Rofougaran R, Pakkar A, Wee W R, McDonnell P J
Doheny Eye Institute, University of Southern California School of Medicine, Los Angeles, USA.
J Refract Surg. 1997 Sep-Oct;13(6):568-70. doi: 10.3928/1081-597X-19970901-13.
Dichlorotriazinyl aminofluorescein (DTAF) has been used to stain corneal stromal collagen as part of in vivo experimentation. Toxicity of this drug, if present, might alter the observed wound healing. To determine if this drug has any deleterious effect on keratocytes, we evaluated it in vitro.
Human keratocytes in 96 well plates were exposed to different concentrations of DTAF (10e-7, 10e-6, 10e-5, 10e-4, 10e-3, 10e-2, and 10e-1 mg/ml of media). Exposure times of 1 and 24 hours at each concentration of DTAF were evaluated. The cell number was measured 1 and 3 days after exposure to the drug using a coulter-counter and a hemocytometer.
The proliferation of keratocytes after 24 hours of exposure to the drug was inhibited in a dose dependent manner by DTAF, but 1 hour exposure of keratocytes to the drug did not inhibit keratocyte proliferation.
These results suggest that DTAF has inhibitory effects on human keratocyte proliferation after 24 hours of exposure, while exposure limited to 1 hour does not induce such a change.
二氯三嗪基氨基荧光素(DTAF)已被用于对角膜基质胶原蛋白进行染色,作为体内实验的一部分。如果这种药物存在毒性,可能会改变观察到的伤口愈合情况。为了确定这种药物对角细胞是否有任何有害影响,我们在体外对其进行了评估。
将96孔板中的人角膜细胞暴露于不同浓度的DTAF(每毫升培养基中分别为10的-7次方、10的-6次方、10的-5次方、10的-4次方、10的-3次方、10的-2次方和10的-1次方毫克)。评估在每个DTAF浓度下1小时和24小时的暴露时间。在接触药物1天和3天后,使用库尔特计数器和血细胞计数器测量细胞数量。
DTAF以剂量依赖的方式抑制了角膜细胞在接触药物24小时后的增殖,但角膜细胞接触药物1小时并未抑制其增殖。
这些结果表明,DTAF在暴露24小时后对人角膜细胞增殖有抑制作用,而暴露时间限制在1小时则不会引起这种变化。