Differentiation of human endometrial mesenchymal cells to epithelial and stromal cells by seeding on the decellularized endometrial scaffold.

This study aimed to construct the endometrial-like structure by co-culturing of human mesenchymal endometrial cells and uterine smooth muscle cells in the decellularized scaffold. After decellularization of the human endometrium, cell seeding was performed by centrifugation of human mesenchymal endometrial cells with different speeds and times in 15 experimental subgroups. Analysis of residual cell count in suspension was done in all subgroups and the method with the lower number of suspended cells was selected for subsequent study. Then, the human endometrial mesenchymal cells and the myometrial muscle cells were seeded on the decellularized tissue and cultured for 1 wk; then, differentiation of the seeded cells was assessed by morphological and gene expression analysis. The cell seeding method by centrifuging at 6020 g for 2 min showed the highest number of seeded cells and the lowest number of residual cells in suspension. In the recellularized scaffold, the endometrial-like was seen with some protrusions on their surface and the stromal cells had shown spindle and polyhedral morphology. The myometrial cells almost were homed at the periphery of the scaffold and mesenchymal cells penetrated in deeper parts similar to their arrangement in the native uterus. The more expression of endometrial-related genes such as SPP1, MMP2, ZO-1, LAMA2, and COL4A1 and low-level expression of the OCT4 gene as a pluripotency marker confirmed the differentiation of seeded cells. Endometrial-like structures were formed by the co-culturing of human endometrial mesenchymal cells and smooth muscle cells on the decellularized endometrium.

NFR2/ABC transporter axis in drug resistance of breast cancer cells.

Breast cancer is one of the most serious malignancies among women, accounting for about 12% of all cancers. The inherent complexity and heterogeneity of breast cancer results in failure to respond to treatment in the advanced stages of the disease. Breast cancer is caused by several genetic and environmental factors. One of the significant factors involved in the development of breast cancer is oxidative stress, which is generally regulated by nuclear factor erythroid 2-related factor 2 (NRF2). The level of NRF2 expression is low in healthy cells, which maintains the balance of the antioxidant system; however, its expression is higher in cancer cells, which have correlation characteristics such as angiogenesis, stem cell formation, drug resistance, and metastasis. Drug resistance increases with the upregulation of NRF2 expression, which contributes to cell protection. NRF2 controls this mechanism by increasing the expression of ATP-binding cassettes (ABCs). Considering the growing number of studies in this field, we aimed to investigate the relationship between NRF2 and ABCs, as well as their role in the development of drug resistance in breast cancer.

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

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.

The protective effect of vitamin E on rats' ovarian follicles following an administration of diazinon: An experimental study.

BACKGROUND: Diazinon (DZN) is an organophosphate insecticide that has been widely utilized in agriculture all over the world and caused many negative effects on different species such as plants and animal species, especially on a human. OBJECTIVE: The aim of the present study was to evaluate the protective effect of vitamin E on rats' ovarian follicles following an administration of diazinon. MATERIALS AND METHODS: A total of 30 adult female Wistar rats were divided into five groups: a control group (without any intervention), sham group (received only pure olive oil, as solvent), experimental group I (DZN+olive oil, 60 mg/kg), experimental group II (vitamin E, 200 mg/kg), and experimental group III (DZN: 60 mg/kg+vitamin E: 200 mg/kg). All drugs were injected intraperitoneally, except vitamin E which was administrated by gavage. The animals were scarified after two weeks and left ovary was used to measure proliferation of ovarian follicles. Tissues were analyzed by the PCNA technique and viewed with an optical microscope for evaluating cells proliferation. RESULTS: The result of the present study revealed that the number of proliferative cells in the experimental group I decreased significantly in contrast to the control group in secondary and Graffian follicles (p < 0.001). The administration of vitamin E plus DZN significantly increased proliferative cells compared to the DZN group (p < 0.001). Primordial follicles showed that all study groups were lacking PCNA positive cells, which means no expression of PCNA in these follicles. The results of this study showed that primary follicles in all study groups had a few and scattered PCNA positive cells with no significant difference between the groups (p > 0.05). CONCLUSION: Results showed that DZN reduced proliferation in secondary and Graffian follicles and vitamin E increased it. The results of this study suggested that vitamin E by its antioxidant activity was able to improve the DZN-induced ovarian toxicity.

The Effect of Diazinon on Cell Proliferation and Apoptosis in Testicular Tissue of Rats and The Protective Effect of Vitamin E.

BACKGROUND: Diazinon (DZN) is an organophosphate pesticide, and nowadays this pesticide is mostly used in agriculture. In this study, we analyzed the effects of DZN and vitamin E (Vit E) on apoptosis and the proliferation of germ cells in rat testis. MATERIALS AND METHODS: In this experimental study, 30 male Wistar rats were divided into five groups (n=6 per group) consisting of control, sham (received olive oil), experimental group i (60 mg/kg DZN), experimental group ii (60 mg/kg DZN and 200 mg/kg Vit E), and experimental group iii (200 mg/kg Vit E). After six weeks, left testis of rats was removed for the detection of proliferative cell nuclear antigen (PCNA) and terminal deoxynucleotidyl transferase end-labeling (TUNEL). RESULTS: Compared with the control group, DZN in the experimental group i decreased the number of PCNA-positive cells and increased the number of TUNEL-positive cells (P<0.001). Vit E improved detrimental changes by the decrease in the rate of apoptosis and the increase in the proliferation of testicular germ cells (P<0.001). CONCLUSION: Vit E can decrease the number of TUNEL-positive cells and increase the number of PCNA-positive cells by the neutralization of the toxicity caused by DZN in the testicular tissue.

Single dose effect of diazinon on biochemical parameters in testis tissue of adult rats and the protective effect of vitamin E.

BACKGROUND: Diazinon (DZN) is an organophosphate pesticide that widely used for agricultural pest control all over the world. DZN affects target organs including reproductive system by inhibiting the activity of acetylcholinesterase and inducing oxidative stress. Vitamin E (α-tocopherol) is a strong antioxidant which inhibits free radicals, and probably can reduce lipid perxidation effectively in biological systems. OBJECTIVE: The present study, aimed to evaluate the effects of DZN on malondialdehyde (MDA) and glutathione (GSH) levels in testis of rats and protective effect of vitamin E. MATERIALS AND METHODS: In this experimental study, thirty adult male Wistar rats (200-250 gr) were divided into 5 groups (n= 6): control group (did not receive any material), sham group (received only pure olive oil), experimental group 1 (DZN, 60 mg/kg), experimental group 2 (Vit E, 200 mg/kg) and experimental group 3 (DZN+Vit E, with the same dose). All groups were sacrificed after 6 weeks and right testis was used to measure the MDA and GSH levels. The amount of MDA was determined by the thiobarbituric acid assay and 5, 5-Dithio-bis (2nitrobenzoic acid) DTNB-recycling protocol was used for GSH assay. RESULTS: The results showed that DZN increased MDA level (p<0.001) and reduced GSH level (p<0.001). Administration of DZN plus vitamin E decreased the MDA level (p<0.001) and increased GSH level (p=0.001). CONCLUSION: DZN induced lipid peroxidation in the testis of rats. Vitamin E by its antioxidant activity was able to improve the toxic effect of DZN.