For several years, In vivo ocular toxicity tests were used to evaluate the hazard potential of drugs and chemicals that are applied to eye. In ethical terms, eye irritation tests can cause discomfort and pain in laboratory animals. Since 3R principle has been published, several alternative methods have been developed, validated and accepted by regulatory authorities to evaluate the potential toxicity of chemicals and cosmetics. On March 11 st 2013, the commercial sales of any types of cosmetics and personal care products tested on animals, were banned in the European Union. Since then, alternative in vitro toxicity tests are used for the safety evaluation of the cosmetics. For the evaluation of the ocular toxicity of chemicals and cosmetics, the In vitro restructured human ocular tissue models are used as an alternative methods. Some of these models are preferred more because of their morphological and physiological similarity to the human eye. In this review, both in vitro and in vivo ocular toxicity tests will be mentioned; newly developed and validated test methods will be evaluated with a comprehensive perspective.