Evangelia Siska - Academia.edu (original) (raw)

Papers by Evangelia Siska

<p>(A) Schematic representation of a Glucose Biosensor (GB) gene used for the genetic modif... more <p>(A) Schematic representation of a Glucose Biosensor (GB) gene used for the genetic modification of MSCs. The gene encodes a chimeric protein consisting of a signal peptide allowing protein secretion followed by Cyan Fluorescent Protein (CFP), a glucose binding domain and Yellow Fluorescent Protein (YFP). (B) Optical and YFP fluorescence microscopy of GB/hTERT MSCs (scale bar: 100μm). (C) Flow cytometry of GB/hTERT MSCs for mesenchymal CD29, 73, 90, 105 markers. Unstained cells were used as control. (D) Staining of control or differentiated GB/hTERT cells with Alizarine Red/Oil Red O indicative of osteogenic/adipogenic differentiation, respectively (scale bar: 100μm).</p

<p>(A) Immunoblots for hTERT in control, GB and GB/hTERT MSC extract using an anti-hTERT an... more <p>(A) Immunoblots for hTERT in control, GB and GB/hTERT MSC extract using an anti-hTERT antibody. GAPDH was used as a loading control (B) Relative telomerase activity in GB and GB/hTERT MSCs at passage 20. (C) Cumulative population doublings of GB and GB/hTERT MSCs over time. GB MSCs reached replicative senescence at passage 28. (D) mRNA levels of Oct-4, Sox-2 and Nanog pluripotency markers in GB and GB/hTERT MSCs (***: p<0.001, **: p<0.01). Results in (B) and (D) are shown as mean ± SD of 3 independent experiments.</p

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PLOS ONE, 2017

Diabetes is a chronic disease characterized by high levels of blood glucose. Diabetic patients sh... more Diabetes is a chronic disease characterized by high levels of blood glucose. Diabetic patients should normalize these levels in order to avoid short and long term clinical complications. Presently, blood glucose monitoring is dependent on frequent finger pricking and enzyme based systems that analyze the drawn blood. Continuous blood glucose monitors are already on market but suffer from technical problems, inaccuracy and short operation time. A novel approach for continuous glucose monitoring is the development of implantable cell-based biosensors that emit light signals corresponding to glucose concentrations. Such devices use genetically modified cells expressing chimeric genes with glucose binding properties. MSCs are good candidates as carrier cells, as they can be genetically engineered and expanded into large numbers. They also possess immunomodulatory properties that, by reducing local inflammation, may assist long operation time. Here, we generated a novel immortalized human MSC line co-expressing hTERT and a secreted glucose biosensor transgene using the Sleeping Beauty transposon technology. Genetically modified hMSCs retained their mesenchymal characteristics. Stable transgene expression was validated biochemically. Increased activity of hTERT was accompanied by elevated and constant level of stem cell pluripotency markers and subsequently, by MSC immortalization. Furthermore, these cells efficiently suppressed PBMC proliferation in MLR transwell assays, indicating that they possess immunomodulatory properties. Finally, biosensor protein produced by MSCs was used to quantify glucose in cell-free assays. Our results indicate that our immortalized MSCs are suitable for measuring glucose concentrations in a physiological range. Thus, they are appropriate for incorporation into a cell-based, immune-privileged, glucose-monitoring medical device.

Journal of Hospital Infection, 2017

Iodine-Lithium-alpha-Dextrin (ILαD) against Staphylococcus aureus skin infections: A comparative ... more Iodine-Lithium-alpha-Dextrin (ILαD) against Staphylococcus aureus skin infections: A comparative study of in vitro bactericidal activity and cytotoxicity between ILαD and Povidone-Iodine.

Cytotherapy, Jul 25, 2017

Human mesenchymal stem cells (MSC) are important tools for several cell-based therapies. However,... more Human mesenchymal stem cells (MSC) are important tools for several cell-based therapies. However, their use in such therapies requires in vitro expansion during which MSCs quickly reach replicative senescence. Replicative senescence has been linked to macromolecular damage, and especially oxidative stress-induced DNA damage. Recent studies on the other hand, have implicated telomerase in the cellular response to oxidative damage, suggesting that telomerase has a telomere-length independent function that promotes survival. Here, we studied the DNA damage accumulation and repair during in vitro expansion as well as after acute external oxidative exposure of control MSCs and MSCs that overexpress the catalytic subunit of telomerase (hTERT MSCs). We showed that hTERT MSCs at high passages have a significant lower percentage of DNA lesions as compared to control cells of the same passages. Additionally, less damage was accumulated due to external oxidative insult in the nuclei of hTERT o...

Αριστοτέλειο Πανεπιστήμιο Θεσσαλονίκης (ΑΠΘ). Σχολή Επιστημών Υγείας. Τμήμα Ιατρικής. Τομέας Βιολογικών Επιστημών και Προληπτικής Ιατρικής. Εργαστήριο Βιολογικής Χημείας, 2020

alpha-dextrin (ILαD) against Staphylococcus aureus skin infections: a comparative study of in-vit... more alpha-dextrin (ILαD) against Staphylococcus aureus skin infections: a comparative study of in-vitro bactericidal activity and cytotoxicity between ILαD and povidone-iodine",

Surgical Innovation, 2017

Background. To investigate the early and late antiadhesive effect and any changes of fibrin matri... more Background. To investigate the early and late antiadhesive effect and any changes of fibrin matrix regulation enzymes on rat peritoneum, after local administration of bevacizumab. Methods. Rats were subjected to cecal abrasion. Bevacizumab (5 mg/kg) against placebo was given intraperitoneally. On the 2nd, 14th, and 28th postoperative days adhesions were scored, and tissue plasminogen activator (tPA), plasminogen activator inhibitor-1 (PAI-1), matrix metalloproteinase-9 (MMP-9), degree of fibrosis, and angiogenesis were measured in abrased cecum and in intact parietal peritoneum. Results. Bevacizumab significantly reduced adhesions up to 15% on the 2nd, 52.5% on the 14th, and 55% on the 28th postoperative day, and significantly increased tPA concentrations in peritoneum. PAI-1 was decreased, and a significantly higher tPA/PAI-1 ratio along with an increase of MMP-9 was measured at all time points. Fibrosis and angiogenesis were significantly lower on the 14th and 28th postoperative days. Conclusions. Local bevacizumab administration has a strong early and late antiadhesive action on rat peritoneum, mediated by changes in the tPA/PAI-1 and MMP balance in favor of fibrinolysis up to 28 days after operations.

Background. Human mesenchymal stem cells (MSC) are important tools for several cell-based therapi... more Background. Human mesenchymal stem cells (MSC) are important tools for several cell-based therapies. However, their use in such therapies requires in vitro expansion during which MSCs quickly reach replicative senescence. Replicative se-nescence has been linked to macromolecular damage, and especially oxidative stress-induced DNA damage. Recent studies on the other hand, have implicated telomerase in the cellular response to oxidative damage, suggesting that telomerase has a telomere-length independent function that promotes survival. Methods. Here, we studied the DNA damage accumulation and repair during in vitro expansion as well as after acute external oxidative exposure of control MSCs and MSCs that overexpress the catalytic subunit of telomerase (hTERT MSCs). Results. We showed that hTERT MSCs at high passages have a significant lower percentage of DNA lesions as compared to control cells of the same passages. Additionally, less damage was accumulated due to external oxidative insult in the nuclei of hTERT overexpressing cells as compared to the control cells. Moreover, we demonstrated that oxidative stress leads to diverse nucleus malformations, such as multillobular nuclei or donut-shaped nuclei, in the control cells whereas hTERT MSCs showed significant resistance to the formation of such defects. Finally, hTERT MSCs were found to possess higher activities of the basic antioxidant enzymes, superox-ide dismutase and catalase, than control MSCs. Discussion. On the basis of these results, we propose that hTERT enhancement confers resistance to genomic damage due to the amelioration of the cell's basic antioxidant machinery.

Histology and histopathology, Jan 30, 2016

Mesenchymal stromal cells (MSC) have been suggested to have beneficial effects on animal models o... more Mesenchymal stromal cells (MSC) have been suggested to have beneficial effects on animal models of traumatic brain injury (TBI), owing to their neurotrophic and immunomodulatory properties. Adipose tissue-derived stromal cells (ASCs) are multipotent MSC that can be harvested with minimally invasive methods, show a high proliferative capacity, low immunogenicity if allogeneic, and can be used in autologous or heterologous settings. In the present study ASCs were genetically labelled using the Sleeping Beauty transposon to express the fluorescent protein Venus. Venus+ASCs were transplanted intra-cerebroventricularly (ICV), on a rat TBI model and their survival, fate and effects on host brain responses were examined at seven days post-injury (7dPI). We provide evidence that Venus+ASCs survived, migrated into the periventricular striatum and were negative for neuronal or glial lineage differentiation markers. Venus+ASCs stimulated the proliferation of endogenous neural stem cells (NSCs)...

European Polymer Journal, 2017

International Journal of Polymeric Materials and Polymeric Biomaterials, 2016

Frontiers in Neuroscience, 2015

Polyglutamine diseases are fatal neurological disorders that affect the central nervous system. T... more Polyglutamine diseases are fatal neurological disorders that affect the central nervous system. They are caused by mutations in disease genes that contain CAG trinucleotide expansions in their coding regions. These mutations are translated into expanded glutamine chains in pathological proteins. Mutant proteins induce cytotoxicity, form intranuclear aggregates and cause neuronal cell death in specific brain regions. At the moment there is no cure for these diseases and only symptomatic treatments are available. Here, we discuss novel therapeutic approaches that aim in neuronal cell replacement using induced pluripotent or adult stem cells. Additionally, we present the beneficial effect of genetically engineered mesenchymal stem cells and their use as disease models or RNAi/gene delivery vehicles. In combination with their paracrine and cell-trophic properties, such cells may prove useful for the development of novel therapies against polyglutamine diseases.