Anatomical Sciences Journal
Year:2012 | Volume:9 | Issue:37|Papers Count:11

Purpose: The aim of this study was to evaluate the effect of NT-3 on the decrease of the differentiation time of bulge stem cells of rat hair follicle from neuron like cells.Materials and Methods: The bulge region of the rat whisker was isolated from and cultured in DMEM/F12 supplemented with epidermal growth factor (EGF) in 3 groups for 7, 8 and 9 days .Then 10 ng/ml NT-3 was added to each group for 3 days. Finally, cell differentiation was assessed by immunocytochemistry using βⅢ-tubulin antibody and the result was compared with control groups.Results: According to our results hair follicle bulge stem cells expressed CD34 and Nestin in 7-9 and 10-12 first days after cultivation respectively. By using NT-3 duration differentiation process the cells showed positive reaction to βⅢ-tubulin antibody.Conclusion: The results indicate that NT-3 can affect on differentiation speed even in less than 10 days.

Purpose: To investigate of the effects of a poly L-lactic acid (PLLA) nanofiber scaffold on proliferation of frozen-thawed neonate mouse spermatogonial stem cells.Materials and Methods: Spermatogonial cells were isolated from neonatal 3-6-day-old NMRI mice testes by two steps enzymatic digestion and differential plating. The isolated spermatogonial cells were divided into four culture groups: 1) fresh spermatogonial cells, 2) fresh spermatogonial cells seeded onto PLLA 3) frozen-thawed spermatogonial cells, 4) frozen-thawed spermatogonial cells seeded onto PLLA. Cells in all groups were cultured in DMEM supplemented with 5% FCS and 10 ng/ml GDNF for 3 weeks. Diameter and number of clusters which were determined during the culture and semi-quantitative RT-PCR were carried out at the end of 3rd week for all culture groups. Presence of spermatogonia at the culture was determined by reverse transcription polymerase chain reaction (RT-PCR) for several important spermatogonial markers (PLZF, Oct4, GFRα-1, VASA, ITGA6 and ITGB1). The significancy of the data was analyzed using Repeated Measures and ANOVA tests.Results: The findings indicated that the viability rate of the fresh cell (control 1 and exprimental 1) and the frozen cells after thawing (control 2 and exprimental 2) were 89.25±2.2 and 63±3.56, respectively and the differences were significant (p<0.001). In vitro culturing of spermatogonial cells on PLLA significantly increased the formation of cell clusters in comparison with those of the control groups (p£0.001). Although the differences of the diameters of clusters in the fresh cell groups were not significant, culturing of frozen-thawed cells on PLLA significantly decreased their diameters (p£0.01). There was a significant down-regulation of spermatogonial genes in the frozenthawed groups after three weeks of culture.Conclusions: The spermatogonial cells seeding on PLLA can increase in vitro cluster formation of neonate fresh and frozen-thawed spermatogonial cells.

Purpose: Low level laser therapy is suggested as a new therapeutic method in diabetic wound healing. This survey aimed to evaluate the effects of low level laser on human fibroblasts cultured in high glucose cultures. Materials and Methods: The human skin fibroblasts were cultured under standard condition. The cells were cultured in high glucose culture medium (15mM/L) for a week and two weeks before laser irradiation. The irradiation was carried out with Helium-Neon (He-Ne) laser unit 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. The releasing rates of Interleukin 6 and Fibroblast growth factor were evaluated by ELISA (enzyme-linked immunoabsorbent assay). Data were analyzed by student t test or Mann-Whitney-U test.Results: All three doses of 0.5 (p=0.049), 1 J/cm2 (p=0.027) had a stimulatory effect on releasing rate of IL-6 of the fibroblasts cultured in culture medium with high glucose concentration in comparison with hyper glycemic non irradiated cells. Low level laser therapy (LLLT) with energy density of 2 J/cm2 induced the release of bFGF of the fibroblasts cultured in culture medium with high glucose concentration for one and two weeks (p=0.047 and p=0.04 respectively).Conclusion: This study showed that LLLT has stimulatory effects on the release of Interleukin 6 and basic fibroblast growth factor from cultured human fibroblasts in high glucose medium in one week and two-week tests.

Purpose: The Increase of oxidative stress production a causes after oxygen free radicals one of the factors involved in male infertility. In this study the role of leptin in product reactive oxygen species in seminal plasma in adult rat were evaluatedMaterials and Methods: Sixty-five male rats were randomly divided into four groups. The control group receives normal saline and other animal, in experimental groups 2, 3 and 4 were given daily intraperitoneal leptin injections of 5, 10, or 30 mg/kg body weight for 7, 15 and 42 at all. Reactive Oxygen Species Levels and Expression of leptin were measured respectively with DCFH-A and immunohistochemical and sperm parameters were assessed by computer-assisted semen analysis (CASA). Two ways ANOVA with Tukey post hoc test were conducted to compare mean of groups. P-value<0.05 was considered significant.Result: The results show that sperm count was higher in treated groups than that of control group. These differences were significant (p£0.05). The interaction between treatment and time showed significant effect on ROS variations.After immune staining leptin were found in the seminiferous, tubules and Leptin receptor were found in the interstitium.Conclusion: the results of this study indicate that leptinadministration by increasing levels of oxygen free radicals, may lead toincrease of sperm count and the number of abnormal sperm in the experimental groups.

Purpose: The present study was conducted to evaluate the effect of different concentrations of morphine on staurosporine-induced neurite outgrowth in PC12 cells.Materials and Methods: PC12 cells were cultured in RPMI1640 culture medium supplemented with 0.2% BSA. Cells were divided into three groups; Ι, ΙΙ and ΙΙΙ, culture in the presence of 50, 100 and 214 nM staurosporine respectively. In each group cells were treated with different concentrations (10-12, 10-10, 10-8, 10-6, 10-4 and 0.0 M) of morphine; treatments 1, 2, 3, 4, 5 and 6 respectively. The percentage of cytotoxicity was performed by lactate dehydrogenase assay. The percentage of death cell was assessed by Hoechst/PI differential staining. Total neurite length was measured.Results: In all groups, the percentage of cytotoxicity as well as the percentage of cell death in treatments 4 and 5 was higher and in treatments 1 and 2 was lower than that of treatment 6 (p<0.05). In group ΙΙΙ, total neurite length in treatments 1 and 2 was higher than that of treatment 6 and in group I, total neurite length in treatments 4 and 5 was lower than treatment 6 (p<0.05). Opioid receptor antagonist (naltrexone) was caused reversed the effects of morphine (p<0.05).Conclusion: Our results suggest that morphine at low concentrations should improve staurosporine-induced neurite outgrowth in PC12 cells.

Purpose: the aim of thos survey is establishment of an animal model through which a rapid distortion of the hair cell of cochlea administering amikacin and furosemide and using distortion product otoacoustic emission (DPOAE) the functioning of rat’s ear would be assessed.Materials and Methods: Forty-eight male Sprague dawley rats (with average of weight 200-250g, 12 weeks post natal) were randomly divided into six equal groups. Except the control group the rest received 0.5mg/g, 0.75mg/g, 1mg/g, 1.25mg/g, and 1.5mg/g, of subcutaneous amikacin respectively. 30 minutes later every rat received 0.1mg/g of furosemide intrapritoneally. DPOAE of rats was measured prior to these injections and 72 hours later. Then tissue section of the rat’s cochlea was prepared.Data were analyzed by means of SPSS (version 17) and ANOVA test and paired sample T test at the significant level of p£0.05.Results: All the cases had a significant decrease in their DPOAE with the frequencies 2KHz-8KHz (p£0.05). The most change in DPOAE was in rats which had received 1mg/g–1.5mg/g amikacin. Histological studies approved distortion of hair cell even within the apical turn.Conclusion: To establish a deafness model due to losing hair cells, it is recommended to use a combination of 1mg/g amikacin and 0.1mg/g furosemide. Besides, DPOAE can be used to approve deafness.

The heat shock proteins (HSPs) are a group of homologous proteins encoded with a multigene family. They can be divided into five groups based on both size and function; including HSP27, HSP60, HSP70, HSP90 and HSP100. Expression of HSPs can be constitutive and can increased when cells undergo metabolic and/or heat stress. HSPA2/Hsp70-2 is a kind of HSP70 and constitutively expressed in testis. This protein is essential for sperm maturation and is associated with infertility. HSPA2 expression has been shown to occur in two phases in human testis; 1) meiosis as an element of the synaptonemal complex and 2) spermiogenesis durinat sperm maturation and is considesered as an index for matured sperm. Several functions have been proposed for HSPA2 protein in human testis including; protein folding, formation and desynapsis of synaptonemal complex, repair of DNA strand breaks, plasma membrane remodeling, and the removal of excess cytoplasm. Background literature suggests that HSPA2 play a key role in spermatogenesis. Therefore, the aim of this biological- medicine study was the evaluation of the role and effect of HSPA2 in male infertility.

Variations in the arterial pattern of the upper limb are abundant and diverse and have been observed frequently either in cadaver dissection or in clinical practice and reports. In the most of these reports, radial or ulnar arteries were commonly involved, which can be damaged during surgical procedures. This report presents a case of a rare variation in arising branches of brachial artery. During routine anatomical dissection of a cadaver about 65-years-old Caucasian male, in anatomy department of Birjand University of Medical Sciences a variation in ulnar artery was observed. This variant artery arose from the lower third of the brachial artery, passed subcutaneously in the forearm. In the hand, this artery coursed over the flexor retinaculum and anastomosed with the superficial palmar branch of the radial artery to create the superficial palmar arch. Common interosseus and ulnar recurrent branches, arose from the radial artery, instead of, the ulnar artery in cubital fossa.As the radial and the ulnar arteries are widely used in cardiac catheterization, coronary angiography, arterial injections in the forearm, as well as cardiac surgery and kidney transplantation, hence, awareness of their variations are important clinically.