Cell and Tissue Journal

Abstract Aim: This study was aimed to examine the effects of Stachys lavandulifolia extract (SL) on ethanol-induced testicular damages.
Material and methods: In the current study, 24 male Wistar rats were divided into 4 groups: control, ethanol (5 g kg -10) and ethanol (5 g kg -1) + SL (500 mg kg-1) group and SL (500 mg kg-1). After the treatment for 4 weeks, mean seminiferous tubule diameter (MSTD) and Johnsen’s mean testicular biopsy score (MTBS) criteria were used for histological evaluation. In addition, sperm parameters including sperm count and motility were analyzed.
Results: Our data indicate that ethanol administration caused a significant decrease in the sperm count and motility in the ethanol group but SL ameliorated this reduction in SL + ethanol group. Moreover, both MSTD and MTBS values were decreased in the ethanol-treated rats. However, administration of ethanol together with SL resulted in significant improvements in both of these values.
Conclusion: In conclusion, our study revealed that the reproductive toxicity was caused by ethanol may be prevented by SL treatment
 

Abstract Aim: In this study, the histology effect of caffeine administration was evaluated on retina and PAX6 expression in rat neonates.
Material and Methods: A total of 24 pregnant rats were divided into three groups. Intraperitoneal injection of saline in the control group and also 50 mg/kg and 100 mg/kg caffeine injection from the ninth day of gestation was done. At 20 or 21 embryonic day, neonates on the first and fifth days were euthanized anesthetized and the eyes were isolated and fixed. The neonates were examined for abnormalities. The Retina Histological and morphometric evaluations, eye tissue fluorescence changes, and PAX6 gene expression were investigated by Real time PCR.
Results: The results showed that the effects of maternal caffeine treatment on the retina in neonates are associated with atrophic changes and cell counts of ganglion cell layer, retinal thickness and thickness of the inner plexiform layer, in the animals were treated with caffeine, were significantly reduced (p < 0.05). Also caffeine induced behavioral changes in mothers and tissue fluorescence and significantly increased (P <0.05) the PAX6 gene expression in the groups receiving caffeine.
Conclusion: Administration of caffeine during pregnancy can induce histopathological changes in organogenesis and developing in rat neonate’s retina.
 

Abstract Aim: In this study, we use combination of enzymatic and explant methods, for isolation and culture of ADMSCs.
Materials and Method: Adipose tissues dissected out from the abdominal region of male Wistar rats. Tissues were minced into small pieces (1-2 mm), and then treated with trypsin-EDTA 0.25% for 30 minutes at 37 ̊C in shaker-incubator. Enzymatic treated tissues were centrifuged and floating parts were cultured. Statistical analysis of the cells number was investigated using GraphPad Prismv6 software.
Results: Results showed that ADMSCs isolated with our methods have growth characteristics similar to conventional methods. ADMSCs were maintained up to 10th passage and exhibited a homogeneous population in terms of appearance and morphology. We demonstrated that ADMSCs by these combination methods have mesenchymal characteristic markers (positive for CD90, CD44, CD73, CD105 and negative for CD35, CD45, CD11b).
Conclusion: In the current study, we showed that the combination of enzymatic and explant methods creates a simple, inexpensive and reproducible method for isolation and culture of ADMSCs.
 

Abstract Aim: In the current study, the protective effect of quercetin (as an antioxidant) was investigated on the ovarian tissue and fertility of female rats exposed to cyclophosphamide.
Material and Methods: 24 Wistar rats were divided into four six-membered groups. The first group was treated with cyclophosphamide (30 mg/kg) and the second group was treated with tween solution. The third and fourth groups administered quercetin (50 and 100 mg/kg) with cyclophosphamide (30mg/kg) intraperitonealy for 30 days. The left ovaries of the rats were removed and after sectioning and staining using Hematoxylin-Eosin were examined using light microscopy. In addition, 16 rat were treated in the same way in 4 groups (one male and three female rat in each group), then the mice mating. After birth, growth indexes were studied. All data were analyzed using one-way ANOVA.
Results: Both doses of quercetin significantly increased the number of primordial follicles and diameter of the primary follicles, the number of blood vessels, the number of offspring and their growth indexes, while decreased the number of atretic graffian follicles,. In addition, a significant increase in the diameter of primordial follicles was observed in rats received 100 mg/Kg quercetin compared to rats received only cyclophosphamide.
Conclusion: Quercetin can reduce the side effects of cyclophosphamide on ovarian tissue and improve fertility indexes.
 

Abstract Aim: In the present investigation, the effects of grape seed (Vitis Vinifera) alcoholic extract were examined on skin Histomorphometric wound healing changes in diabetic Wistar rats.
Material and methods: forty eight male wistar rats were divided into four groups: negative control, positive control, the first experimental and the second experimental. In all of treated rats, a scar in the length of three centimeter was created on the left of the vertebral column. The wound healing was evaluated microscopic as well as macroscopic. The methods Mann – Whitney, Kruskal – Wallis were used for data analyses. The used software was SPSS ver. 16. 
Results: agglutination in diabetic groups was longer than normal, while the process in the group treated with V. vinifera was quicker than the control and also first experimental groups, respectively.
Conclusions: the obtained results showed that V. vinifera ointment accelerate the wound healing in normal and diabetic samples.

Abstract Aim: In present study the hepato-protective effects of Tragopogon graminifolius hydro-ethanolic extract (THE) was investigated in an experimental CCl₄- induced hepatotoxicity model.
Material and methods: Six groups (n =6) of male Wistar rats were treated with mixture of 2ml/kg BW of sterilized olive oil and CCl₄ (1:1), along with 200, 400 and 800 mg/kg BW of THE, respectively. Toxicant group received sterilized olive oil 2ml/kg and CCl₄ (1:1) mixture. After 96 hours, liver tissue pathological and serum biochemical parameters (ALT, AST and ALP) studies were done in CCL₄+THE different dosages treated groups in comparison with control.
Results: Results showed that receiving CCL₄ caused acute inflammation and necrosis in rat liver. THE treated groups significantly showed reduction in all of liver tissue biochemical and histological parameters (p<0.05).
Conclusion: It is concluded that T. graminifolius extract has protective chemical compounds such as antioxidants and flavonoids which are able to protect the liver from damages against CCl₄ poisoning by oxidative stresses confrontation.

Abstract Aim: The aim of this study was to determine whether varicocele can cause   any changes in number and size of ovarian follicles.
Materials and methods: In this study, 30 adult female Wistar albino rats in 3 groups (n=10) consisting of: 1. control, 2. Sham and 3. varicocele are prepared. Varicocele group were operated and left ovarian vein was respectively closed. After 2 months the animal scarified and the ovaries taken for histological process. Statistical analysis was performed using the one way ANOVA and Turkey’s tests were used for post hoc multiple comparisons, was considered statistically significant.
Results: There is no significant differences in rat and ovary weights before and after varicose induction between groups. The number of primordial follicles in varicocele group increased significantly. The number of primary, secondary and graffian follicles in varicocele group decreased significantly. The size of secondary follicles in varicocele group decreased significantly (p < /em><0.05).
Conclusions:  Finally the results of this study shows that varicocele may lead to increased female infertility through various factors, which includes changes in follicular growth.

Abstract Aim: The aim of this study was to evaluate the anti-diabetic effects of Tragopogon graminifolius hydroethanolic leaf's extract (TGE) on blood glucose and insulin level in diabetes male rats. Material and methods: In this experimental study 42 Wistar male rats were randomly divided in 6 groups: control, diabetic (streptozotocin60 mg/kg, i.p), experimental diabetic groups (treatment with TGE , 200, 400 and 800 mg/kg, i.p) and diabetic treatment with metformin (500mg/kg, gavaged) for 10 days. The blood glucose was examined daily by glucometer. At the end of examination blood samples and pancreatic tissue were collected for insulin evaluation and histological study. All data are expressed as mean±SEM and statistical significance was considered at p < strong>Results: Treatment with the 800 mg/kg TGE decreased significantly blood glucose in diabetic rats compared with control group (p < 0.05). In addition, administration of 400 and 800 mg/kg of TGE could significantly decreased plasma glucose levels compared with metformin group. The insulin blood levels increased significantly in treated groups compared with diabetic group (p < 0.05). Conclusion: The TGE has flavonoid and phenolic compositions  and its prescription might be decrease blood glucose and increase blood insulin levels in diabetic rats. 

Abstract Aim: This study aimed to evaluate the histomorphometric and immunohistochemical parameters of the repair of femoral bone defect using bone marrow stromal cells on gelatin – chitosan membrane in adult Albino Wistar rats. Materials and Methods: In this experimental study, sixty male Albino wistar adult rats were equally divided into five groups as follows: Control group that received no treatment after bone defect. Sham group that after bone defect, the culture medium was injected locally at the site of defect. Gelatin-chitosan group that membrane was used into bone defect. Cell group that nonautolog BMSCs were injected locally into defect .Cell-GC group that cell transplantation with chitosan - gelatin membrane were used into the bone defect. Results: The mean area of trabeculae in groups of membranes and cells significantly increased when compared to the control group.The mean number of osteocytes and cells in the bone defect in cell group  significantly increased when compared to the control group.No significant difference were found in chitosan - gelatin and sham groups compared to control group but the mean number of osteocytes significantly decreased in BMSCs with gelatin-chitosan scaffold group compared to control group (p < 0.001) Conclusion: BMSC transplantation and gelatin-chitosan scaffold are effective in repair of bone defect. However, the use of BMSCs with gelatin-chitosan scaffold is not effective in repair of  femoral bone defect.

Abstract Aim: The aim of this study was to investigate the preventive role of vitaminE (Vit.E) on spermatogenesis indexes in rats following exposure to Bisphenol A. Materialand and Methods: Adult male wistar rats with the mean body weight of 231±10g were randomly divided into 4 groups (n=6): control, BPA (250mg/kg/day), Vit.E (150mg/kg/day) and BPA + Vit.E.  Oral treatment was performed three times a week till 56 days. At the end of the treatment, the rats were killed and their left testis were weighed, fixed and stained using Heidenhain's Azan  methed. Histological and morphometrical analysis of spermatogenesis was carried out. Data were analyzed using One Way ANOVA and  the means were considered significantly different  at p < strong>Results: Testis weight (p < 0.01) significantly decreased in the BPA group compared to the control. A significant decrease (p < 0.001) in the mean diameter of seminiferous tubules and the germinal epithelium thickness and the indexes of tubular differentiation, spermiogenesis and meiosis were also observed in the testicular tissue of rats treated with BPA compared to the control ones. In the BPA +Vit.E group, the mentioned parameters increased significantly to the control level. Conclusion: Vitamin E can compensate for the undesired effects of BPA on spermatogenesis and therefore could be considered as a therapeutic supplement in the case of BPA toxicity.  

Abstract Aim: In this research morphological, cell culture, immunochemical and inductive properties of rat’s claw dermal cells were investigated. Material and Methods: First, the toes and fingers of rat were amputated at the point of the last joint and the amputated parts were processed for histological studies. Moreover, the dermal tissue located beneath the claw’s plate was dissected and cultured in vitro. Cultured cells at passage two or three were transferred on the slide and treated with α-smooth muscle actin (ASMA) antibody. Finally the cultured cells were stained with DiI and implanted into the vibrissae semi-follicles and ears. Results: The rat’s claw dermal cells in terms of morphological, cell culture and expression of ASMA demonstrated similarity with dermal papilla cells of hair follicles. These cells showed a bipolar spindle-shaped morphology having an extracellular matrix rich in glycosaminoglycans. Majority of the cells expressed ASMA in cytoplasm with a higher intensity in lamelopodia.  However, with respect to hair follicular dermal papilla cells, the claw dermal cells were not able to induce an ectopic epidermis to form the skin appendage.  Conclusion: The inductive capability which has been seen in hair follicle’s dermal papilla cells can not be generalized to dermal cells present in other skin appendages, including claw. Based on the results presented here, the inductive factors exist in hair follicle dermal papilla cells is not expressed in the claw dermal cells.

Abstract Aim: Sea cucumber has been paid attention in treatment of bone defects due to possessing bioactive compound such as chondroitin sulfate. There were done successful researches in the field of application of mesenchymal stem cells for treatment of bone injury recently. Therefore, this study performed to define the osteogenic and adipogenic potency of Persian Gulf sea cucumber alcoholic extract on differentiation rat bone marrow derived stem cells into osteogenic and adipogenic. Material and Methods: In this experimental study, bone marrow stromal cells were isolated by flushing from male Wistar rat and demonstrated stemness by immunocytochemistry. The cells in 9 experimental group were exposed to various concentration (3.5, 6.25, 12.5, 25, 50, 100, 200, 400, 800 µg/ml) of sea cucumber alcoholic extract and cytotoxicity of sea cucumber alcoholic extract determined by MTT assay. Eventually, after 21 day, osteogenic and adipogenic differentiation were evaluated with alizarin red, oil red staining and alkalin phosphatase kit. Data were analyzed by SPSS software, ANOVA and the level of p ≤ 0.05 was considered significant. Result: An appropriate dosage for treatment of mesenchymal stem cells was determined less than 50 µg/ml. Also, oil red staining showed that sea cucumber alcoholic extract does not have capacity of adipogenic induction while alizarin red staining and alkaline phosphatase revealed that concentration of 25 µg/ml sea cucumber alcoholic extract was most efficient concentration into osteogenic differentiation.  Conclusion: Sea cucumber extract was able to differentiate rat bone marrow mesenchymal stem cells into osteogenic lineage.

Abstract Aim: The aim of present study was to demonstrate that the obstruction of CSF flow in hydrocephalic Texas (HT-x) rat and the resulting changes in its protein components can cause abnormal cortical development.
Material and Methods: Cortical cells were taken from E18 and E21 Wistar fetuses and cultured for 24 hours in Neurobasal medium. Two groups of experiments were explored. These cells were treated with heated and non-heated CSF collected from E20 and E21fetuses. The cells were cultured for a further 48 hours.At the end of this time, proliferation assays and morphological analysis were performed
Results: Prenatal affected CSF had an inhibitory effect on the proliferation of normal primary fetal cortical cells. Heating the CSF removed this effect.
Conclusion: CSF contains important factors that can stimulate both proliferation and cell differentiation and that some of these are likely to be proteins. Disruption of CSF flow can cause the cortical deficiencies in patients with hydrocephalous.

Abstract Aim: Diabetes as a metabolic disorder affects different tissues and organs, among them the female reproductive tract such as uterus may be a vulnerable organ. This study was undertaken to study the effects of hyperglycemia and insulin therapy on uterus structure, estrous cycle and serum levels of pituitary and ovarian hormones in Wistar rats.
Material  and methods: Twenty-four female Wistar rats (180-200g) were randomly divided in: control, sham (Citrate buffer i.p), hyperglycemic (Streptozotocin; 60 mg/kg, i.p) and hyperglycemic+insulin (insulin 20 IU/Kg/day; s.c). At the end of experiment (36 Days) and in diestrus stage, all rats were weighted and blood samples were collected. Thereafter, their right uterus horn was taken out, weighted and then the middle third part of uterine horn was sampled and fixed (Bouein). After histological preparation, the histological and histomorphometrical examination of uterus was performed.
Results: Although in comparison with control and hyperglycemic+insulin groups, in hyperglycemic rats body weight, uterus weight and volume, endometrium and myometrium volume were dramatically decreased (p < 0.001) and the serum level of progesterone significantly increased (p < 0.001), vascular dilation and infiltrating cells resembling plasma cell was also observed. No significant difference was seen between control and insulin treated group for the above parameters.
Conclusion: Insulin deficiency, oxidative stress and alteration in the HPG axis which occurs in hyperglycemic condition, may affect the uterus structure. The observed structural changes might well play a significant role in the reproductive difficulties observed during hyperglycemia

Abstract Aim: In cholestasis changing bile acids and salts and opioids is associated with trauma, fibrosis and necrosis liver tissue. For examination of liver truma advance, this study shows time effects of closing bile duct on rat liver histological changes.
Material and Methods: Cholestasis was induced in male Wistar rats by bilateral ligation of bile duct and cutting. Animals were divided to four groups (Sham, 7, 13 and 21 days after cholestasis). Rats were killed under anesthesia and immediately their livers were dissected. The specimens were processed routinely and were sectioned into 5 microns thickness slices. The sections were stained by Hematoxiline-Eosin (H&E) method and then studied using optical microscope.
Results: Seven days after cholestasis, hepatocyte necrosis and cell disarrangements were observed in liver sections. Thirteen days after cholestasis, necrotic cells were increased and wrinkle chromatic nuclei were seen. Twenty-one days after cholestasis widespread necrosis was seen. Nuclei condensation and bill duct reproduction were observed. Also cell wall disappeared and tissue was out of order.
Conclusion: Data showed that after disease progression and accumulation of bill salts and acids, expansion of necrosis and fibrosis were rapid, and creation of cirrhosis is a good reason for rapid recognition and treatment of the cholestasis.
 
 

Abstract Aim: Ovariectomy decreases epithelial thickness of endometrium, number of cells and mucosal glands and changes other layers of uterus. Quercetin as a flavonoid has several properties, including free radical scavenger and antioxidant. This study has considered quercetin effects on uterus tissue in ovariectomized rats.
Material and Methods: In this study, 25 female Wistar rats were divided in 5 groups (5 samples per group): control, ovariectomized, ovariectomized-treated quercetin, non ovariectomized-treated quercetin, and sham. 15mg/kg bw/day was intraperitonellay injected for 15 days. At the end, after rat euthanization by Ether and removing their uterus, uterus tissues were fixed in formalin saline (10%). The tissue sections were prepared and studied using light microscope after H & E staining.
Results: The thickness of endometrial layer in ovariectomized rats decreased significantly and changed to cubic cells. Cells and glands numbers were significantly decreased in comparison with control. Endometrium thickness, cell and glands numbers significantly increased in ovariectomized-treated quercetin.
Conclusion: The results showed that quercetin can improve changes arising from ovariectomy in uterus tissue as antioxidant, when ovarian hormones are absent.
 

Abstract Aim: Para-Nonylphenol is an environmental pollutant which can generate free radicals causing tissue damage. The aim of this study was to investigate the effect of vitamin E as a strong antioxidant on kidney tissue in rats treated with para-Nonylphenolusing’stereological technique.
Material and Methods: Twenty fourmaleWistar rats with the mean body weight of 198±20 g were randomly divided into 4 groups (n=6), control, vitamin E (100mg/kg/day), Para-Nonylphenol (250mg/kg/day) and Para-Nonylphenol+ vitamin E, and treated orally for 56 days. Finally, the rats were weighted and their right kidneys were taken out, fixed, sectioned, processed and stained using Heidenhain̕& azan method. The kidney tissue was then studied using stereological methods. Data were analyzed using one way ANOVA and means difference was considered significant at p < 0.05.
Results: The mean kidney weight, the total volume of kidney, the volume of cortex and medulla, the volume of proximal and distal tubule lumen, interstitial tissue and the volume of glomerulus and tuft significantly increased in Para-Nonylphenol group compared to the control ones, while the volume of proximal and distal tubules epithelium and the volume of Bowman’s space reduced significantly.
Conclusion: Vitamin E could compensate many adverse effects of Para-Nonylphenol on the kidney tissue. Consequently, vitamin E could be used as a potential antioxidant (supplement) against NP intoxication.
 

Abstract Aim: Previous studies using LOH technique on BDII rats showed four distinct Allelic imbalance on chromosome 15. In this research we have studied region 2 including Allelic imbalance by bioinformatic approachs and FISH (Florescence in situ hybridization) technique.
Material and Methods: The  confirmed tumors by an expert pathologist were used for cell culture. Metaphase chromosomes were prepared by conventional methods. The corresponding genes labeled probe for hybridization was poured onto slides. The labeled probe was then subjected to detected phase. Slides were studied under a fluorescence microscope using software CW4000 Laica.
Results: Using some of the database we could register 104 genes. But most of them did not have a clear function. According of FISH results, Dlgap5, Fermt2 and Socs4 had amplification and Lgals3 had copy number reduction.
Conclusion: according to our results, the Dlgap5 and Socs4 are probably involve in EAC

Abstract Aim: The effects of mobile phones waves investigated on rat ovary.
Materials and Methods: 28 rats with weight of 200±20g and age of 80-90 days selected and divided into four groups including control, sham, exposed number 1 and 2. For two weeks and one month, experimental 1 and 2 groups were exposed to mobile calls 12 times a day and each time for 10 minutes. The sham group was exposed to non-calling switched-on mobile phones. The concentration of hormones measured using ELISA method. The number of follicles calculated with physical dissector technique.
Results: The results showed no significant difference in the ovarian weight and number of primary follicles and corpus luteum. In different groups the number of secondary follicles decreased significantly in the exposed and sham groups compared to the control group. The number of Graffian follicles decreased significantly in the exposed group number1and sham in comparison to the control group while the number of atretic follicles increased significantly in both exposed groups in compared to the control and sham groups. The level of LH increased significantly in exposed group number1 in compared to the control group while the level of FSH increased significantly in the exposed group number 2 when compared to the control and sham groups. The level of estrogen and progesterone increased significantly in the exposed groups in compared to the control and sham groups.
Conclusion: The mobile phone waves may increase ovarian follicles atresia, disrupt the regulation of hormonal secretion and affect the fertility rate.
 

Abstract Aim: This study is examination skin wound healing after using of lotus corniculatus L. aerial part extract in comparison with Phenytoin ointment in rat.
Materials and methods: After anesthesia of 80 Wistar male rats, a rectangular wound was produced at back each of which. Healing woud steps was studied in 8 treated groups: control (A), 1% Phenytoin cream (B), 10% extract ointment (C), 20% extract ointment (D), 40% extract ointment (E), 1% Phenytoin cream + 10% extract ointment (F), 1% Phenytoin cream +20% extract ointment (G), 1% Phenytoin + 40% extract ointment (H). Medication and measuring wounds were done using digital images analysis for 21 days. Samples were collected from wound healing sites for microscopy after 3, 7, 14 and 21 days. Wound healing was determined for all of treated groups. Data were analyzed using SPSS18.
Results: Healed wound area mean in E and H groups was lower than comparing to other groups (p < 0.01). Also based on histopathological results, E and H groups showed more healing wound in comparison with other groups.
Conclusion: According to anti-inflammation, anti-microbial and healing effects of Lotus corniculatus in comparison with 1% Phenytoin, it is believed that the plant compounds are more effective in full thick skin wound healing.