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Öğe Preparation and characterization of graphene-based 3D biohybrid hydrogel bioink for peripheral neuroengineering(Jove, 2022) Zorba Yıldız, Aslı Pınar; Darıcı, Hakan; Yavuz, Burçak; Abamor, Emrah Şefik; Özdemir, Ceren; Yasin, Müge Elif; Bağırova, Melahat; Allahverdiyev, Adil; Karaöz, ErdalPeripheral neuropathies can occur as a result of axonal damage, and occasionally due to demyelinating diseases. Peripheral nerve damage is a global problem that occurs in 1.5%-5% of emergency patients and may lead to significant job losses. Today, tissue engineering-based approaches, consisting of scaffolds, appropriate cell lines, and biosignals, have become more applicable with the development of three-dimensional (3D) bioprinting technologies. The combination of various hydrogel biomaterials with stem cells, exosomes, or bio-signaling molecules is frequently studied to overcome the existing problems in peripheral nerve regeneration. Accordingly, the production of injectable systems, such as hydrogels, or implantable conduit structures formed by various bioprinting methods has gained importance in peripheral neuro-engineering. Under normal conditions, stem cells are the regenerative cells of the body, and their number and functions do not decrease with time to protect their populations; these are not specialized cells but can differentiate upon appropriate stimulation in response to injury. The stem cell system is under the influence of its microenvironment, called the stem cell niche. In peripheral nerve injuries, especially in neurotmesis, this microenvironment cannot be fully rescued even after surgically binding severed nerve endings together. The composite biomaterials and combined cellular therapies approach increases the functionality and applicability of materials in terms of various properties such as biodegradability, biocompatibility, and processability. Accordingly, this study aims to demonstrate the preparation and use of graphene-based biohybrid hydrogel patterning and to examine the differentiation efficiency of stem cells into nerve cells, which can be an effective solution in nerve regeneration.Öğe The effects of the heat shock protein 90 inhibitor 17-allylamino-17-demethoxygeldanamycin, cannabinoid agonist WIN 55,212-2, and nitric oxide synthase inhibitor n?-nitro-l-arginine methyl ester hydrochloride on the serotonin and dry skin-induced itch(Karger, 2021) Todurga Seven, Zeynep Gizem; Tombultürk, Fatma Kübra; Gökdemir, Selim; Özyazgan, SibelIntroduction: In many types of itch, the interaction between immune system cells, keratinocytes, and sensory nerves involved in the transmission of itch is quite complex. Especially for patients with chronic itching, current treatments are insufficient, and their quality of life deteriorates significantly. Objective: In this study, we aimed to investigate the role of the heat shock protein 90 (Hsp90) inhibitor 17-allylamino-17-demethoxygeldanamycin (17-AAG), cannabinoid agonist WIN 55,212-2, and nitric oxide (NO) synthase inhibitor N?-nitro-L-arginine methyl ester hydrochloride (L-NAME) in pruritus. Methods: We created a serotonin (5-HT)-induced (50 ?g/?L/mouse, i.d.) acute and acetone-ether-water (AEW)-induced chronic itching models. 17-AAG (1, 3, and 5 mg/kg, intraperitoneally [i.p.]), WIN 55,212-2 (1 mg/kg, i.p.), and L-NAME (1 mg/kg, i.p.) were applied to Balb/c mice. Results: We found that 17-AAG suppressed the scratches of mice, depending on the dose. The itch behavior was reduced by WIN 55,212-2, but L-NAME showed no antipruritic effect at the administered dose. The combined application of these agents in both pruritus models showed synergism in terms of the antipruritic effect. Our results showed that NO did not play a role in the antipruritic effect of WIN 55,212-2 and 17-AAG. Increased plasma IgE levels with AEW treatment decreased with the administration of 17-AAG (5 mg/kg, i.p.) and WIN 55,212-2. Conclusion: These results demonstrate that Hsp90 may play a role in the peripheral pathway of pruritus, and cannabinoid agonists and Hsp90 inhibitors can be used together in the treatment of pruritus.Öğe Topical application of metformin accelerates cutaneous wound healing in streptozotocin-induced diabetic rats(Springer Link, 2021) Tombultürk, Fatma Kübra; Todurga-Seven, Zeynep Gizem; Hüseyinbaş, Önder; Özyazgan, Sibel; Turgut, Ulutin; Kanigur-Sultuybek, GönülBackground: Diabetic chronic wound, which is one of the diabetic complications caused by hyperglycemia, characterized by prolonged inflammation has become one of the most serious challenges in the clinic. Hyperglycemia during diabetes not only causes prolonged inflammation and delayed wound healing but also modulates the activation of nuclear factor-kappa B (NF-?B) and the expression of matrix metalloproteinases (MMPs). Although metformin is the oldest oral antihyperglycemic drug commonly used for treating type 2 diabetes, few studies have explored the molecular mechanism of its topical effect on wound healing. Therefore, we aimed to investigate the molecular effects of topical metformin application on delayed wound healing, which's common in diabetes. Methods and results: In this context, we created a full-thickness excisional wound model in Wistar albino rats and, investigated NF-?B p65 DNA-binding activity and expression levels of RELA (p65), MMP2 and MMP9 in wound samples taken on days 0, 3, 7, and 14 from diabetic/non-diabetic rats treated with metformin and saline. As a result of our study, we showed that topically applied metformin accelerates wound healing by suppressing NF-?B p65 activity and diminishing the expression of MMP2 and MMP9. Conclusions: Diabetic wounds treated with metformin healed even faster than those in the control group that mimicked standard wound healing.Öğe Anesthesia and postoperative complications in sleeve gastrectomy operations performed in morbid obesity surgery(KUWAIT MEDICAL ASSOC, 2021) Gencer, Muzaffer; Sipahi, MesutObjectives: Sleeve gastrectomy is one of the most popular bariatric procedures today. Following laparoscopic interventions, common complications such as bleeding, organ damage, respiratory problems and emboli development are observed. The objective of this study was to evaluate our experiences with anesthesia and postoperative complications in sleeve gastrectomy operations performed in morbid obesity surgery. Design: Prospective study Setting: Operation room Subjects: Sixty adult patients who provided informed consent and who underwent laparoscopic sleeve gastrectomy from November 2015 to January 2019 (N=60). Intervention: Data collection, sleeve gastrectomy operation Main outcome measures: Postoperative complications were evaluated under headings: bleeding, respiratory problems, prolonged mechanical ventilation, prolonged hospital stay, emboli development and mortality. Results: None of the patients had mortality. Mean age was 36.2 (range: 18-59) years. Of the 60 morbid obese patients, 86.6% were female. Mean preoperative body mass index was 48.7 (range: 37-60) kg/m(2). The integrity of the anastomosis was controlled with a 36 French orogastric tube. The mean operating time was 43 minutes (range: 34-72) Two patients (3.3%) were re-intubated due to hypercarbia. Only two patients had bleeding on the postoperative 1st day. Mean length of hospital stay was 4.8 days. Conclusions: We believe there are fewer risks in the laparoscopic sleeve gastrectemy technique if there is careful preoperative patient preparation and evaluation with a multidisciplinary approach, appropriate perioperative anesthesia management, successful coordination with the surgical team and postoperative care.Öğe Ultrasonografi rehberliğinde interskalen brakial pleksus blok uygulamasının solunum fonksiyonları ve arteriyel kan gazı üzerine etkisi(2020) Gencer, MuzafferAmaç: Çalışmamızda ultrason rehberliğinde interskalen brakial pleksus blok uygulamasının solunum fonksiyonları ve arteriyel kan gazı analizi üzerine etkisini araştırmayı amaçladık. Gereç ve Yöntem: Çalışma, omuz bölgesi ve üst kol cerrahisi planlanan, ASA I-II, cerrahi süresi 2 saati geçmeyen, pulmoner disfonksiyonu olmayan, toplam 64 hasta ile yapıldı. Cerrahi sırasında anestezi sağlamak amaçlı USG-rehberliğinde interskalen brakial pleksus blok işlemi öncesi baseline (T0) ve blok sonrası 20. dk’ da (T20) kalp hızı, non-invaziv sistolik, diyastolik ve ortalama arter basıncı ölçülerek kaydedildi. Spirometri ve arteriyel kan gazı analizi yapılarak interskalen brakial pleksus blok uygulamasının solunum fonksiyonlarına ve arteriyel kan gazı üzerine etkisi değerlendirildi. Ayrıca blok uygulaması ile ilgili veriler kaydedildi. Bulgular: Blok sonrası 20.dk’ da kalp hızında daha belirgin olmak üzere (p =0,003), sistolik (p =0,375), diyastolik (p =0,608) ve ortalama arteriyel basınçta (p =0,496) baseline (T0) değerlere göre düşme gözlendi. Blok işlemi sonrası 20.dk’ da yapılan ikinci (T20) spirometrik ölçümde, baseline (T0) ile karşılaştırıldığında, vital kapasite (VC) (4,86-3,68, p <0,001), zorlu vital kapasite (FVC), zorlu ekspiratuar volüm 1.sn (FEV1) (4,24-3,40, p <0,018), Sniff PmaxPeak (6,18-5,06) (p <0,001) ve ort. PaO2 (86,40-79,24, p <0,024)’ de önemli oranda azalma gözlendi. Bununla birlikte ortalama PaCO2 (38,62-42,34) (p <0,001) artış anlamlı düzeyde değildi. Blok sonrası, 12 hastada Horner sendrom bulguları, 6 hastada hipotansiyon-bradikardi gözlendi. Dört hastada ise ses kısıklığı gelişti. Sonuç: İnterskalen brakial pleksus bloğu tek taraflı hemidiyafragmatik pareziye neden olarak solunum fonksiyonlarını ve arteriyel oksijen basıncını etkiler ve bu sebeple pulmoner hastalığı veya kardiyak patolojisi olan hastalar için risklidir.Öğe Kalsitriol ve doksorubisin kombinasyonunun MCF7 üzerine olası anti kanser etkilerinin araştırılması(DergiPark, 2018) Bildiren, Özge; Cevatemre, Buse; Ay, Ebru Nur; Özen, Güneş; Hepokur, Ceylan; Erkısa Genel, Merve; Ulukaya, EnginAmaç: Bu çalışmada amacımız, belirli bir dozun üzerinde kardiyak yan etkileri oldukça fazla olan ve meme kanseri tedavisinde kullanılan kemoterapötiklerden doksorubisinin, tedavide kullanım miktarlarını azaltmayı ve etkinliğini arttırmayı sağlayan bir maddeyi saptamaktı. Etken madde olarak anti proliferatif etkisi olduğu düşünülen vitamin D analoglarından biri olan kalsitriol seçilmiş ve doksorubisin ile kombine tedavisinin, insan meme kanseri hücre hattı MCF-7 üzerine sitotoksik etkisinin saptanması amaçlanmıştır. Gereç ve Yöntem: MCF-7 insan meme kanseri hücre hattı kalsitriol ile muamele edilerek, gerçek zamanlı olarak, x-CELLigence cihazında 72 saat inkübasyona bırakıldı ve kalsitriolün anti-proliferatif optimum doz tespiti zamana bağlı hücre indeksi grafiği The xCELLigence Real-Time Cell Analysis (RTCA) software programı kullanılarak yapıldı. Kalsitriol optimum dozu ve doksorubisinin farklı dozlarının kombinasyonu MCF-7 hücre kültürü ile muamele edilerek sitotoksisite tayini için Sulforhodamine-B (SRB) uygulaması ve spektrofotometrik ölçüm uygulandı. Spektrofotometrik ölçüm sonuçları anlamlılık sonuçları için Student’s t-Testi ile değerlendirildi. Bulgular: MCF-7 hücrelerinin antiproliferative optimum kalsitriol doz tespiti zamana bağlı hücre indeksi grafiği RTCA software programı kullanılarak yapılmıştır. Sitotoksisite tayini için uygulanan SRB yöntemi sonucu elde edilen spektrofotometrik ölçüm sonuçları GraphPad Prism programı kullanılarak Student’s t-testi ile istatistiksel olarak değerlendirilmiştir. Kalsitriol optimum dozu 250 nM tespit edilmiştir. Doksorubisinin farklı dozları (1,84 µM-0,92 µM), kalsitriol (250 nM) ve kalsitriol olmaksızın MCF-7 hücre hattı ile sitotoksik etki saptaması için muamele edilmiştir. 0,46 µM doksorubisin ve optimum kalsitriol kombinasyonunun sitotoksik açıdan diğer dozlara göre anlamlı olduğu (p=0,0087) fakat doksorubisin kullanımında doz azaltımını sağlayacak kadar etkin olmadığı saptanmıştır. Sonuç: Doksorubisinin, kalsitriol ile kombine kullanımının günümüzde kullanılan dozlar üzerinde azaltıcı yönde anlamlı bir etkisinin olmadığını saptanmıştır. Vitamin D ve doksorubisin birlikte kullanımının meme kanserinde fayda sağlamayacağını söylemek için ise henüz erkendir. İlerleyen çalışmalarda vitamin D’nin farklı analogları ile çeşitli çalışmalar yapılabilir.Öğe NFKB1 rs28362491 and pre-miRNA-146a rs2910164 SNPs on E-Cadherin expression in case of idiopathic oligospermia: a case-control study(Shahid Sadoughi Univ Medical Sciences, 2018) Tunçdemir, Matem; Yenmiş, Güven; Tombultürk, Fatma Kübra; Arkan, Hülya; Soydaş, Tuğba; Tek, Raşit Burak; Altıntaş, Özlem; Özkara, Hamdi; Kanıgür-Sultuybek, GönülBackground: A notable proportion of idiopathic male infertility cases is accompanied by oligozoospermia; and yet, the molecular mechanisms of fertilization problem underlying this defect are still unclear. Epithelial cadherin has been involved in several calcium-dependent cell-to-cell adhesion events; however, its participation in gamete interaction has also not been fully investigated. Objective: The aim was to investigate the changes in the expression of E-cadherin, based on the frequency of Single nucleotide polymorphisms in Nuclear Factor Kappa-B 1 and pre-mir-146a in oligospermic men. Materials and Methods: In this case-control study, semen and blood samples of 131 oligospermic men as the case group and 239 fertile healthy men as the control group were analyzed. Variants single nucleotide polymorphisms rs28362491 and rs2910164 were performed using polymerase chain reaction-restriction fragment length polymorphism method and E-cadherin expression were determined by immunoprecipitation studies. Results: ins/ins genotype of rs28362491 was determined as a risk factor for idiopathic oligospermia by 1.73 times (p=0.0218), whereas no significant differences were found between the groups concerning pre-mir-146a rs2910164 polymorphism (p=0.2274 in case of GC genotype and p=0.9052 in case of GG genotype). Combined genotype analysis results did not show any notable differences between the multiple comparisons of 28362491-rs2910164 in oligospermic men and control groups. In addition, E-cadherin expression of oligospermic men with ins/ins genotype was significantly lower than patients with del/ins genotype (p=0.0221). E-cadherin expression level was low in oligospermic men with respect to the control group in presence of ins/ins genotype of NFKB1 gene. Conclusion: These results suggest that ins allele prevents binding of surface proteins to spermatozoa, leading to a low affinity of sperm-oocyte interaction in oligospermic men.Öğe Effects of Lucilia sericata on wound healing in streptozotocin-induced diabetic rats and analysis of its secretome at the proteome level(Sage Publications Ltd, 2018) Tombultürk, Fatma Kübra; Kasap, Murat; Tunçdemir, Matem; Polat, Erdal; Sirekbasan, Serhat; Kanlı, Ali İsmet; Kanıgür-Sultuybek, GönülThe use of Lucilia sericata larvae on the healing of wounds in diabetics has been reported. However, the role of the excretion/secretion (ES) products of the larvae in treatment of diabetic wounds remains unknown. This study investigated whether application of the ES products of L. sericata on the wound surface could improve the impaired wound healing in streptozotocin-induced diabetic rats. Additional analysis was performed to understand proteome content of L. sericata secretome to understand ES contribution at the molecular level. For this purpose, full-thickness skin wounds were created on the backs of diabetic and control rats. A study was conducted to assess the levels of the ES-induced collagen I/III expression and to assay nuclear factor B (NF-B) (p65) activity in wound biopsies and ES-treated wounds of diabetic rat skin in comparison to the controls. The expression levels of collagen I/III and NF-B (p65) activity were determined at days 3, 7, and 14 after wounding using immunohistological analyses and enzyme-linked immunosorbent assay technique. The results indicated that treatment with the ES extract increased collagen I expressions of the wound control and diabetic tissue. But the increase in collagen I expression in the controls was higher than the one in the diabetics. NF-B (p65) activity was also increased in diabetic wounds compared to the controls, whereas it was decreased in third and seventh days upon ES treatment. The results indicated that ES products of L. sericata may enhance the process of wound healing by influencing phases such as inflammation, NF-B (p65) activity, collagen synthesis, and wound contraction. These findings may provide new insights into understanding of therapeutic potential of ES in wound healing in diabetics.Öğe Regulation of MMP 2 and MMP 9 expressions modulated by AP-1 (c-jun) in wound healing: improving role of Lucilia sericata in diabetic rats(Springer-Verlag Italia Srl, 2019) Tombultürk, Fatma Kübra; Soydaş, Tuğba; Saraç, Elif Yaprak; Tunçdemir, Matem; Coşkunpınar, Ender; Polat, Erdal; Sirekbasan, Serhat; Kanigur-Sultuybek, GönülAimsLucilia sericata larvae have been successfully used on healing of wounds in the diabetics. However, the involvement of the extraction/secretion (ES) products of larvae in the treatment of diabetic wounds is still unknown. Activator protein-1 (AP-1) transcription, composed of c-jun and c-Fos proteins, has been shown to be the principal regulator of multiple MMP transcriptions under a variety of conditions, also in diabetic wounds. Specifically, MMP-2 and MMP-9's transcriptions are known to be modulated by AP-1. c-jun has been demonstrated to be a repressor of p53 in immortalized fibroblasts. The aim of the present study is to investigate the effects of L. sericata ES on the expression of AP-1 (c-jun), p53, MMP-2, and MMP-9 in wound biopsies dissected from streptozotocin induced diabetic rats.MethodsThe expression levels of MMP-2, MMP-9, c-jun and p53 in dermal tissues were determined at days 0, 3, 7 and 14 after wounding, using immunohistochemical analysis and quantitative real-time PCR.ResultsThe treatment with ES significantly decreased through inflammation-based induction of MMP-2 and MMP-9 expression levels in the wounds of diabetic groups, compared to control groups at the third day of wound healing. At the 14th day, there were dramatic decreases in expression of c-jun, MMP-9, and p53 in ES-treated groups, compared to the diabetic group (P<0.001, P<0.05 and P<0.01, respectively).ConclusionES products of L. sericata may enhance the process of wound healing in phases of inflammation, proliferation, and re-epithelization, essentially via regulating c-jun expression and modulating MMP-2 and MMP-9 expressions.Öğe Determination of antibiotic impurities in good manufacturing practices-grade cell therapy medicinal products(Taylor & Francis Inc, 2020) Öztel, Olga Nehir; Korkmaz, Seval; Karaöz, ErdalBackrounds: According to the regulations of the health autorities, cell-based therapy products must be manufactured in good manufacturing production (GMP) facilities, fulfilling the required GMP standards. Products developed under the high quality control (QC) necessarity need to be approved for some QC tests. One of the main residual test is antibiotic test and this test should be validated. The aim of this study is to validate and determine the methods of detection of the antibiotic residue in the final product. Methods: Liquid Chromatography Tandem-Mass Spectrometry (LC-MS/MS) methods were used for the main steps of the production procedure, as well as the final products. Pharmaceutical Grade penicillin G and streptomycin sulfate were used as positive controls. Results: The results suggest that penicillin is broken down during cell culture and streptomycin is eliminated at the first washing step of the final product manufacture. It is shown in this study that LC-MS/MS method is one of the convenient method to test residual anibiotics and can be used to detect the antibiotic residues in cellular therapy products. Discussion: Since the antibiotic residues are eliminated in the final product and also it could be suggested that the methodology we followed is sufficiently safe and final product is pure.Öğe Silver nanoparticle/capecitabine for breast cancer cell treatment(Pergamon-Elsevier Science Ltd, 2019) Hepokur, Ceylan; Kariper, İshak Afşin; Mısır, Sema; Ay, Ebru Nur; Tunoğlu, Servet; Ersez, Mediha Süleymanoğlu; Zeybek, Ümit; Kuruca, Serap Erdem; Yaylım, İlhanThis study aimed to evaluate antiproliferative and proapoptotic effects of Capecitabine bonded silver particles on human breast cancer cells (MCF-7). Different sizes of Ag NPs (in sizes 5, 10, 15, 30 nm) were synthesized. The characterization of silver and drug-bonded silver nanoparticles was performed through UV-VIS, FTIR, and SEM analysis. Silver and drug-bonded silver nanoparticles were measured by zetasizer. Antiproliferative and proapoptotic effects of capecitabine, silver and drug-bonded silver nanoparticles were evaluated using XTT, Anneksin V, respectively. According to the results, silver nanoparticles of 10 nm size have shown the lowest toxic effect. Drug-bonded nanoparticles significantly increased the number of early and late apoptotic cells on MCF-7 cells.
