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

Yang-Sin Lee; Hong Gyu Park; Sung Whan Kim; Jeong-Heon Cha; Yun-Jung Yoo

doi:10.5051/jkape.2009.39.3.331

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Lee, Park, Kim, Cha, and Yoo: Induction of IL-8 and reactive oxygen species in periodontal ligament cells by Aggregatibacter actinomycetemcomitans

Original Article

The Journal of the Korean Academy of Periodontology

The Journal of the Korean Academy of Periodontology 2009; 39(3): 331-337.

Published online: 30 September 2009

DOI: https://doi.org/10.5051/jkape.2009.39.3.331

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

Yang-Sin Lee¹, Hong Gyu Park, Sung Whan Kim^2,³, Jeong-Heon Cha^1,^2,³, Yun-Jung Yoo^1,^2,³

¹Department of Oral Biology, College of Dentistry, Yonsei University, Korea.

²Department of Applied Life Science, The Graduate School, Yonsei University, Korea.

³BK21 project, Oral Science Research Center, and Research Center for Orofacial Hard Tissue Regeneration, College of Dentistry, Yonsei University, Korea.

Correspondence: Yun-Jung Yoo. Department of Oral Biology, College of Dentistry, Yonsei University, 134 Sinchon-dong, Seodaemoon-gu, Seoul, 120-752, Korea. yu618@yuhs.ac, Tel: 02-2228-3060, Fax: 02-2227-7903

Received 13 June 2009 Accepted 10 August 2009

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-acetylcysteine, an antioxidant of ROS, down-regulated the production of IL-8 induced by A. actinomycetemcomitans.

Conclusions

These results suggest that A. actinomycetemcomitans induces IL-8 production and ROS may act as a mediator in this process.

Keywords: Aggregatibacter actinomycetemcomitans, interleukin-8, periodontal ligament, reactive oxygen species

Figures and Tables

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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