Sezen YILMAZ SARIALTIN, Mahmoud ABUDAYYAK, Engin YENİLMEZ, Can Özgür YALÇIN

Evaluation of Cytotoxic Effects of 2-Monochloropropanediol (2-MCPD) on Mouse TM3 Leydig and TM4 Sertoli Cells in vitro

Aim: 2-Monochloropropanediol (2-MCPD), 3-monochloropropanediol (3-MCPD), their esters and glycidyl fatty acid esters (GE)are important food contaminants originating mainly during food processing. Recently, the detection of these contaminantsin different foods and vegetable oils, especially in palm oil, margarines, and processed foods such as pastries and cakes andin baby foods raises concerns about their safety. According to the European Food Safety Authority (EFSA) scientific report published in May 2016, foods and vegetable oil contaminated with 3-MCPD, 2-MCPD and their fatty acid esters and GE werereported as harmful to human health. However, limited data on 2-MCPD toxicity preclude a complete assessment of itstoxicity. For this reason, the cytotoxic, apoptotic effects and deoxyribonucleic acid (DNA) damage potential of 2-MCPD wereevaluated on TM3 Leydig and TM4 Sertoli cells. Material and method: The cell viability was evaluated by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT)and neutral red uptake (NRU) assays. Annexin V-FITC/PI double staining kit was used to indicate the type of cell death by flowcytometry. The alkaline comet assay was used to estimate the genotoxic effect.Results: The cell viability assays showed that 2-MCPD was not cytotoxic within the tested concentrations up to 1000 µM onboth cell lines. These sub-toxic concentrations of 2-MCPD (1-1000 µM) also did not induce DNA damage both on TM3 Leydigand TM4 Sertoli cells.Conclusion: Although results demonstrate that tested concentrations of 2-MCPD (1-1000 µM) were neither cytotoxic norapoptotic but may cause cell death and DNA damage at higher concentrations. On the other hand, those high concentrationsare far from the daily exposure doses.

2-Monokloropropandiol (2-MCPD)’nin Sitotoksik Etkilerinin Fare TM3 Leydig ve TM4Sertoli Hücreleri Üzerinde in vitro Değerlendirilmesi

Amaç: 2-monokloropropandiol (2-MCPD), 3-monokloropropandiol (3-MCPD) ve esterleri ile glisidil yağ asidi esteri (GE), gıda veya gıda işlem kaynaklı kontaminantlar olarak adlandırılmaktadır. Bu gıda kontaminantlarının başta palmiye yağı olmak üzere bitkisel yağlar, margarinler, pasta ve kek gibi işlem görmüş gıdalarda ve bebek mamalarında tespit edilmesi, güvenlikleri konusundaki ilgiyi arttırmıştır. Avrupa Gıda Güvenliği Otoritesi’nin (European Food Safety Authority, EFSA) Mayıs 2016 yılında yayınlamış olduğu bilimsel görüşe göre bitkisel yağ ve işlenmiş gıdalarda bulunan GE, 3-MCPD, 2-MCPD ve bunların yağ asidi esterlerinin insan sağlığı için zararlı olduğu raporlanmıştır. Ancak 2-MCPD toksisitesi hakkında yeterli veri olmadığı için tam bir değerlendirme yapılamamıştır. Bu amaçla 2-MCPD’nin, TM3 Leydig ve TM4 Sertoli hücre hatları üzerinde in vitro sitotoksik, apoptotik etkileri ve deoksiribonükleik asit (DNA) hasarı yapma potansiyeli değerlendirilmiştir.Gereç ve Yöntem: Hücre canlılığı, 3-(4,5-dimetiltiyazol-2-il)-2,5-difeniltetrazolyum bromid (MTT) ve nötral kırmızısı alım (NRU) yöntemleri ile değerlendirilmiştir. Akış sitometrisi ile hücre ölüm tipinin belirlenmesinde aneksin V-FITC/PI ikili boyama kiti kullanılmıştır. Genotoksisite değerlendirmesinde komet yöntemi uygulanmıştır. Bulgular: Hücre canlılık yöntemleri sonucu 2-MCPD’nin test edilen konsantrasyonlarda 1000 µM’a kadar sitotoksik olmadığı tespit edilmiştir. Aynı zamanda 2-MCPD’nin, sub-toksik konsantrasyonlarda (1-1000 µM) her iki hücre hattında da (TM3 Leydig ve TM4 Sertoli hücreleri) DNA hasarını indüklemediği bulunmuştur. Sonuç: 2-MCPD, her ne kadar test edilen konsantrasyonlarda (1-1000 µM) sitotoksik ve apoptotik etki göstermese de daha yüksek konsantrasyonlarda hücre ölümüne ve DNA hasarına neden olabilir. Ancak bu yüksek konsantrasyonlar günlük maruziyet dozlarından çok uzaktır.

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