Elahe Mahdipour

Investigation of the effects of curcumin on serum cytokines in obese individuals: A randomized controlled trial

Background. Obesity is a disorder often accompanied by a heightened state of systemic inflammation and immunoactivation. The present randomized crossover trial aimed to investigate the efficacy of curcumin, a bioactive polyphenol with established anti-inflammatory and immunomodulatory effects, on the serum levels of a panel of cytokines and mediators in obese individuals. Methods. Thirty obese individuals were randomized to receive curcumin at a daily dose of 1 g or a matched placebo for 4 weeks. Following a 2-week wash-out period, each group was assigned to the alternate treatment regimen for another 4 weeks. Serum samples were collected at the start and end of each study period. Serum levels of IL-1α, IL-1β, IL-2, IL-4, IL-6, IL-8, IL-10, VEGF, IFNγ, EGF, MCP-1, and TNFα were measured using a multiplex Biochip Array Technology based method. Results. Mean serum IL-1β (P = 0.042), IL-4 (P = 0.008), and VEGF (P = 0.01) were found to be significantly reduced by curcumin therapy. In contrast, no significant difference was observed in the concentrations of IL-2, IL-6, IL-8, IL-10, IFNγ, EGF, and MCP-1. Conclusions. The findings of the present trial suggested that curcumin may exert immunomodulatory effects via altering the circulating concentrations of IL-1β, IL-4, and VEGF.

Heterocyclic amine-modified polyethylenimine as gene carriers for transfection of mammalian cells

Branched polyethylenimine (PEI) is extensively used as a polycationic non-viral vector for gene delivery. Polyplexes formed with PEI are believed to be released from endocytotic vesicles by the osmotic burst mechanism in the rate-limiting step in transfection. Increasing the buffering capacity of PEI derivatives in the endosomal pH range of 4.5-7.5 should enhance transfection efficiency. In this study, PEI was derivatized by covalently attaching heterocyclic amine moieties (piperazine, pyridine and imidazole rings with pKa values from 5.23 to 6.04) through amide bonds. PEI derivatives with 50% of the primary amines on PEI exhibited increased buffering capacity, increased transfection activity and decreased cytotoxicity in murine neuroblastoma (Neuro-2a) cells. The relative effectiveness in enhancing transfection efficiency was piperazine>pyridine>histidine, but each type of amine was the most effective under a particular set of conditions. Modified vectors could significantly improve transfection efficiency in murine mesenchymal stem cells. PEI25 derivatized at 50% with histidine administered as polyplexes in the tail veins of mice resulted in remarkably enhanced luciferase gene expression in the expected organ distribution and much lower toxicity than underivatized PEI25.

Serum Inflammatory Cytokines and Depression in Coronary Artery Disease

Background: Severe depression may be accompanied by immune dysregulation and is also associated with increased risk of coronary artery disease (CAD). Objectives: We investigated serum levels of 10 cytokines and their relationship with depression in patients with cardiovascular diseases as well as healthy subjects in northeast of Iran. Patients and Methods: The study was carried out on 462 subjects (120 healthy subjects and 342 candidates undergoing angiography). The healthy subjects were referred for routine annual checkups or pre-employment examinations; they did not have clinically evident CAD. A questionnaire was used to obtain demographic data and the Beck depression inventory (BDI) was applied to assess depression. The Evidence Investigator® platform was used for cytokines assays for IL-1α, IL-1β, IL-2, IL-4, IL-6, IL-8, IL-10, TNF-α, MCP-1 and IFN-γ, using sandwich chemiluminescent method. The statistical analysis was performed using SPSS version 11.5. Results: The mean age was 53.3 ± 11.5, 54.8 ± 11.3, and 59.5 ± 11.3 in healthy, angiography (-), and angiography (+) subjects, respectively (P < 0.05). There were significant differences in serum levels of IL-4, IL-6, IL-10, and MCP-1 cytokines, comparing subjects with CAD and healthy persons (P < 0.05). When all subjects were divided to with and without depression regardless of their cardiovascular status, there was a significant difference in serum levels of IL-8 and IL-6 between the groups (P < 0.05). When the subgroup with features of CAD was selected and divided to those with and without depression, there was also a significant difference in serum levels of IL-8 and TNF-α (P < 0.05). Conclusions: The positive interaction between depression and CAD was probably mediated by inflammatory mechanisms.

Hybrid chitosan-ß-glycerol phosphate-gelatin nano-/micro fibrous scaffolds with suitable mechanical and biological properties for tissue engineering.

Scaffold‐based tissue engineering is considered as a promising approach in the regenerative medicine. Graft instability of collagen, by causing poor mechanical properties and rapid degradation, and their hard handling remains major challenges to be addressed. In this research, a composite structured nano‐/microfibrous scaffold, made from a mixture of chitosan–ß‐glycerol phosphate–gelatin (chitosan–GP–gelatin) using a standard electrospinning set‐up was developed. Gelatin–acid acetic and chitosan ß‐glycerol phosphate–HCL solutions were prepared at ratios of 30/70, 50/50, 70/30 (w/w) and their mechanical and biological properties were engineered. Furthermore, the pore structure of the fabricated nanofibrous scaffolds was investigated and predicted using a theoretical model. Higher gelatin concentrations in the polymer blend resulted in significant increase in mean pore size and its distribution. Interaction between the scaffold and the contained cells was also monitored and compared in the test and control groups. Scaffolds with higher chitosan concentrations showed higher rate of cell attachment with better proliferation property, compared with gelatin‐only scaffolds. The fabricated scaffolds, unlike many other natural polymers, also exhibit non‐toxic and biodegradable properties in the grafted tissues. In conclusion, the data clearly showed that the fabricated biomaterial is a biologically compatible scaffold with potential to serve as a proper platform for retaining the cultured cells for further application in cell‐based tissue engineering, especially in wound healing practices. These results suggested the potential of using mesoporous composite chitosan–GP–gelatin fibrous scaffolds for engineering three‐dimensional tissues with different inherent cell characteristics.

Fibroblast Growth Factor 1-Transfected Adipose-Derived Mesenchymal Stem Cells Promote Angiogenic Proliferation

The aim of this study was to investigate, for the first time, the effects of using adipose-derived mesenchymal stem cells (AD-MSCs) transfected with an episomal plasmid encoding fibroblast growth factor 1 (FGF1) (AD-MSCsFGF1), in providing the microenvironment required for angiogenic proliferation. The isolated rat AD-MSCs were positive for mesenchymal (CD29 and CD90) and negative for hematopoietic (CD34 and CD45) surface markers. Adipogenic and osteogenic differentiation of the AD-MSCs also occurred in the proper culture media. The presence of FGF1 in the conditioned medium from the AD-MSCsFGF1 was confirmed by Western blotting. G418 and PCR were used for selection of transfected cells and confirmation of the presence of FGF1 mRNA, respectively. Treatment with the AD-MSCFGF1-conditioned medium significantly increased the NIH-3T3 cell proliferation and human umbilical vein endothelial cell (HUVEC) tube formation compared to conditioned medium from nontransfected AD-MSCs (p < 0.001). In conclusion, the AD-MSCsFGF1 efficiently secreted functional FGF1, which promoted angiogenic proliferation. Using AD-MSCsFGF1 may provide a useful strategy in cell therapy, which can merge the beneficial effects of stem cells with the positive biological effects of FGF1 in various disorders, especially tissue defects, neurodegenerative, cardiovascular and diabetes endocrine pathologies, which remain to be tested in preclinical and clinical studies.

Cytokine profiles in overweight and obese subjects and normal weight individuals matched for age and gender

Background Obesity is associated with a state of systemic inflammation, mediated by adipose tissue-derived cytokines that may also have metabolic effects, including an effect on insulin resistance. The aim of this study was to compare the serum profile of pro- and anti-inflammatory cytokines in obese and non-obese subjects. Methods A total of 242 subjects who were either overweight or obese (body mass index [BMI] ≥ 25 kg/m2) and non-obese subjects (body mass index <25 kg/m2), were recruited in Mashhad in northeastern Iran. The concentrations of serum interleukin-1α, -1β, -2, -4, -6, -8 and -10 (IL-1α, IL-1β, IL-2, IL-4, IL-6, IL-8 and IL-10), were measured in all subjects, together with serum vascular endothelial growth factor, interferon-γ, epidermal growth factor, monocyte chemoattractant protein-1 and tumour necrosis factor-α. Results The groups differed significantly with respect to measures of adiposity and fasted lipid profile. Serum pro-inflammatory cytokines interferon-γ and interleukin-1α, and anti-inflammatory cytokines, interleukin-10, and epidermal growth factor were significantly different between obese and non-obese individuals, as was serum high-sensitivity C-reactive protein. Multivariate regression showed that waist circumference was significantly and independently related to serum monocyte chemoattractant protein-1concentrations (P = 0.001). Conclusion Despite significant differences in several cytokines between the groups, only monocyte chemoattractant protein-1appeared to be independently related to a measure of adiposity in this population sample from Iran.

Regulatory crosstalk between Hox genes and miRNAs during angiogenesis

Angiogenesis, or the growth of new blood vessels, is critical to both health and disease. Several genes are implicated in the regulation of angiogenesis, such as genes encoding growth factors, cytokines, and the extracellular matrix, as well as transcription factors. Among them, Hox genes exert master regulatory effects on several aspects of angiogenesis. The mechanism by which Hox genes are controlled is not well understood. However, recently microRNAs (miRNAs) have been introduced as candidates for Hox gene regulation. This review provides a comprehensive description of Hox genes and miRNAs involved in angiogenesis and presents possible regulatory interactions between them.

A Feasible Method for the Isolation of Mesenchymal Stem Cells from Menstrual Blood and Their Exosomes

BACKGROUND Mesenchymal stem cells (MSCs) are currently the most promising candidates in regenerative medicine. Nonetheless, there are several limitations associated with the MSC tissue source such as infrequent and invasive bone marrow sampling methods. To overcome these limitations, we have procured MSCs from the menstrual blood (MenSCs) as a non-invasive source using a straightforward and cost-effective method. Moreover, we isolated MenSCs-derived exosomes as a safe and highly effective approach to exert the paracrine effects of MSCs. METHODS MSCs were isolated from menstrual blood through two different culture methods: ficoll-isolated mononuclear cells (MNCs) and whole blood culture. These cells were characterized by their plastic adherence, flow cytometry analysis of the surface markers and the differentiation potential. The exosomes were isolated from conditioned media using ultracentrifugation and characterized by different microscopy techniques, western blotting, and ELISA. RESULTS Both Methods resulted in the rapid isolation of cells with MSC properties. However, the cellular yield of the whole blood culture method was remarkably more than MNCs culture. MenSCs also produced a substantial amount of extracellular vesicles (EVs) possessed the minimum criteria for exosome definition. CONCLUSION The results suggest that direct culture of whole menstrual blood cells is a high throughput, straightforward and cost-effective method for MenSCs isolation using no special growth factors. Moreover, these cells are a good producer of exosome which can offer a cell-free therapy approach.

Dicer expression is impaired in diabetic cutaneous wound healing

Diabetes, as a fast growing non-communicable disease, is one of the major health problems of the twenty-first century. Several complications such as cardiovascular disease and renal failure are accompanying diabetes. The chronic cutaneous wound is another diabetes complication, results from the reduced body healing potential. At genome level, diabetic wound environment displays disorganized gene functions emphasizing the critical role of gene regulatory networks in the control of chronic wound repair. MicroRNAs, major regulators of gene activity, have been shown to be impaired in several pathological conditions such as chronic wounds. A reason behind that can be sought in the impaired activity of enzymes involved in the development and production of miRNAs. In current study, streptozocin-induced diabetic rats and non-diabetic controls were used to study the effect of diabetes on Dicer presence in wound environment. Unwounded skin of diabetic animals showed significantly lower level of Dicer expression compared with non-diabetic animal-derived skin. However, at day 7 post-wounding, diabetic animal-derived wounds contained a higher level of Dicer expression compared with non-diabetic ones. Parallel to these findings, the granulation tissue formation and wound closure are impaired in diabetic wounds. This study highlights the dysregulated presence of Dicer at different stages of the diabetic cutaneous wound.

Determine exogenous human DDAH2 gene function in rabbit bone marrow-derived endothelial progenitor cells in vitro.

The in vitro amplification of endothelial progenitor cells (EPCs) is an important method because of its role in gene transferring and regenerative medicine. In this study, we isolated rabbit bone marrow–derived EPCs to further manipulation and overexpression of dimethylarginine dimethylaminohydrolase (DDAH) in EPCs. Isolated EPCs were cultured, expanded in endothelial basal medium. Morphology of EPCs and expression levels of surface markers detected using immunocytochemistry staining and through the use of flow cytometery. Endothelial progenitor cells were transfected with plasmid vectors expressing human DDAH2 (DDAH2‐EPCs). Three days after gene transfer, positive transfected‐EPCs proliferation and DDAH activity were assayed. We observed colonies conformation and endothelium‐like morphology gradually in the third week of culture. Characterization results revealed positive expression of EPC surface markers CD106, Flk‐1, vWF, and CD34 using few identification techniques. Overexpression of DDAH2 increased citrulline production after 96 hours of transfection, 235.34 ± 0.69 vs 95.26 ± 5.76 ng/mL; P = .023. These results suggest that cell population with EPC characteristics can be simply isolated from rabbit bone marrow and successfully engineered to overexpress exogenous gene. In this study, we offer a feasible method to isolate and identify EPCs from bone marrow. In addition, an efficient transfection with a plasmid vector (without risk of interference) can be constructed a hybrid structure with EPC and DDAH2 gene to examine their function in vitro.