Anatomical Sciences Journal
Year:2011 | Volume:9 | Issue:34|Papers Count:8

Objective: Examining of the effect of hydrostatic pressure on in vitro maturation of mouse oocytes.Materials and Methods: Female NMRI mice 6-8-week-old was selected. Antral follicles with diameter of 500mm were isolated from ovaries. Then follicles were divided into control and experiment groups. In experiment group, follicles were transferred to pressure chamber and subjected to 20 mm pressure for 30 min. Follicles without pressure exposure used as control. Each follicle cultured individually in 100 ml a-MEM supplemented with 5% FBS, 100 mIU/ml rFSH, 10 ng/ml rEGF and 7.5 IU/ml HCG for in vitro maturation, under mineral oil. Follicles from two groups were cultured for 48 h and evaluated in vitro maturation of oocyteby the percentage of oocytes in GV, GVBD and MII. Viability of oocytes and cumulus cells was assessed with nuclear vital staining (propidium iodide and bisbensamide).Results: The results showed that, exposure to hydrostatic pressure improved the oocyte in vitro maturation. After 24 h the percentage of GVBD and metaphase (MII) oocytes increased in hydrostatic pressure treated follicles compared to control (P<0.05). After 48 h, the percentage of metaphase (MII) oocytes increased in hydrostatic pressure treated follicles compared to control (P<0.05). Furthermore, a significant increase in the cumulus cells was observed (P<0.05). Hydrostatic pressure had no effect on oocyte viability.Conclusion: It seems that hydrostatic pressure can be a cell death inducer in cumulus cells. Hydrostatic pressure may play a role in oocyte maturation and fertilization by improving in releasing and mediating signals to oocyte by increasing cell death in cumulus cells.

Purpose: This study aimed to evaluate the effects of low level laser on human fibroblasts which were cultured in high glucose cultures.Materials and Methods: The human skin fibroblasts were maintained in Dulbecco's Modified Eagle's Medium (DMEM) in a humidified air-5% CO2 atmosphere. The cells were cultured. Irradiation was carried out with Helium-Neon (He-Ne) laser unit with power output of 1.5 mW and the diameter of irradiating beam of 1.7 cm on the case groups. In the next step the cells were cultured in high glucose culture medium (15mM/L) for one week before laser irradiation. The irradiation was carried out with the same laser unit with power output of 1.5 mW and the diameter of irradiating beam of 1.7 cm on the case groups. Control cells did not receive laser. The cells were irradiated with three doses of 0.5, 1 and 2 J/cm2 on three consecutive days. Cell proliferation was evaluated by dimethylthiazol-diphenyltetrazolium bromide (MTT) assay. Data were analyzed by student t test or Mann-Whitney-U test.Results: Laser treatments of 0.5 (p-value=0.002) and 1 (p-value=0.046) had a stimulatory effect and three irradiations of 2 J/cm2 had a negative effect (p-value=-0.011) on proliferation rate of fibroblasts which were cultured in normal culture medium compared to non irradiated cells. All three doses of 0.5 (p-value=0.042), 1 (p-value≈0.00) and 2 J/cm2 (p-value≈0.00) had a stimulatory effect on proliferation rate of fibroblasts which were cultured in culture medium with high glucose concentration (15mM/L) in comparison with hyper glycemic non irradiated cells.Conclusion: Low level laser irradiation with helium-neon unit on cultured human fibroblasts in normal and high glucose mediums increased their growth and proliferation.

Purpose: The aim of this study was to compare xenograft scaffold and prous Hydroxyapatite/Tricalcium phosphate for the repair of bony defect.Material and methods: 5 New Zealand rabbits with the average weight of 2 kg were used. The rabbits were anesthetized with ketamine 60 mg/kg and xylazin 6 mg/kg After creating a critical-sized segmental defect (2×3 cm) in tibia bones, an implant of HA/TCP with osteoblasts in the hole of left tibia and an implant of xenograft with osteoblasts in the hole of right tibia was inserted. At the end of the second month, the animals were sacrificed. To follow up the new bone formation, x-ray images were taken at 8th week post-operation. To confirm the tissue repairment, the histology of repaired defects was evaluated with H&E staining after decalcification.Results: The present study demonstrates that the critical-size segmental defect of tibia can be repaired with both synthetic HA/TCP and xenograft implants.Conclusion: xenograft implants showed better osteointegration compared to HA/TCP phosphate.

Purpose: The evaluation of epididymal sperm ultrastructure in chronic alcohol-consuming rats.Materials and Methods: 16 male mature Wistar rats with the same age of 10 weeks were categorized into two different groups. Control group included 8 rats allowed free access to rat chow and water. Experimental group included 8 rats with free access to rat chow and 5% ethanol in the same volume (50 cc daily) as controls that received water. After 30 days, epididymal spermatozoa from two groups were aspirated for sperm electron microscopic study.Results: No ultrastructural changes were observed in control group. In experimental animals, most of spermatozoa showed several alternations in their ultrastructures. Anomalies such as abnormal nuclear chromatin density, swollen area, rupture and lysis of plasmalemma, persistence of numerous cytoplasmic droplets, mitochondrial swelling and vaculization, absence of axonemal microtubules, complete degeneration of axoneme, deletion of one or more outer dense fibers as well as absence of tail plasmalemma were seen in majority of the alcohol-treated spermatozoa.Conclusions: Spermatozoa from alcohol consuming rats show spectrum of anomalies in their head, middle and principle piece of tail. These may be one of the possible causes of subfertility or infertility due to alcohol consumption.

Purpose: The aim of the present study was to investigate transplantation of MSCs and theirderived Neural like progenitors (NLPs) into the spinal cord after injury and evaluate the survival, migration, differentiation properties of these cells in Rat spinal cord.Materials and Methods: NLPs were derived from MSCs (induced by bFGF (Fibroblast growth factor b), hEGF (Human Epidermal growth factor) and RA (Retinoic Acid)), and analyzed by flowcytometry and immuno fluorescence staining. MSCs and NLPs injected into model animals vein and collagen scaffold implanted into injured site. Behavioral testing was performed weekly for 5 weeks.Results: Improvement of transplanted animals evaluated after 5 weeks. Unfortunately, Substantial changes were not observed among the rats after the transplantation. Immuno fluorescence staining analysis using human nuclei and BrdU antibodies confirmed survival and migration of hMSCs and NLPs into the injury site. Transplanted cells were found to adjacent segments located rostro-caudaly to the injury epicenter.Conclusion: Our findings indicate that hMSCs and NLPs couldn't facilitate recovery from spinal cord injury. However, these cells can express specific neuronal markers in injured site. There are many questions to be answered regarding this mechanism.

Purpose: Study of the effects of melatonin on the lacrimal gland and corneal epithelium.Materials & Methods: In this study 20 white mice with the age of 11 months and weight of 20-23 g were divided equally in to the control and the experimental groups at random. The experimental group were daily injected intraperitoneally 10 mg/kg of melatonin dissolved in physiological serum for two weeks. The mice of control group were received only physiological serum. After the end of the injection period, the mice were killed with ether and their lacrimal glands and eyes were removed and processed using light microscope. Morphometric analysis was carried out to find the thickness of corneal epithelium, then, the data was evaluated with SPSS soft ware and T Test. Results: The thickness of corneal epithelium in experimental group compared to control group was increased significantly (P<0.001) and different sizes of inclusions were observed in the lumen of acini.Conclusion: Melatonin has different effects on the lacrimal gland and corneal epithelium.

Purpose: The aim of the present study was to evaluate the colocalization of GnIH and GnRH neurons in the ewe preoptic area (POA) of hypothalamus during the phases of estrous cycle.Materials and Methods: Three ewes in three phases of proestrus, estrus and luteal of estrous cycle (n=9) were selected and based on their estrous cycle phase, the number of GnIH and GnRH neurons and percent of these two neuropeptides colocalization in POA were estimated by using immunohistochemistry method.Results: GnIH and GnRH neurons were present in the POA during the different phases of ewe estrous cycle and their colocalization was observed. The number of GnIH neurons in luteal phase was substantially more than proestrus and estrus phases (P<0.01). However, there was no significant difference between the numbers of GnRH neurons of the POA in three estrous phases (P>0.05). In the luteal phase, percent of GnRH neurons that coexpress GnIH were significantly less than proestrus and estrus phases (P=0.008).Conclusion: GnIH expression in neurons of POA of ewe in luteal phase is more than follicular phase, but there was no significant difference in expression of GnRH neurons in the POA during the estrous phases. Also during all phases of the estrous cycle, GnRH neurons coexpress GnIH and GnIH neurons coexpress GnRH. These findings showed the direct effect of GnIH neurons on GnRH neurons in POA and indirect control of LH secretion.

Purpose: The aim of this study was to investigate the effects of using Retinoic Acid during pregnancy on the spermatogenesis, testosterone and gonadotropin hormones in mature male rat.Materials and Methods: Pregnant rats were injected by 10, 20 and 30 mg/kg Retinoic acid (alltrans Retinoic Acid) intraperitoneally on the 8.5, 10.5, 12.5 and 14.5 days after copulation. An equivalent amount of the vehicle was similarly injected to corresponding control rats. Then animals did child birth. On time of maturity (10 weeks), from male animals in all of groups blood to determined the amount of hormones and the reproductive organs were separated. Morphometerical and Histological changes were studied in epididym and testis among experimental and control groups. The results were evaluated by using one way ANOVA and Tukey- test.Results: Results indicateed that there were significant decrease in LH and FSH hormones, seminiferous tubules diameter, number of spermatocyte, spermatid and leydig cells experimental groups compared with the control group especially in animals which received 30 mg/kg of retinoic acid (p£0.05). Also RA is the cause of decrease of the epididymis and deferent weight, and sperms in the epididymis, significantly (p£0.05).Conclusion: Using of Retinoic Acid during pregnancy decreases the normal spermatogenesis and steroidogenesis in mature male rats.