CCL28 effects on periodontal pathogens.

BACKGROUND: Chemokines are small proteins that signal to and attract cells of the immune system; they are vital components in the modulation of immunity and wound healing. A newly described chemokine was reported to have antibacterial and antifungal activity. This chemokine, chemokine (C-C motif) ligand 28 (CCL28; also called mucosae-associated epithelial chemokine), is secreted by mucosal epithelial cells and is found in saliva and in breast milk. The objective of this study was to test whether CCL28 has antibacterial activity against two anaerobic periodontal pathogens: Porphyromonas gingivalis and Actinobacillus actinomycetemcomitans. METHODS: We used a bacterial viability test, in which two fluorescent dyes are bound differentially to living and killed bacteria. We tested the bacteria at concentrations of 2 x 10(7)/ml, exposing them to CCL28 at concentrations from 0.04 to 10 microM. RESULTS: CCL28 was effective at killing both organisms. After 1 hour of exposure to the chemokine under appropriate oxygen conditions, the percentage of living organisms was reduced significantly for each species. We estimated the 50% effective concentration to be approximately 0.7 microM for P. gingivalis and approximately 2.0 microM for A. actinomycetemcomitans (N = five experiments each). We confirmed these observations using standard bacterial plating methods. CONCLUSION: Because this chemokine is secreted into the saliva, a reduction in salivary flow (as in xerostomia) may diminish the oral self-defense mechanisms by also reducing the exposure of bacteria to the antibacterial action of CCL28.

Efficacy of two contemporary single-cone filling techniques in preventing bacterial leakage.

This in vitro study evaluated the sealing efficacy of three root-filling systems/techniques in preventing bacterial leakage. Instrumented single-rooted root segments were filled with (1) warm vertical compaction with gutta-percha/AH Plus; (2) single-cone technique with ActiV GP; and (3) single-cone technique with Gutta-Flow. A dual-chamber leakage model using S. mutans as a microbial marker was used for leakage evaluation. Bacterial penetration was monitored over a 100-day period. Leakage was recorded when turbidity was observed in the lower chamber. Gutta-percha warm vertical compaction exhibited the best seal with bacterial leakage observed in only 16.7% of the specimens between 59 and 100 days. All ActiV GP specimens leaked between 7 and 100 days; 50% of the Gutta-Flow specimens leaked between 22 and 100 days. The two contemporary single-cone techniques did not insure a durable apical seal against bacterial leakage. A warm vertical compaction technique using thermoplasticized gutta-percha and AH Plus sealer appears to be more effective in minimizing bacterial leakage.

Sublethal concentrations of Au (III), Pd (II), and Ni(II) differentially alter inflammatory cytokine secretion from activated monocytes.

Many transition metals have been viewed collectively as nonspecific biological toxins in cells, which has limited investigation into their possible therapeutic effects. In the current study, the effects of Au(III), Ni(II), and Pd(II) on the differential secretion of cytokines from monocytes has been investigated. This is critical to understanding any therapeutic potential of these metals, their allergenicity, or the clinical effects of current metal therapies such as chrysotherapy. Lethal concentrations (defined as > 50% suppression of mitochondrial succinate dehydrogenase (SDH) activity) of metals were determined by dose-response curves with the use of 72 h exposures to human THP-1 monocytes. Then, secretion of TNFalpha, IL1beta, and IL6 were measured after the monocytes were exposed to sublethal concentrations of metals, with or without stimulation by lipopolysaccharide. The concentrations of Au(III), Pd(II), and Ni(II) required to suppress SDH activity by 50% were found to be 255, 270, and 90 microM, respectively. No sublethal concentration of any metal alone caused secretion of the cytokines. However, LPS-induced cytokine secretion was significantly and differentially altered by sublethal concentrations of each metal. Differential responses were highly dependent on metal concentration and involved both suppression and potentiation of the LPS activation. In the case of Ni(II), potentiation of TNFalpha, IL1beta, and IL6 ranged from 200% for TNFalpha to over 1200% for IL6. Metals such as Au(III), Pd(II), and Ni(II) differentially alter cytokine expression from monocytes. These results imply that metals have more specific effects on cell signaling than previously assumed. These results also are important in explaining multiple clinical effects often seen with chrysotherapy, identifying potential new avenues for metal therapy, and understanding the inflammatory effects of metals such as nickel.

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.