Cytotoxicity and genotoxicity assessment of Calcium Lactate as food additive in HUVEC cell

Abstract

Abstract Background and objective: Annually millions of people in the around world suffer from food-borne diseases through consumption of contaminated food and water. Increasing outbreaks of transmitted diseases through foodstuff indicate the increasing of public health problems in low- and middle-income countries and as well as effect consumers' health can be involved in the economies of the countries. Therefore, the use of chemical compounds and the presence of microorganisms in food can play an important role in the development of food-borne diseases. Cancers one of the most common diseases associated with the consumption of chemical compounds which the second rating mortality in the developed and third rating in developing countries. Recently, chemical compounds (food additives) have been introduced as one of the most important factors in the development of these diseases. In this study potential cyto-genotoxicity of CL on HUVECs and interaction with serum albumin (SA) was investigated. Materials and Methods: The cytotoxicity effect of dietary food additive such as calcium lactate in HUVEC cell line was assessed using MTT assay and the genotoxicity effect of the aformentioned additives was evaluated using flowcytometry, DNA damage and chromatin fragmantation methods. Also, spectroscopic techniques such as spectrophotometric and spectrofluorimetric were used to determination the interaction of calcium lactate with serum albumin.

Results: The results of MTT test for the determine the cell viability of HUVEC cells after 24 and 48 hours treated with different concentration of calcium lactate showed cell viability in different concentrations of this additive reduced, but in some concentrations was not significant. The assessment of the survival of calcium lactate after 24 and 48 hours showed that the survival of the treated cells was significant at 4 and 3 mM concentrations (P <0.05). The occurrence of early and secondary stages of apoptosis within treated HUVECs with this additive was evaluated using flow-cytometry procedure. Based results DNA ladder and DAPI staining assays, it can be deduced that this food additive did not lead in DNA or chromatin fragmentation whitin 48 hours. The attained results showed that treated cells with various concentrations of calcium lactate of food additive showed cytotoxic effects but not genotoxic. The results of spectophotometry technique exhibited increased in absorbance of bovine serum albumin (1mM) upon addition of 10 to 70 µm calcium lactate, which confirmed interaction between this additive and serum albumin. Also, the experimental data of intrinsic fluorescence showed that the quenching of albumin by lactate was due to the complex formation of calcium lactate and bovine serum albumin. Conclusio: Calcium lactate decreased HUVEC cells growth in a dose- and time- dependent manner but did not lead apoptosis and necrosis, DNA or chromatin fragmentation. Also, calcium lactate, can interaction with bovine serum albumin. Key Words: Calcium lactate, HUVEC cells, Cytotoxicity, Genotoxicity, Interaction, Bovine serum albumin