Comparative fluorophotometric evaluation of the ocular surface retention time of cross-linked and linear hyaluronic acid ocular eye drops on healthy dogs

Purpose: Evaluate and compare the retention time on the canine ocular surface of crosslinked hyaluronic acid (X-HA), linear hyaluronic acid (L-HA) and saline solution using fluorescent compounds (fluorescein sodium salt, Alexa Fluor 488 cadaverine and Alexa Fluor 488 maleimide).

Methods: 0.9% saline solution (SAL) was combined with fluorescein sodium salt. X-HA and L-HA were covalently modified using Alexa Fluor 488 reactive moieties. Eye drops were applied to 70 eyes of 35 dogs that were previously assessed and determined to have a normal ocular surface. Employing a blue light filter (450-490 nm), digital images were captured from instillation to 180 min. Images were analyzed to assess the percent of the total ocular area covered with green fluorescence at various time points.

Results: X-HA exhibited a dual phase behavior: A broad microgel coverage first, followed by accumulation in tear film meniscus and medial canthus in the second phase, remaining in contact with the ocular surface up to 180 min. Coverage with L-HA and SAL eye drops quickly migrated to the tear meniscus. No traces of the fluorescent compounds were observed by 45 min in eyes treated with SAL solution compound and, by 120 min, eyes treated with L-HA.

Conclusions: X-HA exhibited a significantly increased ocular surface contact time with the ocular surface compared with L-HA and SAL. Not only could this indicate extended lubrication time but also supports the potential use of this compound as a method for topical sustained-release drug application.