PMID: 15161851May 27, 2004

Experimental and numerical studies of adenovirus delivery to outflow tissues of perfused human anterior segments

Investigative Ophthalmology & Visual Science
C R EthierW D Stamer


To investigate the efficacy of two different methods of adenoviral transfer of genes to trabecular meshwork (TM) and Schlemm's canal (SC) cells in cultured human anterior segments, using both experimental and numerical analyses. Replication-deficient adenoviruses having coding sequence for beta-galactosidase (beta-gal) under the control of the cytomegalovirus promoter were used. Efficiency of gene transfer over time was verified by infecting cultured human TM cells and assaying for beta-gal activity. Next, ostensibly normal paired human eyes were prepared by standard techniques and perfused for 2 to 5 days to measure baseline facilities. Eyes were then infected by one of two methods: standard transcorneal puncture, or injection into a 1 mm diameter silastic segment of supply tubing immediately upstream of the perfusion dish. In both cases, the nominal total dose was 2 x 10(8) viral particles. Five days after viral injection, eyes were harvested and fixed, and wedges from each of four quadrants were examined histologically. Sections were assayed for beta-gal activity and/or stained with toluidine blue. In a parallel study, flow and viral transport within perfused anterior segments were numerically simulated for conditions that a...Continue Reading


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Related Concepts

Adenoviruses, Human
Anterior Eyeball Segment Structure
In Silico
Defective Viruses
Shuttle Vectors
Organ Culture Techniques
Trabecular Meshwork Structure
Gene Expression Regulation, Enzymologic

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