Comparison of the efficacy of piascledine and transforming growth factor ß1 on chondrogenic differentiation of human adipose-derived stem cells in fibrin and fibrin-alginate scaffolds.

OBJECTIVES: The aim of this study was to compare the chondrogenic induction potential of Piascledine and TGF-ß1 on adipose-derived stem cells (ADSCs) in fibrin and fibrin-alginate scaffolds. MATERIALS AND METHODS: Human subcutaneous adipose tissues were harvested from three patients who were scheduled to undergo liposuction. Isolated ADSCs were proliferated in a culture medium. Then, the cells were seeded in fibrin or fibrin-alginate scaffolds and cultured for 14 days in a chondrogenic medium containing Piascledine, TGF-ß1, or both. The rate of cell proliferation and survival was evaluated by using MTT [3-(4, 5-dimethylthiazolyl-2)-2, 5-diphenyltetrazolium bromide] assay and the rate of the expression of type II collagen, aggrecan, and type X collagen genes was evaluated by real-time polymerase chain reaction (real-time PCR) method. RESULTS: The MTT results showed that Piascledine is able to enhance the proliferation and survival of ADSCs in fibrin scaffolds in comparison to other groups (P<0.05). Real-time PCR evaluation revealed that the expression of type II collagen was higher in TGF- ß1groups, but the expression of aggrecan was higher in TGF-ß1 alone or along with Piascledine in fibrin-alginate scaffolds. Furthermore, the expression of type X collagen was lower in Piascledine alone or along with TGF-ß1 in fibrin scaffold. CONCLUSION: Piascledine can enhance the proliferation and differentiation of ADSCs in fibrin scaffolds.

Cadmium Testicular Toxicity in Male Wistar Rats: Protective Roles of Zinc and Magnesium.

Cadmium (Cd) is a highly toxic element, which may cause toxicity to most organs in the body. Zinc (Zn) and magnesium (Mg) are essential minerals with probable benefits on Cd harmful effects. Finding an efficient and non-pathological treatment against Cd toxicity seems promising. Fifty adult rats were divided into ten experimental groups of five rats each. The Cd group was treated with 1 mg Cd/kg and the control group received 0.5 cm3 normal saline. The other eight groups received Zn (0.5 and 1.5 mg/kg) and Mg (0.5 and 1.5 mg/kg) either alone or in combination with 1 mg Cd/kg through IP injection for 3 weeks. Testis malondialdehyde (MDA), sperm parameters, and testis histopathology were investigated. Cd reduced sperm parameters and increased testis MDA. Moreover, Cd exposure caused a significant histological damage in testis of male rats. However, Zn or Mg treatment prevented and reversed Cd toxic alterations in testis. These findings suggest that co-administration of Zn or Mg could improve cadmium testicular toxicity in male Wistar rats.

Effect of Thymoquinone on P53 Gene Expression and Consequence Apoptosis in Breast Cancer Cell Line.

BACKGROUND: Nigella sativa has been a nutritional flavoring factor and natural treatment for many ailments for so many years in medical science. Earlier studies have been reported that thymoquinone (TQ), an active compound of its seed, contains anticancer properties. Previous studies have shown that TQ induces apoptosis in breast cancer cells but it is unclear the role of P53 in the apoptotic pathway. Hereby, this study reports the potency of TQ on expression of tumor suppressor gene P53 and apoptosis induction in breast cancer cell line Michigan Cancer Foundation-7 (MCF-7). METHODS: MCF-7 cell line was cultured and treated with TQ, and 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay was carried out for evaluating the half-maximal inhibitory concentration (IC50) values after 24 h of treatment. The percentage of apoptotic cells was measured by flow cytometry. Real-time polymerase chain reaction (PCR) was performed to estimate the messenger RNA expression of P53 in MCF-7 cell line at different times. RESULTS: The IC50 value for the TQ in MCF-7 cells was 25 µM that determined using MTT assay. The flow cytometry and real-time PCR results showed that TQ could induce apoptosis in MCF-7 cells, and the P53 gene expression was dramatically up-regulated by ascending time, respectively. Hence, there was significant difference in 48 and 72 h. CONCLUSIONS: Our results demonstrated that TQ could induce apoptosis in MCF-7 cells through up-regulation of P53 expression in breast cancer cell line (MCF-7) by time-dependent manner.

Impact of sperm chromatin evaluation on fertilization rate in intracytoplasmic sperm injection.

BACKGROUND: Sperm DNA in human beings and most vertebrates is packed by protamines into highly compact form of chromatin. There are many staining methods to assess sperm chromatin. Three different methods of staining were used simultaneously in this study and the goal was to determine which of these sperm tests has a relation with fertilization rate in intracytoplasmic sperm injection (ICSI). MATERIALS AND METHODS: Thirty couples who referred to Yamagata University Hospital (Yamagata, Japan) for ICSI were included in this study. The greater part of semen was prepared for ICSI. The remaining part was used for staining with aniline blue, acridine orange, and chromomycin A3 (CMA3). For evaluation of abnormal morphology and abnormality of head, Papanicolaou-stained smears were used. The analysis of data was done using Spearman coefficient of correlation and logistic regression model. Receiver operator characteristic (ROC) curve was used for discrimination of CMA3 staining power to identify ICSI rates. RESULTS: Percentage of CMA3 positivity, unlike those of aniline blue and acridine orange, showed significant negative correlation with fertilization rate. Moreover, the percentage of CMA3 positivity showed a positive correlation with the percentage of abnormal morphology and abnormality of head. By dividing patients into CMA3 <48% and CMA3> 48% groups, the area under the curve was 0.646. CONCLUSIONS: CMA3 staining (protamine deficiency) could be considered as a useful tool for evaluation of male fertility prior to infertility treatment.

The effects of protamine deficiency on ultrastructure of human sperm nucleus.

BACKGROUND: Chromomycin A3 (CMA3) staining is one of the staining methods for detecting protamine deficiency in sperm nucleus. CMA3 is a fluorochrome that competes with protamines for binding to DNA double helix. It has been shown in our previous studies that percentage of CMA3 positive spermatozoa in semen has a close significant relationship with the fertilization rate in in vitro fertilization (IVF). The aim of this study was to examine the ultrastructural differences between sperms in patients who had high fluorescent percentages of yellow or red in CMA3 staining (protamine deficient) with patients with low fluorescent percentages. MATERIALS AND METHODS: Semen samples are taken from five patients with high fluorescent percentages and five patients with low fluorescent percentages. Then the samples are passed for the different steps of preparing for electron microscopy. After the sectioning and mounting on grids, they are investigated under the transmission electron microscope. RESULTS: Sperms in patients with low percentages of positive spermatozoa often have a normal appearance. Sperms in high fluorescent samples frequently have unpacked chromatin. Furthermore acrosomes of these sperms are thinner or disturbed. Also sometimes there are irregularities in sperm head membrane. CONCLUSION: Protamine deficiency in sperm nucleus can cause ultrastructural anomalies in sperm chromatin such as unpacking of it. It also is concomitant with acrosome and sperm membrane disturbances.