Browsing by Author "Mammadov, E"

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    Obtaining Stem Cell Spheroids from Foreskin Tissue and the Effect of Corchorus olitorius L. on Spheroid Proliferation
    Becer, E; Soykut, G; Kabadayi, H; Mammadov, E; Çalis, I; Vatansever, S
    Objectives: Mesenchymal stem cells are self-renewing stem cells. The human foreskin has potential to be used as a source of stem cells. The aim of the study was to obtain spheroid formation of human foreskin cells (hnFSSCs) isolated from newborn human foreskin tissue. In addition, the apoptotic and proliferative effects of a traditional plant, Corchorus olitorius L. (C. olitorius), on hnFSSC spheroids were investigated. Materials and Methods: After a routine circumcision procedure the cells were isolated and cultured in suitable medium. The plant leaves was extracted with ethanol and their composition was analyzed by liquid chromatography coupled with mass spectrometry (LC-MS/MS). The foreskin stem cells were characterized immunocytochemically by CD45, CD34, and CD90 antibodies. hnFSSC spheroids were formed using the hanging drop technique. Immunofluorescence staining was used on the obtained spheroids to determine the distribution of caspase-3 and Ki-67 after being treated with C. olitorius extract for 48 h. Results: Immunostaining analysis showed that hnFSSCs were positive for CD45 and CD34 and negative for CD90. According to LC-MS/MS C. olitorius was rich in flavanols and hydrocinnamic acid derivatives. Although the spheroids obtained were loose and floating, the cells interacted with each other. Caspase-3 activity was higher in the control group than in the extract-treated group and Ki-67 was higher in the extract-treated group than in the control group, suggesting that the plant might have the capacity to increase stem cell proliferation due to its rich polyphenolic content. Conclusion: The results suggest that hnFSSCs and spheroids might be used in stem cell generation, tissue repair and renewal as human foreskin tissue has potential to be used as a stem cell source. C. olitorius also increased proliferation of hnFSSCs, showing that polyphenols might increase proliferation of stem cells.
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    In Vitro Cultivation, Characterization and Osteogenic Differentiation of Stem Cells from Human Exfoliated Deciduous Teeth on 3D Printed Polylactic Acid Scaffolds
    Islam, A; Mammadov, E; Kendirci, R; Aytac, E; Cetiner, S; Vatansever, HS
    Background: Tissue engineering mainly focuses on creating appropriate conditions for the regeneration of tissues. Scaffolds, signal molecules, and stem cells interact with each other and compose the essential components of this field. Objectives: This study aimed at investigating the osteogenic induction ability of PLA Poly Lactic Acid (PLA) scaffolds and comparing the osteogenic differentiation behavior of Stem Cells from Human Exfoliated Deciduous Teeth (hSHEDs) in standard culture medium and on PLA scaffolds. Methods: The current clinical experimental study was conducted between April 2016 and October 2016 at the Near East University cell culture laboratory located in North Cyprus. The pulp tissues of deciduous teeth (non-decayed and in the absence of abscess, fistula or periapical lesion) were sampled from 10 healthy children aged between 6 and 11 years. The isolated hSHEDs were divided to 4 groups. The control group/Group1 consisted of cells, which were cultivated in standard culture medium, and Group2 cells were differentiated into an osteogenic lineage using osteogenic differentiation medium. Group 3 represented the non-differentiated group, which was transferred onto three dimensional (3D) printed PLA scaffolds and Group 4 cells were differentiated to the osteogenic lineage and transferred onto 3D printed PLA scaffolds. All groups were analyzed immunohistochemically and by immune-labeling, and were evaluated semi-quantitatively using the HSCORE. Results: Cultivation of hSHEDS on PLA scaffolds was assessed for 14 and 21 days; osteogenic differentiation was detected both histochemically and immunohistochemically. Generally, Osteocalcin (OCN) immunoreactivities were higher than Osteonectin (ON) immunoreactions in all groups. Despite higher OCN immunoreactivities, the intensities of OCN between 14 days and 21 days in group 4 (497.3 +/- 0.57% and 486.7 +/- 5.77%, respectively) were similar (P > 0.05). While the intensity of ON was 280.0 +/- 10% in group 4, in group 2 the intensity of ON was 206.7 +/- 5.77%, and on the 14th day the results were statistically significant (P < 0.0001). Conclusions: Poly lactic acid is a suitable scaffold material for osteogenic induction of the hSHEDs. The expression patterns of both markers showed that a 14-day cultivation period is adequate for hSHEDs with/without PLA scaffolds to differentiate into osteoblasts.
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    The Effect of Quince Seed Mucilage on Human Foreskin Stem Cell Proliferation and Self-Renewal Potential
    Mammadov, B; Mammadov, E; Becer, E; Vatansever, HS
    BACKGROUND/AIMS: Quinceseed mucilage (QSM) is used in Iranian folk medicineto treat wounds and burns. Mucilage is rich in polysaccharides and proteins. Approximately 80% of breastfeeding women experience nipple pain and soreness, often applying homemade QSM to treat nipple cracks. There are limited studies on the cytotoxic effects of QSM on fibroblast formation. The present study investigated the proliferative effects of QSM on mesenchymal stem cells isolated from newborn foreskin (hnFSSCs). MATERIALS AND METHODS: Following a standard circumcision procedure, cells were isolated and cultured in suitable media to support growth. The quince seed gel was prepared and pulverized by drying. Foreskin stem cells were immunocytochemically characterized using CD45, CD34, and CD90 antibodies. The cytotoxic effect of quince seed gel on hnFSSCs was determined using the MTT assay. The cells were then treated with quince seed gel for 24 h, and immunohistochemical staining for Ki-67, c-Myc, OCT3/4, and Sall4 was performed. RESULTS: Immunohistochemical analysis revealed that hnFSSCs were positive for CD, CD90, and CD45 and weakly positive for CD34. The MTT results showed that quince seed gel treatment at 100 mu g/mL for 24 h was the most appropriate concentration and duration compared with the positive control. QSM-treated cells showed significantly higher immunoreactivity for Ki-67 (H-score: 266.5 +/- 12.6), OCT3/4 (H-score: 239 +/- 8), and Sall4 (H-score: 243.8 +/- 7.5) in comparison with the control group (p<0.05). In contrast, c-Myc (H-score: 226 +/- 18.8) immunoreactivity was moderate in both groups, with no significant difference (p>0.05). CONCLUSION: Our results suggest that QSM can support the maintenance of self-renewal and pluripotency properties in human foreskin- derived stem cells.