We studied alkaline phosphatase (ALP) activity, interleukin (IL)-6 production, and morphologic characteristics of cultured PDLFs that were isolated from periodontally healthy sites (H-PDLFs) and diseased sites (D-PDLFs) in humans. Quantitative analyses of 84 genes that are related to inflammation were performed using real-time polymerase chain reaction arrays. RESULTS A mineralizing medium induced a significant increase of ALP in H-PDLFs, but no significant enzymatic changes were detected in D-PDLFs after such treatment. The protein and gene expression of IL6 showed a significant upregulation in D-PDLFs, which also demonstrated a significant upregulation of 54% of genes in the inflammatory gene arrays. CONCLUSIONS To our knowledge, these results represent the first biologic evidence that D-PDLFs retain uniquely inflammatory phenotypes that could maintain localized destructive signals in periodontitis. The overexpression of proinflammatory cytokines by PDLFs could amplify local inflammation by the continuous triggering of immune responses. In addition, the location of these cells could be critical in the progression of the inflammatory front into the deeper tissues.

In vitro osteogenic potential of an experimental calcium silicate-based root canal sealer.

OBJECTIVE This in vitro study compared the cytotoxicity and osteogenic potential of an experimental calcium silicate-based sealer with an epoxy resin-based sealer (AH Plus; Dentsply Caulk, Milford, DE) and a zinc oxide-eugenol-based sealer (Pulp Canal Sealer; SybronEndo, Orange, CA). METHODS Disks prepared from the respective sealer and from Teflon (negative control) were placed in direct contact with a MC3T3-E1 osteogenic cell line at 6 weekly intervals after immersion in a culture medium. Succinic dehydrogenase activities were evaluated using 3-(4,5-dimethylthiazole-2-yl)-2,5-diphenyl tetrazolium bromide (MTT) assay. Extracts from these sealers after the 6-week immersion period were investigated also by MTT assay. Aged sealers were then switched to an osteogenic medium for examination of the alkaline phosphatase activity and mineralization of extracellular matrices produced by the differentiated cells. RESULTS All sealers exhibited severe toxicity after 24 hours, after which toxicity decreased gradually over the experimental period except for Pulp Canal Sealer, which remained severely toxic. Toxicity of the extracts derived from the sealers was concentration dependent, with those derived from the experimental sealer being the least cytotoxic at a 1:10 dilution. Minimal alkaline phosphatase activity and no bone formation were seen with Pulp Canal Sealer. The production of alkaline phosphatase was less intense for the experimental sealer at 7 days. However, both AH Plus and the experimental sealer did not inhibit mineralization of the extracellular matrix after 28 days. CONCLUSION The experimental calcium silicate-based sealer may be regarded as minimally tissue irritating and does not interfere with bone regeneration even when it is inadvertently extruded through the apical constriction.

Corrosion of machined titanium dental implants under inflammatory conditions

The effects of hyperglycemia, altered cell function, or inflammatory mediators on implant corrosion are not well studied; yet, these effects are critical to implant biocompatibility and osseointegration. Because implant placement is burgeoning, patients with medically compromising systemic conditions such as diabetes are increasingly receiving implants, and the role of other inflammatory diseases on implant corrosion also needs investigation. In the current study, the corrosion properties of commercially available, machined titanium implants were studied in blood, cultures of monocytic cells, and solutions containing elevated dextrose concentrations. Implant corrosion was estimated by open circuit potentials, linear polarization resistance, and electrical impedance spectroscopy (EIS) for 26 h. In selected samples, THP1 monocytic cells were activated for 2 h with Lipopolysaccharide prior to implant exposure, and IL-1beta secretion was measured to assess the affect of the implants on monocyte activation. Implants under conditions of inflammatory stress exhibited more negative E(corr) values, suggesting an increased potential for corrosion. Linear polarization measurements detected increased corrosion rates in the presence of elevated dextrose conditions over PBS conditions. EIS measurements suggested that implants underwent surface passivation reactions that may have limited corrosion over the short term of this test. This result was supported by cyclic polarization tests. IL-1beta secretion was not altered under conditions of corrosion or implant exposure. The results suggest that inflammatory stress and hyperglycemia may increase the corrosion of dental endosseous titanium-based implants, but that longer, more aggressive electrochemical conditions may be necessary to fully assess these effects.

Ni(II) ions dysregulate cytokine secretion from human monocytes

Nickel-containing alloys are used in dentistry because of their low cost, but poor corrosion behavior increases their risk of causing adverse biological responses. Intraorally, nickel-containing alloys accumulate bacterial plaque that triggers periodontal inflammation via toxins such as lipopolysaccharide (LPS). Recent evidence suggests that in monocytes, Ni(II) amplifies LPS-induced secretion of several cytokines that mediate periodontal destruction. Thus, we investigated the effects of Ni(II), with or without LPS, on the secretion of a broader array of cytokines from monocytes. We then measured monocytic expression of two proteins, Nrf2 and thioredoxin-1 (Trx1), that influence the regulation of cytokine secretion. Cytokine arrays were used to measure the effects of 0-50 microM Ni(II) on cytokine secretion from human THP1 monocytes, with or without LPS activation. Immunoblots were used to estimate Nrf2 and Trx1 levels. Our results indicate that both Ni(II) alone and Ni(II) with LPS have broad-based effects on cytokine secretion. Ni(II) increased Nrf2 levels by threefold, and LPS amplified the effects of Ni(II) by 10-fold. Trx1 levels did not change under any condition tested. Our results suggest that Ni(II)-induced changes in cytokine secretion by monocytes are diverse and may be influenced by Nrf2 but are not likely influenced by changes in whole-cell Trx1 levels.

Corrosion of phosphate-enriched titanium oxide surface dental implants (TiUnite) under in vitro inflammatory and hyperglycemic conditions.

Endosseous dental implants use is increasing in patients with systemic conditions that compromise wound healing. Manufacturers recently have redesigned implants to ensure more reliable and faster osseointegration. One design strategy has been to create a porous phosphate-enriched titanium oxide (TiUnite) surface to increase surface area and enhance interactions with bone. In the current study, the corrosion properties of TiUnite implants were studied in cultures of monocytic cells and solutions simulating inflammatory and hyperglycemic conditions. Furthermore, to investigate whether placement into bone causes enough mechanical damage to alter implant corrosion properties, the enhanced surface implants as well as machined titanium implants were placed into human cadaver mandibular bone, the bone removed, and the corrosion properties measured. Implant corrosion behavior was characterized by open circuit potentials, linear polarization resistance, and electrical impedance spectroscopy. In selected samples, THP1 cells were activated with lipopolysaccharide prior to implant exposure to simulate an inflammatory environment. No significant differences in corrosion potentials were measured between the TiUnite implants and the machined titanium implants in previous studies. TiUnite implants exhibited lower corrosion rates in all simulated conditions than observed in PBS, and EIS measurements revealed two time constants which shifted with protein-containing electrolytes. In addition, the TiUnite implants displayed a significantly lower corrosion rate than the machined titanium implants after placement into bone. The current study suggests that the corrosion risk of the enhanced oxide implant is lower than its machined surface titanium implant counterpart under simulated conditions of inflammation, elevated dextrose concentrations, and after implantation into bone.