The effects of bone marrow-derived mesenchymal stem cells on ovalbumin-induced allergic asthma and cytokine responses in mice

Objective(s): Allergic Asthma is an inflammatory disease of the lungs that is characterized by increased infiltration of leukocytes into the airways, limiting the respiratory function. Studies suggest that a defective general regulatory system against inflammation could be a significant factor in allergic asthma. It has been shown that Mesenchymal stem cells (MSCs) have a cellular immunosuppressive therapeutic potential for inflammatory disorders. We investigated whether administration of MSCs during allergen challenge would affect the underlying mechanisms in allergic airways inflammation. Materials and Methods: Fifty mice were used in five control and experimental groups; the experimental mice sensitized by intraperitoneal injection of OVA and aluminum hydroxide emulsion on days 0, 7, and 14, were then challenged intranasally with OVA or sterile PBS on days 14, 25, 26, and 27. Before allergen challenge on day 14, experimental mice received tail vein injection of MSCs in PBS, whereas control mice received PBS alone. Cytokine and IgE analyses were carried out using lung washes as well as serum samples.Results: Our, results showed that MSCs significantly reduced total cells and eosinophilia and serum OVA-specific IgE concentration in OVA-sensitized and challenged mice. Also, results showed that MSCs markedly inhibited expressions of Th2 and Th17 cytokines and elevated levels of Treg cytokines. Conclusion: we found that administration of MSCs could be used as a potential therapeutic approach for allergic asthma

Functional recovery of sciatic nerve through inside-out vein graft in rats

AbstractObjectivePresent study aimed at further comprehensive functional, histomorphometrical and immunohistochemical assessment of peripheral nerve regeneration using rat sciatic nerve transection model.MethodsThe 10-mm rat sciatic nerve gap was created in rats. In control group nerve stumps were sutured to adjacent muscle and in treatment group the gap was bridged using an inside-out vein graft. In sham-operated group the nerve was manipulated and left intact. All animals underwent walking track analysis test 4, 8, and 12 weeks after surgery. Subsequently, muscle mass measurement was performed to assess reenervation, histological examination to observe the sciatic nerve regeneration morphologically and Immunohistochemistry to detect Schwann cells using anti S-100. Results were analyzed using a factorial ANOVA with two between-subjects factors. Bonferroni test for pairwise comparisons was used to examine the effect of treatments.ResultsFunctional analysis of myelinated nerve fibers showed that nerve function improved significantly in the time course in treatment group. However, quantitative morphometrical analysis of myelinated nerve fibers showed that there was no significant difference between 8 and 12 weeks in treatment group. Muscle weight ratio was bigger and weight loss of the gastrocnemius muscle was ameliorated by inside-out vein grafting. The position of positive immunohistochemical reactions further implied that regenerated axons and Schwann cell-like cells existed after vein grafting was performed, and was accompanied by the process of myelination and structural recovery of regenerated nerves.ConclusionFunctional analysis of peripheral nerve repair is far more reliable than quantitative morphometrical analysi

Fungicidal effect of Origanum vulgare essential oil against Candida glabrata and its cytotoxicity against macrophages

Introduction: Candida glabrata is a yeast fungus regularly isolated from patients with impaired immunity who receive a routine antifungal therapy. Drug-resistant strains of C. glabrata have been emerged in recent years. The aim of this study was to examine the therapeutic efficacy Origanum vulgare essential oil (OVEO) against drug-resistant strains of C. glabrata and its cytotoxic effect on macrophages.Methods: Specimens were collected from mucosal surfaces of the oral cavity of medically approved oropharyngeal candidiasis (OPC) in HIV-positive patients and volunteered healthy individuals using sterile swabs or mouthwashes. In vitro antifungal susceptibility testing was done using microdilution and disc diffusion methods. Chemical composition of OVEO was determined using gas chromatography mass spectrometry. The cytotoxic effect of essential oil on macrophages was examined using tetrazolium dye (MTT).Results: Minimum inhibitory concentration (MIC) range of OVEO in healthy individuals and OPC patients was 150-200 and 150-250 &mu;g/mL, respectively. OVEO efficiently inhibited growth of resistant isolates. In isolates obtained from HIV patients, both MIC50 and MIC90 of OVEO were 200 &mu;g/mL while in healthy individuals were 150 and 200 &mu;g/mL, respectively. Moreover, OVEO induced significant reduction in proliferation of murine RAW264.7 and peritoneal macrophages in concentrations higher than 100 and 300 &mu;g/mL, respectively. Main constituents of OVEO were thymol (27.3), &gamma;-terpinene (20.7) and carvacrol (16.1).Conclusion: OVEO could be used as a fungicidal agent against fungal infections caused by azole-resistant C. glabrata. A combination therapy along with standard antifungals is suggested to avoid its cytotoxic effects.</p

Morphological Features of Cell Death Through Microscopic View

Cells are active components in their environment and constantly adjusting their performance to improve extracellular milieu changing. This approach is reflected their tends of maintaining intracellular homeostasis. When the cells encounter stress or pathologic stimuli, they can undergo a new manner (adaptation) and new steady state for achieving viability and function. If the external stress is exceeded or adaptive capability continues, cell injury develops (1, 2). Cells injury is engaged in a process that is reversible or irreversible. Injury is reversible when the stimuli are limited and removed. Only in the persistent stress or pathologic stimuli a reversible phase converted into irreversible phase (point-of-noreturn) and causes cell death(3). However, cell death is the most important step in embryogenesis, organ development, hemostasis and also the evolution of disease in any organ (1). The most classifications of mammalian cell death are apoptosis and necrosis. Apoptosis occurs naturally in many situations and helps to eliminate the cells that lost their efficiency. Cells undergoing apoptosis show biochemical events lead to cell changes (morphology) and death (4, 5). These features include cell blebbing, shrinkage, chromatin condensation and nuclear fragmentation. Unlike necrosis, which is a pathologic cell death, apoptotic cells release cell fragment called apoptotic bodies (1, 6). This image was taken with light microscopes (Nikon, TE2000 inverted microscopes), peripheral blood monocytes during 13 days culture initially change into macrophage-like cells and then due to lack of nutrition materials and a high level of toxic substance, to undergo cell death. The morphological changes as mention above (Fig1 legend) approved apoptosis process and cell death. However, sometimes the end stage of apoptosis accompanied by necrosis and it is not possible to distinguish them from the microscopic view

Development of MF-59 adjuvant using chitosan derivatives to influenza vaccine

Background & Objective: The influenza virus causes hundreds of deaths in the world. Despite the availability of some authorized vaccines for all age groups and its approved beneficial effects, the influenza is still a major problem for public health. Materials & Methods: In this study, the adjuvant of MF59 was developed by methylglycol-chitosan. The physico-chemical characteristics and immune response stimulation of novel adjuvant were measured in combination with the influenza virus. Results: The new adjuvant was produced with a nano-droplet diameter of 156 and Zeta potential of 29. The pH of adjuvant was not significantly changed for two years. Immunization of mice with developed adjuvant makes more HAI titer than MF59 adjuvant. Conclusion: The favorable characteristics of physico-chemical properties and lack of local side effects of developed adjuvant as well as its strong humoral immunity can introduce it as a novel MF59 adjuvants

Effect of mesenchymal stem cells on allergic asthma in mouse model

Background: Allergic Asthma is an inflammatory disease of the respiratory system that is well known by increased inflammatory cells in the airways and causes difficulty in respiration. The prevalence of allergic asthma is increasing worldwide, and it has become a significant cause of health challenge especially in developed countries. Inhaled β2-agonists and Inhaled or oral corticosteroids are common medications for treating the disease, but they cannot be used for long periods of time because of frequently occurring side effects and they can’t change the main pathogenesis of the problem. Deficiency in regulatory system against inflammation could be an important factor in allergic asthma. Mesenchymal stem cells (MSCs) have potential of cellular immunosuppressive therapy of inflammatory disorders. The aim of present study was to evaluate the effects of MSC therapy on mechanisms of allergic asthma in mice model. Methods: This experimental study was conducted from August 2014 to March 2015. The animals were housed and maintained in Biotechnology Center of Urmia University, Iran. Mice were sensitized by intra-peritoneal injection of ovalbumin (OVA) and aluminum hydroxide emulsion and then were challenged intra-nasally with OVA. Before allergen challenge on day 14, experimental mice received tail vein injection of MSCs in PBS. Regulatory T cells of spleen, cytokines and IgE analysis were carried out using lungs wash as well as serum samples. Results: Our results showed that MSCs significantly reduced total cells and eosinophilia, serum OVA-specific IgE concentration in OVA-sensitized and challenged mice. Also results showed that MSCs markedly inhibited expressions of Th2 cytokines and elevated levels of Treg cells and Treg cytokines. Conclusion: In the present study, we demonstrated the inhibitory effect of MSCs on airway inflammation using mice model of allergic asthma. The mice were sensitized with OVA and compared to the results of dexamethasone administration. Our results demonstrated that administration of MSCs could be used as a potential therapeutic approach for the allergic asthma

Comparison The Effects of Two Monocyte Isolation Methods,Plastic Adherence and Magnetic Activated Cell Sorting Methods,on Phagocytic Activity of Generated Dendritic Cells

Objective: It is believed that monocyte isolation methods and maturation factors affect the phenotypic and functional characteristics of resultant dendritic cells (DC). In the present study, we compared two monocyte isolation methods, including plastic adherence-dendritic cells (Adh-DC) and magnetic activated cell sorting- dendritic cells (MACS-DC), and their effects on phagocytic activity of differentiated immature DCs (immDCs).Materials and Methods: In this experimental study, immDCs were generated from plastic adherence and MACS isolated monocytes in the presence of granulocyte-macrophage colony-stimulating factor (GM-CSF) and interleukin 4 (IL-4) in five days. The phagocytic activity of immDCs was analyzed by fluorescein isothiocyanate (FITC)-conjugated latex bead using flow cytometry. One way ANOVA test was used for statistical analysis of differences among experimental groups, including Adh-DC and MACS-DC groups.Results: We found that phagocytic activity of Adh-DC was higher than MACS-DC, whereas the mean fluorescence intensity (MFI) of phagocytic cells was higher in MACS-DC (p<0.05).Conclusion: We concluded that it would be important to consider phagocytosis parameters of generated DCs before making any decision about monocyte isolation methods to have fully functional DCs

Investigating the Effects of All-trans Retinoic Acid on Histopathology of Pancreas of streptozotocin -Induced Diabetes in C57BL/6 mice

Backgrounds & Objectives: Type 1 diabetes is an auto-immune disease and caused by insufficient insulin production by the body. All-trans Retinoic Acid (ATRA) is an antioxidant, anti-cancer and anti-inflammatory agent. The objective of the present study was to investigate the effects of ATRA on histopathology of pancreas in diabetic mice. Material & Methods: Diabetes was induced by multiple low-dose of streptozotocin injection (40 mg/kg/day for 5 consecutive days) in male C57BL/6 mice. After induction of diabetes, mice were treated with ATRA (20 mg/kg/day i.p.) for 21 days. On the last day, pancreases were isolated and stained with hematoxylin &eosin (H&E) and Gomeri aldehyde fuchsin (GAF) for histological analyses (the number of islets and β cells, diameter of islets) of pancreas. Results: ATRA treatment in streptozotocin-induced diabetic mice was increased the mean diameter of islets and the number of islets and beta cells compared to the diabetic group. (p<0/05). Conclusion: The administration of ATRA improved pancreas tissue during destruction of the pancreatic beta-cells in STZ-induced type 1 diabetes in mice

Ginger ameliorates reproductive toxicity of formaldehyde in male mice: Evidences for Bcl-2 and Bax

Introduction: Root of dietary ginger considerably improves the activity of antioxidant enzymes in reproductive system and reduces the signs of cell damage in testis tissues. The present study conducted numerous sperm, hormonal, bio-chemical analysis and gene expression in order to evaluate the reproductive damages caused by exposure to formaldehyde (FA) and to investigate the ameliorative properties of co-administration of FA and Zingiber officinale (Ginger) in mice model. Methods: Forty-eight male NMRI mice were randomized into 6 groups of 8 animals each, including control group, control sham (received distilled water by gavage), FA group (10 mg/kg twice per day), intraperitoneally (i.p) and 3 FA groups (10 mg/kg i.p) + Ginger (500, 1000 and 2000 mg/kg/d by gavage, respectively). Sperm parameters, sexual hormones, antioxidant activity and expression of Bax and Bcl-2 genes were analyzed after 35 days. Results: FA significantly diminished sperm parameters, sexual hormones and antioxidant enzymes (P < 0.05). Also, the expression of Bcl-2 and Bax genes had significant (P < 0.05) increase and decrease, respectively in FA group. Co-administration of Ginger extract significantly recovered the above parameters. Conclusion: Co-administration of Ginger extract ameliorates reproductive damages of FA by its androgenic, antioxidant and anti-apoptotic properties. Hence, it might be beneficial in these patients

Generation of Mature Monocyte-Derived Dendritic Cells in the Presence of Heparin and Monocyte Conditioned Medium: Phenotypic and Functional Comparison

ABSTRACT Background: Dendritic cells (DC) induce tumor or pathogen-specific T cell responses in humans. Several laboratories have developed culture systems, including maturation factors for human DC from peripheral blood monocytes. We comprehensively compared standard maturation stimulus, an autologous monocyte-conditioned medium (MCM), with heparin for their ability to promote uniformly mature DC that elicit T cell responses. Methods: A short (4-day) priming of plastic adherent monocytes with granulocyte-macrophage colony stimulating factor (GM-CSF) and IL-4 with or without heparin was followed by 48-hour incubation in MCM to generate fully mature and stable DC. Phenotypic and functional analyses were carried out using anti-CD14 and anti-CD83 monoclonal antibodies, and mixed lymphocyte reaction, respectively. Results: We found that fully matured DC with a large amount of cytoplasm and copious dendritic projections were visible at the end of culturing period in the presence of MCM, heparin and MCM plus heparin. Thus, DC generated with these maturation factors are nonadherent and have typical satellite morphology. Flow cytometric analysis using anti-CD14 (monocyte marker) and anti-CD83 (mature DC marker) revealed that expression of CD14 decreased in MCM plus heparin-treated DC, and the expression of CD83 was increased when heparin and MCM used as a maturation factor. Functionally, MCM and MCM plus heparin-treated DC showed stronger mixed leukocyte reaction than heparin alone. Conclusion: These results support the use of the MCM with heparin as maturation factor that could result in functionally mature monocyte-derived DC in comparison to either MCM or heparin alone. Iran. Biomed. J. 15 (3): 79-84, 201