Journal of the Korean Ophthalmological Society 2005;46(1):133-143.
Published online January 31, 2005.
Effect of Dexamethasone and Diclofenac on the Proliferation of Human Corneal Keratocytes.
Ji Eun Lee, Seung Uk Lee, Jong Hwan Kim, Jong Soo Lee
1Department of Ophthalmology, Pusan National University, College of Medicine, Busan, Korea. jongsool@pusan.ac.kr
2Duck-Chon Kim's Eye Clinic, Busan, Korea.
덱사메타존과 디클로페낙이 각막실질세포에 미치는 영향
이지은1,이승욱1,김종환2,이종수1
Department of Ophthalmology, Pusan National University, College of Medicine, Busan, Korea1 Duck-Chon Kim`s Eye Clinic, Busan, Korea2
Correspondence:  Ji-Eun Lee, M.D.1
Abstract
PURPOSE
To evaluate the inhibitory effect of dexamethasone and diclofenac on the proliferation of cultured human keratocytes, and to investigate the apoptotic response and the cellular morphologic changes associated with dexamethasone and diclofenac in vitro. METHODS: Human corneal keratocytes were exposed to 0.05, 0.1, 0.2, 0.4, 0.8, 1.6, and 3.2 mM concentration of dexamethasone and diclofenac for 4, 24, and 48 hours. MTT based calorimetric assay, flow cytometric analysis, fluorescent micrograph, inverted phase-contrast micrograph, and electron microscopy were used to evaluate the results. RESULTS: The inhibitory effect of human keratocyte proliferation increased at higher concentrations and longer exposure times of dexamethasone and diclofenac (p<0.05). In flow cytometry, the maximal apoptotic response developed at 0.4 mM concentration of dexamethasone and 1.6 mM concentration of diclofenac after 4 hours. Apoptotic cells were demonstrated in fluorescent micrograph. Dexamethasone-treated cases showed a more damaged appearance, more swollen rather than spindle shaped, with greater detachment from the bottom of the dish and the chromatin of the nuclear remnant condensed along the nuclear periphery with cytoplasmic vesication and cytoplasmic blebs formation, and partial disruption of the nuclear membrane compared with diclofenac. CONCLUSIONS: The apoptotic response of dexamethasone and diclofenac is associated with the inhibitory effect of human corneal keratocyte proliferation. For inhibition of cellular proliferation of human corneal keratocytes, dexamethasone may be more effective at lower concentration and shorter exposure time than diclofenac.
Key Words: Apoptosis;Dexamethasone;Diclofenac;Flow cytometry;Human corneal keratocyte


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