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Öğe Are Intervertebral Disc Tissue Cells Damaged When Attempting to Prevent Thrombus Formation Using Dabigatran, A New Oral Anticoagulant?(Turkish Neurosurgical Soc, 2019) Kaplan, Necati; Karaarslan, Numan; Yılmaz, İbrahim; Yasar Şirin, Duygu; Akgün, Feride Sinem; Çalışkan, Tezcan; Özbek, HanefiAIM: To investigate the effect of dabigatran, a new oral anticoagulant, on human primary cell cultures isolated from intact intervertebral disc tissue. MATERIAL and METHODS: Cell cultures were prepared from tissues obtained from six cases who had undergone surgery due to spinal trauma. Dabigatran, an active pharmacological agent, was applied to intact annulus fibrosus (AF)/nucleus pulposus (NP) primary cell cultures from the study group. After performing cell viability, toxicity, and proliferation tests on all cultures in the control and study groups, the surface morphologies of the samples were evaluated. Subsequently, chondroadherin (CHAD), cartilage oligomeric matrix protein (COMP), and matrix metalloproteinase (MMP)-13 and -19 expressions were measured via a real-time polymerase chain reaction (RT-PCR). Data were analyzed statistically. RESULTS: In the proliferation assays performed on the 20th day of the study, cells in the dabigatran-supplemented group were reported to have lost 46.37% more viability than those in the control group. Expressions of all genes examined except MMP-13 were evaluated in the control group by time, but in contrast to the control group results, COMP and MMP-19 gene expressions decreased in the dabigatran-treated group. No CHAD or MMP-13 expression was noted in these cultures. CONCLUSION: The potential for a systemically applied drug to accumulate in tissue and negatively affect surrounding tissues and microstructures must be emphasized.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; Şirin, Duygu Yaşar; Kaplan, Necati; Çalışkan, Tezcan; 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 Soc, 2019) Karaarslan, Numan; Yılmaz, İbrahim; Özbek, Hanefi; Yasar Şirin, Duygu; Kaplan, Necati; Akyuva, Yener; 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 Complications of 200 cervical anterior surgery cases and the management of these complications in light of the literature(2019) Kaplan, Necati; Hacıoğlu Kasım, Fatma Bahar; Özger, Özkan; Karaarslan, NumanAim: The aim of this retrospective study was to evaluate the possible complications and the complication management of cervicalanterior discectomies and fusions in light of the literature.Materials and Methods: The study population consisted of patients who presented to the clinic with neck pain and/or arm pain, lossof strength, and sensory disturbances who were operated on after a lack of response to conservative/medical treatment. This studyincluded 200 cervical discopathy and/or cervical spondylosis cases. The literature review was performed according to the PreferredReporting Items for Systematic Reviews and Meta?analyses guidelines without language or country restrictions.Results: The most common complication was dysphagia. The complications also included dural tears, cerebrospinal fluid leakage,graft extrusions, neurological deterioration, postoperative hematomas, and recurrent laryngeal nerve injuries. These were found tobe consistent with the literature.Conclusion: In order to minimize the incidence of complications, the preoperative clinical examinations and radiological imaging ofeach patient should be examined carefully, and the appropriate surgical planning should be performed. It is also important to complywith the rules of asepsis and antisepsis, make sure the surgical time is as short as possible, and perform a dissection based on thepatient’s anatomy with the appropriate surgical equipment. In addition, it is important to wash the surgical area frequently, drain thesystem at the end of the operation, close the tissues in accordance with anatomical integrity, and perform close clinical follow-ups.Öğe Delivering Growth Factors through a Polymeric Scaffold to Cell Cultures Containing both Nucleus Pulposus and Annulus Fibrosus(Turkish Neurosurgical Soc, 2019) Akyuva, Yener; Kaplan, Necati; Yılmaz, İbrahim; Özbek, Hanefi; Şirin, Duygu Yaşar; Karaarslan, Numan; Ateş, ÖzkanAIM: To design a novel, polyvinyl alcohol (PVA)-based polymeric scaffold that permits the controlled release of insulin-like growth factor 1 (IGF-1)/bone morphogenetic protein (BMP)-2 following intervertebral disc administration. MATERIAL and METHODS: The drug delivery system was composed of two different solutions that formed a scaffold within seconds of coming into contact with each other. Swelling, pH, and temperature tests and analysis of the controlled release of growth factors (GFs) from this system were performed. The release kinetics of the GFs were determined through enzyme-linked immunosorbent assay (ELISA). Cell proliferation and viability were monitored with microscopy and analyzed using an MTT assay and acridine orange/propidium iodide (AO/PI) staining. Chondroadherin (CHAD), hypoxia inducible factor-1 alpha (HIF-1 alpha), and collagen type II (COL2A1) gene expressions were determined with quantitative real-time polymerase chain reaction (qRT-PCR) analysis to show the effects of IGF-1/BMP-2 administration on annulus fibrosus cell (AFC)/nucleus pulposus cell (NPC) cultures. For the statistical evaluation of the obtained data, experimental groups were compared with a post hoc Tukey's test following an analysis of variance. RESULTS: The scaffold allowed for the controlled release of IGF-1 and BMP-2 in different time intervals. It was observed that as the application time increased, the number of cells and the degree of extracellular matrix development increased in AFC/NPC cultures. AO/PI staining and an MTT analysis showed that cells retained their specific morphology and continued to proliferate. It was observed that HIF-1 alpha and CHAD expression increased in a time-dependent manner, and no COL2A1 expression in the AFC/NPC cultures was observed. CONCLUSION: The designed scaffold may be used as an alternative method for intervertebral disc administration of GFs after further in vivo studies. Such prototype scaffolds may be an innovative technology in targeted drug therapies after reconstructive neurosurgical interventions.Öğe Does Nimodipine, a Selective Calcium Channel Blocker, Impair Chondrocyte Proliferation or Damage Extracellular Matrix Structures?(Bentham Science Publ Ltd, 2019) Kaplan, Necati; Yılmaz, İbrahim; Karaarslan, Numan; Kaya, Yasin Emre; Şirin, Duygu Yaşar; 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 alpha) 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 Epidemiological investigation of 673 patients who resorted to the emergency department for mild head trauma complaints(2019) Özger, Özkan; Kaplan, Necati; Karaarslan, NumanAim: Mild head trauma (MHT) or mild traumatic brain injury (MTBI) is an injury whose incidence is increasing in emergency services.This retrospective study carried out an epidemiological evaluation of patients with MHT, who underwent head computed tomography(HCT) with a 15-point score on the Glasgow Coma Scale (GCS).Material and Methods: This study retrospectively evaluated 673 patients with MHT, who were examined by the department ofneurosurgery in the emergency department of Istinye University, Canakkale Anatolian Hospital between 2015 and 2019. The caseswere evaluated because of age, gender, cause of trauma, HCT findings, duration of admission to the emergency department, andother body traumas associated with head trauma.Results:390 (57.95%) patients were male, while 283 (42.05%) were female. The mean age and standard deviation were calculatedas 23.72 ± 24.87 years. Of the 673 cases, 494 (73.40%) were admitted to the emergency department due to non-high falls. Aftertrauma, 642 (95.39%) patients were admitted to the emergency department within the admitted to the emergency department withinthe first two hours after injury. 656 (97.48%) of the patients were treated in the emergency department. 105 (15.60%) patients hada scalp incision and underwent a small surgical procedure. The most common accompanying body trauma detected was that ofthe maxillofacial region in 26 (3.86%) patients. HCT pathology was detected in 20 (2.97%) patients. These pathologies included; 14(2.08%) non-surgical intracerebral hemorrhage, 2 (0.30%) skull base fractures, 1 (0.15%) traumatic subdural hematoma, 1 (0.15%)traumatic epidural hematoma, 1 (0.15%) pneumocephalus and 1 (0.15%) cerebral edema.Conclusion: Head trauma is an important issue in this country. Brain CT may not be necessary in patients with a GCS score of 15.After a short observation, if patients live near the medical center, they can be sent home to return the next day for further evaluation.Öğe Evaluation of the Effect of Daptomycin, a Glycopeptide Agent, on Intact Intervertebral Disc Tissue(Turkish Neurosurgical Soc, 2019) Kaplan, Necati; Yılmaz, İbrahim; Karaarslan, Numan; Yasar Şirin, Duygu; Şimşek, Abdullah Talha; Çalışkan, Tezcan; Özbek, HanefiAIM: To evaluate the effects of pre- and intra-operatively administered daptomycin (DAP) on the intact human primary intervertebral disc tissue cells. MATERIAL and METHODS: Primary cell cultures were established using tissues obtained through decompressive laminectomy, traumatic intervertebral disc herniation excision, and posterior transpedicular stabilization. Non-drug-administered samples were used as a control group. The samples treated with DAP formed the study group. Molecular assays for proliferation and gene expression were performed. The obtained data were evaluated statistically, and results with a value of p<0.05 were accepted as significant. RESULTS: While no reduction was observed in the proliferation, the gene expression of intact intervertebral disc tissue cells was time-dependently decreased compared to the control group, and these results were reported to be statistically significant. CONCLUSION: This study observed the effect that a pharmaceutical preparation, which was used on intervertebral disc tissue before and after the operation, had on normal, healthy, and intact tissue. It concludes that alterations in the expression of genes involved in the anabolic and/or catabolic process, even in adjacent healthy tissue, may slow down the healing process of the damaged tissue or cause undesired cell differentiation.Öğe Investigation of the Effects of Methylphenidate, an Amphetamine Derivative, on Intervertebral Disc Tissue Cell Cultures and Matrix Structures(Turkish Neurosurgical Soc, 2019) Kaya, Yasin Emre; Karaarslan, Numan; Yasar Şirin, Duygu; 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 beta) and matrix metalloproteinase (M MP) -7 and M MP-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 Pregabalin treatment for neuropathic pain may damage intervertebral disc tissue(Spandidos Publ Ltd, 2018) Karaarslan, Numan; Yılmaz, İbrahim; Şirin, Duygu Yaşar; Özbek, Hanefi; Kaplan, Necati; Kaya, Yasin Emre; Ateş, ÖzkanThe aim of the present study was to determine whether pharmaceutical preparations with pregabalin (PGB) as an active ingredient, which are widely prescribed by clinicians, exert toxic effects on human primary nucleus pulposus (NP) and annulus fibrosis (AF). Primary human cell cultures were obtained from intact (n=6) and degenerated (n=6) tissues resected from the two groups of patients. Different doses of PGB were applied to these cultures and cells were subjected to molecular analyses at 0, 24 and 48 h. Cell vitality, toxicity and proliferation were assessed using a spectrophotometer. The expression of chondroadherin (CHAD), a (member of the NP-specific protein family), hypoxia-inducible factor-1 alpha (HIF-1 alpha) and type II collagen (COL2A1) was measured using reverse transcription-quantitative polymerase chain reaction. The results revealed that cell intensity increased in a time-dependent manner and cell vitality continued in the cultures without pharmaceuticals. Cell proliferation was suppressed in the PGB-treated cultures independent from the dose and duration of application. PGB was demonstrated to suppress the expression of CHAD and HIF-1 alpha. In contrast, COL2A1 gene expression was not revealed in any experimental group. The present study utilized an in vitro model and the PGB active ingredient used herein may not be representative of clinical applications; however, the results demonstrated that PGB has a toxic effect on NP/AF cell cultures containing primary human intervertebral disc tissue. In summary, the use of pharmacological agents containing PGB may suppress the proliferation and differentiation of NP/AF cells and/or tissues, which should be considered when deciding on an appropriate treatment regime.Öğe The association between different molecular weights of hyaluronic acid and CHAD, HIF-1 alpha, COL2A1 expression in chondrocyte cultures(Spandidos Publ Ltd, 2018) Şirin, Duygu Yaşar; Kaplan, Necati; Yılmaz, İbrahim; Karaarslan, Numan; Özbek, Hanefi; Akyuva, Yener; Mahiroğulları, MahirThe aim of the present study was to investigate the effects of three different formulations of hyaluronic acid (HA): Low molecular weight (MW) Sinovial One((R)), medium MW Viscoplus((R)) and high MW Durolane((R)), on chondrocyte proliferation and collagen type II (COL2A1), hypoxia-inducible factor 1 alpha (HIF-1 alpha) and chondroadherin (CHAD) expression in primary chondrocyte cultures. Standard primary chondrocyte cultures were established from osteochondral tissues surgically obtained from 6 patients with gonarthrosis. Cell morphology was evaluated using an inverted light microscope; cell proliferation was determined with a MTT assay and confirmed with acridine orange/propidium iodide staining. Levels of CHAD, COL2A1 and HIF-1 alpha expression were assessed using specific TaqMan gene expression assays. The results demonstrated the positive effect of HA treatment on cell proliferation, which was independent from the MW. COL2A1 expression increased in the medium and high MW HA treated groups. It was observed that HIF-1 alpha expression increased in the high MW treated group alone. CHAD expression increased only in the medium MW HA treated group. Evaluation of gene expression revealed that levels of expression increased as the duration of HA application increased, in the medium and high MW HA treated groups. In terms of increased viability and proliferation, a longer duration of HA application was more effective. Taken together, it may be concluded that the administration of medium and high MW HA may be a successful way of treating diseases affecting chondrocytes in a clinical setting.
