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    Assessment of the Impact of Curcumin on Cell Cultures Derived from the Primary Intervertebral Disc Tissue in Humans
    (Maltepe Üniversitesi, 2023) Albayrak, Mehmet; Yılmaz, İbrahim; Yüzbaşı, Muharrem Furkan; Akalan, Hande; Şirin, Duygu Yaşar; Karaarslan, Numan; Özbek, Hanefi
    Aim: Degenerative disc disease in the lumbar spine is widely observed. Degenerative disc diseases are among the causes of low back pain in older age. Modern drug discovery studies have aimed to identify potential molecules that target multiple pathways with a safer profile against degeneration. This study aimed to evaluate the effects of curcumin, a natural phenolic compound, on primary cell cultures prepared using intervertebral disc (IVD) tissues resected during the surgeries of patients with lumbar disc herniation. Materials and Methods: Primary cell cultures were prepared using human IVD tissues of eight patients. Untreated groups served as the control and curcumin-treated groups as the study sample. In-vitro cytotoxicity analyses were performed in all groups. Acridine orange (AO)/propidium iodide (PI) and Janus Green B staining were performed to evaluate cell surface morphologies. One-way analysis of variance and Tukey HSD, a multiple comparison test, were used to assess the obtained data. Results: Proliferation slightly increased as of 24 h in the curcumin-treated samples, but decreased in the 48 and 72 hour curcumin-treated samples compared to the control samples. The obtained results were statistically significant (p
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    The effects of rivaroxaban, an oral anticoagulant, on human IVD primary cultures
    (Termedia Publishing House Ltd, 2022) Caliskan, Tezcan; Akalan, Hande; Yilmaz, Ibrahim; Karaarslan, Numan; Sirin, Duygu Yasar; Özbek, Hanefi
    Introduction: The present study aimed to investigate the potential effects of rivaroxaban, an oral anticoagulant that inhibits the effects of factor Xa, on intact intervertebral disc tissue cells and the extracellular matrix (ECM). Material and methods: Rivaroxaban was applied to primary human cell cultures prepared from tissues of the intervertebral disc. Comparative molecular analyses were performed on non-drug-treated control group samples. Descriptive statistics were presented as the mean +/- standard deviation. An analysis of variance test was performed to determine whether there were significant differences in the mean across the groups. When differences across groups were observed, Tukey's honestly significant difference post-hoc test was used for multiple pairwise comparisons. The significance of the obtained data was determined statistically. The alpha significance value was < 0.05. Results: The cells in the control group and in the rivaroxaban-treated group were viable, healthy, and proliferated (p < 0.05). However, the expression levels of the chondroadherin gene (CHAD), cartilage oligo matrix protein (COMP), matrix metalloproteinase (MMP)-13, and MMP-19 genes were changed (p < 0.05). Conclusions: Although rivaroxaban does not suppress cell proliferation due to morphological, biological, and biochemical changes in the intervertebral disc tissue, it may change the expression of genes that are related to ECM maintenance.
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    Effects of Tocilizumab on Intervertebral Disc Degeneration, Cell Senescence and Inflammation via BMP-2, Hif-1α, IL-1β and SOX9
    (Asian Network Scientific Information-Ansinet, 2023) Yilmaz, Ibrahim; Akalan, Hande; Sirin, Duygu Yasar; Karaarslan, Numan; Kasim, Emin; Ozbek, Hanefi; Ates, Ozkan
    Background and Objective: Immunosuppressive tocilizumab (TCZ), which is frequently used in the treatment of rheumatoid arthritis, can have many side effects as well as an uncontrolled inflammatory response. This study aimed to evaluate the effect of tocilizumab (TCZ) administered to intervertebral disc (IVD) tissues in vitro on the proinflammatory cytokines and proteins of degeneration, senescence and inflammation-related signaling pathways at the pharmaco-molecular level. Materials and Methods: Primary cell cultures were prepared using human IVD tissues obtained during lumbar microdiscectomy. Untreated groups served as the control and TCZ-treated groups as the study sample. Analyses were performed using a commercial kit, supravital and fluorescent dyes. Changes in bone morphogenetic protein (BMP)-2, hypoxia-inducible factor (Hif)1-alpha (Hif1-alpha), interleukin (IL)-1 beta (IL-1 beta) and sex-determining region Y (SRY)-box 9 (SOX9) protein expressions were evaluated using western blotting. An alpha value of less than 0.05 was considered significant. Results: Proliferation decreased in the samples treated with TCZ (10 mu g mLG1 ) on day 15 (p<0.05). Protein expressions of BMP-2, Hif-1 alpha, IL-1 beta and SOX9, which play a vital role in anabolic and catabolic pathways, changed in samples treated with TCZ (10 mu g mLG1 ). Conclusion: This change was statistically significant (p<0.05). Therefore, results concluded that the inflammation, extracellular matrix degradation and nucleus pulposus degeneration after disc herniation are controlled by BMP-2, Hif-1 alpha, IL-1 beta and SOX9.
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    Is favipiravir a potential therapeutic agent in the treatment of intervertebral disc degeneration by suppressing autophagy and apoptosis?
    (Turkish Neurosurgical Soc, 2022) Yılmaz, İbrahim; Akalan, Hande; Şirin, Duygu Yaşar; Karaarslan, Numan; Özbek, Hanefi; Ateş, Özkan
    AIM: To evaluate the effects of favipiravir (FVP) on cell viability and cytotoxicity in human degenerated primary intervertebral disc (IVD) tissue cell cultures. Furthermore, the protein expressions of hypoxia-inducible factor 1 alpha (HIF-1 alpha), nuclear factor-kappa-b (NF-kappa B), and interleukin-1 beta (IL-1 beta) were also examined. MATERIAL and METHODS: Untreated cell cultures served as the control group, named group 1. Cell cultures treated with FVP served as the study group, named group 2. Pharmacomolecular analyses were performed in all groups at 0, 24, 48, and 72 hours (h). Obtained data were evaluated statistically. RESULTS: Cell proliferation was suppressed in the FVP-treated samples compared to the control group samples at 24 and 72 h, and this was statistically significant (p<0.05). Decreased or increased protein expression levels of HIF-1 alpha, NF-kappa B, and IL-1 beta in FVP-treated samples may be an indication of suppression in anabolic events as well as proliferation in IVD cultures. FVP administration showed that AF/NP cells in a culture medium may induce a strong inflammatory response to FVP. This strong inflammatory response is likely to cause slowed proliferation. It may also be a trigger for many catabolic events. NF-kappa B expression increased within the first 24 h and then decreased rapidly. Based on the data obtained, it may be suggested that the rapidly increasing NF-kB may have stimulated the expression of many antiproliferative genes. CONCLUSION: The suppression of IL-1 beta and NF-kB protein expressions in IVD cells treated with FVP is important in the treatment of IVD degeneration (IDD). If the protein expression of HIF-1 alpha could be increased along with the suppression of IL-1 beta and NF-kB, FVP would perhaps be a promising pharmacological agent in the treatment of IDD.