Handan Sevim

May toxicity of amiodarone be prevented by antioxidants? A cell-culture study

BackgroundAtrial Fibrillation is the most common arrhythmia encountered following cardiac surgery. The most commonly administered drug used in treatment and prophylaxis is amiodarone which has several toxic effects on major organ functions. There are few clinical data concerning prevention of toxic effects and there is no routinely suggested agent. The aim of this study is to document the cytotoxic effects of amiodarone on cell culture media and compare the cytoprotective effects of commonly used antioxidant agents.MethodsL929 mouse fibroblast cell line was cultured and 100,000 cells/well-plate were obtained. First group of cells were treated with increasing concentrations of amiodarone (20 to 180 μM) alone. Second and third group of cells were incubated with one-fold equimolar dose of vitamin C and N-acetyl cysteine prior to amiodarone exposure. The viability of cells were measured by MTT assay and the cytoprotective effect of each agent was compared.ResultsThe cytotoxicity of amiodarone was significant with concentrations of 100 μM and more. The viabilities of both vitamin C and N-acetyl cysteine treated cells were higher compared to untreated cells.ConclusionsVitamin C and N-acetyl cysteine are commonly used in the clinical setting for different purposes in context of their known antioxidant actions. Their role in prevention of amiodarone induced cytotoxicity is not fully documented. The study fully demonstrates the cytoprotective role of both agents in amiodarone induced cytotoxicity on cell culture media; more pronounced with vitamin C in some concentrations. The findings may be projectile for further clinical studies.

Bone marrow derived mesenchymal stem cells ameliorate inflammatory response in an in vitro model of familial hemophagocytic lymphohistiocytosis 2

BackgroundFamilial hemophagocytic lymphohistiocytosis 2 (FHL2) is the most common familial type of hemophagocytic lymphohistiocytosis with immune dysregulation. FHL2 patients have mutations in the perforin gene which cause overactivation and proliferation of cytotoxic T lymphocytes and natural killer cells. Perforin is the key component of the cytolytic granule response function of cytotoxic T lymphocytes and natural killer cells. Perforin dysfunction causes a cytotoxic immune deficiency with a clinical outcome of uncontrolled and continuous immune stimulation response. This excessive stimulation leads to continuous systemic inflammation and, ultimately, multiorgan failure. Radical therapy is hematopoietic stem cell transplantation which is limited by the availability of a donor. Exacerbations of inflammatory attacks require a palliative immunosuppressive regimen. There is a need for an alternative or adjuvant therapy to maintain these patients when immunosuppression is ineffective or a donor is not available. Beneficial actions of mesenchymal stem cells (MSCs) have been shown in autoimmune diseases in clinical trials and are attributed to their immune-modulatory properties. This study aimed to assess the immune-modulatory effect of MSCs in an in-vitro model of FHL2.MethodsWe generated a targeted mutation in the perforin gene of NK92 cells to create an in-vitro FLH2 model using Crispr/Cas technology. A coculture setup was employed to assess the immunomodulatory efficacy of MSCs.ResultsEngineered NK92 clones did not show PRF1 mRNA expression and failed to secrete perforin upon phorbol myristate acetate–ionomycin stimulation, providing evidence for a valid FHL2 model. Coculture media of the engineered cells were investigated for the abundance of several cytokines. Coculture with MSCs revealed a reduction in major proinflammatory cytokines and an induction in anti-inflammatory and immunomodulatory cytokines compared to the parental NK92 cells.ConclusionsThis study shows the ameliorating effect of MSCs as an adjuvant immune modulator toward the therapy of FHL2 patients. MSCs are supportive therapy candidates for FHL2 patients under circumstances where prolonged immunosuppression is required to gain time before allogeneic hematopoietic stem cell transplantation.

In Vitro Cytotoxicity of Maxillofacial Silicone Elastomers: Effect of Nano-particles

PURPOSE Silicone elastomers are generally used for maxillofacial extraoral prostheses. The purpose of this in vitro study was to evaluate the cytotoxicity of different kinds of nanoparticles added to two types of maxillofacial elastomers. MATERIALS AND METHODS A-2000 and A-2006 silicone elastomers were used. The silicone specimens were divided into eight groups according to the presence of additional nanoparticles (n = 18). The following represents the groups in the study: Group A: A-2000 silicone (control group); Group B: A-2006 silicone (control group); Group C: A-2000 silicone and the addition of titanium dioxide (TiO2 ); Group D: A-2006 silicone and the addition of TiO2 ; Group E: A-2000 silicone and the addition of fumed silica; Group F: A-2006 silicone and the addition of fumed silica; Group G: A-2000 silicone and the addition of silaned silica; Group H: A-2006 silicone and the addition of silaned silica. A paired sample t-test was used to analyze the cytotoxicity of each group after 24, 48, and 72 hours. RESULTS Based on the results of the 24-hour analysis, the biocompatibility values of the (A-2006) fumed silica group were higher than those of the control groups. There was no statistically significant difference in A-2006 and A-2000 groups. The cytotoxicity values of the control groups and TiO2 (A-2000 silicone) elastomer groups increased at all test times; however, the cytotoxicity values of the TiO2 (A-2006), fumed silica (A-2006), silaned silica (A-2006), fumed silica (A-2000), and silaned silica (A-2000) groups increased significantly only from 24 to 48 hours. CONCLUSION Nanoparticles of TiO2 , fumed silica, and silaned silica added to a commercial silicone-based elastomer used for fabrication of maxillofacial prostheses are nontoxic.

Cytotoxicity of long-term denture base materials

Introduction: The aim of this study was to evaluate the cytotoxic effect of various denture base materials following four different aging periods. Methods: In total, 48 disc-shaped specimens per each group were prepared: Group I: acrylic resin polymerized in cool water and heated up to 100°C over 45 min and boiled for 15 min; Group II: acrylic resin polymerized under pressure in 40°C–45°C water bath for 10 min; Group III: autopolymerized hard relining resin Cold Liner Rebase; Group IV: autopolymerized hard relining resin Truliner; Group V: soft relining resin DentuSil. Then the specimens were stored in water for 24 h or 15 days, or thermocycled 2500 times or 10,000 times. Cytotoxicity was evaluated with 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay using L929 cells after 72-h cell incubation. Cell viability percentages were counted and statistical analyses were performed. The results were also evaluated according to ISO standard 10993-5. Results: All materials showed similar cell viability percentages following 24-h water storage and 2500 and 10,000 thermal cycles. Following 15-day water storage, a statistically significant difference was observed between the materials. Comparisons of the aging periods for each material showed statistically significant differences. Groups III and IV showed moderately cytotoxic effect following 15-day water storage. The remaining groups showed slightly cytotoxic or non-cytotoxic effect. Discussion: Polymerizing acrylic resins under pressure can be an alternative to conventional polymerizing to ensure a faster denture repair while providing similar cell viability values. Heat-cured acrylic resins provide higher cell viability than hard chairside lining materials in a 15-day period.

The effects of mesenchymal stem cells on the structure and contractile force of the carotid artery in a rat aneurysm model

Background/aim: An aneurysm is a pathological enlargement of an artery characterized by the thinning of the elastic fiber layer in the tunica media. Because the aneuritic artery wall is weakened, these vessels can rupture and cause serious bleeding. Surgical methods are often used for the treatment of aneurysms. However, cell-based therapies are less invasive and potentially safer alternatives. In this study, the therapeutic efficacy of rat adipose tissue-derived mesenchymal stem cells (MSCs) was investigated in a new carotid artery aneurysm model.Materials and methods: Arteries were pretreated with elastase to create aneurysms. Gelatin matrices containing MSCs were applied to the outer surface of the elastase-treated carotid artery sections.Results: Healing of the aneuritic arteries for which MSC applications were performed was significantly better than in the aneuritic group. The histological structure of the vessels was largely reconstituted, and the contractile force of the MSC-treated group was similar to the untreated healthy group.Conclusions: Application of MSCs facilitates the healing of aneurysms. Hereby, MSC application could be a promising approach for clinical applications after further validation processes are concluded.

Ozonated Olive Oil with a High Peroxide Value for Topical Applications: In-Vitro Cytotoxicity Analysis with L929 Cells

ABSTRACT Previous studies have shown that ozonated vegetable oils have been used topically for healing of cutenous wounds. The aim of this study is to evaluate the dose dependent use of ozonated olive oil with high peroxide value (OZ) on the viability of cells for preventing side effects in topical applications. To the best of our knowledge, there are no reports investigaing the effect of peroxide value of ozonated olive oil associated with its cytotoxic activity on mouse non-neoplastc fibroblast cell lines (L929). Therefore, the present study was carried out by using OZ alone and/or in combination with glycerol and olive oil. In our study OZ was prepared by using pure olive oil. Both olive oil and glycerol are non-toxic and can be mixed with OZ uniformly. The cytotoxic activity of samples against L929 fibroblasts was assessed using the tetrazolium salt 3-[4,5-dimethylthiazol-2-yl]-2,5 diphenyltetrazolium bromide (MTT) assay. The peroxide value of synthesized OZ was found to be in the range of 2700–2900 mEq O2/kg oil. The OZ/olive oil group did not show any cell death at all concentrations tested (p > 0.05) however OZ/glycerol group showed statistically significant reductions in viability at higher concentrations (p = 0.004–0.006) compared to the control group. Conclusively, using OZ/olive oil with a peroxide value of 2700–2900 mEq O2/kg oil for short-term incubation was non-cytotoxic to the L929 fibroblast cell line.

Effect of artificial saliva with different pH levels on the cytotoxicity of soft denture lining materials

Background The aim of this study was to evaluate the cytotoxic effects of 9 different soft denture liners on the viability of L-929 mouse fibroblast cells at different incubation periods by storing them in artificial saliva (AS) with different pH levels. Methods 96 disk samples from each lining material were prepared and divided into 4 groups: GI: No treatment; GII: Stored in artificial saliva with pH 3 for 21 days; Group III: Stored in artificial saliva with pH 7 for 21 days; and Group IV: Stored in artificial saliva with pH 14 for 21 days. The cytotoxicity of the extracts to cultured mouse fibroblasts (L-929) was measured by MTT (tetrazolium salt 3-[4,5-dimethylthiazol-2-yl]-2,5-dipHnyltetrazolium bromide) assay. Data were analyzed using 1-way analysis of variation (ANOVA). Results It was found that for the pH 3 values of New Truliner, Trusoft, Mollosil Plus, Dentusil, TDV, and HydroCast®; for the pH 7 values of Ufi Gel P and Elite plus; and for the pH 14 values of HydroCast®, there was a noncytotoxic effect during both the 24-hour and 48-hour incubation periods. In the control group 48-hour incubation period, HydroCast®, TDV, Mollosil, 24-hour incubation period Elite plus, for pH 3 values; Elite Plus 24-hour incubation period, for pH 7 values Trusoft 48-hour incubation period there was a moderately cytotoxic effect. Conclusions This in vitro study revealed that storage in artificial saliva with different pH levels can affect the cytotoxicity of soft lining materials.

Bone Marrow Mesenchymal Stem Cells to Improve Damaged Heart Function after Modified Myocardial Infarction Rat Model

Akut miyokard infarktüsü ve kalp yetmezliği; tedavideki gelişmelere rağmen günümüzde kardiyovasküler mortalite ve morbiditenin en önemli nedenlerindendir. Bu çalışmada, modifiye edilmiş iskemik kalp modeli oluşturulan Wistar albino sıçanlarda, kemik iliğinden elde edilen mezenşimal kök hücreler ile rejenere olan miyokardiyal alanlar, elektrokardiyografik (EKG) ve histopatolojik analizler yapılarak gözlenmiştir. Kök hücre uygulama grubunda damar ve hücre sayısında diğer gruplara göre zamana bağlı olarak artış gözlenmiştir. Ayrıca bu grupta EKG verilerinden P dalgası, QRS kompleksi, T dalgası amplitüdleri diğer gruplara göre farklılık göstermezken, T dalgasının süresi ve kalp atım sayısında önemli derecede azalma gözlenmiştir (p≤0,001). Referans grubuna göre kontrol grubunda S-T çökmesinde artış gözlenirken, kök hücre uygulama grubunda azalma gözlenmiştir.