Publications by authors named "S Zavareh"

38 Publications

An efficient protocol for decellularization of the human endometrial fragments for clinical usage.

Prog Biomater 2021 May 21. Epub 2021 May 21.

Anatomy Department, Faculty of Medical Sciences, Tarbiat Modares University, 14115-111, Tehran, Iran.

The present study was aimed to compare different decellularization protocols for human endometrial fragments. The freeze-thaw cycles in combination with treatment by Triton X-100 and four concentrations of sodium dodecyl sulfate (SDS; 0.1, 0.5, 1, and 1.5%) with two exposure times (24 and 72 h) were applied for tissues decellularization. After analysis the morphology and DNA content of tissues the group with better morphology and lower DNA content was selected for further assessments. The nucleus by Acridine orange and extracellular matrix (ECM) using Masson's trichrome, Alcian blue, and periodic acid-Schiff staining were studied. The amount of tissues collagen types I and IV, fibronectin, glycosaminoglycans (GAGs), and elastin was analyzed by Raman spectroscopy. The ultrastructure and porosity of decellularized scaffold were studied by scanning electron microscopy (SEM). The MTT assay was applied for assessments of cytotoxicity of scaffold. The treated group with 1% SDS for 72 h showed the morphology similar to native control in having the minimum level of DNA and well preserved ECM. Raman spectroscopy results demonstrated, the amount of collagen types I and IV, GAG, and fibronectin was not significantly different in decellularized scaffold compared with native group but the elastin protein level was significantly decreased (P < 0.001). SEM micrographs also showed a porous and fiber rich ECM in decellularized sample similar to the native control. This combined protocol for decellularization of human endometrial tissue is effective and it could be suitable for recellularization and clinical applications in the future.
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http://dx.doi.org/10.1007/s40204-021-00156-5DOI Listing
May 2021

The Effect of Sodium Selenite on Expression of Mitochondrial Transcription Factor A during Maturation of Mouse Oocyte.

Avicenna J Med Biotechnol 2021 Apr-Jun;13(2):81-86

Department of Anatomy, Faculty of Medical Sciences, Tarbiat Modares University, Tehran, Iran.

Background: The aim of the present study was to investigate the effect of Sodium Selenite (SS) supplemented media on oocyte maturation, expression of mitochondrial transcription factor A (TFAM) and embryo quality.

Methods: Mouse Germinal Vesicle (GV) oocytes were collected after administration of Pregnant Mare Serum Gonadotropin (PMSG); in experimental group 1, oocytes were cultured and then subjected for maturation in the absence of SS, and in experimental group 2, they were matured in the presence of 10 of SS up to 16 . The control group included MII oocytes obtained from the fallopian tubes after ovarian stimulation with PMSG, followed by human chorionic gonadotropin. Then, the expression of in MII oocytes in all three groups was investigated using real-time RT-PCR. The fertilization and embryo developmental rates were assessed, and finally the quality of the blastocysts was evaluated using propidium iodide staining.

Results: The oocyte maturation rate to MII stage in SS treated group was significantly higher than non-treated oocytes (75.65 . 68.17%, p<0.05). Also, the rates of fertilization, embryo development to blastocyst stage as well as the cell number of blastocyst in SS supplemented group were higher than other experimental group (p<0.05). There was a significant decrease in gene expression in both groups compared to the group with obtained oocytes (p<0.05). Moreover, there was a significant increase in gene expression in oocytes that matured in the presence of SS compared to that of the group without SS (p<0.05).

Conclusion: Supplementation of oocyte maturation culture media with SS improved the development rate of oocytes and embryo and also enhanced expression in MII oocytes which can affect the mitochondrial biogenesis of oocytes.
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http://dx.doi.org/10.18502/ajmb.v13i2.5526DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC8112142PMC
May 2021

Effect of Mouse Ovarian Vitrification on Promoter Methylation of Inhba and Inhbb in Granulosa Cells of Follicles.

Cryo Letters 2021 Mar-Apr;42(2):67-72

School of Biology, Damghan University, Damghan; Institute of Biological Sciences, Damghan University, Damghan, Iran.

Background: Cryopreservation can induce cellular, genomic, and epigenetic abnormalities.

Objective: To analyse the impact of ovarian vitrification on follicular development and its epigenetic effect on promoter methylation of Inhba and Inhbb in granulosa cells.

Materials And Methods: Mouse ovaries were divided into control, toxicity, and vitrified groups. The growth and development of follicles were examined. After in vitro culture of follicles, DNA was extracted from isolated granulosa cells and treated with sodium bisulfite. The promoter methylation of Inhba and Inhbb was analyzed by direct PCR sequencing.

Results: Vitrification reduced the growth of follicles; however, antral cavity formation was not influenced negatively. Vitrification reduced the percentage of 5-methylcytosine in the Inhba promoter, while CpG sites in the promoter of Inhbb remained unmethylated.

Conclusion: Vitrification had adverse effects on follicle growth and the epigenetics of granulosa cells. The results of the current study show that vitrification methods of ovary need more improvement.
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May 2021

Epigenetic effects of Bisphenol A on granulosa cells of mouse follicles during in vitro culture: An experimental study.

Int J Reprod Biomed 2021 Feb 21;19(2):129-136. Epub 2021 Feb 21.

School of Biology, Damghan University, Damghan, Iran.

Background: Bisphenol A (BPA), a synthetic endocrine-disrupting chemical, is a reproductive toxicant. Granulosa cells have significant roles in follicle development, and KIT ligand (KITL) and Anti-Müllerian hormone (AMH) are essential biomolecules produced by them during folliculogenesis.

Objective: Due to the widespread use of BPA and its potential epigenetic effects, this study examined the impact of BPA on promoter methylation of and genes in mouse granulosa cells.

Materials And Methods: Preantral follicles were isolated from ovaries of immature mice and cultured for eight days. Then, follicles were treated with 50 and 100 μM of BPA, and 0.01% (v/v) ethanol for 24 and 72 hr. Growth and degeneration of follicles and antrum formation were analyzed. The granulosa cells were isolated mechanically, and their extracted DNA was treated with sodium bisulfite. The promoter regions of the and were analyzed with PCR and sequencing.

Results: BPA did not change follicle survival and antrum formation significantly (p = 0.41). However, the culture in the presence of 100 μM BPA had an inhibitory effect on growth. Before BPA treatment, the CpG of the and promoters were unmethylated and partially methylated, respectively. While the percent of 5mC in the promoter reduced at 100 μM of BPA, it did not alter the promoter methylation.

Conclusion: BPA at higher concentrations has an inhibitory effect on follicle growth. Moreover, it seems that the epigenetic impact of BPA restricts to the demethylation of CpG sites.
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http://dx.doi.org/10.18502/ijrm.v19i2.8471DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7922291PMC
February 2021

The effect of sodium selenite on apoptotic gene expression and development of cultured mouse oocytes in comparison with obtained oocytes.

Vet Res Forum 2020 15;11(4):377-383. Epub 2020 Dec 15.

Prevention of Metabolic Research Disorder Center, Research Institute for Endocrine Disorder, Shahid Beheshti University of Medical Science, Tehran, Iran.

maturation (IVM) of oocytes is widely used in assisted reproduction technologies. The present study aimed to improve the oocyte maturation and its development through enriching the culture media with sodium selenite (SS). Moreover, the effects of SS on the expression of the oocytes apoptosis-related genes were assessed. In this study, male and female NMRI mice were used and after collecting their germinal vesicle (GV) oocytes, they were cultured with SS (experimental group) and without SS (control group). Collected metaphase II oocytes (MII) from the fallopian tube were considered as group. After culture, the oocytes were assessed in terms of nuclear maturation. The MII oocytes were inseminated and the development was examined until the blastocyst stage. Also, oocytes were subjected to the molecular analysis for evaluating the expression of BAX, BCL2, P53, and BAD genes using the real-time RT-PCR. The maturation rate was significantly increased in the SS supplemented group compared to the control one. The developmental rate of the embryos was significantly higher for both of the and SS supplemented groups rather than the control one, however, no significant difference was seen between these rates of the experimental and groups. Real-time RT-PCR did not show any significant differences in the expression of the apoptosis-related genes for all of the studied groups. The p53 gene was not expressed in any of groups. Sodium selenite improved the oocyte developmental competence but did not change the expression of the apoptosis-related genes in MII oocytes.
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http://dx.doi.org/10.30466/vrf.2018.93471.2255DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7904118PMC
December 2020