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Bu çalışma, hiperfosfatemili hastaların tedavisinde sıklıkla reçete edilen ve fosfat bağlayıcı grubunda yer alan sevelamer hidroklorür ve kalsiyum karbonat ajanlarının genotoksik ve/veya sitotoksik potansiyelini karşılaştırmalı olarak ortaya çıkarmak amacıyla yürütülmüştür. Çalışmalar in vitro koşullarda insan periferal lenfositlerle mikronukleus (MN) ve tek hücre jel elektroforez (Komet) testleri yapılmıştır. Ayrıca nokta mutasyonu ve çerçeve kayması mutasyonlarını uyarıp uyarmadığını belirlemek amacıyla Salmonella typhimurium'un mutant suşlarıyla (TA98 ve TA100) geri mutasyon testi (Ames testi) gerçekleştirilmiştir. İnsan periferal lenfositleri 20 ve 44 saat, komet testinde ise 1 saat boyunca 1/2, 1, ve 2 mg/mL konsantrasyonlardaki sevelamer hidroklorür ve kalsiyum karbonat ile muamele edilmiştir. Ames/Salmonella bakteriyel ters mutasyon testinde ise sevelamer hidroklorür ve kalsiyum karbonatın, 2 mg/mL, 1 mg/mL, 0,5 mg/mL, 0,25 mg/mL, 0,125 mg/mL konsantrasyonları kullanılmıştır.Bu çalışmadaki test maddeleri sevelamer hidroklorür ve kalsiyum karbonatın mikronukleus oluşumu hafif artış yönünde uyardığı ancak bu artışın kontrole göre anlamlı olmadığı saptanmıştır. Komet testinin bulgularına göre kullanılan test maddeleri bir istisna (2 mg/mL, sevelamer hidroklorür, 48 saat) hariç önemli DNA hasarlarına neden olmamıştır. Ames testinde ise mutajenik olmadığı ancak zayıf bir sitotoksik etkiye sahip olduğu bulunmuştur. Her iki test maddesi, özellikle de sevelamer hidroklorür, NBI'yi kontrole göre anlamlı şekilde azaltmıştır. Yani sevelamer hidroklorürün zayıf bir sitotoksisitesinden söz edilebilir. Ayrıca iki test maddesi birbirlerine karşılaştırıldığında sevelamer hidroklorürün genotoksisite ve sitotoksisite açısından kalsiyum karbonattan daha reaktif olduğu gözlenmiştir.Anahtar Kelimeler: Sevelamer hidroklorür, Kalsiyum Karbonat, Mikronukleus, Tek Hücre Jel Elektroforezi, AMES/Salmonella Bakteriyel geri mutasyon testi. This study was conducted to compare the genotoxic and / or cytotoxic potential of two agents included in the phosphate binding group, Sevelamer hydrochloride and Calcium carbonate, which are frequently prescribed in the treatment of patients with hyperphosphatemia. Micronucleus (MN) and single cell gel electrophoresis (comet assay) tests were performed on human peripheral lymphocytes. In addition, the Ames Salmonella/microsome mutagenicity assay was performed with mutant strains of Salmonella typhimurium (TA98 and TA100) to determine whether they stimulate substitutions and frame shift mutations. Human peripheral lymphocytes were treated with sevelamer hydrochloride and calcium carbonate at concentrations of 1/2, 1, and 2 mg / mL for 20 and 44 hours for micronucleus assay and 1 hour in the comet assay. In the Ames / Salmonella bacterial reverse mutation test, sevelamer hydrochloride and calcium carbonate concentrations of 2 mg / mL, 1 mg / mL, 0.5 mg / mL, 0.25 mg / mL and 0.125 mg / mL were used.In this study, sevelamer hydrochloride and calcium carbonate stimulated slight increase in micronucleus formation but this increase was not significant compared to the results of the control group. According to the findings of the comet test, sevelamer hydrochloride and calcium carbonate did not cause significant DNA damage with one exception (2 mg / mL, sevelamer hydrochloride, 48 hours). In the Ames test, it was found that the drugs were devoid of mutagenic activity in bacterial assay, but had a weak cytotoxic effect. Both sevelamer hydrochloride and calcium carbonate, particularly SH, significantly reduced NBI compared to the control group. Thus, a weak cytotoxicity effect can be mentioned. Furthermore, sevelamer hydrochloride was more reactive in terms of genotoxicity and cytotoxicity than calcium carbonate when the two agents were compared.Key Words: Sevelamer Hydrochloride, Calcium Carbonate, Micronucleus, Single Cell Gel Electrophoresis, Ames Salmonella/microsome mutagenicity assay. 120

“Single cell gel electrophoresis (SCGE)”, also called “Comet Assay”, is a sensitive, reliable and rapid technique for quantifying and analyzing DNA damage in individual cells. The comet assay is widely used in living cells, researches and the applications on comet assay is becoming broader day by day. To date, the comet assay has been used for a variety of applications, including genotoxic and cytotoxic agent analyses, environmental toxicology, cancer research, and radiation biology. Briefly, in comet assay, fully frosted microscope slides were first covered with 0.5% normal melting point agarose (NMA) and air-dried at the room temperature then cells were mixed with 0.5% low melting point agarose (LMA) at 37oC to form a cell suspension which was spread onto the slide surface and let it solidified. A third layer of 0.5% low melting point agarose was then added and again allowed to solidify. After preparing the three layer agarose the slides were immersed in lysing solution at least for an hour. The slides were then placed in an electrophoresis tank which contained neutral or alkaline buffer solution and kept in there for a short while prior to electric field application. After electrophoresis, the slides were washed with neutralization solution or PBS and stained with a DNA-specific fluorescent dye and analyzed using a fluorescent microscope. [Archives Medical Review Journal 2010; 19(4.000): 236-]