Production of hepatocyte-like cells from human umbilical vein mesenchymal stem cells

The human umbilical vein, as a readily available stem cell source, is a good alternative to harvest mesenchymal stem cells. Human umbilical cord vein mesenchymal stem cells have recently been isolated and have demonstrated the ability to differentiate into various cell types such as fat, bone, cartilage and neuronal cells. In this study, we have investigated whether human umbilical cord vein mesenchymal stem cells are also able to differentiate into hepatocyte-like cells. Hepatic differentiation was performed with a 2-step protocol and the use of hepatocyte growth factor and oncostatin M for cell culture. During four weeks of induction, most cells displayed a cuboidal morphology. Immunological analysis indicated that umbilical cord vein mesenchymal stem cells-derived hepatocyte-like cells expressed liver-specific protein markers such as albumin and cytokeratin-18. The hepatocyte-like cells also displayed several characteristics of hepatocytes, including expression of transthyretin, glucose 6-phosphatase, cytokeratin-8,18, alpha-fetoprotein, hepatocyte nuclear factor-3β and albumin. The result of indocyanine green cell uptake, as a test substance to evaluate hepatocyte-like cell function, was positive for differentiated cells. Glycogen storage was examined by periodic acid-Schiff staining. Accumulation of intracellular glycogen was detected in the hepatocyte-like cells. Based on these observations, we have concluded that umbilical cord vein mesenchymal stem cells are endowed with hepatogenic potential and may provide a stem cell source to be used as cell therapy for liver diseases

Hydrostatic pressure improves in-vitro maturation of oocytes derived from vitrified-warmed mouse ovaries

Background: Cryopreservation has limited successes and in-vitro maturation is used to improve its results. Hydrostatic pressure (HP) plays an important role in follicular development. Objective: This study was designed to examine the effects of HP on in-vitro maturation of oocytes and cell death in cumulus cells derived from vitrified-warmed mouse ovaries. Materials and Methods: Preovulatory follicles were harvested from non-vitrified and vitrified-warmed 6-8 week-old female NMRI mouse ovaries and randomly assigned to following groups: non-vitrified (control), non-vitrified with HP exposure (treatment I), vitrified-warmed (treatment II) and vitrified-warmed with HP exposure (treatment III). The follicles of treatments I and III were subjected to HP (20 mmHg) for 30 min and after that all groups were cultured for 24h and assessed for in-vitro maturation of oocytes. The viability and apoptosis of cumulus cells and oocytes were assessed using supravital nuclear staining and TUNEL assay, respectively. Results: Oocytes harvested follicles in both control and treatment II had a significantly lower percentage of metaphase II oocytes (MII) than the treatment I and III (23.5±3.1, 15.03±4.6 and 32.7±3.2, 25.5±4.6; respectively) (p<0.05). Viability of the cumulus cells reduced in treatment I, II and III (83.4, 83.3 and 77.7%) compared to control (86.9%), (p<0.05). The apoptotic index in cumulus and oocyte complexes in treatments I and III (10.7±0.8 and 15.3±0.8) was higher than in control and treatment II (6.7±0.5 and 9.7±0.5) (p<0.05). Conclusion: These results demonstrate that HP had a mild effect on cell death incidence in cumulus cells without any effect on oocyte. However, it can be used as a mechanical force to improve in-vitro maturation of oocytes derived from vitrified-warmed mouse ovaries

Antibacterial Activity of Silver Nanoparticle and L-carnitine Advantages on Mixed Vaginitis Caused by Candida albicans/ Escherichia Coli in Mice Models: An Experimental Study: Effects of Silver Nanoparticle and L-carnitine on Mixed Vaginitis

Mix vaginitis refers to at least two potential pathogenic microbes in the vagina. Recently, the popularity of nanoparticles is increasing; these materials have been widely used as an antimicrobial agent in the treatment of chronic infections in which silver nanoparticles (AgNPs) are more widely considered. We aimed to establish a mixed vaginitis model in adult mice with Candida albicans and Escherichia coli, then evaluated the effect of AgNPs and L. carnitine (LC) to treat the vaginitis. In our study, the microdilution method and minimum biofilm inhibitory concentration were used for the antimicrobial activity of AgNPs. Vaginitis was made by intra-vaginal inoculation of 107 CFU/ml of both E. coli/C. albicans in adult NMRI mice. Mice were classified into 8 groups: (1) healthy mice without any treatment, (2) mice were infected intravaginally with equal volumes of C. albicans and E. coli suspensions, (3) healthy mice that received daily intraperitoneal injection of 250 mg/kg LC for two weeks, (4) infected mice that treated with a daily injection of 250 mg/kg LC for two weeks, (5) healthy mice that received daily intravaginal inoculation of 250 ppm of AgNPs for two weeks, (6) infected mice treated with daily intravaginal inoculation of 250 ppm AgNPs for two weeks, (7) healthy mice that received daily intravaginal inoculation of 250 ppm AgNPs and a daily injection of 250 mg/kg LC for two weeks, and (8) mice treated with daily intravaginal inoculation of 250 ppm AgNPs and a daily injection of 250 mg/kg LC for two weeks. All treatments with AgNPs and LC were daily for two weeks. A vaginal smear was taken throughout the experiment, and tissue sections were prepared using the hematoxylin-eosin method. The results showed that the 50% inhibitory concentration (IC-50) of AgNPs for E. coli, C. albicans, and their mixture was 96.84, 11.23, and 35.67 ppm, respectively, and their IC- 90 values were 201.77, 105.51, and 173.13 ppm, respectively. MBIC-90 % of AgNPs for E. coli, C. albicans, and the mixture of them were 500, 125, and 250 ppm, respectively. The estrus cycle in treated mice was similar to intact mice, and the order of their vaginal tissue sections confirmed the treatment of mixed vaginitis. In conclusion, co-administration of AgNPs and LC may eliminate the adverse effect of AgNPs and mixed vaginitis

Possible involvement of calcium channels and plasma membrane receptors on Staurosporine-induced neurite outgrowth

Staurosporine as a protein kinases inhibitor induced cell death or neurite outgrowth in PC12 cells. We investigated the involvement of calcium channel and plasma membrane receptors on staurosporine inducing neurite outgrowth in PC12 cells. PC12 cells were preincubated with NMDA receptor inhibitors (1.8 mM ketamine and 1μM MK801, treatment 1) or L-Type Calcium channels (100 μM nifedipine and 100 μM flavoxate hydrochloride, treatment 2) or calcium-calmoduline kinasses (10 μM trifluoprazine, treatment 3) and nifedipine, MK801, flavoxate hydrochloride and ketamine (treatment 4 or without pretreatments (control). Then, the cells were cultured in RPMI culture medium containing 214nM staurosporine for induction of neurite outgrowth. The percentage of Cell cytotoxicity and apoptotic index was assessed. Total neurite length (TNL) and fraction of cell differentiation were assessed. After 24h, the percentage of cell cytotoxicity were increased in treatments 1, 2 and 4 compared with control (p<0.05). After 6h, apoptotic index was similar between all treatments. After 12h, apoptotic index were increased in treatment 4 compared with control (p<0.05). After 24h, apoptotic index were increased in treatments 1, 2 and 4 compared with control (p<0.05). TNL were decreased in treatments 1, 2 and 4 compared with control in different times of assessment (6, 12 and 24 h) (p<0.05). The fraction of cell differentiation were decreased in treatments 1, 2 and 4 compared with control (p<0.05). It can be concluded that the possible involvement of L-type calcium channel and the N-methyl D-aspartate receptor on staurosporine-induced neurite outgrowth process in PC12 cells

Protective Effect of Royal Jelly against Renal Damage in Streptozotocin Induced Diabetic Rats

Background: Royal jelly has been shown to have antioxidant and antidiabetic effects. The objective of this study was to evaluate the protective effect of RJ against kidney damage in streptozotocin induced diabetic rats. Methods: Thirty two male Wistar rats were divided randomly into four groups (n=8 per group). Normal control and diabetic control groups received 1cc/day distilled water, normal RJ-treated and diabetic RJ-treated groups received 100mg RJ/kg body weight daily. Diabetes was induced by intraperitoneal injection of streptozotocin. At the end of the experiment, urine and kidney samples were collected for biochemical and histopathological analysis. Results: The results showed that diabetes could increase levels of urine urea, total protein and albumin significantly, and could decrease the levels of creatinine and uric acid in urine. In the kidney tissue homogenates, catalase activity and antioxidant power were significantly lower, whereas malondialdehyde levels were significantly higher in diabetic group when compared with control group. Diabetic rats showed severe histological changes in kidney tissues. Treatment of diabetic rats with RJ improved significantly all of these parameters. Conclusion: The present study revealed that treatment with RJ resulted in significant improvement in histopathological alterations in kidney tissue and urine parameters of diabetic rats. This could be due to its antioxidant activity and the ability of RJ for scavenging the free radicals released in diabetes. These findings suggest that RJ has protective effects on kidneys affected by diabetes mellitus

Hydrostatic Pressure Affects In Vitro Maturation of Oocytes and Follicles and Increases Granulosa Cell Death

Objective: This study examines the effects of hydrostatic pressure on in vitro maturation (IVM) of oocytes derived from in vitro grown follicles.Materials and Methods: In this experimental study, preantral follicles were isolated from 12-day-old female NMRI mice. Each follicle was cultured individually in Alpha Minimal Essential Medium (α-MEM) under mineral oil for 12 days. Then, follicles were induced for IVM and divided into two groups, control and experiment. In the experiment group follicles were subjected to 20 mmHg pressure for 30 minutes and cultured for 24-48 hours. We assessed for viability and IVM of the oocytes. The percentage of apoptosis in cumulus cells was determined by the TUNEL assay. A comparison between groups was made using the student’s t test.Results: The percentage of metaphase II oocytes (MII) increased in hydrostatic pressure-treated follicles compared to controls (p<0.05). Cumulus cell viability reduced in hydrostatic pressure-treated follicles compared to controls (p<0.05). Exposure of follicles to pressure increased apoptosis in cumulus cells compared to controls (p<0.05).Conclusion: Hydrostatic pressure, by inducing apoptosis in cumulus cells, participates in the cumulus oocyte coupled relationship with oocyte maturation

Polycystic Ovary Induction in Mouse by Testosterone Enanthate

Background &Objective: Polycystic ovary is the most common cause of infertility in Women. Animal models are required for understanding the pathogenesis of polycystic ovary. The objective of this study then was to develop an animal model for inducing the polycystic ovaries using testosterone enanthate.Materials & Methods: In this study, for inducing the polycystic ovary phenotype, female rats about12-14 days-old were injected daily with testosterone enanthate for 2 and 4 weeks (experiment groups: 1 and 2), while the control groups (1 and 2) were injected only with vehicle.The ovaries from both groups were fixed and then were used for histological studies.Results: Testosterone enanthate treatment causes the histological changes in mouse ovary and significantly increased the percentage of preantral and cystic follicles and decreased the percentage of antral follicles in the experiment group, comparing with the control group (P<0.05).Conclusion: It concluded that testosterone enanthate can induces polycystic ovary in mouse

The Effect of Repaglinide on Polycystic Ovary Induced by Testosterone Enanthate in NMRI Mice

Background & Objective: Poly Cystic Ovarian Syndrome (PCOS) is the most common cause of infertility due to anovulation in women. One of the treatments of this syndrome is using Insulin sensitive drugs. The main objective of this project is to analyze the effect of Repaglinide on the histomorphology of polycystic ovary. &nbsp;Materials & Methods: In this study, PCOS is induced by the injection of Testosterone Enanthate in immature mice aged five to seven weeks old for four weeks. Then, the mice were divided in to two groups of control and case. The control group had no injection. In the case group, each mouse received 30 &micro;gr Repaglinide. Body weight, the ratio of ovary weight to body weight, and ovarian diameter were measured and the changes in histomorphology of the ovary, characteristics of ovarian tissue, and the number of follicles were investigated. T- Test was used in this study (T-test, P&pound;0.05. ( Results: The result revealed that Repaglinide had a reducing effect on body weight, the ratio of ovary weight to body weight, and the diameter of ovary (P&pound;0.05). Repaglinide also caused a decrease in Tunica albuginea changes, hyperthecosis, the number of degenerated oocyte, granolusa cell, picnosis and hyper vascularization. The average of primordial, primary, pre-antral, cystic, and atretic follicles are significantly decreased in the case group compared to the control group (P&pound;0.05). Conclusion: Repaglinide can compensate the PCOS ovarian damages. Further studies are recommended in this field

A Histomorphological Comparative Effects of Metformin, Pioglitazone, Repaglinide and Acarbose on Polycystic Ovary in Mice

Background & Objective: Polycystic ovary syndrome (PCO) is one of the most common causes of infertility due to anovulatory in women. The use of insulin-sensitizing agents was one of the treatments for this syndrome. The main purpose of this study was to evaluate the effects of Metformin, Pioglitazone, Ripaglinide and Acarbose on histomorphological changes of polycystic ovaries. Materials & Methods: In this study, Polycystic ovaries were induced by injection of testosterone enanthate (TE) in immature female mice, for four weeks, then they had been divided into five groups; the control, Metformin, Pioglitazone, Repaglinide and Acarbose groups. Body weight, the ratio of ovary-to-animal weight, ovarian diameter, histomorphological changes of ovaries and characteristics of ovarian tissues were studied. Tukey test was used to calculate these parameters (P≤0.05). Results:TE significantly increased the percentage of cystic Follicular and decreased follicular growth compared to treatment groups. The body weight, the ratio of ovary-to-animal weight and ovarian diameter, in all groups showed a significant decrease compared to the control group (p<0.05). In Metformin and Pioglitazone treated group, the number of degenerated oocytes, pyknotic-granlosa cells and vascularization were decreased and luteinization can be seen only in these groups. There is a significant reduction in mean growth of primordial, primary, pre-antral, cystic and atretic follicles in the treatment group. However, the mean number of pre-antral follicle showed a remarkable increase. The average number of antral follicles increased in metformin and pioglitazone groups (p<0.05). The results showed that Metformin and Pioglitazone groups cause a meaningful decrease in the ratio of ovary-to-animal weight (p<0.05). Conclusion: According to these results, Metformin and Pioglitazone have the same effects and can compensate for the damages due to PCOs. These drugs can develop follicular growth. Repaglinide can compensate for the damages in some cases, and, Acarbose has a negative effect on follicular growth

The Study of Retinoic Acid Effects on Testicular Development of NMRI Mice

Background & Objectives: Retinoids are important molecules that regulate crucial processes of development in all vertebrates. In this article, we study the effect of retinoic acid on testis development. Materials & Methods: A group of newborn NMRI mice was chosen to receive intraperitoneally injections of 25 mg/Kg.b.w of retinoic acid and the control group had no injection. After 21 days, the male animals were isolated and sacrificed in 60 days postpartum, and testes were removed from their bodies. Apparent characteristics of the testes of seven mice from each group were observed. In the light microscopic study, these epithelial cells of the tubules were counted and the diameter of seminiferous tubules were measured with statistical T-test analysis and were compared with control group. Results: The findings showed that the weight of the testes in animals that were affected by retinoic acid did not change significantly compared to control group. In germ epithelium of testis, the thickness of the epithelium and the number of the spermatogonia, round and elongated spermatids has decreased significantly compared to the control group (P<0.05). Conclusion: According to the findings of this article, using the retinoic acid after birth has influence on testis development and its seminiferous tubules epithelium. Retinoic acid has an adverse effect on the cell divisions in seminiferous epithelium, therefore the number of affected germ cells decrease. Therefore, the use of vitamin A and its synthetic derivatives like retinoic acid for pregnant women should be done with caution