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    Effect of bone marrow and adipose tissue-derived mesenchymal stem cells on the natural course of corneal scarring after penetrating injury
    (Academic Press Ltd- Elsevier Science Ltd, 2016) Demirayak, Bengi; Yüksel, Nurşen; Çelik, Onur Sinan; Subaşı, Cansu; Duruksu, Gökhan; Ünal, Z. Seda; Yıldız, Demir Kürşat; Karaöz, Erdal
    In the present study, we investigate and compare the efficacy of bone marrow- and adipose tissue-derived mesenchymal stem cell (MSCs) in corneal wound healing. A penetrating injury was created in the right corneas of Wistar rats (n = 40). Ten microliters of phosphate-buffered solution (PBS) containing 2 x 10(5) green fluorescent protein (GFP) labeled bone-marrow-derived MSCs to group 1 (n = 15), 10 mu l of PBS containing 2 x 10(5) GFP-labeled adipose-tissue-derived MSCs to group 2 (n = 15), 10 mu l PBS was injected into anterior chamber in group 3 (n = 10, control). Corneal opacity scoring, in vivo confocal microscopy, and histopathological evaluation were done at the end of 8 weeks. Immunofluorescence sections were evaluated to detect transplanted cells. Immune staining was performed to measure the expression levels of keratocan, aldehyde dehydrogenase (ALDH) and CD34. The gene expression levels of tumor necrosis factor (TNF-alpha), the interleukin 6 receptor (IL-6R), interleukin 12b (IL-12b), and transforming growth factor beta (TGF-beta 1) was measured on corneas. The establishment of stem cells in the corneas of the transplanted groups was confirmed by immunofluorescence staining. The expression of keratocan, ALDH, and CD34 increased in the transplanted groups (p < 0.05). The density of keratocytes increased significantly in both transplanted groups according to the in vivo confocal microscopy data (p < 0.05). The expression of TNF-alpha, IL-6R, and IL-12b decreased significantly in the transplanted groups (p < 0.05). Based on our findings, we consider that allogeneic stem cells facilitate the regeneration of corneal stroma and can be a cell source for stromal repopulation in diseased cornea. (C) 2016 Elsevier Ltd. All rights reserved.
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    The effect of recombinant tyrosine hydroxylase expression on the neurogenic differentiation potency of mesenchymal stem cells
    (Korean Spinal Neurosurgery Soc, 2018) Turaç, Gizem; Duruksu, Gökhan; Karaöz, Erdal
    Objective: Tyrosine hydroxylase (TH) is a rate-limiting enzyme in dopamine synthesis, making the enhancement of its activity a target for ensuring sufficient dopamine levels. Rat bone marrow mesenchymal stem cells (rBM-MSCs) are known to synthesize TH after differentiating into neuronal cells through chemical induction, but the effect of its ectopic expression on these cells has not yet been determined. This study investigated the effects of ectopic recombinant TH expression on the stemness characteristics of rBM-MSCs. Methods: After cloning, a cell line with stable TH expression was maintained, and the proliferation, the gene expression profile, and differentiation potential of rBM-MSCs were analyzed. Analysis of the cells showed an increment in the proliferation rate that could be reversed by the neutralization of TH. Results: The constitutive expression of TH in rBM-MSCs was successfully implemented, without significantly affecting their osteogenic and adipogenic differentiation potential. TH expression improved the expression of other neuronal markers, such as glial fibrillary acidic protein, beta-tubulin, nestin, and c-Fos, confirming the neurogenic differentiation capacity of the stem cells. The expression of brain-derived neurotrophic factor (BDNF) and ciliary neurotrophic factor (CNTF) significantly increased after the chemical induction of neurogenic differentiation. Conclusion: In this study, the expression of recombinant TH improved the neuroprotective effect of MSCs by upregulating the expression of BDNF and CNTF. Although the neuronal markers were upregulated, the expression of recombinant TH alone in rBM-MSCs was not sufficient for MSCs to differentiate into neurogenic cell lines.
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    Effects of VEGF(+) mesenchymal stem cells and platelet-rich plasma on inbred rat ovarian functions in cyclophosphamide-induced premature ovarian insufficiency model
    (Humana Press Inc, 2019) Vural, Birol; Duruksu, Gökhan; Vural, Fisun; Görgüç, Merve; Karaöz, Erdal
    Premature ovarian insufficiency (POI), a fertility disorder affecting women under 40years of age, is characterized by early loss of ovarian function. This study was aimed to maintain ovarian function in POI animal models by mesenchymal stem cells (MSCs) transplantation with/without the supplementation of platelet-rich plasma (PRP). Adipose tissue-derived MSCs were isolated from inbred rats (Fisher-344), and constitutive expression of both VEGF and GFP were maintained by transfection with plasmids, pVEGF and pGFP-N. PRP was derived from the blood of healthy untreated rats. A total of 60 rats were divided into 5 groups of 12 rats in each. First group was kept as untreated-control (Control), and POI model was induced in Fisher-344 rats by cyclophosphamide in the next four groups. Second group was kept as sham-operated-control (Sham). MSC, PRP and MSC+ PRP-treated groups were transplanted following the validation of POI model in rats. After 2months following the transplantation, anti-mullerian-hormone (AMH) and oestradiol (E-2) blood levels were measured. Follicles were evaluated after hematoxylin-eosin staining, and the immunofluorescence staining and gene expression analyses were performed to show the ovarian regeneration. The follicular count was improved after MSC- and MSC+PRP-treatment to 63% of Control-group and significantly higher than that in Sham-group, but a significant increase was not observed in PRP-group. Higher AMH and E-2 levels were measured in MSC+PRP than in Sham-group, and CXCL12, BMP-4, TGF-beta and IGF-1 expressions were also increased. This study showed MSCs +/-PRP transplantation after POI supports recovery of the follicular count and function. For ovarian recovery, a single administration of PRP was found not sufficient. Although MSC treatment increased follicular regeneration, better results were obtained in the co-transplantation of MSCs and PRP. These results might be promising for follicular regeneration in POI patients.