Comparative study of the osteogenic potential of mesenchymal stem cells derived from different sources

Mesenchymal stem cells (MSCs) can regenerate missing tissues and treat diseases. Hence, the current work aimed to compare the proliferation rate and the osteogenic differentiation potential of bone marrow MSCs (BMSCs), gingival MSCs (GMSCs) and submandibular MSCs (SMSCs). MSCs derived from bone marrow, gingiva and submandibular salivary gland were isolated and cultured from rats. The proliferation capacity was judged by MTT proliferation Assay. Osteogenic differentiation was assessed by Alzarin red stain and quantitative RT-PCR was performed for Runx-2 and MMP-13. The highest significant proliferation was estimated in the BMSCs compared to GMSCs and SMSCs (p-value was < 0.01). All studied cell types formed mineralized nodules as stained with Alizarin Red stain at the 3rd passage of differentiation. However, BMSCs seemed to generate the highest level of mineralization compared to GMSCs and SMSCs. RT-PCR revealed that the expression of Runx-2 and MMP-13 mRNAs was significantly increased in the BMSCs compared to GMSCs and SMSCs (p-value was < 0.01). BMSCs displayed maximum osteogenesis results followed by the GMSCs and lastly by the SGSCs. Thus, it could be recommended that GMSCs can be used as a second choice after BMSCs when bone tissue reconstruction is needed

Homing and reparative effect of intra-articular injection of autologus mesenchymal stem cells in osteoarthritic animal model

<p>Abstract</p> <p>Background</p> <p>This work aimed to study the homing evidence and the reparative effect of mesenchymal stem cells (MSCs) in the healing process of induced osteoarthritis in experimental animal model (donkeys).</p> <p>Methods</p> <p>Twenty-seven donkeys were equally divided into 3 groups based on the observation period after induction of arthritis (3, 6 and 9 weeks) to achieve different degrees of osteoarthritis. Each group was subdivided into three subgroups of three animals each based on the follow-up period (1, 2 and 6 months) after treatment. The induction was done through intra-articular (IA) injection of 2 ml of Amphotericin-B in both carpal joints. MSCs were harvested in a separate procedure, labeled with green fluorescent protein (GFP) using monster GFP vector and suspended in hyaluronic acid for IA injection. Treatment approaches consisted of cell-treatment using MSCs suspended in 3 ml of hyaluronic acid (HA) for the right carpal joint; and using the same amount of (HA) but without MSCs for the left contralateral carpal joint to serve as a control. Animals were assessed clinically and radiologically before and after treatment. Synovial fluid was also evaluated. Histopathologically; articular cartilage structural changes, reduction of articular cartilage matrix staining, osteophyte formation, and subchondral bone plate thickening were graded. Data was summarized using median and percentile for scores of histopathologic grading. Comparison between groups was done using non-parametric Mann Whitney test.</p> <p>Results</p> <p>The reparative effect of MSCs was significant both clinically and radiologically in all treated groups (P < 0.05) compared to the control groups. Fluorescence microscopy of sections of the cell-treated joints of all animals indicated that the GFP-transduced injected cells have participated effectively in the reparative process of the damaged articular surface and have integrated within the existing articular cartilage. The cells were associated with the surface of the cartilage and, were also detected in the interior.</p> <p>Conclusions</p> <p>Homing was confirmed by the incorporation of injected GFP-labeled MSCs within the repaired newly formed cartilage. Significant recovery proves that the use of IA injection of autologous MSCs is a viable and a practical option for treating different degrees of osteoarthritis.</p

INCREASED URINARY 8-OXO-7,8-DIHYDRO-2′-DEOXYGUANOSINE EXCRETION IN A SAMPLE OF EGYPTIAN CHILDREN WITH BETA THALASSEMIA MAJOR: MARKER FOR LIPID PEROXIDATION-INDUCED DNA DAMAGE

Objective: The objective of this research was to evaluate oxidative stress status in children with β-thalassemia major.Methods: Our study was conducted in children with β-thalassemia aged from 5 to 15 years. Investigate the urinary excretion of human 8-oxo-7,8- dihydro-2′-deoxyguanosine, which will be analyzed by enzyme-linked immunosorbent assay. To investigate serum levels of antioxidant enzymes include glutathione s-transferase (GST) and catalase (CAT).Results: We found a significant elevation of the urinary 8-oxo-7,8-dihydro-2′-deoxyguanosine level with p=0.001 compared to control group, a significant reduction of both GST and CAT p=0.05 and 0.03, respectively, compared to control group. There was a significant negative correlation between urinary 8-oxo-7,8-dihydro-2′-deoxyguanisine and CAT level, r=−0.378, p=0.016, hemoglobin - r=−0.610, p=0.001, hematocrit (%) - r=−0.478, p=0.002, while a significant positive correlation between urinary 8-oxo-7,8-dihydro-2′-deoxyguanisine and alanine aminotransferase - r=0.547, p=0.001, and serum ferritin - r=0.391, p=0.013. There was a significant negative correlation between CAT and serum ferritin - r=−0.320, p=0.44.Conclusion: We conclude that the strongly increased urinary excretion 8-oxo-7,8=dihydro-2′-deoxyguanisine indicates elevated lipid peroxidation induced DNA damage in internal organs such as the liver. These highly pro mutagenic lesions may contribute to the increased risk of thalassemia patients to develop hepatocellular carcinoma

URINARY MARKERS OF OXIDATIVE DNA DAMAGE IN TYPE 1 DIABETIC CHILDREN: RELATION TO MICROVASCULAR COMPLICATIONS

  Objective: Type 1 diabetes mellitus (T1DM) is a widespread metabolic disease, which frequently carries with it a significant impact on human health. Oxidative damage and tissue inflammation have been claimed to be a typical pathogenic component for the progression of diabetic complications. We aim in this study to explore the relation of urinary 8-oxo-7,8-dihydro-2′-deoxyguanosine (8-oxodG) (as a marker of nucleic acid oxidation) to microvascular complications in T1DM.Methods: A case–control study, enrolling 45 T1DM children and an equivalent number of healthy subjects, was performed. Full clinical examination and anthropometric measurement were performed to all subjects. Urinary assessment for 8-oxodG and albumin was done in addition to blood sampling for lipid profile and glycated Hb (HbA1c) assay. Complete ocular examination for assessment of diabetic retinopathy (DR) was also done.Results: Levels of urinary 8-oxodG, serum cholesterol, triglycerides, and low-density lipoprotein in cases were significantly higher than non-diabetics; these levels were likewise higher in uncontrolled T1DM patients in comparison with well-controlled T1DM subjects. Urinary 8-oxodG and HbA1c were significantly higher in diabetic patients with albuminuria and DR compared to patients without complications. Significant positive correlation was found between 8-oxodG with HbA1c (r=0.8, p&lt;0.01), diastolic blood pressure (DBP) (r=0.4, p=0.02), and cholesterol (r=0.4, p=0.05).Conclusion: Urinary 8-oxodG was found to be a reliable marker for assessing oxidative DNA damage in T1DM and can be used in the determination of microvascular complications related to diabetes

Adiponectin: an adipocyte-derived hormone, and its gene encoding in children with chronic kidney disease

BACKGROUND: The prevalence of cardiovascular disease (CVD) and inflammation is high in patients with chronic kidney disease (CKD). Adiponectin (ADPN) is an adipocytokine that may have significant anti-inflammatory and anti-atherosclerotic effects. Low adiponectin levels have previously been found in patients with high risk for CVD. METHODS: On seventy eight advanced CKD (stages 4 and 5) pediatric patients undergoing maintenance hemodialysis( MHD) or conservative treatment (CT) the following parameters were studied: body mass index, left ventricular mass index(LVMI), serum adiponectin , cholesterol, HDL-cholesterol, high sensitivity C-reactive protein (hs CRP),interleukin 6(IL6) and single-nucleotide polymorphisms (SNPs) in the ADIPOQ gene at positions 45, and 276. Seventy age-and gender-matched healthy subjects served as control subjects. RESULTS: Markedly (P = 0.01) elevated plasma adiponectin levels were observed in CKD patients, especially CT patients, compared to control subjects. The wild type of ADIPOQ 45T > G (T) allele is the main gene for patients and controls. MHD and CT patients had significantly higher frequency of the TT genotypes of +276G > T gene (P = 0.04) compared with control subjects. A significant positive correlation was observed between plasma adiponectin and IL6 level, whereas negative correlations were found between adiponectin level, cholesterol, HDL cholesterol and hs CRP. In a stepwise backward multiple regression model only IL6 (P = 0.001) was independently associated with plasma adiponectin levels. The adiponectin gene the 276 GT+TT genotypes were associated with a higher level of adiponectin . CONCLUSIONS: The present study demonstrated that ADPN is related to several metabolic and inflammatory CV risk factors in a manner consistent with the hypothesis that this protein might have a protective role against these factors. We observed an association between the +276G>T SNP in the adiponectin gene and CKD in children. Genetic variation of +276 gene seemed to have a positive impact on circulating adiponectin levels in CKD patients

Impact of Genetic Polymorphism of Myeloid Differentiation Primary Response Gene 88, Enhancer of Zeste Homolog 2, and B-cell Lymphoma 2 like 11 in Patients with Diffuse Large B Cell Lymphoma Treated with Rituximab, Cyclophosphamide, Doxorubicin, Vincristin

BACKGROUND: Despite the growing landscape of genetic drivers in Diffuse Large B-cell Lymphoma, yet their clinical implication is still unclear and R-CHOP regimen remains a “one size fits all” therapy. We aimed in this study to examine the prevalence of EZH2, BCL211 and MYD 88 genetic polymorphisms in DLBCL patients and correlate the results with various clinical and survival outcomes. METHODS: Genotyping of MYD88 (rs387907272 T/C), EZH2 (rs3757441 C/T), and BCL2L11 (rs3789068 A/G) polymorphisms were conducted using real time polymerase chain reaction analysis in a total of 75 DLBCL patients. RESULTS: Most of our cases carried the wild TT genotype of MYD88 gene (64%), the mutant TT genotype of EZH2 gene (52%) and the wild AA genotype of BCL2L11 gene (48%). Regarding cell of origin, Germinal Centre (GC) phenotype was present in 56% of cases while 44% expressed the Post-GC (PGC) phenotype. Poor response outcome to first line R-CHOP was significantly correlated with the mutated CC genotype of MYD 88 (p=0.02), while better response to R-CHOP was significantly associated with younger age &lt;50 years (p &lt;0.0001), good PS (p=0.046), normal LDH level (p=0.003), earlier stage (p &lt;0.0001), good IPI score (p=0.009), absence of extranodal disease (p &lt;0.0001) and absence of bulky disease (p=0.004). The median PFS and the 2 year OS were significantly higher in younger age, earlier stage, good IPI score, absence of extranodal disease, absence of bulky disease and in GC phenotype. CONCLUSIONS: Our results emphasized that the mutated genotype of MYD 88 gene polymorphism is significantly associated with poor response to R-CHOP therapy

Efficacy of Mesenchymal Stem Cells in Suppression of Hepatocarcinorigenesis in Rats: Possible Role of Wnt Signaling

<p>Abstract</p> <p>Background</p> <p>The present study was conducted to evaluate the tumor suppressive effects of bone marrow derived mesenchymal stem cells (MSCs) in an experimental hepatocellular carcinoma (HCC) model in rats and to investigate the possible role of Wnt signaling in hepato-carcinogenesis.</p> <p>Methods</p> <p>Ninety rats were included in the study and were divided equally into: Control group, rats which received MSCs only, rats which received MSCs vehicle only, HCC group induced by diethylnitroseamine (DENA) and CCl_<b>4</b>, rats which received MSCs after HCC induction, rats which received MSCs before HCC induction. Histopathological examination and gene expression of Wnt signaling target genes by real time, reverse transcription-polymerase chain reaction (RT-PCR) in rat liver tissue, in addition to serum levels of ALT, AST and alpha fetoprotein were performed in all groups.</p> <p>Results</p> <p>Histopathological examination of liver tissue from animals which received DENA-CCl₄only, revealed the presence of anaplastic carcinoma cells and macro-regenerative nodules type II with foci of large and small cell dysplasia. Administration of MSCs into rats after induction of experimental HCC improved the histopathological picture which showed minimal liver cell damage, reversible changes, areas of cell drop out filled with stem cells. Gene expression in rat liver tissue demonstrated that MSCs downregulated <it>β-catenin</it>, proliferating cell nuclear antigen (<it>PCNA</it>), <it>cyclin D </it>and <it>survivin </it>genes expression in liver tissues after HCC induction. Amelioration of the liver status after administration of MSCs has been inferred by the significant decrease of ALT, AST and Alpha fetoprotein serum levels. Administration of MSCs before HCC induction did not show any tumor suppressive or protective effect.</p> <p>Conclusions</p> <p>Administration of MSCs in chemically induced HCC has tumor suppressive effects as evidenced by down regulation of Wnt signaling target genes concerned with antiapoptosis, mitogenesis, cell proliferation and cell cycle regulation, with subsequent amelioration of liver histopathological picture and liver function.</p

Diagnostic significance of hsa_circ_0000146 and hsa_circ_0000072 biomarkers for Diabetic Kidney Disease in patients with type 2 diabetes mellitus

Background: Diabetic Kidney Disease (DKD) is a significant challenge in healthcare. However, there are currently no reliable biomarkers for renal impairment diagnosis, prognosis, or staging in DKD patients. CircRNAs and microRNAs have emerged as noninvasive and efficient biomarkers. Methods: We explored Cannabinoid receptor 1 (CNR1), C reactive protein (CRP), hsa_circ_ 0000146 and 0000072, and hsa-miR-21 and 495 as diagnostic biomarkers in DKD. The serum concentrations of CRP and CNR1 were measured using ELISA. Rt-qPCR was used to evaluate the expression levels of CNR1, circRNAs, and miRNAs in 55 controls, 55 type 2 diabetes mellitus patients, and 55 DKD patients. Their diagnostic value was determined by their ROC curve. KEGG pathway was used to predict the functional mechanism of the circRNA's target genes. Results: DKD patients exhibited a significant increase in CRP and CNR1 levels and the expression of miR-21 and 495. The expression levels of circ_0000146 and 0000072 decreased in DKD patients. ROC analysis revealed that circRNAs and miRNAs alone or CNR1 and CRP have significant diagnostic potential. The functional prediction results showed the involvement of hsa_circ_0000146 and 0000072 in various pathways that regulate DKD. Conclusions: Therefore, the examined circRNAs and miRNAs may represent a novel noninvasive biomarker for diagnosing and staging DKD

Signaling mechanisms of a water soluble curcumin derivative in experimental type 1 diabetes with cardiomyopathy

BACKGROUND: Curcumin exhibits anti-diabetic activities, induces heme-oxygenase-1 (HO-1) and is an inhibitor of transcriptional co-activator p300. A novel water soluble curcumin derivative (NCD) has been developed to overcome low invivo bioavailability of curcumin. We evaluated the effect of the NCD on signaling mechanisms involved in cardiomyocyte hypertrophy and studied whether its action is mediated via inducible HO-1. MATERIALS AND METHODS: Rats were divided into controls, controls receiving NCD, diabetic, diabetic receiving NCD, diabetic receiving pure curcumin, diabetic receiving HO inhibitor, zinc protoporphyrin IX (ZnPP IX) and diabetic receiving NCD and ZnPP IX. NCD and curcumin were given orally. After 45 days, cardiac physiologic parameters, plasma glucose, insulin, glycated hemoglobin (GHb), HO-1 gene expression and HO activity in pancreas and cardiac tissues were assessed. Gene expression of p300, atrial natriuretic peptide (ANP) and myocyte enhancer factor 2 (MEF2A and MEF2C) were studied. RESULTS: NCD and curcumin decreased plasma glucose, GHb and increased insulin levels significantly in diabetic rats. This action may be partially mediated by induction of HO-1 gene. HO-1 gene expression and HO activity were significantly increased in diabetic heart and pancreas. Diabetes upregulated the expression of ANP, MEF2A, MEF2C and p300. NCD and curcumin prevented diabetes-induced upregulation of these parameters and improved left ventricular function. The effect of the NCD was better than the same dose of curcumin

Relation of Asthma Control with Quality of Life among a Sample of Egyptian Asthmatic School Children

BACKGROUND: Asthma is considered a chronic health illness that not only resulted in physical symptoms but also emotional effects. It is; therefore, so important to assess the quality of life of asthmatic patients besides their level of disease control. AIM: To determine the correlation of asthma control with the health-related quality of life (HRQOL) of asthmatic children in Egypt. METHODS: One hundred and twenty-eight asthmatic Egyptian children were enrolled in the study. They were subjected to asthma severity grading, asthma control questionnaire (ACQ) and pediatric asthma quality of life questionnaire (PAQLQ). Studied cases were taken from 6 primary and preparatory schools, Giza governorate. RESULTS: The mean child control score was significantly higher in not well-controlled asthmatics compared to well-controlled asthmatics (p &lt; 0.005). The not well controlled asthmatic children showed significantly lower activity limitation score, symptoms score, and overall asthmatic score compared to controlled asthmatic children (p &lt; 0.05). The severity of asthma shows significant positive correlation with symptoms score, emotional function score and overall asthmatic score (p &lt; 0.05). CONCLUSION: The quality of life for the asthmatic children is strongly correlated with the level of asthma control and severity