Abstract
Purpose: :
Viscoelastic substances are widely used in ophthalmic surgery. Sodium hyaluronate is the most popular agent. For use in glaucoma surgery we investigated its effect on cultured human Tenon fibroblasts. The goal was to find a substance to inhibit fibroblast proliferation and to maintain the functional bleb after trabeculectomy. Scarring should be prevented and the bleb in the subtenon space should be maintained for some time. In filtering glaucoma surgery, conjunctival fibroblasts play a major role in the post-operative wound healing process. With this series of cell culture experiments, we aimed to determine those concentrations of sodium hyaluronate, also in combination with mitomycin C, that are safe for a possible treatment of filtering blebs during or after surgery.
Methods: :
Monolayer human Tenon fibroblasts were cultured in 96-well plates. Sodium hyaluronate (Healon from AMO, Uppsala, Sweden, 10 mg/ml stock solution) was diluted in DMEM medium in a concentration range from 0.1 mg to 0.01 mg/ml and added to the wells. Mitomycin C was dilutet at 9, 90 and 180 mM. After 24 hours surviving cells were assessed with the vital stain crystal violet. The optical density values were read in an ELISA reader at 550 nm wavelength.
Results: :
Sodium hyaluronate caused a dose dependent enhancement of the cell proliferation activity. Mitomycin C treated cells showed the well-known inhibition of cell proliferation. The inhibition of cell proliferation induced by mitomycin C was decreased by a combined treatment with sodium hyaluronate. At a concentration of 90 mM mitomycin C an inhibition of proliferation of 15% was completely reversed by the addition of 0.1 mg sodium hyaluronate (p < 0.05).
Conclusions: :
Sodium hyaluronate did not inhibit proliferation of human Tenon fibroblasts, but showed a cell proliferating effect. The cell inhibiting effect of mitomycin C was also inhibited by sodium hyaluronate. It is unclear whether the effect is caused by the active agent sodium hyaluronate or by those substances, which are always included in the vial by the manufacturer. Wound healing control with the use of sodium hyaluronate seems not to be possible and other strategies have to be found.
Keywords: cell survival • wound healing