Farklı rezin bazlı yapıştırma simanlarının zamana bağlı sitotoksisitelerinin xCELLigence yöntemi ile karşılaştırmalı olarak incelenmesi

Abstract

Kompozit ve seramik esaslı estetik restorasyonların yapıştırılmasında kullanılan kompozit rezin esaslı yapıştırma simanlarının klinik kullanımı son yıllarda giderek artmaktadır. Rezin simanların fiziksel ve mekanik özellikleri kadar biyouyumluluğunun değerlendirilmesi de çok önemlidir.Bu çalışmanın amacı, kendinden adeziv rezin simanların, sitotoksik etkisinin, L929 hücrelerinin canlılığı üzerinde, gerçek zamanlı hücre analizi ile değerlendirilmesidir.Altı farklı kendinden adeziv rezin simanlarının sitotoksisiteleri (BifixSE; G-Cem; NX3; Maxcem elite; RelyX U200 ve Smart Cem 2 ) gerçek zamanlı hücre analizi (RT-CA, xCELLigence; Roche Applied Science, Germany) ile test edildi. Her rezin siman grubu için 24 örnek (5mm çapında ve 2mm yüksekliğinde) hazırlandı. Ekstraksiyon solüsyonu olarak yapay tükürük solüsyonu tercih edildi. Örnekler 1, 4 ve 7 gün yapay tükürük solüsyonunda bekletildi. 100 µL L-929 fibroblast hücreleri 48 kuyucuklu platelere ekildikten sonra xCELLigence sistemi ile hücrelerin çoğalması monitörize edildi. Hücreler büyüme (log) fazına geldikten sonra rezin simanlardan salınan kompanentleri içeren ekstraksiyon solüsyonundan (1/1 ve 1/2 dilüsyonlarda) 10 µL eklendi ve 1'er saat aralıklarla monitörize edildi.Gruplar arasındaki fark varyans analizini (ANOVA testi) takiben, post hoc TUKEY testi ile araştırıldı.Sonuç olarak bu çalışmada test edilen tüm rezin simanlar istatistiksel olarak anlamlı derecede toksik etki gösterdi. Fakat sadece Bifix SE' nin 1.gün uygulaması ve GCem' in 1. gün ½ lik konsantrasyonu toksik etki göstermedi. Tüm grupların 7 günlük ekstraktları 1. ve 4. gün ekstraktlarına göre daha fazla toksik etki gösterdi. En yüksek sitotoksik etki Maxcem Elite'in 7. gün ekstraktında gözlendi. Ayrıca Maxcem Elite'in 7. gün uygulamasında hiç hücre canlılığı gözlenmedi.Çalışmamız, xCELLigence yönteminin rezin simanların sitotoksisitesini belirlemede kolay ve hızlı bir yöntem olduğunu gösterdi. Biyolojik risklerin daha iyi anlanması ve biyouyumluluğun geliştirilmesi için, rezin simanların sitotoksisitesi ile ilgili daha fazla çalışma yapılması gerekmektedir.Anahtar kelimeler: Kendinden adeziv rezin siman, sitotoksisite, hücre kültürü, xCELLigence

In recent years, resin-based adhesive cements are widely used in cementation composite and ceramic based esthetic restorations in recent years. Evaluation of biocompatibility is as important as the physical and mechanical properties of resin cements. The aim of this study is to evaluate the cytotoxic effects of six different self adhesive resin cements on the cell viability of L929 fibroblast cells with real-time cell analyzer.The cytotoxicity of self adhesive resin cements including; (BifixSE; G-Cem; NX3; Maxcem elite; RelyX U200 and Smart Cem 2 ) were analyzed with real-time cell analyzer (RT-CA, xCELLigence; Roche Applied Science, Germany). For each resin cement group, 24 cylinders (5 mm diameter and 2 mm height) were fabricated. Artificial saliva was used as an extracting solution. All specimens were stored in the artificial saliva for 1, 4, and 7 days. After seeding 100 µL of the L-929 fibroblast cell suspensions into the wells (20,000 cells/well) of the E-plate 48, the proliferation of the cells was monitored by the xCELLigence system. When the cells were in the log growth phase, the cells were exposed to 10 µL of extracting solution containing the components released by the resin cements (1/1 and 1/2 dilutions) and monitored every 1 hour.The statistical analyzes among the groups were evaluated using ANOVA and followed by post hoc Tukey test.As a conclucision, in this study all tested resin cements showed statistically significant cytotoxic effect (p<0.05). However, Bifix SE group and 1/2 dilutions of GCem group presented no cytotoxic effect at 1st day (p>0.05). Extracts of all groups at 7th day indicated higher cytotoxic effect than the extracts of those at 1st and 4th day. The most intense cytotoxic effects were caused by Maxcem Elite at 7th day. Furthermore, no cell viability was dedicated in the well of Max cem Elite at 7th day.The data suggests that the xCELLigence live cell analysis system can be used as a rapid and easy assessment method to determine the cytotoxicity of resin cements in cell-based in vitro assays. Further studies about the cytotoxicity of resin cements would be helpful to better understand the biological risks and to improve the biocompatibility of the resin cements.Key words: Self adhesive resin cement, cytotoxicity, cell culture, xCELLigenceEVALUATION of THE TIME DEPENDENCE CYTOTOXICITY OF DIFFERENT RESIN BASED ADHESIVE CEMENTS BY XCELLIGENCE METHODSedef TOPÇUOĞLUDoctorate Thesis, Department of ProsthodonticsSupervisor: Doç. Dr. N. Tülin POLAT2012, 88 pagesIn recent years, resin-based adhesive cements are widely used in cementation composite and ceramic based esthetic restorations in recent years. Evaluation of biocompatibility is as important as the physical and mechanical properties of resin cements. The aim of this study is to evaluate the cytotoxic effects of six different self adhesive resin cements on the cell viability of L929 fibroblast cells with real-time cell analyzer.The cytotoxicity of self adhesive resin cements including; (BifixSE; G-Cem; NX3; Maxcem elite; RelyX U200 and Smart Cem 2 ) were analyzed with real-time cell analyzer (RT-CA, xCELLigence; Roche Applied Science, Germany). For each resin cement group, 24 cylinders (5 mm diameter and 2 mm height) were fabricated. Artificial saliva was used as an extracting solution. All specimens were stored in the artificial saliva for 1, 4, and 7 days. After seeding 100 µL of the L-929 fibroblast cell suspensions into the wells (20,000 cells/well) of the E-plate 48, the proliferation of the cells was monitored by the xCELLigence system. When the cells were in the log growth phase, the cells were exposed to 10 µL of extracting solution containing the components released by the resin cements (1/1 and 1/2 dilutions) and monitored every 1 hour.The statistical analyzes among the groups were evaluated using ANOVA and followed by post hoc Tukey test.As a conclucision, in this study all tested resin cements showed statistically significant cytotoxic effect (p<0.05). However, Bifix SE group and 1/2 dilutions of GCem group presented no cytotoxic effect at 1st day (p>0.05). Extracts of all groups at 7th day indicated higher cytotoxic effect than the extracts of those at 1st and 4th day. The most intense cytotoxic effects were caused by Maxcem Elite at 7th day. Furthermore, no cell viability was dedicated in the well of Max cem Elite at 7th day.The data suggests that the xCELLigence live cell analysis system can be used as a rapid and easy assessment method to determine the cytotoxicity of resin cements in cell-based in vitro assays. Further studies about the cytotoxicity of resin cements would be helpful to better understand the biological risks and to improve the biocompatibility of the resin cements.Key words: Self adhesive resin cement, cytotoxicity, cell culture, xCELLigence