Improvement of heart failure by human amniotic mesenchymal stromal cell transplantation in rats

Background: Recently, stem cells have been considered for the treatment of heart diseases, but no marked improvement has been recorded. This is the first study to examine the functional and histological effects of the transplantation of human amniotic mesenchymal stromal cells [hAMSCs] in rats with heart failure [HF]

Methods: This study was conducted in the years 2014 and 2015. 35 male Wistar rats were randomly assigned into 5 equal experimental groups [7 rats each] as 1- Control 2- Heart Failure [HF] 3- Sham 4- Culture media 5- Stem Cell Transplantation [SCT]. Heart failure was induced using 170 mg/kg/d of isoproterenol subcutaneously injection in 4 consecutive days. The failure confirmed by the rat cardiac echocardiography on day 28. In SCT group, 3×106 cells in 150 microl of culture media were transplanted to the myocardium. At the end, echocardiographic and hemodynamic parameters together with histological evaluation were done

Results: Echocardiography results showed that cardiac ejection fraction in HF group increased from 58/73 +/- 9% to 81/25 +/- 6/05% in SCT group [p value < 0.001]. Fraction shortening in HF group was increased from 27/53 +/- 8/58% into 45/55 +/- 6/91% in SCT group [p value < 0.001]. Furthermore, hAMSCs therapy significantly improved mean diastolic blood pressure, mean arterial pressure, left ventricular systolic pressure, rate pressure product, and left ventricular end-diastolic pressure compared to those in the HF group, with the values reaching the normal levels in the control group. A marked reduction in fibrosis tissue was also found in the SCT group [p value < 0.001] compared with the animals in the HF group

Conclusion: The transplantation of hAMSCs in rats with heart failure not only decreased the level of fibrosis but also conferred significant improvement in heart performance in terms of echocardiographic and hemodynamic parameters

Neuronal differentiation of rat hair follicle stem cells: the involvement of the neuroprotective factor seladin-1 [DHCR24]

The seladin-1 [selective Alzheimer disease indicator-1], also known as DHCR24, is a gene found to be down-regulated in brain region affected by Alzheimer disease [AD]. Whereas, hair follicle stem cells [HFSC], which are affected in with neurogenic potential, it might to hypothesize that this multipotent cell compartment is the predominant source of seladin-1. Our aim was to evaluate seladin-1 gene expression in hair follicle stem cells. In this study, bulge area of male Wistar rat HFSC were cultured and then characterized with Seladin-1 immunocytochemistry and flow cytometry on days 8 to 14. Next, 9-11-day cells were evaluated for seladin-1 gene expression by real-time PCR. Our results indicated that expression of the seladin-1 gene [DHCR24] on days 9, 10, and 11 may contribute to the development of HFSC. However, the expression of this gene on day 11 was more than day 10 and on 10th day was more than day 9. Also, we assessed HFSC on day 14 and demonstrated these cells were positive for beta-? tubulin, and seladin-1 was not expressed in this day. HFSC express seladin-1 and this result demonstrates that these cells might be used to cell therapy for AD in future

immunohistochemistry and toluidine blue roles for helicobacter pylori detection in patients with gastritis

Background: helicobacter pylori, which is associated with many upper gastrointestinal diseases, is found in half of the population of the world. Several special stains and immunohistochemistry stain for H. pylori are available. The need for and usefulness of immunohistochemical [IHC] technique has been debated for years. Toluidine blue is a simple stain for microbiological studies and is easily available in laboratories. Therefore, this study was conducted to compare hematoxylin and eosin [HandE], Giemsa and toluidine blue staining with immunehistochemistry for detection of H. pylori in patients with gastritis and also to correlate the results of these staining methods with pathological grading

Methods: we reviewed 54 consecutive gastric biopsy specimens stained by HandE and Giemsa as well as by toluidine blue and immunohistochemistry stains for H. pylori

Results: H. pylori was positively identified by IHC in 43 [79.63%] patients, while positive samples were found in 18 [33.33%], 24 [44.44%] and 33 [61.11%] patients using HandE, Giemsa and toluidine blue staining methods. Our results showed that classical histological staining methods are not sensitive enough to identify low numbers or coccoid forms of organism, while toluidine blue and immunohistochemistry play an important role in detection of H. pylori infection

Conclusion: toluidine blue has been proved to be much more reliable than HandE and Giemsa in detection of H. pylori. In addition, in post treatment biopsies and in biopsies with unexplained chronic active gastritis without histological evidence of H. pylori should have immunohistochemistry done to detect possible low density or coccoid form of organisms. Iran. Biomed. J. 17 [1]: 36-41, 2013

role of biodegradable engineered nanofiber scaffolds seeded with hair follicle stem cells for tissue engineering

The aim of this study was to fabricate the poly caprolactone [PCL] aligned nanofiber scaffold and to evaluate the survival, adhesion, proliferation, and differentiation of rat hair follicle stem cells [HFSC] in the graft material using electrospun PCL nanofiber scaffold for tissue engineering applications. The bulge region of rat whisker was isolated and cultured in DMEM: nutrient mixture F-12 supplemented with epidermal growth factor. The morphological and biological features of cultured bulge cells were observed by light microscopy using immunocytochemistry methods. Electrospinning was used for production of PCL nanofiber scaffolds. Scanning electron microscopy [SEM], 3-[4, 5-di-methylthiazol- 2-yl]-2, 5-diphenyltetrazolium bromide [MTT] assay, and histology analysis were used to investigate the cell morphology, viability, attachment and infiltration of the HFSC on the PCL nanofiber scaffolds. The results of the MTT assay showed cell viability and cell proliferation of the HFSC on PCL nanofiber scaffolds. SEM microscopy images indicated that HFSC are attached, proliferated and spread on PCL nanofiber scaffolds. Also, immunocytochemical analysis showed cell infiltration and cell differentiation on the scaffolds. The results of this study reveal that PCL nanofiber scaffolds are suitable for cell culture, proliferation, differentiation and attachment. Furthermore, HFSC are attached and proliferated on PCL nanofiber scaffolds

Delivery of epidermal neural crest stem cells [EPI-NCSC] to hippocamp in Alzheimer's disease rat model

Alzheimer's disease [AD] is characterized by progressive neuronal loss in hippocamp. Epidermal neural crest stem cells [EPI-NCSC] can differentiate into neurons, astrocytes and oligodendrocytes. The purpose of this study was to evaluate the effects of transplanting EPI-NCSC into AD rat model. Two weeks after induction of AD by injection of Amyloid-beta 1-40 into CA1 area of rat hippocamp, Y-maze and single-trial passive avoidance tests were used to show deficit of learning and memory abilities. EPI-NCSC were obtained from the vibrissa hair follicle of rat, cultured and labeled with bromodeoxyuridine. When Alzheimer was proved by behavioral tests, EPI-NCSC was transplanted into CA3 area of hippocamp in AD rat model. The staining of EPI-NCSC markers [nestin and SOX10] was done in vitro. Double-labeling immunofluorescence was performed to study survival and differentiation of the grafted cells. We showed that transplanted EPI-NCSC survive and produce many neurons and a few glial cells, presenting glial fibrillary acidic protein. Total number of granule cells in hippocamp was estimated to be more in the AD rat model with transplanted cells as compared to AD control group. We observed that rats with hippocampal damage made more errors than control rats on the Y-maze, when reward locations were reversed. Transplanted cells were migrated to all areas of hippocamp and the total number of granule cell in treatment group was equal compared to control group. Transplantation of EPI-NCSC into hippocamp might differentiate into cholinergic neurons and could cure impairment of memory in AD rat model

Role of endogenous vitamin E in renal ischemic preconditioning process: differences between male and female rats

Antioxidants such as alpha-tocopherol [vitamin E] and beta-carotene [vitamin A] play an important role in protective effect of repeated brief periods of ischemia, namely ischemic preconditioning [IPC]. Values of these antioxidants were investigated and compared after induction of ischemia reperfusion [IR] and kidney IPC in both male and female rats. Forty eight Wistar rats were divided randomly into six groups of 8: groups A and B [male and female controls, respectively], group C [male IR or IR cases], group D [female IR cases] and groups E and F [male and female IPC cases, respectively]. In groups C and D, ischemia was induced by clamping of left renal arteries for 45 min. In groups E and F, rats underwent four cycles of 4 min of arterial clamping and 11 min of de-clamping before final 45 min ischemia induction. Afterward, serum was collected to assess the blood urea nitrogen, creatinine and vitamins E and A values. Renal tissues were obtained for histological assessments. alpha-tocopherol levels in male and female rats showed a significant increase in IPC compared with IR group [P<0.01] and also in female IPC compared with male IPC group. beta-carotene levels had no significant variations. Histological evaluation showed that IR-induced renal injuries were less in female rats. Also, protective effects of IPC were more in female rats [P<0.01]. Renal IPC reduced damages in both male and female rats, but tissue injuries in females were decreased much more along with the increase of endogenous vitamin E

Neuropathological changes in brain cortex and hippocampus in a rat model of Alzheimer's disease

Alzheimer's disease [AD] is a neurodegenerative disorder with progressive loss of cognitive abilities and memory loss. The aim of this study was to compare neuropathological changes in hippocampus and brain cortex in a rat model of AD. Adult male Albino Wistar rats [weighing 250-300 g] were used for behavioral and histopathological studies. The rats were randomly assigned to three groups: control, sham and beta-amyloid [Abeta] injection. For behavioral analysis, Y-maze and shuttle box were used, respectively at 14 and 16 days post-lesion. For histological studies, Nissl, modified Bielschowsky and modified Congo red staining were performed. The lesion was induced by injection of 4 microL of Abeta [1-40] into the hippocampal fissure. In the present study, Abeta [1-40] injection into hippocampus could decrease the behavioral indexes and the number of CA1 neurons in hippocampus. Abeta injection CA1 caused Abeta deposition in the hippocampus and less than in cortex. We observed the loss of neurons in the hippocampus and cerebral cortex and certain subcortical regions. Y-maze test and single-trial passive avoidance test showed reduced memory retention in AD group. We found a significant decreased acquisition of passive avoidance and alternation behavior responses in AD group compared to control and sham group [P<0.0001]. Compacted amyloid cores were present in the cerebral cortex, hippocampus and white matter, whereas, scattered amyloid cores were seen in cortex and hippocampus of AD group. Also, reduced neuronal density was indicated in AD grouper ovariectomy

Prenatal mercuric chloride exposure causes developmental deficits in rat cortex

Introduction: Environmental pollution with heavy metals such as mercury is a major health problem. Growing studies on the field have shown the deleterious effects of mercury on human and nonhuman nervous system, especially in infants, however the effects of prenatal exposure to mercuricchloride on cortical development are not yet well understood. The aim of this study was to investigate the effect of prenatal exposure to mercuric chloride on morphological characteristics of brain cortex. Methods: Mercuric chloride [2 mg/kg] or normal saline were injected [I.P.] to 36 Sprague – dawley rats in the 8th, 9th or 10th day of gestation. The embryos were surgically removed in the 15th day of gestation, and brain cortices were studied by histological techniques. Results: Histological studies showed that embryos of mercuric chloride treated rats hadcortical neuronal disarrangement withdifferent orientations of nuclei, increased diameter of cortex, increased mitosis of cells, increased cell death, decreased cellular density and increased intracellular space. Conclusion: These findings suggest some micro structural abnormalities in cortical regions after prenatal exposure to mercuric chloride. These structural abnormalities may underliesome neurologic disturbances following mercury intoxication

Effects of total light deprivation on dorsal lateral geniculate nucleus of male neonate rats

This study examines the effects of total light deprivation on the developing lateral geniculate nucleus, the primary integration centre for visual information Sprague-Dawley rats were reared for one month in a dark room from 7th postnatal day before eye opening. A group of rats was taken back into normal condition for 15 days, and then perfused. Coronal sections of LGN were prepared and stained with Cresyl Violet and Cytochrome Oxidase to investigate the number of neurons, volume and length, as well as neuronal activity level. The results showed that LD for one month causes progressive loss of neurons and decreases neuronal activity level in the LGN. It can be concluded that during early postnatal development of the rats' visual system, light deprivation causes structural and functional changes in LGN

Effects of tamoxifen on morphological and ultrastructural aspects of developing hippocampus of rat

Tamoxifen treatment induced cell death in the hippocampus formation of the prenatal and postnatal rat. The present study delineates the effect of tamoxifen on developing hippocampus in prenatal, postnatal and full term neonate rats received certain doses of the partial antagonist tamoxifen. After perfusion and fixation, the brains were removed and processed for light and electron microscopy. The morphology, ultrastructure and the density of the neurons in different ages [E22, P1, P7 and P21] and in different areas of developing hippocampus including cornu ammonis [CA1 and CA3], dentate gyrus and subiculum were studied. These findings showed that in tamoxifen-treated groups, the cell number of pyramidal neurons of CA1 and subiculum significantly decreased comparing to control groups in E22, P1 and P7 but not in third weeks. The mitochondria of the above mentioned groups also showed a dilated feature with less cristae than control group and most of them were greatly enlarged and swollen into spherical shapes rather than the normal ovoid or rod shape. The present study shows that prenatal exposure to tamoxifen alters neurogenesis in developing rat hippocampus. These results demonstrated the non-neuroprotective roles of tamoxifen

Transplantation of olfactory mucosa improve functional recovery and axonal regeneration following sciatic nerve repair in rats

Olfactory ensheathing glia [OEG] has been shown to have a neuroprotective effect after being transplanted in rats with spinal cord injury. This study was conducted to determine the possible beneficial results of olfactory mucosa transplantation [OMT] which is a source of OEG on functional recovery and axonal regeneration after transection of the sciatic nerve. In this study, 36 adult female Sprague-Dawley rats were used. The sciatic nerve was transected in 24 rats and immediately repaired by sciatic-sciatic anastomosis, and randomly divided equally into two groups. The experimental group received the OMT at the transected site and the control group received the respiratory mucosa transplant. In another twelve rats as sham-operated animals, the sciatic nerve was exposed but no transection was made. DiI retrograde tracing was injected in the gastrocnemius muscle two months after surgery to allow visualization of the extent of axonal regeneration. Functional recovery was also assessed at 15, 30, 45 and 60 days after surgery using walking track analysis and sciatic function index [SFI] calculations. The total number of DiI labeled motorneurones in the ventral horn [L4-L6] and the SFI scores were significantly higher in the group of rats that received olfactory mucosa rather than respiratory mucosa. The outcome indicates that olfactory mucosa is a useful treatment to improve nerve regeneration in mammals with peripheral nerve injury

beneficial effect of [-]-epigallocatechin-3-gallate in an experimental model of Alzheimer's disease in rat: a behavioral analysis

Progressive cognitive decline is one of the hallmark symptoms of Alzheimer's disease [AD] which can be modeled by beta-amyloid injection into specific regions of brain. Since epigallocatechin-3-gallate [EGCG] is a potent antioxidant agent which its role against oxidative stress and inflammation has been shown in prior studies, we tried to determine whether EGCG administration protects against beta-amyloid-induced memory and coordination impairment in rats. Animals [male Wistar rats] were divided into four groups: sham operated, EGCG-pretreated sham operated [sham + EGCG], untreated lesion [lesion], and EGCG-pretreated lesion [lesion + EGCG]. Animals in lesion, lesion + EGCG, and sham + EGCG groups received sterile saline or saline plus EGCG [10 mg/kg] intraperitoneally one day pre-surgery and every other day for three weeks. The lesion was induced one day after EGCG pretreatment by injection of 4 microl of sterile saline or water containing 2 nmol/microl beta-amyloid [1-40] into the hippocampal fissure. For behavioral analysis, psychomotor coordination [PMC] index and spontaneous alternation behavior were assessed using Rota-rod Treadmill and Y-maze, respectively at the third week post-lesion. We found that beta-amyloid [1-40] injection into hippocampus can decrease these behavioral indexes in lesion group in comparison with sham group which is similar to behavioral changes in AD. On the other hand, pretreatment with EGCG can improve the PMC index and spatial Y-maze alternation in the lesion + EGCG group in comparison with lesion group. We concluded that EGCG can be effective in restoring beta-amyloid-induced behavioral derangements in rats regarding coordination and memory abilities