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Alternatives to Laboratory Animals - ATLA

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A Reassessment of the In Vitro RBC Haemolysis Assay with Defibrinated Sheep Blood for the Determination of the Ocular Irritation Potential of Cosmetic Products: Comparison with the In Vivo Draize Rabbit Test

Eloísa Nunes Alves, Rosaura de Farias Presgrave, Octávio Augusto França Presgrave, Fernanda Peres Sabagh, João Carlos Borges Rolim de Freitas and Alexandre Pinto Corrado

We examined the correlation between results obtained from the in vivo Draize test for ocular irritation and in vitro results obtained from the sheep red blood cell (RBC) haemolytic assay, which assesses haemolysis and protein denaturation in erythrocytes, induced by cosmetic products. We sought to validate the haemolytic assay as a preliminary test for identifying highly-irritative products, and also to evaluate the in vitro test as alternative assay for replacement of the in vivo test. In vitro and in vivo analyses were carried out on 19 cosmetic products, in order to correlate the lesions in the ocular structures with three in vitro parameters: (i) the extent of haemolysis (H50); (ii) the protein denaturation index (DI); and (iii) the H50/DI ratio, which reflects the irritation potential (IP). There was significant correlation between maximum average scores (MAS) and the parameters determined in vitro (r = 0.752–0.764). These results indicate that the RBC assay is a useful and rapid test for use as a screening method to assess the IP of cosmetic products,and for predicting the IP value with a high level of concordance (94.7%). The assay showed high sensitivity and specificity rates of 91.6% and 100%, respectively.

Full text pdf 36(3), 275–284