Background: Tear film instability, hyperosmolarity, ocular surface inflammation, apoptosis and neuro-sensory abnormalities are causes of dry eye. Lubricant target tear film instability, the most effective agent for tear film stabilization being Sodium Hyaluronate 0.1 -0.3%. Sodium Hyaluronate is suggested to be protective for epithelium in dry eye. To test this hypothesis, this study was performed in vitro with commercially available solutions containing hyaluronic acid (HA) in concentrations ranging from 0.1 to 0.3%. To evaluate the desiccation protection capability of different Sodium Hyaluronate 0.1 to 0.3%, we employed a reproducible in vitro cell culture system. Methods: Conjunctival (Chang 1-5c-4) and corneal cells (pRSV-T 2.040) were cultivated under standard conditions. Under confluent cell growth cells, conjunctival epithelial cell line Chang 1-5c-4 and the corneal cell line 2.040 pRSV-T were wetted for 20 min with five commercial ophthalmic solutions and one agent in trial phase (sample 1: 0.1% HA; sample 2: 0.3% HA; sample 3: 0.15% HA, 2% dexpanthenol, sodium chloride; sample 4: 0.1% HA; sample 5: 0.2% HA; sample 6: unknown, PBS as negative control, unsupplemented medium as positive control). After 20 minutes cells were exposed to continuous air flow for 0, 15, 30 and 45 minutes. Assessment of viable cells was performed by alamarBlue® assay and LIVE/DEAD® Viability/ Cytotoxicity Kit. Results: It has been shown that both cell lines showed different response to protection by the tested solutions. Greatest protection was observed at 15 minutes with most agents. Best results in protection from desiccation was assessed with sample 2 even at maximum exposure time at 45 minutes. Sample 2 showed an average survival rate of 91% at 45 minutes exposure time, whereas no significant amount of vital cells were detected after application with sample 6. Sample 6 was the only substance that presented with early significant cell loss at 0 and 15 minutes by 35%. Conclusions: Higher concentration of Hyaluronate acid with 0.3% and an ionic composition close to the normal tearfluid seem to provide the best protective effect against desiccation in experimental dry eye.