PurposeThe aim of this study was to design a new nanocomposite that would have high cytotoxicity against invasive breast cancer cells and minimum side effects on normal cells.MethodsAn albumin nano-carrier for delivery of CuNPs was developed. The ACuNPs formation was characterized by TEM, DLS and UV-Vis, fluorescence and circular dichroism spectroscopy. The cytotoxic efficacy of the ACuNPs against human breast cancer cells (MDA-MB 231) and normal cells (MCF-10A) was compared using a standard MTT assay. The mechanism of cell death induced by ACuNPs was considered by inverted and fluorescent microscopy, flow cytometry and gel electrophoresis. The effects of compounds on ROS generations in MDA-MB 231 cells were also studied.ResultsIt was found that the resulted ACuNPs with a diameter of 62.7 nm and zeta potential of about -10.76 mV, are suitable for extravasation into tumor cells. In ACuNPs, the 90% of the secondary structure and almost all the tertiary structure of albumin remained intact. Comparing to CuNPs, ACuNPs could significantly suppress the viability of cancer cells while they were less toxic on normal cells. Compared with the untreated cells, the MDA-MB 231 cell line showed higher levels of ROS production after treatment with ACuNPs. The increase in ROS production after 24 hours indicated that ACuNPs induce apoptosis.ConclusionsThe ACuNPs characteristics such as intact structure of albumin, high toxicity against cancer cells comparing to normal cells and apoptosis induction as the mechanism of cell death, revealed that this nanocomposite is a good candidate to be used as a chemotherapeutic agent against invasive breast cancer cells.
Cryopreservation is known to induce oxidative stress in spermatozoa. Although melatonin has powerful antioxidant properties, little is known about its effects on human sperm quality during cryopreservation. The present study was undertaken to investigate the effects of melatonin treatment on human sperm parameters essential for fertilization. We first evaluated the effects of various concentrations of melatonin (0-15 mM) on human sperm parameters such as motility, viability and levels of intracellular reactive oxygen species during cryopreservation in order to identify an optimal dose with the greatest effects for further studies. Liquefied semen samples were then divided into three aliquots: cryopreserved without melatonin (control), cryopreserved with 3 mM melatonin and fresh groups. After being thawed, samples were evaluated for motility, viability, membrane integrity, intracellular reactive oxygen species levels, caspase-3 activity and AKT phosphorylation. Treatment of spermatozoa with the various concentrations of melatonin significantly increased their motility and viability and decreased their intracellular reactive oxygen species levels compared with the control group. The optimal melatonin concentration (3 mM) significantly decreased the intracellular reactive oxygen species levels, caspase-3 activity and the percentage of both dead and apoptotic-like sperm cells and increased the vitality, progressive motility and total motility and AKT phosphorylation compared with the control group. Thus, melatonin exerts protective effects against cryodamage during human spermatozoa cryopreservation and may exert its effects via the PI3K/AKT signaling pathway.
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