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

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

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

The effect of Rat mesenchymal stem cells and its soluble factors on peripheral blood neutrophil function

Background & aim: Mesenchymal stem cells (MSCs) are a population of adult stem cells which is an appropriate source for therapeutic purposes. The aim of this study was to investigate the effects of rat mesenchymal cells and soluble factors on the function of peripheral blood neutrophils. Methods:This experimental study was conducted on rat mesenchymal stem cells. Mesenchymal cells obtained from bone marrow of the femur and Tybaof 6-8 week rats and were cultured in DMEM. After maturation, the mesenchymal cells and supernatant at ratios of 1:4, 1:2 and 3:4 were adjacent with peripheral blood neutrophil phagocytosis. Subsequently, the respiratory burst of neutrophils, the yeast phagocytosis and nitroblue tetrazolium test was evaluated for revival. The Data were analyzed by t-tests, ANOVA and Tukey's test (p ˂0.05). Results:The rates of phagocyted neutrophil treated with MSCs compared to controls were decreased.This reduction was not statistically significant (p >0.05).The phagocitic cell in the rats of the treated group with supernatant compared to the control group in all three ratios of 1:4, 1:2, 3:4 increased significantly (p>0.05). by the increase in the ratio was observed (P>0.05). Respiratory burst of neutrophils treated with mesenchymal stem cells compared to the control group significantly decreased. Respiratory burst was increased in the groups treated with cell supernatant at ratios of 1:2 only (P>0.05). Conclusion:Mesenchymal cell-cell interaction with neutrophils was remarkable for therapeutic strategies in diseases associated with neutrophil function in response to physiological and pathological cell therapy with MSCs

Changes in some pro-and anti-inflammatory cytokines produced by bovine peripheral blood mononuclear cells following foot and mouth disease vaccination

Interleukin (IL)-17 is exclusively produced by CD4 helper T-cells upon activation. It most often acts as a pro-inflammatory cytokine, which stimulates the release of pro-inflammatory cytokines IL-6, IL-8, TNF-α, and granulocyte-macrophage colony-stimulating factor (GM-CSF). In this study, we studied the in-vitro IL-17 response to specific antigens and a variety of mitogens and compared the IL-17 response to IL-2, IL-4, IL-5, IL-6, IL-10, and IFN-γ responses. We used a foot and mouth disease (FMD) vaccine as specific antigens and mitogens (phytohemagglutinin [PHA], pokeweed mitogen [PWM], and concanavalin A [Con A]) to stimulate peripheral blood mononuclear cells (PBMCs) of vaccinated calves. Cell culture supernatant was harvested and analyzed for cytokines, using commercially available bovine ELISA kits. The mitogens induced a significant increase in IL-17 production. IL-17 was produced at high levels in response to the T cell-stimulated mitogens, PHA, and Con A, and at low levels in response to PWM mitogens. In contrast, level of the produced IL-17 cytokines in response to the FMDV antigens was lower as compared to those produced by mitogens. The FMDV antigens and mitogens significantly increased IL-17 production. There was not a correlation between IL-17 production and type-1 cytokine, IFN-γ, and IL-2, while there was a correlation between type-2 cytokine, IL-4, and IL-5 at either cytokine level produced by PBMCs stimulated by FMDV antigens. Moreover, there was an interaction between IL-17 and IL-6, that is, as IL-6 cytokine level elevated or diminished, IL-17 cytokine level increased or decreased, as well

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

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

CuO nanoparticles induce cytotoxicity and apoptosis in human K562 cancer cell line via mitochondrial pathway, through reactive oxygen species and P53

Objective(s): This study focused on determining cytotoxic effects of copper oxide nanoparticles (CuO NPs) on chronic myeloid leukemia (CML) K562 cell line in a cell-specific manner and its possible mechanism of cell death. We investigated the cytotoxicity of CuO NPs against K562 cell line (cancerous cell) and peripheral blood mononuclear cell (normal cell). Materials and Methods: The toxicity was evaluated using cell viability, oxidative stress and apoptosis detection. In addition, the expression levels of P53, Caspase 3, Bcl-2, and Bax genes in K562 cells were studied by reverse transcription polymerase chain reaction (RT-PCR) analysis. Results: CuO NPs exerted distinct effects on cell viability via selective killing of cancer cells in a dose-dependent manner while not impacting normal cells in MTT assay. The dose-dependent cytotoxicity of CuO NPs against K562 cells was shown through reactive oxygen species (ROS) generation. The CuO NPs induced apoptosis was confirmed through acridine orange and propidium iodide double staining. Tumor suppressor gene P53 was up regulated due to CuO NPs exposure, and increase in Bax/Bcl-2 ratio suggested mitochondria-mediated pathway is involved in CuO NPs induced apoptosis. We also observed that Caspase 3 gene expression remained unchanged up to 24 hr exposure.                                                                             Conclusion: These molecular alterations provide an insight into CuO NPs-caused inhibition of growth, generation of ROS, and apoptotic death of K562 cells