J Korean Acad Periodontol.  2009 Sep;39(3):331-337. 10.5051/jkape.2009.39.3.331.

Induction of IL-8 and reactive oxygen species in periodontal ligament cells by Aggregatibacter actinomycetemcomitans

• Lee YS
• Park HG
• Kim SW
• Cha JH
• Yoo YJ

• KMID: 2212147
• DOI: http://doi.org/10.5051/jkape.2009.39.3.331

Abstract

PURPOSE
Interleukin (IL)-8 is one of pro-inflammatory cytokines. Reactive oxygen species (ROS) are reduced metabolites of O2. Aggregatibacter actinomycetemcomitans is one of representative periodontopathogens. To investigate the role of A. actinomycetemcomitans in IL-8 expression of periodontal ligament (PDL) cells, we estimated the production of IL-8 and ROS in A. actinomycetemcomitans treated PDL cells.
METHODS
The IL-8 production was determined by enzyme-linked immunosorbent assay. The ROS production was estimated using H2DCFDA and FACS.
RESULTS
A. actinomycetemcomitans increased the production of IL-8 and ROS at 10, 100, and 500 multiplicity of infection. N-cetylcysteine, an antioxidant of ROS, down-regulated the production of IL-8 induced by A. actinomycetemcomitans.
CONCLUSION
These results suggest that A. actinomycetemcomitans induces IL-8 production and ROS may act as a mediator in this process.

Keyword

Aggregatibacter actinomycetemcomitans; interleukin-8; periodontal ligament; reactive oxygen species

MeSH Terms

Cytokines
Enzyme-Linked Immunosorbent Assay
Fluoresceins
Interleukin-8
Interleukins
Periodontal Ligament
Reactive Oxygen Species
Cytokines
Fluoresceins
Interleukin-8
Interleukins
Reactive Oxygen Species

Figure

• 

  Figure 1 Induction of IL-8 production in periodontal ligament cells treated with A. actinomycetemcomitans. Periodontal ligament cells were treated with A. actinomycetemcomitans at 10, 100, and 500 MOI for 5 h(A) and 24 h (B). The concentration of IL-8 in culture supernatants was estimated by ELISA. * Significantly different from none-treated cells (P < 0.05).

• 

  Figure 2 Induction of ROS in periodontal ligament cells treated with A. actinomycetemcomitans. Periodontal ligament (PDL) cells were treated with A. actinomycetemcomitans (Aa) at 10, 100, and 500 MOI for 24 h. ROS induction in PDL cells were estimated by FACS (A). PDL cells were treated with A. actinomycetemcomitans at MOI 500 for 5 h and 24 h and ROS induction in PDL cells were estimated by FACS (B).

• 

  Figure 3 Effect of N-acetylcysteine on A. actinomycetemcomitans induced IL-8 production in periodontal ligament cells. Periodontal ligament cells were pretreated with N-acetylcysteine (NAC, 15 mM) for 30 min and then treated with A. actinomycetemcomitans (Aa) at 500 MOI for 5 h (A) and 24 h (B). The concentration of IL-8 in culture supernatants was determined by ELISA. * Significantly different from none-treated cells (P < 0.05). ** Significantly different from A. actinomycetemcomitans-treated cells (P < 0.05).

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