The Effect of Mesenchymal Stem Cells Derived Microvesicles on the Treatment of Experimental CCL4 Induced Liver Fibrosis in Rats

BACKGROUND AND OBJECTIVES: The release of microvesicles (MVs) from mesenchymal stem cells (MSCs) has been implicated in intercellular communication, and may contribute to beneficial paracrine effects of stem cell-based therapies. We investigated the effect of administration of MSC-MVs on the therapeutic potential of carbon tetrachloride (CCL₄) induced liver fibrosis in rats.METHODS: Our work included: isolation and further identification of bone marrow MSC-MVs by transmission electron microscopy (TEM). Liver fibrosis was induced in rats by CCl4 followed by injection of prepared MSC-MVs in injured rats. The effects of MSC-MVs were evaluated by biochemical analysis of liver functions, RNA gene expression quantitation for collagen-1α, transforming growth factor β (TGF-β), interleukin-1β (IL-1β), vascular endothelial growth factor (VEGF) by real time reverse transcription PCR (RT-PCR) techniques. Finally histopathological examination of the liver tissues was assessed for all studied groups.RESULTS: BM-MSC-MVs treated group showed significant increase in serum albumin levels, VEGF quantitative gene expression (p < 0.05), while it showed a significant decrease in serum alanine transaminase (ALT) enzyme levels, quantitative gene expression of TGF-β, collagen-1α, IL-1β compared to CCL₄ fibrotic group (p < 0.05). Additionally, the histopathological assessment of the liver tissues of BM-MSC-MVs treated group showed marked decrease in the collagen deposition & improvement of histopathological picture in comparison with CCL₄ fibrotic group.CONCLUSIONS: Our study demonstrates that BM-MSC-MVs possess anti-fibrotic, anti-inflammatory, and pro-angiogenic properties which can promote the resolution of CCL₄ induced liver fibrosis in rats.

Combination of Obestatin and Bone Marrow Mesenchymal Stem Cells Prevents Aggravation of Endocrine Pancreatic Damage in Type II Diabetic Rats

One of the new promising therapies in treatment of diabetes mellitus is mesenchymal stem cells (MSCs) which have an interesting therapeutic potentiality based on their paracrine effect and transdifferentiation potentiality. Also obestatin improves the generation of functional β cells/islet-like cell clusters in vitro, suggesting implications for cell-based replacement therapy in diabetes. So the aim of this study was to evaluate the effect of combination of both MSCs and obestatin on an experimental model of type II diabetes mellitus (T2DM). Sixty male rats were divided into; group I (control group), group II (T2DM group) induced by administration of high fat diet (HFD) and injection of streptozotocin (STZ) in low dose, group III (T2DM treated with MSCs), group IV (T2DM treated with obestatin), group V (T2DM treated with MSCs and obestatin). Fasting blood glucose, C-peptide, insulin and lipid profile were measured. HOMA-IR and HOMA-β were calculated. Pancreatic expression of insulin, glucagon like peptide -1 (GLP-1) and pancreatic duodenal homeobox 1 (Pdx1) mRNA levels were measured. In addition pancreatic histological changes, insulin and Bax were analyzed by immunohistochemical examination of islets of Langerhans. Diabetic rats showed significant increase in HOMA-IR, serum glucose and lipid profile levels with significant decrease in insulin, HOMA-β, GLP-1 and Pdx1 levels. MSCs and obestatin caused significant improvement in all parameters with more significant improvement in combined therapy. The protective effects afforded by MSCs and obestatin may derive from improvement of the metabolic profile, antiapoptosis and by increase in pancreatic GLP-1and Pdx1 gene expression.

Cardioprotective Effects of Wharton Jelly Derived Mesenchymal Stem Cell Transplantation in a Rodent Model of Myocardial Injury

BACKGROUND: Whartons jelly-derived mesenchymal stem cells are a valuable alternative source that possess multipotent properties, easy to obtain and available in large scale compared to BMMSCs. We investigated the possibility of cardiac function improvement post isoproterenol induced cardiac injury in a rat model following human WJMSCs transplantation. MATERIALS AND METHODS: MSCs were extracted and cultured from cord WJ, characterized by morphology, Immunophenotyping and differentiation to osteoblast and adipocytes. WJMSCs were labeled with PKH2 linker dye. Wistar rats were divided into control group, ISO group (injected with 2 doses of isoproterenol) to induce myocardial injury and ISO group transplanted with labelled WJMSCs. ECG, electrocardiographic patterns, cardiac marker enzymes, tracing of labeled MSCs and immunohistochemical analysis of myocardial cryosections were studied. RESULTS AND CONCLUSIONS: WJ derived MSCs were expanded for more than 14 passages while maintaining their un-differentiated state, were positive for MSC markers and were able to differentiate into adipocyte and osteoblast. We demonstrated that intravenously administered WJMSCs were capable of homing predominently in the ischemic myocardium. Cardiac markers were positively altered in stem cell treated group compared to ISO group. ECG and ECHO changes were improved with higher survival rate. WJMSCs could differentiate into cardiac-like cells (positive for cardiac specific proteins) in vivo. WJMSCs infusion promoted cardiac protection and reduced mortality, emphasizing a promising therapeutic role for myocardial insufficiency.

Comparative Evaluation for Potential Differentiation of Endothelial Progenitor Cells and Mesenchymal Stem Cells into Endothelial-Like Cells

Understanding the mechanisms of vascular remodeling could lead to more effective treatments for ischemic conditions. We aimed to compare between the abilities of both human Wharton jelly derived mesenchymal stem cells (hMSCs) and human cord blood endothelial progenitor cells (hEPCs) and CD34+ to induce angiogenesis in vitro. hMSCs, hEPCs, and CD34+ were isolated from human umbilical cord blood using microbead (MiniMacs). The cells characterization was assessed by flow cytometry following culture and real-time PCR for vascular endothelial growth factor receptor 2 (VEGFR2) and von Willebrand factor (vWF) to prove stem cells differentiation. The study revealed successful isolation of hEPCs, CD34+, and hMSCs. The hMSCs were identified by gaining CD29+ and CD44+ using FACS analysis. The hEPCs were identified by having CD133+, CD34+, and KDR. The potential ability of hEPCs and CD34+ to differentiate into endothelial-like cells was more than hMSCs. This finding was assessed morphologically in culture and by higher significant VEGFR2 and vWF genes expression (p<0.05) in differentiated hEPCs and CD34+ compared to differentiated hMSCs. hEPCs and CD34+ differentiation into endothelial-like cells were much better than that of hMSCs.

Comparative study between the effects of human CD34 and rat bone marrow mesenchymal stem cells on amelioration of CCl[4] induced liver fibrosis

Human umbilical cord blood [UCB] cells and rat bone marrow mesenchymal stem cells [BM-MSCs] have many advantages as grafts for cell transplantation. The aim of this study was to investigate the impact of UCB cells and BM-MSCs on reversal of hepatic injury and revival of hepatic function in a rat model of carbon tetrachloride [CCl[4]]-induced liver fibrosis

Novel prognostic biomarkers for HCC progression in Egyptian patients

Objectives: To evaluate values of Cyclin D and Cdk4 in HCC, chronic hepatitis C, HCV related liver cirrhosis and healthy controls, their clinico-radiological correlations and prognosis of HCC

Methods: Group 1: Fifty patients with HCC, Group 2.Fifty patients with chronic hepatitis C with or without cirrhosis and Group 3: Thirty healthy controls were enrolled. All patients were positive for hepatitis C virus [HCV] antibody and confirmed by HCV RNA. Calculation of Barcelona-Clinic Liver Cancer [BCLC] staging system, MELD and Child-Pugh scores. mRNA for cyclin Dl and Cdk4 were analyzed by quantitative RT-PCR

Results: The mean Cyclin Dl and Cdk4 values were higher in HCC group compared with the other two groups [p value= 0.001]. In HCC group, the mean Cdk4 and cyclin Dlvalues were significantly higher among HCC patients with multiple hepatic focal lesion [HFL] [p value= 0. 0001, and003 respectively] compared with those with single lesion. A significant correlation between size of [HFL], alpha-Fetoprotein[AFP] and mean Cdk4 value [p value= 0.028, 0.0001 respectively]

Conclusions: Significant values of cyclin Dl and Cdk4 were found in HCC, compared to normal and chronic hepatitis C and correlated to the number, size of HFL and AFP level. Thus, the assessment of cyclin Dl and Cdk4 may provide a novel strategy for prognostication and targeted therapy of HCC

Retraction: Adipose Stem Cells as Alternatives for Bone Marrow Mesenchymal Stem Cells in Oral Ulcer Healing

This article has been retracted at the authors' request.

Interferon [IFN]-mediated antiviral activity against the hepatititis C [HCV] through microRNAs [microRNAs]

MicroRNAs are a class of small non-coding RNA molecules that function through post-transcriptional regulation of gene expression by a process termed RNA interference [RNAi], and that also have to prominence as critical regulators in a wide array of mechanisms of cell physiology. The study will attempt to evaluate the expression of several microRNAs in peripheral blood mononuclear cells [PBMCs] from patients with chronic hepatitis C [CHC] at 12 hours after the first injection of pegylated interferon in comparison with healthy controls. forty patients with chronic hepatitis C virus infection [CHCV], their age ranged between [20-56] years, selected from the National Hepatology and Tropical Medicine Research Institute were included in this study, after 12 hours of the first interferon injection, and twenty healthy individuals were included to serve as controls. All the patients and controls were subjected to the following history, clinical examination, abdominal ultrasonography and collection of samples for routine laboratory investigations. CBCs and Taqman quantitative RT-PCR for MicroRNAs expression analysis of miR- 128a, miR 196a, miR- 196b, miR 296. Our study revealed that the microRNAs had a higher levels of expression in cases of CHCV infection. Our study concluded that there's a highly significant increase in expression levels of IFN-induced microRNAs were observed in patients of microRNAs-128a, 196a, 196b, 296. The future use of miR inhibitors or mimics and / or siRNAs might be useful for the development of diagnostic and therapeutic strategies aimed at the recovering of protective innate responses in HCV infections

Adipose Stem Cells as Alternatives for Bone Marrow Mesenchymal Stem Cells in Oral Ulcer Healing

BACKGROUND AND OBJECTIVES: Adipose tissue is now recognized as an accessible, abundant, and reliable site for the isolation of adult stem cells suitable for tissue engineering and regenerative medicine applications. METHODS AND RESULTS: Oral ulcers were induced by topical application of formocresol in the oral cavity of dogs. Transplantation of undifferentiated GFP-labeled Autologous Bone Marrow Stem Cell (BMSCs), Adipose Derived Stem Cell (ADSCs) or vehicle (saline) was injected around the ulcer in each group. The healing process of the ulcer was monitored clinically and histopathologically. Gene expression of vascular endothelial growth factor (VEGF) was detected in MSCs by Reverse Transcription-Polymerase Chain Reaction (RT-PCR). Expression of VEGF and collagen genes was detected in biopsies from all ulcers. Results: MSCs expressed mRNA for VEGF MSCs transplantation significantly accelerated oral ulcer healing compared with controls. There was increased expression of both collagen and VEGF genes in MSCs-treated ulcers compared to controls. CONCLUSIONS: MSCs transplantation may help to accelerate oral ulcer healing, possibly through the induction of angiogenesis by VEGF together with increased intracellular matrix formation as detected by increased collagen gene expression. This body of work has provided evidence supporting clinical applications of adipose-derived cells in safety and efficacy trials as an alternative for bone marrow mesenchymal stem cells in oral ulcer healing.

Effect of pentoxifylline on serum hyaluronic acid in patients with non-alcoholic fatty liver disease

Liver fibrosis is the excessive accumulation of extracellular matrix proteins that occurs in most types of chronic liver diseases. Non-alcoholic fatty liver disease [NAFLD] can be associated with progressive hepatic fibrosis. In this study we evaluated the effect of pentoxifylline [PTX] on serum hyaluronic acid [HA] levels as a marker of fibrosis. In this study we included 30 subjects [14 males and 16 females], divided into three groups. The NAFLD group included 20 patients with fatty livers as shown by ultrasound examination. Patients were randomised into a placebo group of 10 patients who received a placebo, and a pentoxifylline PTX group of 10 patients who received pentoxifylline at 400 mg/day for 6 months. The control group included 10 normal individuals. In the NAFLD group the mean value of the base line serum HA was 133 +/- 150.48, while in the control group it was 33.5 +/- 10.01; the difference between the groups was statistically significant [p < 0.001]. The mean value of the base line serum HA in the PTX treated group was 169.5 +/- 156.19, while after 6 months of treatment it was 59 +/- 44.34, with a statistically significant difference [p = 0.007]. In the placebo treated group the mean value of the base line serum HA was 96.5 +/- 143.004, while after 6 months of treatment it was 59.7 +/- 44.29; this difference was not statistically significant [p = 0.594]. Our showed that, when administered for 6 months, PTX caused a significant decline in HA levels, which may be an index reflecting improvement of hepatic fibrosis. Further investigations should be conducted with a large number of patients to confirm our and correlate this with histological findings

Assessment of heme oxygenase-1 (HO-1) activity in the cavernous tissues of sildenafil citrate-treated rats / 亚洲男科学杂志(英文版)

<p><b>AIM</b>To assess heme oxygenase-1 (HO-1) activity in the cavernous tissue of sildenafil citrate-treated rats.</p><p><b>METHODS</b>One hundred and ninety-two Sprague-Dawley male rats, divided into four equal groups, were investigated. Group 1, the control group, received regular animal chow; group 2 received sildenafil citrate by intragastric tube; group 3 received sildenafil and HO inhibitor (zinc protoporphyrin, ZnPP); and group 4 received sildenafil and nitric oxide synthase (NOS) inhibitor L-nitroarginine methyl ester (L-NAME). Twelve rats from each group were killed after 0.5 h, 1 h, 2 h and 3 h of drug administration. Then HO-1 activity, cGMP levels and NOS enzymatic activity in the cavernous tissues were estimated.</p><p><b>RESULTS</b>In cavernous tissue, HO-1 activity, NOS enzymatic activity and cGMP concentration increased significantly in sildenafil-treated rats compared to other groups throughout the experiment. Rats receiving either HO or NOS inhibitors showed a significant decrease in these parameters. HO-1 cavernous tissue activity and NOS enzymatic activity demonstrated a positive significant correlation with cGMP levels (r = 0.646, r = 0.612 respectively; P < 0.001).</p><p><b>CONCLUSION</b>The actions of PDE5 inhibitor sildenafil citrate in the cavernous tissue are partly mediated through the interdependent relationship between both HO-1 and NOS activities.</p>

Plasma and cardiac oxytocin during rat gestation and postpartum: evaluation of their effects on the isolated perfused rat heart

Our purpose was to determine the dynamic changes in plasma and cardiac oxytocin during gestation, to study the cardiac effects of oxytocin in the isolated perfused rat heart model and whether pregnancy modifies the hormone's action. Cardiac and plasma oxytocin were evaluated at three stages of gestation [6,13 and 20 days] and at 2 days postpartum. In addition, at the same stages, hearts were excised and attached to a Langendorf's apparatus. Heart rate [HR] and left ventricular developed pressure [LVDP] were measured while being exposed serially to same plasma concentrations of oxytocin observed at each studied stage. Compared with non-pregnant controls, plasma and cardiac oxytocin were significantly and respectively decreased by 41.9%; 38% in early, 42.5%; 36.6% in mid and 8,9%; 47.9% in late gestation, to increase again by 178.1% and 81% postpartum. Hearts from pregnant rats had higher HR, LVDP and delta P/delta t[max] than did hearts from non-pregnant animals. Postpartum, hearts showed lower HR by 9.2%, LVDP by 6.9% and delta P/delta t [max] by 13.9% compared to non-pregnant rats. During pregnancy, reproductive hormones may regulate cardiac oxytocin release and consequently its negative chronotropic and inotropic effects. After parturition, an activated oxytocin system may help the body to get rid of the excess volume

Annexin V as index of apoptosis in aorta of STZ induced diabetic rats

Diabetes mellitus [DM] is a chronic disease associated with hyperglycemia and the production of reactive oxidative intermediates and a disturbed antioxidant defense mechanism. Apoptosis or programmed cell death is a fundamental component of tissue differentiation and development, it also plays a central role in DM. The highly regulated mechanism of apoptosis involves an externalization process of phosphatidylserine [Ps]. Annexin V binds with high affinity to Ps-exposing apoptotic cell and can inhibit thereby the procoagulant and proinflammatory activities of the dying cell. Heat shock proteins have been shown to protect organism in vitro and in vivo against oxidative stress which plays a role in apoptosis in DM. HSP72 inhibits mitochondria! cytochrome release and subsequent caspase activation that leads to apoptosis. Fifty male albino rats weighing 170-200gm were used in this study. They were divided in to 5 equal groups, each of 10 rats. Group I: Control group. Group II: Diabetic group. Type 2 diabetes mellitus was induced by intraperitoneal injection of streptozotocin at a dose of 40mg/kg body weight. Group III: Diabetic group, treated with 1 unit insulin injected subcutaneously daily. Group IV: Diabetic group, receiving vit. E 600mg/kg B.Wt injected intramuscularly, 3 times/week, All groups were sacrified one month after the beginning of the experiment. Blood samples were obtained from retro-orbital vein for assessment of glucose and malondialdehyde [MDA], then Animals were sacrificed and aortic tissues were removed,PCR for Annexin V and for HsP72 were done. Compared to control group, fasting serum glucose is significantly higher in the diabetic group [group II] [p<0.05] and it decreased significantly with administration of insulin [group III]. MDA is significantly higher in the diabetic group II compared with control one [p<0.05]. It decreased significantly with administration of insulin [group III] or vitamin E [group IV] in comparison to diabetic group [group II]. Administration of vitamin E and insulin [group V] leads to significant reduction in MDA back to control level. The expression of Annexin V is significantly higher in the diabetic group [group II] compared to control group [p<0.05]. Administration of insulin [group III] or vitamin E [group IV] decreases it significantly. The expression of HSP72 is significantly lower in the diabetic group compared to control one [p<0.05]. It increased significantly with administration of insulin [group III] or vitamin E [group IV]. Serum MDA level rise in STZ induced diabetic rats as a marker of oxidative stress and administration of vitamin E was found to normalize this level, in addition a significant rise in expression of Annexin V as a marker of apoptosis in aorta of STZ induced diabetic rats with decreased expression of HSP72 which may be involved in cellular protection against oxidative stress in DM