مجله سلول و بافت
سال پنجم شماره 4 (زمستان 1393)

In the present study، the effect of different dose of boric acid (B) on viability، morphology and some biochemical characteristic of rat bone marrow mesenchymal stem cells (MSCs) were investigated. MSCs after 3rd passage was treated with 0. 6،3 and 6 nano،micro and milligram/ml of B for 12،24 and 36hrs. The viability was estimated using 3- (4، 5-dimethylthiazol-2-yl) -2، 5-diphenyltetrazolium bromide (MTT) assay and the dose of 6 Nano، micro and milligram/ml and 36hrs was chosen to continue the investigation. The cell proliferation base on colony forming assay and population doubling number (PDN) was studied; also the morphology of the cells was investigated using florescent dye. The concentration of total calcium and the activity of the enzymes Alanine aminotransferase، aspartate aminotransferase، lactate dehydrogenase and alkaline phosphatase was estimated. Data was analyzed using ANOVA. Treatment of the cells with all the doses caused the viability to decrease significantly after 36hrs. Morphology of the nuclei and cytoplasm as well as number and diameter of the colonies showed significant reduction due to the treatments. Whereas treatment with 6mg/ml caused significant decrease of the PDN after 1، 3 and 6days. In addition significant decrease of the activity of Alanine aminotransferase، aspartate aminotransferase and lactate dehydrogenase were observed in response to treatment with all the doses. Treatment with 6ng and µg/ml caused significant increase in the alkaline phosphatase activity and total calcium concentration، whereas 6mg/ml left no effect on total calcium level and alkaline phosphatase activity. Treatment of MSCs with mg dosage of B showed toxicity، but regarding some factors we observed positive results with respect to nano and microgram dosage، therefore more research is recommended.

In this study، the extract of earthworm (Eisenia foetida) (in solvent: olive oil and vaselline) was applied as a novel healing stimulator on suture wound in goldfish (Carassius auratus). Groups were: negative control (Intact skin)، positive control (suture wound with phenytoin)، pseudo-control (sham: suture wound with solvent)، and experimental (suture wound with 10 mg earthworm extract). A 2 cm longitudinal cut was made from head to tail، and then suture was applied followed by daily treatments. After 14 days، blood samples were collected. Macroscopic and microscopic observations were also done. Meanwhile، the content of lipid peroxidation (LPO/MDA) was measured in fish blood sera. surface and microscopic observations of wounds revealed a better result for earthworm extract treatment in suture wound healing when compared with phenytoin. MDA level in goldfish blood sera was lesser in earthworm extract-treated groups compared to other groups. Earthworm extract is a new candidate in aquatic wound healing. Some factors of the extract may enter the blood and ultimately activate the antioxidant systems leading to decrease the content of LPO in fish blood sera.

Thymus transcaucasicus blongs to the family Lamiaceae. With respect to the importance of plant developmental stages in the expanding knowledge of biology، for this research Thymus transcaucasicus which has nutritional، medicinal and economical value was chosen. The flowers and buds in different developmental stages were collected from their natural habitats in Abali and Roodbar، fixed in FAA، stored in ethanol 70، embedded in paraffin wax and sliced with a microtom. Staining was carried out with Hematoxylin and Eosine. Microscopic study of pollens was carried out using asetolys، in order to investigate the pollen surface ornamentation we used scanning electronic microscope (SEM)، and to compare the pollen proteins from Abali and Roodbar، electrophoresis and Bradford techniques were applied. Anther was having four sacs and its wall was made of four layers including: epidermis، endothecium، middle layer and tapetum layer. Tapetum was in secretive form. The pollen at the time of pollination had three nucleus and six colpus with reticulate Exin ornaments. The total amount of protein in Thymus transcaucasicus pollen collected from the Abali area was much more than protein extracted from pollen collected from Roodbar area. Due to the results of this study، in the Thymus transcaucasicus anther wall with four layers and secretive tapetum was observed. The present research is the first study on development and protein content of Thymus transcaucasicu pollen.

In this study، we have used immunohistochemical studies to examine the effects of maternal hypothyroidism on the expression of laminin in the lung alveoli newborn Wistar rats examined. in this study we have used female virgin rat، wistar strain. Rats were divided in two experimental groups (hypothyroid and hyperthyroid treated with thyroxin) and one control group (n=12). 14 days before mating in the experimental groups، the rats respectively were exposed to Propylthiouracil (50 mg/L) as well as thyroxin (1 mg/L) and Propylthiouracil (50 mg/L) simultaneously. After 14 days، blood test was taken from mothers and in case of desired changes in hormone levels، rats were allowed to mate. After delivery، the lung of the 10 days old newborns were used for immunohistochemical studies. There was a significant reduction in laminin expression in hypothyroid group (p= 0. 001) where as no significant difference was observed in hypothyroid group treated with thyroxin compared to control group. Thyroid hormones have important role on laminin expression and development of lung، since maternal hypothyroidism causes large reduction in alveiolar laminin expression. Therefore changes in maternal thyroid hormone can cause large changes in alveolar development in newborns.

In vitro investigation of hydroethanolic extraction of Cirsium vulgare on growth rates of neonate rat neural stem cells. Neural stem cells were isolated from hippocampus of neonatal rat brain. To determine optimal concentration of Cirsium vulgare extraction، isolated neural stem cells were treated with 200، 400، 600، 800 and 1000 µg/ml for 48 h and then cells proliferation rate were evaluated by MTT assay. In addition Sox2 mRNA expression in neural stem cells was evaluated using quantitative real-time PCR. The results of this study showed that the Cirsium vulgare extract caused significant increase in cell proliferative activity and Sox2 mRNA expression when compared with the control group. with respect to the effect of Cirsium vulgare extract on the proliferation rate of neural stem cells، the use of hydroethanolic extraction of Cirsium vulgare can be used for clinical purposes to treat some of the neurodegenerative disorders such as ischemic stroke and spinal cord injury.

Due to the role of silver nanoparticles on elevation of reactive oxygen species، and high sensitivity of spermatozoa to these agents، it is possible that silver nanoparticles reduce the fertility potential of spermatozoa. The aim of present study was to evaluate the effects of silver nanoparticles on sperm parameters and the concentration of reactive oxygen species in mouse serum and seminal fluid. In this experimental study، 24 adult male Syrian mice were divided into a control and three experimental groups. The experimental groups received silver nanoparticles orally at different doses (0. 05، 0. 1 and 0. 2 ml orally) for 5 weeks. Then blood samples were taken from hearts to examine the Malondialdehyde (MDA) concentration by spectrophotometry. In addition، to analyze the sperm parameter cauda epididymis was dissected and swim-out spermatozoa were analyzed. The 3rd experimental group (highest dose) significantly showed the lowest mean of sperm count (16±2. 6)، the lowest percentage of rapid progressive motile spermatozoa (18. 16±5. 45)، the highest rates of abnormal spermatozoa (35. 83±4. 4) and the highest concentrations of MDA in serum and seminal plasma compared to the control and other experimental groups. The silver nanoparticles as a factor which increase the effects of free radicals and reactive oxygen species in serum and seminal fluid caused reduction of sperm parameters.

The purpose of this study was to consider the effect of mild oxidative stress period on the frozen-thawed bull semen performance. In this study، 120 minutes was considered for equilibration of sperm before freezing. One µm of Nitric Oxide (NO) at the 0، 45، 90 and 120 cooling period was added to the diluant before cryopreservation. Sperm motion parameters، plasma membrane integrity، acrosome integrity، viability، apoptosis and mitochondria activity were assessed. Induction of oxidative stress with nitric oxide in the T0 and T45 groups led to significant improvement of total (88. 4 ± 2. 8، 84. 7 ± 2. 7) and progressive motility (50. 4 ± 2. 5، 49. 5 ± 2. 5) when compared to other groups. Percentage of plasma membrane integrity and viability were not different in T0 (74. 4 ± 2. 9 and 85. 6 ± 2. 9) and T45 (75. 6 ± 2. 9 and 84. 6 ± 2. 3) groups، where as it was significantly higher when compared with T90 (63. 6 ± 2. 9 and 69. 6 ± 2. 3) and T120 (61. 5 ± 2. 9 and 54 ± 2. 3). Moreover، the highest significant percentage of spermatozoa with active mitochondria was observed in the T0 (82. 5 ± 3. 1) when compared with T45 (65. 7 ± 3. 1)، T90 (42 ± 3. 1) and T120 (43 ± 3. 1). Acrosome integrity and linearity of sperm were not affected by the oxidative stress treatment time. It seems that applying oxidative stress using 1 µM NO would improve post-thawed bull sperm quality at the T0 and T45 time of cooling.

The purpose of this study was to investigate the effects of leptin on in vitro maturation، fertilization and embryonic development of immature mouse oocytes. Immature oocytes were isolated from NMRI female mice and divided randomly into 4 groups: leptin 0 ng/ml (control)، 10 ng/ml، 50 ng/ml، 100 ng/ml. 24 h later matured (MII) oocytes were identified and fertilized with incubated sperm. Embryonic development was investigated every 24 h using inverted microscope. From isolated oocytes 53/1% in control group، 56/2% in first group،42/4% in second group and 38/3% in third group developed to MII. The rate of GVBD was 22/05% in control group، 31/27% in first group، 40/63% in second group and 36/39% in third group. The physiologic concentration of leptin (10 ng/ml) considerably increased the rate of MII when compared to other groups (p<0. 05)، whereas the GVBD rate was higher in group treated with concentrations more than physiologic dose. When different concentration of leptin was applied after IVF، no significant effects were observed between four treatment groups in different stages of embryonic development.

Mangifera indica is an evergreen tree belongs to the Anacardiaceae family، in Iran it is found in the Hormozgan and Baluchistan provinces. Till now، no research has been carried out regarding the structural characteristic and development of this species; therefore in the present study structure of vegetative organs and developmental characteristics of reproductive stages was investigated. The vegetative organs were cut and fixed in glycerin and ethanol. The young flowers and buds were sampled at different developmental stages. Samples were fixed in FAA and sliced using a microtome. Staining was carried out with Eosin and Haematoxylin. The primary and secondary structures of root، shoot، petiole، leaf، inflorescence axil and shoot apical meristem showed similarity to dicotyledons. Generative organs contained androecium with 5 stamens which one was reproductive while others were sterile. The anther had 2 parts and 4 pollen sacs. The tapetum layer was secretary type and pollens were three porated. The gynocecium had a carpel with one cameotropous ovule and one tegument. The placentation was of a basal type. The secretary tissues were seen in vegetative organs of the plant. Base on the anatomical findings، we observed a characteristics similarity between this plants and other Angiosperms. With respect to the results of the developmental stages of reproductive organs new information for recognition of this plant was obtained.

The aim of the present study was to stereological evaluate the protective effect of green tea extract on the morphological changes of seminiferous tubules following sodium arsenite exposure in mice. 24 adult male NMRI mice were randomly divided into four groups including control، sodium arsenite (5mg/kg/day)، green tea extract (100 mg/kg/day) and finally sodium arsenite+green tea extract. Mice were (orally) treated for 34 days. At the end، mice were sacrificed and their right testis were taken out، fixed، processed and stained with heidenhain'' azan method. The morphological and changes of testicular tissue was examined using stereological methods. Data were analyzed using one way ANOVA and means difference was considered significant at P<0. 05. Sodium arsenite treatment caused a significant decrease in the mean seminiferous tubules diameter (p<0. 008)، germinal epithelium height (p<0. 001) and the basement membrane thickness (p<0. 011) as well as in the number of spermatocyte (p<0. 003)، round spematid (p<0. 028)، long spermatid (p<0. 011) and sertoli (p<0. 03) cells compared to the control group. The above changes increased to the control level in sodium arsenite+green tea exteract group. The results indicate that green tea extract may be useful in reducing the toxic effects induced by sodium arsenite.