S134 Abstracts / Toxicology Letters 229S (2014) S40–S252 P6: Emerging in vitro models P-2.127 In vitro study on inhibitory effects of infusions made from two kinds of tea on lipid peroxidation induced by malondialdehyde Alireza Ebadollahinatanzi * , Azin Shooli The Institute of Applied Scientific Education of Jehad-e-Agriculture, Imam Khomeini Higher Education Center, Karaj-Tehran, Iran Objectives: Free radicals cause a series of diseases including dia- betes, chronic inflammation and cancers. Nowadays the worldwide efforts have been more focusing on finding the natural compounds based on antioxidants. Since herbal infusions present in kinds of tea may have antioxidant properties therefore in this study anti lipid peroxidation of infusions made from two kinds of tea namely black, and green tea have been evaluated. Methods: Two kinds of the most popular Iranian tea from local vender were provided. Infusion times of 40 and 18 min were exerted for preparation of black and green tea infusions, respec- tively. A concentration range from 5 to 5000 ppm was prepared for each two infusions. Malondialdehyde (50 M) was applied to egg yolk (control) to induce lipid peroxidation. Measurement of Lipid peroxidation (LPO) was based on Dorman method (1995) using thiobarbituric acid (TBA) reagent. The inhibition percents by black and green tea infusions (BTI and GTI) were compared to silymarin extract, SE. Results: Analysis of LPO level by TBA showed that, there is a sig- nificant increase in the level of MDA compared to control group (P < 0.001). The dose of 5000 g/ml of BTI and GTI was determined to be most effective among concentrations of two infusions so that they reduced the increase of MDA by 49.46% and 74.94%, respec- tively. There is no significant difference between GTI and SE in reduction of LPO levels. Conclusion: It is concluded that GTI at high concentration prob- ably has anti lipid peroxidation effect and is equipotent to SE. http://dx.doi.org/10.1016/j.toxlet.2014.06.474 P-2.128 Establishment of rodent and human in vitro systems for the investigation of testicular toxicity Alessandra Gastaldello 1,4,* , Jessica Escoffier 1,4 , Stefan Kustermann 2,4 , Nicole Clemann 2,4 , Serge Nef 1,4 , Luc Stoppini 3,4 1 University of Geneva, Geneva, Switzerland, 2 Non-Clinical Safety, F. Hoffmann-La Roche Ltd., Basel, Switzerland, 3 University of Applied Sciences Western Switzerland, Geneva, Switzerland, 4 Swiss Centre for Applied Human Toxicology (SCAHT), Geneva, Switzerland To date, in vitro models for the detection of testicular toxicity has been scarcely used but in-vivo models are time-consuming and expensive. As testicular toxicity remains a persistent challenge e.g. in pharmaceutical development, there is a strong need of reliable in vitro testicular assays (Saldutti et al., 2013). With the final aim to use human cells, we are working with testicular rat cells for the establishment of in vitro systems that could be valuable models to test testicular toxicity. Testes from 7-day-old rats were dissected in small pieces and then cultured at the air-liquid interface to establish three- dimensional (3D) cultures, or dissociated to obtain a cell suspension for two-dimensional (2D) cultures. Cell composition of these sys- tems was investigated by qRT-PCR and immunofluorescence up to 56 days. The analysis showed that testis cells grew in 2D as multi- layer cultures, at 7 days they were mainly composed by myoid cells, Sertoli cells and Spermatogonia and from 14 days also by Sperma- tocytes. On the other hand, qRT-PCR of 3D cultures showed the presence of mRNAs for genes typical of Spermatocytes and Sper- matidis, which were not present in 2D cultures. Our results suggest that both models contain meiotic cells (Spermatocytes) while the 3D system contains also post-meiotic (Spermatidis) cells. In conclusion, after their final validation, both systems could represent alternative in vitro models to assess tes- ticular toxicity of toxic compounds. Reference Saldutti, L.P., Beyer, B.K., et al., 2013. In vitro testicular toxicity models: opportunities for advancement via biomedical engineering techniques. ALTEX 30, 353–377. http://dx.doi.org/10.1016/j.toxlet.2014.06.475 P-2.129 PON-1 activity as a protective player in nevirapine hepatotoxicity: Data comparison in 2D and 3D cell cultures Aline T. Marinho 1,* , Clara G. Dias 1 , Pedro F. Pinheiro 2 , Alexandra M.M. Antunes 3 , M. Matilde Marques 3 , Emília C. Monteiro 1 , Joana P. Miranda 2,4 , Sofia A. Pereira 1,4 1 Centro de Estudos de Doenc ¸ as Crónicas (CEDOC), Faculdade de Ciências Médicas, Universidade NOVA de Lisboa, Lisbon, Portugal, 2 Instituto de Investigac ¸ ão do Medicamento (iMed.ULisboa), Faculdade de Farmácia, Universidade de Lisboa, Lisbon, Portugal, 3 Centro de Química Estrutural, Instituto Superior Técnico, Universidade de Lisboa, Lisbon, Portugal Paraoxonase1 (PON-1) is a high density lipoprotein (HDL)- associated enzyme produced by the liver. PON-1 has antioxidant capacity, is involved in drug metabolism and detoxification of reactive metabolites and has been proposed as a hepatotoxicity biomarker. Nevirapine (NVP) is an anti-HIV drug associated to hep- atotoxicity through the formation of reactive metabolites, but its effect on PON-1 is not fully understood. The current work aimed to compare the effects of NVP and two of its Phase I metabolites on PON-1 activity in 2D and 3D rat hepa- tocyte cultures, to identify the best model to assess this effect. Hepatocyte cultures (2D and 3D) were exposed to NVP, 2- or 12-hydroxy-NVP (80 g/mL) for 8 days. Unexposed cultures were used as controls. PON-1 activity was measured by quantification of p-nitrophenol formation. PON-1 activity was induced upon 4 and 8 days of treatment with 12-hydroxy-NVP in 2D (p < 0.05) and 3D cultures (p < 0.01), when compared to controls. NVP also increased PON-1 activity (p < 0.001) but only in 3D cultures and to a lower extent than 12- hydroxy-NVP. This effect might be associated to NVP conversion into 12-hydroxy-NVP, which is more effective in 3D-hepatocyte cultures. Interestingly, NVP bioactivation to 12-hydroxy-NVP has been related with NVP-induced hepatotoxicity. Thus, the increase in PON-1 activity might represent a protective cellular response against reactive species. This study supports the usefulness of 3D-hepatocyte cultures to evaluate the effect of compounds, which are toxic upon bio- 4 JPM and SAP co-supervised this work.