Kaczmarek Radosław, Szurman Peter, Misiuk-Hojło Marta, Grzybowski Andrzej
Department of Ophthalmology, Wroclaw Medical University, Wrocław, Poland.
Med Sci Monit. 2009 Aug;15(8):BR227-31.
The control of retinal pigment epithelial cell proliferation is an essential factor in the clinical management of proliferative vitreoretinopathy (PVR). Factors inhibiting PVR without toxic potential are of special interest in ophthalmology. The aim of the study was to investigate the antiproliferative and cytotoxic effects of a preservative-free triamcinolone acetonide (PFTA) suspension on a human retinal pigment epithelial (ARPE19) cell line in vitro.
MATERIAL/METHODS: ARPE19 cells (immortal non-transformed cells from a human donor) were seeded and incubated in vitro with increasing concentrations of PFTA (0.01-1 mg/ml). After 1, 3, and 7 days, cellular proliferative activity was assessed by 5'-bromo-2'-deoxyuridine (BrdU) incorporation into cellular DNA and cell proliferation was determined using the 3-(4,5-dimethylthiazol-2yl)-2,5-diphenyltetrazolium bromide (MTT) assay. To determine cytotoxicity, ARPE19 cells were grown to confluence and subsequently cultured in serum-deficient medium to ensure a static milieu and the MTT test was performed after 24 hours of incubation with PFTA. Cell recovery after transient PFTA exposure was also compared with continuous exposure after 7 days.
PFTA inhibits ARPE19 cell proliferation in a dose-dependent manner. Significant inhibition of cell proliferation was observed on the first day of the study at 1, 0.1, and 0.01 mg/ml PFTA and significant reductions in ARPE19 cells were noted for 1 and 0.1 mg/ml PFTA. Proliferation was resumed in all ARPE19 cultures and was dependent on the initial PFTA concentration. PFTA did not cause a cytotoxic effect.
PFTA influences the proliferation of vital ARPE19 cells in a reversible manner without cytotoxic effect.
视网膜色素上皮细胞增殖的控制是增殖性玻璃体视网膜病变（PVR）临床治疗中的一个重要因素。在眼科领域，无毒副作用的抑制PVR的因素备受关注。本研究旨在体外研究无防腐剂曲安奈德（PFTA）悬浮液对人视网膜色素上皮（ARPE19）细胞系的抗增殖和细胞毒性作用。
材料/方法：接种ARPE19细胞（来自人类供体的永生未转化细胞），并在体外与浓度递增的PFTA（0.01 - 1 mg/ml）一起孵育。在1、3和7天后，通过将5'-溴-2'-脱氧尿苷（BrdU）掺入细胞DNA来评估细胞增殖活性，并使用3-(4,5-二甲基噻唑-2-基)-2,5-二苯基四氮唑溴盐（MTT）法测定细胞增殖。为了确定细胞毒性，将ARPE19细胞培养至汇合，随后在无血清培养基中培养以确保静态环境，并在与PFTA孵育24小时后进行MTT试验。还比较了短暂暴露于PFTA后的细胞恢复情况与7天后持续暴露的情况。
PFTA以剂量依赖性方式抑制ARPE19细胞增殖。在研究的第一天，观察到1、0.1和0.01 mg/ml PFTA对细胞增殖有显著抑制作用，并且1和0.1 mg/ml PFTA组的ARPE19细胞数量显著减少。所有ARPE19培养物中的增殖均恢复，且依赖于初始PFTA浓度。PFTA未引起细胞毒性作用。
PFTA以可逆方式影响重要的ARPE19细胞的增殖，且无细胞毒性作用。
