Effect of Amino Acids and Albumin on Damage Induced by Cyanate in Osteoblast

Park, Kyung Dae; Park, Sung Bae; Choi, Hye Jung; Mun, Kyo Cheol; Kim, Hyun Chul

Abstract

PURPOSE: Cyanate, known as one of the uremic toxins and derived spontaneously from urea, has several effects on the biologic substances including erythropoietin, antioxidant and ceruloplasmin. To find out the protective materials from the hazardous effect of cyanate in osteoblast, we added twenty amino acids, albumin globulin and hemoglobin in the culture media containing osteoblastic cells with cyanate.
METHODS
Osteoblastic ROS 17/2.8 cells, exposed to various concentrations of sodium cyanate, were used to analyze for the cytotoxicity. The cyanate-induced cytotoxicity was assessed by the methylthiazolyldiphenyl-tetrazolium bromide (MTT) assay by measuring the absorbance of the reaction solution at 570 nm. Viability of the treated cells was expressed as A570 of sample/A570 of control. The degree of the carbamylation was measured using trinitrobenzenesulphonic acid. The degree of the carbamylation in amino acid was about 50% in average.
RESULTS
The degree of the carbamylation in albumin was increased depending on the incubation time with cyanate and the concentration of the cyanate. The degree of the carbamylation in globulin and hemoglobin was nearly zero. Asp, Glu, Leu, Trp and Tyr among the twenty amino acids revealed the protective effect against the damage induced by cyanate. And only albumin among the three proteins revealed the protective effect.
CONCLUSION
On the basis of these results, Asp, Glu, Leu, Trp, Tyr and albumin are useful tools for the protection against damages by cyanate carbamylation.