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Eun Kyoung Kim, James C H Tan; Simulating anterior chamber aqueous exchange and drug delivery by single needle perfusion in mice. Invest. Ophthalmol. Vis. Sci. 2014;55(13):5275.
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The mouse eye is a desirable model in which to screen drugs and probe molecular mechanisms relevant to glaucoma. However, the small mouse eye presents technical challenges for measuring intracameral drug delivery and aqueous humor dynamics. Here we simulate and quantify intracameral drug exchange and delivery for single needle live mouse perfusion.
Our apparatus for simulating single needle anterior chamber fluid exchange comprised two microsyringe pumps (one for infusion, one for withdrawal) driving 50 micron microsyringes connected through a Y-connector (0.73ul volume, with minimal dead space) and single 35-gauge needle to an artificial chamber (microcentrifuge tube; volume=60ul), representing the mouse anterior chamber. Connections were via rigid PEEK tubing (0.15 mm inner diameter). Both pumps were connected to independent computer systems with LabChart to monitor flow rates and perfusion pressure. Anterior chamber exchange was performed by withdrawing 50% of artificial chamber fluid at a constant flow rate (3ul/min) then infusing an equal volume of Bradford reagent into the artificial chamber
Bradford reagent concentration relative to reference control after one 50% volume exchange was 19±0.3%; after two exchanges was 43±5.3%; after three exchanges was 55±3.1%; after four exchanges was 64±0.8%; and after five exchanges was 89±1.7%, based on duplicate measurements. Duration of each exchange was 20 minutes. Scaled to the mouse anterior chamber volume, each 50% exchange is expected to take 1 minute 52 seconds with 5 exchanges taking 9 minutes 20 seconds per mouse eye.
We have simulated anterior chamber drug exchange and delivery by single needle cannulation relevant to mice. Intracameral drug concentration that is 90% of the perfused drug is achieved after five 50% volume exchanges, which we estimate to take 10 minutes per mouse eye. We are now confirming these findings in live mice.
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