Dual-purpose laser irradiation and perfusion testing system for in-vitro experiments using cultured trabecular meshwork endothelial cells

摘要

Abstract: The means by which Argon laser trabeculoplasty (ALT) lowers intraocular pressure (IOP) is a matter of debate. Mechanical and biological laser-tissue interaction theories have been proposed. To investigate the effect laser irradiation has upon the aqueous outflow facility of trabecular meshwork (TM) cells, a suitable in-vitro model is required. Therefore the purpose of this study was to design, construct, and validate a laser irradiation and perfusion testing apparatus. The system was designed to utilize cultured TM cells seeded onto filter supports. Outflow facility will be quantified by calculating the hydraulic conductivity of the monolayer. An appropriate filter support was located, and its perfusion characteristics determined using water. Afterwards, the steady state perfusion flow rate of the filter was ascertained to be 0.096 plus or minus 0.008 ml/min when culture medium is used. Following these tests a single, baseline perfusion experiment was conducted using a TM cell monolayer. Analysis of the data produced a baseline hydraulic conductivity of 0.673 plus or minus 0.076 $mu@l/min/mm Hg/cm$+2$/, well within the range found in previous reports. A dual purpose, in vitro-cellular perfusion and laser irradiation testing apparatus has been developed, tested and validates using known baseline cellular perfusion and laser irradiation testing apparatus has been developed, tested, and validated using known baseline cellular perfusion values. Future experiments will be conducted to verify these initial findings, and further experiments will be conducted using Argon laser irradiation. The response of the TM cell monolayer will then be compared to the baseline figures. !21