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  • Küçük Resim
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    Does transcription factor, induced by daptomycin and vancomycin, affect HIF-1α, Chondroadherin, and COL2A1?
    (2018) Karaarslan, Numan; Yilmaz, Ibrahim; Yasar Sirin, Duygu; Ozbek, Hanefi; Kaya, Yasin Emre; Akyuva, Yener; Kaplan, Necati; Dogan, Mustafa; Gumustas, Seyit Ali; Erdem, Ilknur; Ates, Ozkan
    Aim: In this study, it was firstly aimed to investigate the effect of Daptomycin (DAP) on the proliferation in Vancomycin (VCM)- administered primary chondrocyte cultures and non-drug-administered primary chondrocyte cultures. Our second objective was to investigate the effects of DAP and VCM on the NP-specific marker protein chondroadherin (CHAD), which is associated with spinal cord and dorsal column growth, on the transcription factor-1 alpha (HIF-1α), which is induced by hypoxia, and on a type II collagen (COL2A1), which is also known to play a significant role in the development of extracellular matrix, at the pharmaco-molecular level. Material and Methods: Standard human primary chondrocyte cultures were established. DAP and VCM were added to the samples. In all groups, molecular analysis was performed at 0th, 24th and 48th hours. In addition, the surface morphology of the cells was evaluated. Results: Changes in cell morphology and cell death in cultures were observed 24 hours after administration of antibiotics to cell cultures. It was observed that drug administration was associated with the cell viability and that cell viability rate for two antibiotics was similar at the 0th and 48th hours. The expression of three genes decreased at the 24th hour in the experimental group where DAP was administered. Conclusion: Thanks to this molecular-based research, it should not be forgotten that DAP and VCM active pharmacological agents, especially used in the treatment of Methicillin-resistant Staphylococcus aureus induced surgical infections, have a negative effect on human chondrocyte and ECM components.
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    Evaluation of the effect of apixaban on the primary intact intervertebral disc cell cultures
    (2019) Akgun, Feride Sinem; Karaarslan, Numan; Yilmaz, İbrahim; Ozbek, Hanefi; Yasar Sirin, Duygu; Simsek, Abdullah Talha; Kaplan, Necati; Ates, Ozkan
    Aim: Apixaban is a frequently preferred pharmacological agent in clinics to prevent deep vein thrombosis and pulmonary embolism. Such new oral anticoagulants may cause hemorrhage’s in tissues and/or organs or may cause gastrointestinal symptoms without bleeding. It is also reported in the literature that it may lead to mental disorders, unwanted disorders in the urinary tract and skeletal-muscle system. However, when the literature is examined, there are no studies, which are of high-evidential value, evaluating the efficacy of apixaban on healthy, intact intervertebral disc tissue, and matrix-like structures. In this pharmaco-molecular study, it was aimed to investigate the effects of a new oral anticoagulant agent containing the active ingredient apixaban on the intact intervertebral disc tissue cells, extracellular matrix (ECM) structure and to evaluate its positive and / or negative effects on gene expressions of cartilage oligo matrix protein (COMP), chondroadherin (CHAD), and Matrix Metalloproteinase (MMP)s.Material and Methods: The primary cell cultures were prepared from the intact tissues of the patients with the traumatic intervertebral disc herniation. Apixaban was administered to the cultures and molecular analyses were performed for 21 days. The data obtained from the apixaban-administered and non-apixaban-administered samples were evaluated statistically and the significance value was accepted as P 0.05.Results: The changes were observed in the cell proliferation and the expressions of the mentioned genes in the apixaban-administered group. The suppression of COMP value and the increase in MMP-13 value may be indicative of the development of matrix degeneration in the apixaban-administered group, compared to the non-drug-administered control group. Conclusion: The selectivity is one of the most important features of the drugs. However, it should not be forgotten that no drug will only produce the desired effect.
  • Küçük Resim
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    Evaluation of the expression and proliferation of degenerative markers in primary cell cultures obtained from human intervertebral disc tissue
    (2020) Akalan, Hande; Kaya, Yasin Emre; Yilmaz, İbrahim; Karaarslan, Numan; Yasar Sirin, Duygu; Ozbek, Hanefi
    Aim: A major cause of low back pain is disc degeneration. Nevertheless, no specific and reliable markers of the degeneration of the nucleus pulposus (NP) are available. This presented study aimed to examine changes in the expressions of genes in primary cell cultures isolated from intact and degenerated tissues to give insights into the biopathogenesis of intervertebral disc (IVD) tissue.Material and Methods: Tissues of eight patients (n = 8; average age: 41.74 ± 9.86 years) were resected through microdiscectomy, and primary cell cultures were prepared using degenerated disc tissue. The cultured degenerated tissues served as the study group. The samples in the control group comprised the intact tissues of patients (n = 8; average age: 38.68 ± 7.91 years) resected following a trauma. Morphology of the cell surface were evaluated using an inverted light/fluorescent microscopy at 0 and 24 h on days 10 and 21. The expressions of the chondroadherin (CHAD), cartilage oligomeric matrix protein (COMP), interleukin-1 beta (IL-1 beta), and matrix metalloproteinase (MMP)-7 and MMP-19 genes were evaluated using the reverse transcription-quantitative polymerase chain reaction (RT-qPCR). The data obtained were statistically analyzed.Results: The four genes investigated, except COMP (P > 0.05), changed significantly in primary cell cultures isolated from degenerative IVD tissues. This result was statistically significant (P 0.05). The gene expressions in the samples derived from intact IVD tissues changed markedly and these changes were associated with proliferation (P 0.05).Conclusion: Analyzing the changes in gene expression levels associated with IVD should contribute to future studies on the prevention and treatment of such pathologies. The data obtained from the present study will shed light on cellular-based personal targeted therapies through which genetic information can be transmitted to cells.
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    In-vitro evaluation of the effects of tigecycline on annulus fibrosus and nucleus pulposus
    (2020) Kaya, Yasin Emre; Yilmaz, Ibrahim; Karaarslan, Numan; Bilir, Bulent; Yasar Sirin, Duygu; Ozbek, Hanefi
    Aim: This study aimed to examine the effects of tigecycline on primary cell cultures established using intervertebral disc tissues.Material and Methods: Primary Annulus Fibrosus and Nucleus Pulposus cultures were obtained from human intervertebral disc tissue. Untreated samples served as the control group, and treated samples served as the study group. Treated and untreated samples were statistically evaluated using an alpha significance value of 0.05.Results: Proliferation and gene expression decreased in the tigecycline administered cultures when compared to the control group samples (P 0.05). Conclusion: Drugs used in clinics may have side effects other than those indicated in their package insert.
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    Investigation of the effects of tigecycline, a semi-synthetic derivative of minocycline, on chondrocyte cultures
    (2020) Kaya, Yasin Emre; Karaarslan, Numan; Yilmaz, İbrahim; Yasar Sirin, Duygu; Yaman, Onur; Ozbek, Hanefi; Ates, Ozkan
    Aim: Although antibiotics are generally well-tolerated, they may have cytotoxic effects. The present randomized, double-blind, in vitro study aimed to investigate the effects of tigecycline on cartilage tissue cells and the extracellular matrix.Material and Methods: Cartilage tissues of patients (n = 8) were used for the preparation of primary cell cultures. Tigecycline-treated cell cultures served as the study group. Non-treated cell cultures served as the control group. Analyses were performed at 0, 24, 48, and 72 h in both groups. The results obtained were statistically evaluated. The alpha significance value was determined to be 0.05.Results: Proliferation remained unchanged in the tigecycline-treated cell cultures. The gene expressions of the markers involved in anabolic pathways increased in the tigecycline-treated cell cultures. The results obtained were statistically significant (P 0.05). Conclusion: Although tigecycline had no toxic effect on the chondrocyte cell cultures and caused no damage to the extracellular matrix, the present study was performed in an in vitro environment.