title: Comparative toxicity of tetracaine, proparacaine and cocaine evaluated with primary cultures of rabbit corneal epithelial cells.
abstract: Cocaine was first used as a topical anesthetic for the eye by Dr Carl Koller in 1884. It became evident that this agent produced erosion of the corneal epithelium in high doses or with repeated use. Synthetic local anesthetics such as tetracaine and proparacaine were developed which were more potent and less toxic than cocaine, but still produced corneal epithelium defects if used chronically. This investigation was undertaken to compare and rank the cytotoxicity of the most commonly used ocular local anesthetics, tetracaine, proparacaine and cocaine, with primary cultures of rabbit corneal epithelial cells. Cultures were exposed to either low concentrations of local anesthetics for 4-24 hr or to higher concentrations of local anesthetics for 15-120 min. Plasma membrane integrity was evaluated by measuring leakage of the cytosolic enzyme, lactate dehydrogenase, into the medium. Cell shape changes were evaluated by observing morphological changes. Mitochondrial dehydrogenase activity and cell viability were assessed by measuring 3-(4,5-dimethythiazol-2yl)-2,5-diphenyl tetrazolium bromide reduction. The cytotoxicity of the local anesthetics as evaluated by the lactate dehydrogenase leakage cytotoxicity test was ranked as follows: tetracaine (EC50 = 0.96 mM) >> proparacaine (EC50 = 4.4 mM) > cocaine (EC50 = 9.7 mM). The mitochondrial reduction assay seemed to be more sensitive than the lactate dehydrogenase leakage test in predicting toxicity: tetracaine (EC50 = 0.81 mM) >> proparacaine (EC50 = 3.4 mM) > cocaine (EC50 = 7.1 mM). When corneal epithelial cells were treated with local anesthetics, marked morphological changes occurred at concentrations that did not cause a decrease in viability. This was especially true for cocaine-treated cells. Tetracaine and proparacaine have the same anesthetic potency in vivo, although tetracaine is considered to be more irritating than proparacaine. This in vitro study showed that tetracaine was approximately four times more toxic than proparacaine. Cocaine was less toxic in vitro than proparacaine and tetracaine when compared on an equimolar basis, but in vivo it may be more toxic because of the higher concentrations that must be used to obtain the same degree of anesthesia as well as its marked effects on cell morphology.
authors.affiliation: Department of Pharmacology/Toxicology, College of Pharmacy, University of Texas at Austin 78712.
authors.forename: R L
authors.initials: RL
authors.lastname: Grant
authors.forename: D
authors.initials: D
authors.lastname: Acosta
mesh_terms: D000779:Anesthetics, Local / Q000633:toxicity*; D000818:Animals; D002478:Cells, Cultured; D003042:Cocaine / Q000633:toxicity*; D004305:Dose-Response Relationship, Drug; D004728:Endothelium, Corneal / Q000187:drug effects* / Q000201:enzymology / Q000473:pathology; D007770:L-Lactate Dehydrogenase / Q000378:metabolism; D008928:Mitochondria / Q000187:drug effects; D011430:Propoxycaine / Q000633:toxicity*; D011817:Rabbits; D013748:Tetracaine / Q000633:toxicity*; D013778:Tetrazolium Salts / Q000378:metabolism; D013844:Thiazoles / Q000378:metabolism; D013997:Time Factors
chemical_list: D000779:Anesthetics, Local; D013778:Tetrazolium Salts; D013844:Thiazoles; D013748:Tetracaine; C005717:proxymetacaine; D007770:L-Lactate Dehydrogenase; D011430:Propoxycaine; C022616:thiazolyl blue; D003042:Cocaine