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    Evaluation of the expression and proliferation of degenerative markers in primary cell cultures obtained from human intervertebral disc tissue
    (2020) Kaya, Yasin Emre; Akalan, Hande; Yılmaz, İbrahi?m; Karaarslan, Numan; Şirin, Duygu Yaşar; Özbek, 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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    A study of the effects of metformin, a biguanide derivative, on Annulus Fibrosus and Nucleus Pulposus cells
    (2020) Kaya, Yasin Emre; Karaarslan, Numan; Yılmaz, İbrahim; Sirin, Duygu Yaşar; Akalan, Hande
    AIM: To investigate the effects of metformin, a drug used widely for the treatment of type 2 diabetes mellitus, on human primary cell cultures prepared from uninjured segment of disc material intervertebral disk tissues. MATERIAL and METHODS: Primary cell cultures were prepared using the tissues of six patients (three males and three females) who had undergone lumbar microdiscectomy and sequestrectomy. Untreated samples served as the control group, and metformintreated samples served as the experimental group. All the samples were evaluated using an inverted light microscope, acridine orange/propidium iodide staining (AO/PI), and a fluorescence microscope. The cytostatic and cytotoxic effects of metformin, which was administered to the samples using a commercial MTT assay kit, were also evaluated. The data obtained were statistically assessed, and the alpha significance value was accepted as less than 0.05. In addition, for the groups’ changes in the expressions of chondroadherin (CHAD), cartilage oligomeric matrix protein (COMP), interleukin-1β (IL-1β) matrix metalloproteinase 7 (MMP-7), and matrix metalloproteinase 19 (MMP-19), genes related to the extracellular matrix synthesis and degradation were determined using gene-specific TaqMan Gene Expression Assays. RESULTS: The administration of the drug adversely affected nucleus pulposus (NP)/annulus fibrosus (AF) cells and extracellular matrix–like structures. This was statistically significant (p<0.05). CONCLUSION: Clinicians should not disregard the adverse effects of metformin, which is used widely in clinical practice, on the components of intervertebral disk tissues.
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    A STUDY ON THE EFFECTS OF DIRECT FACTOR XA INHIBITORS AND DIRECT THROMBIN INHIBITORS ON HUMAN PRIMARY CHONDROCYTE CULTURES
    (2019) Kaya, Yasin Emre; Akalan, Hande; Yilmaz, Ibrahim; Karaarslan, Numan; Şirin, Duygu Yaşar; Özbek, Hanefi; Ateş, Özkan
    Aim:This study investigates the effects of two direct factor Xa inhibitors, apixaban and rivaroxaban, and a direct thrombin inhibitor, dabigatran, on human primary chondrocytecultures.Materials and Methods:Monolayer cultured chondrocytes were prepared. Cell cultures were treated with dabigatran, apixaban, and rivaroxaban. Cultures without drug treatments served as the control group. Using an inverted light microscope, the cell surface morphology was examined. Cell viability and the toxicity of drugs were evaluated using a commercial assay kit, and the results were confirmed using two nucleic acid binding dyes, acridine orange and propidium iodide. The expressions of cartilage oligomeric protein, matrix metalloproteinase-7, and matrix metalloproteinase-19 were assessed using the real-time polymerase chain reaction analysis. All the analyses were performed within 21 days. The data obtained were statistically evaluated.Results:The administration of the three drugs changed the cell viability, proliferation, and expressions of cartilage oligomeric protein, matrix metalloproteinase-7, and matrix metalloproteinase-19. The results were statistically significant (P<0.05).Conclusion:Results obtained from in vitro studies may not provide accurate and reliable insight for clinical practices. However, clinicians should know that drugs used for the prevention or treatment of diseases may suppress chondrocyte proliferation and damage the extracellular matrix formation.
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    A study on the effects of direct factor XA inhibitors and direct thrombin inhibitors on human primary chondrocyte cultures
    (2019) Kaya, Yasin Emre; Akalan, Hande; Yılmaz, İbrahim; Karaarslan, Numan; Şirin, Duygu Yaşar
    Aim: This study investigates the effects of two direct factor Xa inhibitors, apixaban and rivaroxaban, and a direct thrombin inhibitor, dabigatran, on human primary chondrocytecultures. Materials and Methods:Monolayer cultured chondrocytes were prepared. Cell cultures were treated with dabigatran, apixaban, and rivaroxaban. Cultures without drug treatments served as the control group. Using an inverted light microscope, the cell surface morphology was examined. Cell viability and the toxicity of drugs were evaluated using a commercial assay kit, and the results were confirmed using two nucleic acid binding dyes, acridine orange and propidium iodide. The expressions of cartilage oligomeric protein, matrix metalloproteinase-7, and matrix metalloproteinase-19 were assessed using the real-time polymerase chain reaction analysis. All the analyses were performed within 21 days. The data obtained were statistically evaluated. Results:The administration of the three drugs changed the cell viability, proliferation, and expressions of cartilage oligomeric protein, matrix metalloproteinase-7, and matrix metalloproteinase- 19. The results were statistically significant (P<0.05). Conclusion:Results obtained from in vitro studies may not provide accurate and reliable insight for clinical practices. However, clinicians should know that drugs used for the prevention or treatment of diseases may suppress chondrocyte proliferation and damage the extracellular matrix formation.