April 2011
Volume 52, Issue 14
Free
ARVO Annual Meeting Abstract  |   April 2011
Expression Of Inflammatory Cytokines And Receptors Using An Ex Vivo, Dry-Eye Model Under Hyperosmotic Stress
Author Affiliations & Notes
  • Eun-Soon Kim
    Laboratory of Genome Research, Asan Institute for Life Sciences, Asan Medical Center, Seoul, Republic of Korea
  • Sun Young Kim
    Laboratory of Genome Research, Asan Institute for Life Sciences, Asan Medical Center, Seoul, Republic of Korea
  • Mi Hyun Cheon
    Department of Ophthalmology, University of Ulsan College of Medicine, Asan Medical Center, Seoul, Republic of Korea
  • Dong-Ah Ko
    Department of Ophthalmology, University of Ulsan College of Medicine, Asan Medical Center, Seoul, Republic of Korea
  • Jae-hyung Kim
    Department of Ophthalmology, University of Ulsan College of Medicine, Asan Medical Center, Seoul, Republic of Korea
  • Jae Yong Kim
    Department of Ophthalmology, University of Ulsan College of Medicine, Asan Medical Center, Seoul, Republic of Korea
  • Myoung Joon Kim
    Department of Ophthalmology, University of Ulsan College of Medicine, Asan Medical Center, Seoul, Republic of Korea
  • Hungwon Tchah
    Department of Ophthalmology, University of Ulsan College of Medicine, Asan Medical Center, Seoul, Republic of Korea
  • Footnotes
    Commercial Relationships  Eun-Soon Kim, None; Sun Young Kim, None; Mi Hyun Cheon, None; Dong-Ah Ko, None; Jae-hyung Kim, None; Jae Yong Kim, None; Myoung Joon Kim, None; Hungwon Tchah, None
  • Footnotes
    Support  None
Investigative Ophthalmology & Visual Science April 2011, Vol.52, 1132. doi:
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      Eun-Soon Kim, Sun Young Kim, Mi Hyun Cheon, Dong-Ah Ko, Jae-hyung Kim, Jae Yong Kim, Myoung Joon Kim, Hungwon Tchah; Expression Of Inflammatory Cytokines And Receptors Using An Ex Vivo, Dry-Eye Model Under Hyperosmotic Stress. Invest. Ophthalmol. Vis. Sci. 2011;52(14):1132.

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Abstract

Purpose: : To establish an ex vivo, dry-eye model using a multi-layered culture of human conjunctival epithelial cells (HCECs) under hyperosmotic stress and to investigate the expression of inflammatory cytokines and receptors in the ex vivo, dry-eye model according to the high EGF condition.

Methods: : HCECs were obtained from brain-dead or expired donors and primary mono-layered culture was performed using bronchial epithelial growth medium(BEGM, Clonetics Corp) for one week. Multi-layered culture using air-liquid interface (ALI) media (1:1 mixture of BEGM and DMEM) was done for two weeks. They were cultured in the same concentrations of all supplements, except for EGF, which was used at 0.5 ng/ml. The ex vivo, dry-eye model was exposed for one or two days under hyperosmotic stress (350~450 mOsm/kg), obtained by adding 5M NaCl. The high EGF condition was created by adding 25 ng/mL of EGF to the ALI media. The expressions of inflammatory cytokines and receptors such as EGF, EGFR, IL-6, IL-8, IL-1β, and VEGF were measured using real time PCR (Roche diagnostics). The concentrations of IL-1β, IL-6, IL-8, TNF-α, and VEGF in the supernatant culture media were analyzed using Bio-Plex cytokine asssays (Bio-Rad Laboratories) after one day's exposure. Apoptosis was analyzed using TUNEL staining.

Results: : The control group (312 mOsm/kg) and the hyperosmotic group (400 mOsm/kg) were exposed for one day. The mRNA of the EGF, EGFR, IL-6, IL-8, IL-1β, and VEGF in the usual EGF group were increased respectively in the high EGF group. The average concentrations of IL-1β, IL-6, IL-8, TNF-α, and VEGF were measured in the control group and also increased respectively in the high EGF group.

Conclusions: : The expression of inflammatory cytokines and receptors were increased under hyperosmotic conditions using an ex vivo, dry-eye model. The high EGF might alter the responses of inflammatory cytokines in a condition of severe hyperosmotic stress condition.

Keywords: inflammation • cytokines/chemokines • cornea: tears/tear film/dry eye 
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