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© 1996 Oxford University Press

research-article

Prolonged Adverse Effects of Benzalkonium Chloride and Sodium Dodecyl Sulfate in a Primary Culture System of Rabbit Corneal Epithelial Cells

ROBERTA L. GRANT and DANIEL ACOSTA

Division of Pharmacology and Toxicology, College of Pharmacy, University of Texas Austin, Texas 78712-1074

Received December 4, 1995; accepted April 26, 1996

This investigation was undertaken to determine prolonged adverse effects of benzalkonium chloride (BzCl), a cationic surfactant, and sodium dodecyl sulfate (SDS), an anionic surfactant, after an initial treatment of and subsequent removal from a primary culture system of rabbit corneal epithelial cells. Metabolic integrity and cell growth were evaluated at specified periods after a 1-hr treatment with the surfactants because of their importance in tissue repair. Intracellular calcium ([Ca2+]1) and intracellular pH (pH1) were also measured because of their importance in cellular homeostasis. ATP/ADP ratios were used to assess metabolic integrity, and propidium iodide staining of cells was used to measure relative cell number and cell growth. Digitized fluorescence imaging was used to measure [Ca2+]1 with fura-PE3 and pH1 with 2', 7'-bis(2-carboxyethyl)-5(6)-carboxyfluorescein (BCECF). BzCl induced a concentration-dependent decrease in ATP/ADP ratios 24 hr after its removal from the cultures, whereas SDS had minimal effects on metabolic integrity throughout the 48-hr postexposure measurement period. The proliferative response of cultures treated with SDS, however, was decreased when compared with BzCl-treated cells. BzCl induced an increase in [Ca2+]1, whereas SDS decreased [Ca2+]1 1–3 hr after removal of surfactants. BzCl produced a sustained decrease in pH1 in surviving cells 1–4 hr after its removal, with a return to control values at 24–48 hr. SDS transiently increased pH1 1 hr after its removal and decreased pH1 at the 48-hr post-treatment period. In conclusion, the two surfactants in vitro had distinctly different prolonged effects on corneal epithelial cells, which may suggest that BzCl and SDS differentially affect cellular recovery in vivo.


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