Abstract
Purpose :
Toxicity and efficacy tests of eye drops is complex because the administration times and wash out duration of eyedrop should be considered. In addition, using a pipet tip on the corneal wound healing experiments, the size of the wound is not uniform and a physical injury is caused. We have developed a fully automated high throughput screening system based on the microfluidic cell culture array to evaluate toxicity and efficacy of eyedrops.
Methods :
The present micro fluidic chip used in the experiment is high throughput drug screening system and we can observe 8 step’ concentrations at the same time. These process are operated by fully automatic pneumatic controller that is engaged by the programmed LabVIEW software. We evaluated the toxic effect of benzalkonium chloride (BAK) and wound healing effect of ursolic acid on immortalized human corneal epithelial cell line (HCE-Ts) in this system.
Results :
1. Micro fluidic high throughput screening (μHTDS) system for cytotoxicity of preservatives in eye drop : Cytotoxicity of BAK on HCE-Ts cells was increased depend on treatment time with comparing same dose conditions. Live cells with 10 minutes treatment are quickly decrease from 0.002% comparing to 5 minutes treatment results. The result in this study was similarity between plate experiment of conventional method and μHTDS system. 2. Micro fluidic high throughput screening system for wound healing (proliferation and invasion) model : 64 cell chambers which are arranged 8x8 forms, it means that each 8 concentrations can be observed on 8 chambers at same concentration. 7uM was most effective concentration of ursolic acid on migraion and invasion of HCE-Ts cells.
Conclusions :
We have developed a micro fluidic high throughput drug screening (μHTDS) system for cytotoxicity and efficacy screening of eyedrops with 8 different drug concentrations. Our μHTDS system can obtain more progressed cytotoxicity and efficacy results with minimizing the consumption of reagent, time, and labor. This system can be useful for cytotoxicity and efficacy screening the new drug against diverse cell type instead of human eye.
This is an abstract that was submitted for the 2016 ARVO Annual Meeting, held in Seattle, Wash., May 1-5, 2016.