Indirect coculture of stem cells with fetal chondrons using PCL electrospun nanofiber scaffolds.

In vitro coculture system provides a powerful tool for tissue engineering. In this study, we evaluated the gene expressions of human adipose-derived stem cells (ASCs) on polycaprolactone (PCL) scaffold in coculture model with fetal chondrons. Electrospun PCL scaffolds (900 nm fiber diameter) were created and human infrapatellar fat pad-adipose-derived stem cells (IPFP-ASCs) were seeded on these scaffolds. Scanning electron microscopy (SEM) showed attachment of human IPFP-ASCs to scaffold. IPFP-ASCs on scaffolds were cocultured with fetal chondrons in transwell. Gene expressions were investigated using real-time polymerase chain reaction (real-time PCR). In comparison with control group, the expression level of collagen type 2 and aggrecan were significantly decreased but Indian Hedgehog(IHH) significantly increased (P < 0.05).These findings may interpreted that IPFP-ASCs seeded on PCL scaffold, in cocultures with fetal chondrons are tending toward osteogenesis rather than chondrogenesis.

Evaluation of the melatonin and oxidative stress markers level in serum of fertile and infertile women.

BACKGROUND: Infertility is defined as the inability to achieve the pregnancy within a year of unprotected intercourse. Infertility is a complex issue and different factors such as stress oxidative can be involved in this problem. So, any attempt to neutralize oxidative stress would be helpful in the treatment of infertility. Melatonin is a known scavenger of free radicals. OBJECTIVE: The aim of our study was to evaluate the level of melatonin and its correlation with oxidative biomarkers in fertile and infertile women. MATERIALS AND METHODS: The participants including fertile and infertile women were divided into two groups of 30 people. Blood sampling was performed and sera were collected. The level of Malondialdehyde (MDA), total antioxidant capacity (TAC) and melatonin were detected. Data were analyzed using T-test and their correlation was assessed using Spearman test. RESULTS: Serum melatonin from fertile women was higher than infertile women but the difference was not significant (p= 0.46). MDA level in fertile women was significantly lower than infertile women (p<0.001) and the level of TAC in fertile women was significantly higher than infertile women (p<0.001). Spearman test revealed a significant and direct correlation between melatonin and TAC in fertile and infertile women and a significant but reverse correlation between melatonin and MDA in infertile and fertile women. CONCLUSION: Differences in the level of oxidative stress biomarkers in fertile and infertile individuals have been reported. This study revealed a significant correlation between melatonin and oxidative stress biomarkers, concluding that melatonin level could be involved in infertility.

The effect of BSO-induced oxidative stress on histologic feature of testis: testosterone secretion and semen parameters in mice.

OBJECTIVES: Buthionine sulfoximine (BSO) inhibits synthesis of glutathione as the main intracellular antioxidant. The aim of the present study is to investigate the effect of BSO-induced oxidative stress on histological structure of testis, testosterone secretion and semen parameters. MATERIALS AND METHODS: Thirty male BALB/c mice were divided into 3 groups. In control group, the mice did not receive any chemical. In the experimental group, the mice received 2 mmol/kg BSO for 35 days. In the sham group, the mice received the solvent of BSO (0.9% saline). After the treatment, the mice were sacrificed. Their testes were fixed in Buein's fixative, embedded in paraffin and prepared for histological studies. To assess semen parameters, the sperms were collected from cauda epididymis. Blood samples were used for determination of super oxide dismutase (SOD), malondialdehyde (MDA), glutathione peroxidase (GPX), glutathione (GSH), catalase (CAT) and the serum testosterone level. The data analyzed using ANOVA and Dunnett's tests and SPSS software, version11.5. P- values at 0.05 level considered significant. RESULTS: Data showed that in experimental group in comparison to control group; the concentration of CAT, GPX, SOD,GSH and the total level of testosterone is reduced while MDA level is increased significantly. The number of sperms with progressive motility were decreased (P<0.001) but sperms with abnormal morphology were increased (P<0.001). Histological studies revealed that the values for tubal differentiation index and spermatogenic index in experimental group were reduced (P<0.001). CONCLUSION: It is concluded that exposure to oxidative stress induced by BSO could affect testicular structure and semen parameters.

Serum Levels of Melatonin and Oxidative Stress Markers and Correlation between Them in Infertile Men.

INTRODUCTION: Infertility is the problem of 15% of young couples in different societies. One of the factors that could affect fertility is oxidative stress. Therefore, the aim of the present study is to investigate the level of Melatonin, a free radical scavenger, and its correlation with oxidative biomarkers in infertile men. METHODS: For this purpose, fertile and infertile men in 2 groups, 30 people in each group, were studied. The fertile men were selected from husbands of patients admitted to Alzahra obstetric and gynecology hospital, according to WHO standards. The infertile men were selected from patients referred to infertility ward. Blood sampling from the participants carried out at a specific time, sera collected and the levels of malondialdehyde, total antioxidant capacity and Melatonin were detected in the sera. The data were analyzed using t-test and Sperman's correlation method. RESULTS: Melatonin level in the sera from fertile men were 522 (39.32) ng/L and in infertile men were 511.78 (34.6) ng/L. MDA level in fertile and infertile men were 2.26 (0.34) vs 2.99 (0.44) nmol/ml which was significantly different. The level of TAC in the sera from fertile men were significantly higher than in infertile men. The result obtained for correlation coefficient Spearman's test revealed a significant, strong and direct correlation between Melatonin and TAC and a significant and reverse correlation between melatonin and MDA. CONCLUSION: It is concluded that melatonin could be involved in infertility. In other word, melatonin treatment and antioxidant-rich nutrition could help fertility by combating oxidative stress.

Ultrastructural and morphometrical study of preimplantation endometrium in superovulated mice treated with progesterone or Sildenafil.

BACKGROUND: Endometrial development has an important role in blastocyst adhesion and implantation. During IVF cycles, endometrial development is enhanced by progesterone. OBJECTIVE: The aim of this study was to compare ultrastructural and morphometrical characteristics of mice uterine endometrium in natural cycle with those in superovulated cycles received progesterone or Sildenafil. MATERIALS AND METHODS: In This study, 60 female bulb/c mice were divided into 4 groups: a control and 3 experimental; gonadotropin, gonadotropin+ Sildenafil and gonadotropin+ progesterone. In experimental groups the mice superovulated mated. In the gonadotropin+ progesterone and gonadotropin+ Viagra groups, the mice respectively received 1mg progesterone and 3 mg Sildenafil citrate. Their uterine specimens were prepared for morphometrical and ultrastructural study. Height of the epithelial cells was measured, using motic software. Statistical analysis was performed using ANOVA. RESULTS: Microscopy revealed that in control group the cells had numerous apical microvilli and the height of the cells was 20.52±2.43 µm. In gonadotropin+ progesterone group, the granules were found in basal and apical portions and cellular height were 17.91±2.78 µm which were significantly shorter than in the control and gonadotropin groups (p<0.001). In this group, the apical membrane also contained pinopodes. In gonadotropin +Sildenafil group, the granules were found in both apical and basal portions and the height of the cells were 17.60±2.49 µm which were significantly shorter than in the control and gonadotropin groups (p<0.001). In this group, pinopodes appeared slightly extensive than the other groups. CONCLUSION: It is concluded that superovulatory drugs in mice stimulate endometrial maturation but injection of Sildenafil is nearly more positive. This article extracted from Ph.D. thesis. (Bahman Rashedee).

Study of ultrastructure and apoptosis in the endometrium of women with or without endometriosis.

BACKGROUND: More than 40% of infertilities are due to endometriosis. Ultrustructural and histochemical study of endometrium will help to clarify the etiology of endometriosis. OBJECTIVE: The aim of the present study was to investigate the ultrastructure and occurrence of apoptosis in endometrial cells of women with or without endometriosis. MATERIALS AND METHODS: In the present case-control study, endometrial specimens from 12 women without endometriosis (as control) and 12 women with endometriosis (as case) were examined. Specimens for control group were obtained from the patients that were referred to gynecology hospital for hysterectomy due to various reasons. In case group the endometriosis was diagnosed according to laparoscopy and endometrial samples were taken using pippel biopsy. The specimens from both case and control groups were processed for Transmission Electron Microscopy (TEM), TUNEL reaction technique and morphometric studies. RESULTS: The results show that endometrial epithelium lost its continuity in women with endometriosis and endometrial cells have euchromatic nucleus in comparison to those from non-endometriosis. There were several apoptotic cells in the luminal and glandular endometrial epithelium and stroma from endometrium of control group. However, apoptotic cells were rarely seen in the endometrium from women with endometriosis. The difference in number of apoptotic cells between two groups statically was significant (p<0.001). CONCLUSION: Regarding the ultrastructural characteristics of endometrial epithelial cells and comparison of apoptotic occurrence in control and case groups it is concluded that endometrial cells in endometriosis group have higher potential to survive and possibly implant.