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Öğe A study of the effects of metformin, a biguanide derivative, on annulus fibrosus and nucleus pulposus cells(Turkish Neurosurgical Society, 2020) Kaya, Yasin Emre; Karaarslan, Numan; Yılmaz, İbrahim; Yaşar Şirin, Duygu; Akalan, Hande; Özbek, HanefiAIM: 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-1 beta (IL-1 beta) 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.Öğe A study on side effects of antibiotics used in the treatment of patients with head trauma and legal responsibility of clinicians in terms of the right to health(2019) Karaarslan, Abdulkadir; Şimşek, Abdullah Talha; Doğan, Mustafa; Potoğlu, Bilgehan; Yılmaz, İbrahim; Karaarslan, NumanAim: Patients with head trauma are routinely, and commonly treated with prophylactic antibiotics, which may also be used for the treatment of infection that may be developed during hospital stay. The relevant antibiotic may, however, some side effects and adverse event. The present study aimed to investigate whether the side effects and adverse events of the prophylactic antibiotics were considered as a complication or medical negligence. Material and Methods: Descriptive statistics were used for the evaluation of the data. Results: No studies were found in the literature. Ceftriaxone, ampicillin-sulbactam, and cefazolin sodium were preferred for antibiotherapy. Meropenem or vancomycin was solely administered to patients when observed active pathogens in cultureantibiogram. Clinicians should be cautioned the potential side effects and adverse events of some drugs frequently used in clinics. Conclusion: Otherwise, they may legally be held liable for medical negligence.Öğe A study on the effects of direct factor Xa inhibitors and direct thrombin inhibitors on human primary chondrocyte cultures(Namık Kemal University, 2019) Kaya, Yasin Emre; Akalan, Hande; Yılmaz, İbrahim; Karaarslan, Numan; Yaşar Şirin, Duygu; Özbek, Hanefi; Ateş, ÖzkanAim: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.Öğe Are radio-contrast agents commonly used in discography toxic to the intact intervertebral disc tissue cells?(Wiley, 2019) Karaarslan, Numan; Yılmaz, İbrahim; Özbek, Hanefi; Yaşar Şirin, Duygu; Kaplan, Necati; Çalışkan, Tezcan; Özdemir, Çiğdem; Akyuva, Yener; Ateş, ÖzkanIn the literature, there have been no studies showing clear results on how radio-contrast pharmaceuticals would affect intact disc tissue cells. In this context, it was aimed to evaluate the effects of iopromide and gadoxetic acid, frequently used in the discography, on intact lumbar disc tissue in pharmaco-molecular and histopathological level. Primary cell cultures were prepared from the healthy disc tissue of the patients operated in the neurosurgery clinic. Except for the control group, the cultures were incubated with the indicated radio-contrast agents. Cell viability, toxicity and proliferation indices were tested at specific time intervals. The cell viability was quantitatively analysed. It was also visually rechecked under a fluorescence microscope with acridine orange/propidium iodide staining. Simultaneously, cell surface morphology was analysed with an inverted light microscope, while haematoxylin and eosin (H&E) staining methodology was used in the histopathological evaluations. The obtained data were evaluated statistically. Unlike the literature, iopromide or gadoxetic acid did not have any adverse effects on the cell viability, proliferation and toxicity (P <0.05). Although this study reveals that radio-contrast pharmaceuticals used in the discography, often used in neurosurgical practice, can be safely used, it should be remembered that this study was performed in an in vitro environment.Öğe Are specific gene expressions of extracellular matrix and nucleus pulposus affected by primary cell cultures prepared from intact or degenerative intervertebral disc tissues?(Turkish Neurosurgical Society, 2019) Karaarslan, Numan; Yılmaz, İbrahim; Özbek, Hanefi; Yaşar Şirin, Duygu; Kaplan, Necati; Akyuva, Yener; Gönültaş, Aylin; Ateş, ÖzkanAIM: To determine the gene expression patterns of nucleus pulposus (NP) in cell cultures obtained from degenerated or intact tissues. MATERIAL and METHODS: Whereas 12 of the cases were diagnosed with lumbar disc herniation and had undergone lumbar microdiscectomy, 12 cases had undergone traumatic intervertebral discectomy and corpectomy, along with discectomy after spinal trauma. NP-specific markers and gene expressions of the reagents of the extracellular matrix in the experimental setup were tested at the 0th, 24th, and 48th hours by quantitative reverse transcriptase polymerase chain reaction (qRT-PCR). Visual evaluations were simultaneously made in all samples using invert and fluorescence microscopy. Vitality and proliferation analyses were evaluated by UV spectrophotometer. As a method of statistical evaluation, Spearman was used for categorical variants, and the Pearson correlation was used for variants with numerical and plain distribution. RESULTS: No association was found either between the tissue type and times (r=0.000; p=1.000) or between the region that the tissue was obtained from and hypoxia transcription factor-1 alpha (HIF-1 alpha) gene expression (r=0.098; p=0.245). There was no correlation between cell proliferation and chondroadherin (CHAD) expression or between type II collagen (COL2A1) and CHAD gene expressions. It was found that CHAD and HIF-1 alpha gene expressions and HIF-1 alpha and COL2A1 gene expressions affected cell proliferation. CONCLUSION: Cell culture setups are of paramount importance because they may influence the pattern of changes in the gene expressions of the cells used in these setups.Öğe Do endocrinopathies cause changes in transverse carpal ligament thickness and carpal tunnel area in carpal tunnel syndrome?(Galenos Publishing House, 2020) Karaarslan, Numan; Şimşek, Abdullah Talha; Öznam, Kadir; Bülbül, Ahmet MuratObjective: The purpose of this study was to investigate the effect of transverse carpal ligament (TCL) thickness and the size of carpal tunnel area on clinical findings and electrophysiological changes through the wrist magnetic resonance imaging (MRI). Methods: In this prospective study, the thickness of the TCL and the carpal tunnel areas of preoperative cases diagnosed with carpal tunnel syndrome (CTS) were measured via wrist MRI results. The effect of TCL thickness and carpal tunnel area on endocrinopathies such as diabetes mellitus (DM) and hypothyroidism and the effect of these variables on clinical findings and electrophysiologic changes were evaluated in the light of the literature. Results: TCL thickness and carpal tunnel area were not statistically significant among DM, hypothyroidism and electrophysiological changes (p>0.05). Conclusion: This study include one of the limited researches comparing the carpal tunnel area and TKL thickness in cases with and without endocrinopathy CTS. However, there is a need for further researches that have a greater number of cases, including multi-centered, differentwinged people.Öğe Do we damage nucleus pulposus tissue while treating cerebrovascular ischemic neurological deficits with nimodipine?(2018) Karaarslan, Numan; Yılmaz, İbrahim; Yaşar Şirin, Duygu; Baykız, Derya; Demirkıran, Aykut; Ateş, ÖzkanAim: Nimodipine is used to prevent cerebrovascular-originated ischemic neurological deficits, yet its effects on nucleus pulposus (NP) cells or annulus fibrosus (AF) cells weren’t studied. This study aimed to examine nimodipine’s effects on vitality and proliferation of chondroadherin (CHAD), type II collagen (COL2A1), and hypoxia-inducible factor 1 alpha (HIF 1?) gene expression in human primary NP/AF cells. Material and Methods: NP/AF cell cultures obtained from 6 patients who underwent microdiscectomy were treated with 100 µMolar nimodipine and analyzed at 0, 24, and 48 h. Data were evaluated using one-way ANOVA and post-hoc Tukey HSD with 95% confidence interval. Results: We observed suppressed cell proliferation and increased necrosis in nimodipine-treated NP/AF cell cultures, especially degenerated tissue. COL2A1 gene expression wasn’t detected in any experimental groups. CHAD and HIF 1? expression had timedependent decreases in control. CHAD and HIF 1? expression were found to decrease at 24h, but increased at 48h in degenerated tissue. In nimodipine-applied intact tissues, CHAD expression was stable at 24h but 1.62 times higher than control at 48h. HIF 1? levels were lower than control. Conclusion: In nimodipine-treated degenerated AF/NP cultures, CHAD and HIF 1? expressions had time-dependent decreases. However, after complete RT-PCR data evaluation, no correlation between nimodipine application and gene expression occurred.Öğe Does nimodipine, a selective calcium channel blocker, impair chondrocyte proliferation or damage extracellular matrix structures?(Bentham Science Publishers, 2019) Kaplan, Necati; Yılmaz, İbrahim; Karaarslan, Numan; Kaya, Yasin Emre; Yaşar, Şirin, Duygu; Özbek, HanefiBackground: The study aimed to investigate the effects of the active ingredient, nimodipine, on chondrocyte proliferation and extracellular matrix (ECM) structures in cartilage tissue cells. Methods: Chondrocyte cultures were prepared from tissues resected via surgical operations. Nimodipine was then applied to these cultures and molecular analysis was performed. The data obtained were statistically calculated. Results: Both, the results of the (3-(4,5 dimethylthiazol2-yl)-2,5-diphenyltetrazolium (MTT) assay and the fluorescence microscope analysis [a membrane permeability test carried out with acridine orange/propidium iodide staining (AO/PI)] confirmed that the active ingredient, nimodipine, negatively affects the cell cultures. Conclusion: Nimodipine was reported to suppress cellular proliferation; chondroadherin (CHAD) and hypoxia-inducible factor-1 alpha (HIF-1?) expression thus decreased by 2.4 and 1.7 times, respectively, at 24 hrs when compared to the control group (p < 0.05). Furthermore, type II collagen (COL2A1) expression was not detected (p < 0.05). The risk that a drug prescribed by a clinician in an innocuous manner to treat a patient by relieving the symptoms of a disease may affect the proliferation, differentiation, and viability of other cells and/or tissues at the molecular level, beyond its known side effects or adverse events, should not be forgotten.Öğe Does oseltamivir protect human chondrocyte and nucleus pulposus cells from degeneration by inhibiting senescence and proinflammation mediated by the NLRP3 inflammasome and NF-?B?(Verduci Editore s.r.l, 2022) Yılmaz, İbrahim; Akalan, Hande; Öznam, Kadir; Karaarslan, Numan; Yaşar Şirin, Duygu; Özbek, HanefiOBJECTIVE: Recent drug design studies suggest that inflammation is among the most important factors in the development of both intervertebral disc (IVD) degeneration (IVDD) and osteoarthritis (OA) due to cartilage damage. This study aimed to investigate whether the anti-inflammatory drug oseltamivir has a toxic effect on IVD and cartilage tissue cells. It assessed what effect oseltamivir has on hypoxia-inducible factor (HIF)-1 alpha (HIF1?), which plays an important role in anabolic pathways in IVD and cartilage tissue. In addition, the study analyzed whether oseltamivir could inhibit the release of inflammatory interleukin-1 beta (IL-1?) via the nuclear factor kappa-B (NF-?B) signaling pathway by activating the nucleotide-binding oligomerization domain and leucine-rich repeat protein-3 (NLRP3) inflammasome. MATERIALS AND METHODS: Human lumbar IVD (n = 8) tissues were isolated for annulus fibrosus (AF) and nucleus pulposus (NP) primary cell cultures, and human tibial and femoral cartilage tissues (n = 8) were isolated for primary chondrocyte cultures. Untreated groups served as the control and oseltamivir-treated groups as the study sample. Cell viability and cytotoxicity were evaluated at 0, 24, 48, and 72 h in all groups for changes in HIF-1?, IL-1?, NF-?B, and the NLRP3-inflammasome protein expressions using Western blotting. The ? significance value was < 0.05. RESULTS: In the oseltamivir-treated groups, cell proliferation decreased in both AF/NP cell and chondrocyte cultures obtained from IVD cartilage tissues. After Western blotting analysis, changes were observed in the protein expressions of HIF-1?, IL-1?, NF-?B, and the NLRP3 inflammasome in both AF/NP cells and chondrocytes. The results were statistically significant (p < 0.05). CONCLUSIONS: Oseltamivir treatment may be a promising regenerative strategy to manage IVDD and osteoarthritic cartilage tissues.Öğe Does transcription factor, induced by daptomycin and vancomycin, affect HIF-1?, Chondroadherin, and COL2A1?(Inonu University School of Medicine, 2018) Karaarslan, Numan; Yılmaz, İbrahim; Yaşar Şirin, Duygu; Özbek, Hanefi; Kaya, Yasin Emre; Akyuva, Yener; Kaplan, Necati; Doğan, Mustafa; Gümüştaş, Seyit Ali; Ateş, Özkan; Erdem, İlknurAim: 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-1?), 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.Öğe Effect of naproxen on proliferation and differentiation of primary cell cultures isolated from human cartilage tissue(Spandidos Publications Ltd, 2018) Karaarslan, Numan; Batmaz, Ahmet Güray; Yılmaz, İbrahim; Özbek, Hanefi; Çalışkan, Tezcan; Yaşar Şirin, Duygu; Kaplan, Necati; Öznam, Kadir; Ateş, ÖzkanNon-steroidal anti-inflammatory drugs (NSAIDs) that are applied through oral, injectable or topical routes have been widely used in painful and inflammatory musculoskeletal diseases. The current study aimed to determine whether naproxen, an aryl acetic acid derivative with analgesic and anti-inflammatory effects, has a toxic effect on human chondrocytes. Samples containing monolayer primary chondrocyte cultures were prepared following resection from osteochondral tissues obtained from patients with gonarthrosis. Cell viability, toxicity and proliferation and levels of stage-specific embryonic antigen-1, a precursor to human prechondrocytes, were evaluated spectrophotometrically. The results from the untreated control group were compared with those of the study groups, where naproxen was administered in varying doses (1-1,000 mu M). Surface morphologies of the cells were compared using inverted light and environmental scanning electron microscopy. Treatment groups were compared by analysis of variance with Tukey's honest difference post hoc test. P<0.01 was considered to indicate a statistically significant difference. The research revealed significant changes to proliferation and differentiation of chondrocytes in all treatment groups (P<0.01). Naproxen was demonstrated to suppress chondrocyte proliferation and differentiation, which may be an important factor to consider when prescribing this medication to patients.Öğe Effects of etanercept, a tumor necrosis factor receptor fusion protein, on primary cell cultures prepared from intact human intervertebral disc tissue(Spandidos Publications, 2019) Çalışkan, Tezcan; Şirin, Duygu Yaşar; Karaarslan, Numan; Yılmaz, İbrahim; Özbek, Hanefi; Akyuva, Yener; Kaplan, Necati; Kaya, Yasin Emre; Şimşek, Abdullah Talha; Güzelant, Aliye Yıldırım; Ateş, ÖzkanThe aim of the present study was to investigate the effects of etanercept (ETA), a tumor necrosis factor (TNF) inhibitor, on human cell cultures prepared from intact intervertebral disc tissue. ETA is used as a treatment for cases of rheumatoid arthritis, psoriatic arthritis, axial spondyloarthritis and ankylosing spondylitis accompanied by moderate or severe joint pain. ETA was applied to primary cell cultures [annulus fibrosus and nucleus pulposus (NP) from intact intervertebral disc tissue]. Cell cultures without ETA treatment served as the control group. Morphological and quantitative molecular analyses of the two groups were performed. The number of viable cells and cell proliferation decreased in the ETA-treated cultures as compared with those in the control group. Furthermore, in the treatment group, the chondroadherin gene, an NP-specific marker, was not expressed after 24 h. By contrast, the cartilage oligo matrix protein was expressed 24, 48 and 72 h post-ETA treatment, while its expression was significantly lower than that in the control group. In addition, the expression of interleukin-1 beta, as well as matrix metallopeptidase-7 and -19, was markedly decreased. Overall, the cell proliferation and gene expression in the ETA-treated cells were significantly different from those in the control group (P<0.05). These results suggest that the treatment duration and dosage of TNF inhibitors, which are used to suppress active inflammation, should be considered in the clinical setting. These biological agents may delay the healing of intervertebral disc tissue damage by slowing cell proliferation and altering gene expression via anabolic and catabolic pathways.Öğe Evaluation of empirical antibiotic treatment in culture negative pyogenic vertebral osteomyelitis(Turkish Neurosurgical Society, 2019) Doğan, Mustafa; Şimşek, Abdullah Talha; Yılmaz, İbrahim; Karaarslan, NumanAIM: To investigate the efficacy of empirical antibiotic treatment in culture-negative pyogenic vertebral osteomyelitis (PVO) cases. MATERIAL and METHODS: The records of patients with culture-negative PVO who were treated at infectious diseases and neurosurgery outpatient clinics in the past four years were examined retrospectively. The control group comprised healthy subjects with similar age, gender, and body mass index but without pathology. The comparison of the groups was performed by analysis of variance. Statistical significance was accepted as p < 0.05. RESULTS: No statistically significant difference in the white blood cell count and erythrocyte sedimentation rate was found between the spondylodiscitis and the healthy subject groups when the blood parameters obtained before and after the treatment (p > 0.05). However, a statistical significance was assessed in the results of the comparison for C-reactive protein (p < 0.05). CONCLUSION: In the context of evidence-based medicine and the rational use of antibiotics, it is clear that antibiotics should be preferred according to the culture antibiogram results in the treatment of infectious diseases.Öğe Evaluation of neutrophil-to-lymphocyte ratio as a marker of inflammatory response in spondylodiscitis(2018) Karaarslan, Numan; Yılmaz, İbrahim; Akgün, Feride Sinem; Çalıskan, Tezcan; Doğan, Mustafa; Bilir, Bülent; Ateş, ÖzkanAim: Spondylodiscitis, if not diagnosed on time, can cause morbidity or mortality at high rates. This study was carried out with the aim of testing the hypothesis that “neutrophil-to-lymphocyte ratio can be used” especially in cases where it is difficult to diagnose spondylodiscitis. Material and Methods: This study involved 24 patients admitted to the State Hospital of Ministry of Health and Namik Kemal University for spondylodiscitis between January2014 and June2017. After excluding the cases that did not meet the inclusion criteria (n=6), the remaining cases (n=24) were referred to as the study group. A control group was created from healthy volunteers (n=24) who applied for routine physical checkups at the clinic between the same dates and who were similar in terms of age, sex, and body mass index to the study group. Hemogram parameters of the cases in both groups; white blood cell, C-reactive protein, erythrocyte sedimentation rate, and neutrophil-to-lymphocyte ratio were statistically compared. Results: Patients in the spondylodiscitis group, compared to healthy volunteers had statistically significant neutrophil-to-lymphocyte ratio value. Conclusion: Especially in cases where the diagnosis of spondylodiscitis is not assured, the neutrophil-to-lymphocyte ratio parameter, which is less costly than other diagnostic methods and the analysis results of which can be obtained in a shorter time, may be used to support clinical diagnosis.Öğe Evaluation of the effect of apixaban on the primary intact intervertebral disc cell cultures(2019) Akgün, Feride Sinem; Karaarslan, Numan; Yılmaz, İbrahim; Özbek, Hanefi; Yaşar Şirin, Duygu; Şimşek, Abdullah Talha; Kaplan, Necati; Ateş, ÖzkanAim: 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 withoutbleeding. It is also reported in the literature that it may lead to mental disorders, unwanted disorders in the urinary tract and skeletalmuscle system. However, when the literature is examined, there are no studies, which are of high-evidential value, evaluating theefficacy of apixaban on healthy, intact intervertebral disc tissue, and matrix-like structures. In this pharmaco-molecular study, itwas aimed to investigate the effects of a new oral anticoagulant agent containing the active ingredient apixaban on the intactintervertebral disc tissue cells, extracellular matrix (ECM) structure and to evaluate its positive and / or negative effects on geneexpressions 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 intervertebraldisc herniation. Apixaban was administered to the cultures and molecular analyses were performed for 21 days. The data obtainedfrom the apixaban-administered and non-apixaban-administered samples were evaluated statistically and the significance valuewas accepted as P <0.05.Results: The changes were observed in the cell proliferation and the expressions of the mentioned genes in the apixabanadministered group. The suppression of COMP value and the increase in MMP-13 value may be indicative of the development ofmatrix 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 willonly produce the desired effect.Öğe Evaluation of the expression and proliferation of degenerative markers in primary cell cultures obtained from human intervertebral disc tissue(Annals of Medical Research Publishing Inc, 2020) Kaya, Yasin Emre; Akalan, Hande; Yılmaz, İbrahim; Karaarslan, Numan; Yaşar Şirin, Duygu; Özbek, HanefiAim: 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.Öğe In-vitro evaluation of the effects of tigecycline on annulus firosus and nucleus pulposus(Annals of Medical Research Publishing Inc, 2020) Kaya, Yasin Emre; Yılmaz, İbrahim; Karaarslan, Numan; Bilir, Bülent; Yaşar Şirin, Duygu; Özbek, HanefiAim: 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 signifiance 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.Öğe Investigation of the effect of dipyrone on cells isolated from intervertebral disc tissue(Spandidos Publications, 2019) Akgün, Feride Sinem; Şirin, Duygu Yaşar; Yılmaz, İbrahim; Karaarslan, Numan; Özbek, Hanefi; Şimşek, Abdullah Talha; Kaya, Yasin Emre; Kaplan, Necati; Akyuva, Yener; Çalışkan, Tezcan; Ateş, ÖzkanThe present study aimed to evaluate the effects of dipyrone, an indispensable analgesic, anti-pyretic and anti-spasmodic used in emergency departments, on nucleus pulposus and annulus fibrosus cells in vitro. After surgical biopsy, primary cell cultures were prepared from intact intervertebral disc tissues. Dipyrone was administered to the cultures in the experimental groups except for the control group. The data obtained were statistically evaluated. The proliferation was identified to be suppressed via MTT analysis. The gene expression profile of the intervertebral disc cells in the dipyrone-treated groups was significantly changed. The expression of chondroadherin, cartilage oligo matrix protein, interleukin-1 beta and metalloproteinase (MMP)-19 genes were decreased, but MMP-13 and MMP-7 genes expressions were increased, as determined via reverse transcription-quantitative PCR. AO/PI staining revealed that no apoptotic or other type of cell death was detectable after administration of dipyrone does not mean that the drug is innocuous. The occurrence of cellular senescence and/or the halt of cell proliferation may also be important mechanisms underlying the adverse inhibitory effects of dipyrone. Therefore, prior to administering dipyrone in clinical practice, all possible adverse effects of this drug should be considered.Öğe Investigation of the effects of methylphenidate, an amphetamine derivative, on intervertebral disc tissue cell cultures and matrix structures(Turkish Neurosurgical Society, 2019) Kaya, Yasin Emre; Karaarslan, Numan; Şirin, Duygu Yaşar; Özbek, Hanefi; Kaplan, Necati; Yılmaz, İbrahimAIM: To investigate the effects of methylphenidate (MPH), on intervertebral disc tissue (IVD) cell cultures and extracellular matrix structures. Changes in the expression of some important marker genes involved in anabolic and catabolic mechanisms of IVD extracellular matrix formation were also evaluated. MATERIAL and METHODS: Primary cultures of nucleus pulposus cells (NPCs) and annulus fibrosus cells (AFCs) were isolated from tissues obtained from the operated patients. Cell viability and proliferation were tested, and the cell surface morphologies were evaluated by microscopy. The expressions of the chondroadherin (CHAD), cartilage oligomeric matrix protein (COMP), interleukin-1 beta (IL-1?) and matrix metalloproteinase (MMP) -7 and MMP-19 genes were evaluated using the quantitative real-time polymerase chain reaction (qRT-PCR). A value of p < 0.05 was considered statistically significant. RESULTS: The viability and proliferation of intervertebral disc tissue cells decreased in response to MPH treatment and the expression of the investigated genes also changed. CONCLUSION: The data obtained from in-vitro studies may not directly adaptable to clinical applications. However, the fact that the central nervous system stimulant MPH can suppress proliferation of cells derived from IVD tissue should be considered carefully by clinicians.Öğe Investigation of the effects of tigecycline, a semi-synthetic derivative of minocycline, on chondrocyte cultures(Annals of Medical Research Publishing Inc, 2020) Kaya, Yasin Emre; Karaarslan, Numan; Yılmaz, İbrahim; Yaşar Şirin, Duygu; Yaman, Onur; Özbek, Hanefi; Ateş, ÖzkanAbstract 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.
