Development of In Vitro Methodologies to Investigate Binding by Sodium Hyaluronate in Eye Drops to Corneal Surfaces

Abstract

Purpose:To develop in vitro methods to assess binding by sodium hyaluronate in eye drops to corneal surfaces.Methods:Two different, complementary corneal binding set-ups were developed. In a dynamic in vitro model, confluent corneal epithelial cells (HCE-T) were assembled in chamber slides and a declining channel. A static model was constructed with ex vivo porcine corneas clamped in Franz cells. To test the predictive capacity of models, four different eye drops containing sodium hyaluronate were spiked with tritium-labeled sodium hyaluronate to standardize quantification. In both settings, eye drops were applied for 5 min and physiological conditions were mimicked by flushing with artificial tear fluid. Spreading experiments on HCE-T next to synthetic membranes were used for further characterization.Results:Binding was more pronounced in dynamic HCE-T model. Three of the four eye drops demonstrated sigmoidal elution of sodium hyaluronate, suggesting pronounced binding. One solution eluted distinctly faster, likewise the buffer control. The static method produced a similar ranking but at lower levels. When eye drops in which phosphate buffer was replaced by citrate buffer (i.e., to prevent calcification) were used, binding was not influenced. All eye drops spread immediately when placed on HCE-T and at the same order of magnitude on glass and polyethylene terephthalate surfaces.Conclusion:Dynamic and static models performed on different corneal sources were used to determine sodium hyaluronate binding kinetics in solutions under physiological conditions. These methodologies resulted in a ranking of the capacity of sodium hyaluronate to bind in vitro to corneal surfaces.

Sodium hyaluronate promotes proliferation, autophagy, and migration of corneal epithelial cells by downregulating miR-18a in the course of corneal epithelial injury

“…These results are consistent with previous reports. 31,36 More importantly, our results also confirmed that SH could markedly enhance autophagy in CEI model cells.…”

Using Natural Deep Eutectic Systems as Alternative Media for Ocular Applications

“…To develop ocular drops that properly adhere to the cornea, both the cornea surface and the tear film must be considered and kept stable and consistent over time. Instillation of a formulation can disrupt the tear film over time and lead to cell damage that consequently causes conditions such as dry eye disease [ 35 ]. Surface tension allows us to understand the stability of the tear film and the tear film break-up time, which influences the eye drops’ spreading ability and adherence [ 36 ].…”

“…This means that NADES formulations present hydrophilic properties, which are necessary for their application as ophthalmic solutions. Bock et al [ 35 ] used the sessile drop method to test four commercial eye drops on three different surfaces. A hydrophobic polyethylene terephthalate, such as PTFE, was used, as well as glass, which is hydrophilic, and cell monolayers of human corneal epithelial cells.…”