Oxidative stress as a component of chromium-induced cytotoxicity in rat calvarial osteoblasts
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It has been documented that medical prosthetic alloys release metal ions into surrounding tissues and cause cytotoxicity, but the mechanisms remain undefined. In that regard the cellular oxidative stress may be a common pathway in cellular responses to metal ions. The objective of this study was to approach the hypothesis that oxidative stress mediates chromium-induced cytotoxicity in rat calvarial osteoblasts. Osteoblasts were exposed to different concentrations of Cr6+ or Cr3+ (5–20 μM) in the presence or absence of the antioxidant N-acetyl-cysteine (NAC; 1–5 mM). Cellular viability, differentiation, and intracellular ultrastructural alterations were evaluated by MTT assay, alkaline phosphatase (ALP) activity assay, and transmission electron microscopy. Cellular oxidative stress was evaluated by intracellular reactive oxygen species (ROS) production. ROS production was monitored by the oxidation-sensitive fluorescent probe 2′7′-dichlorofluorescin diacetate (DCFH-DA). A time- and concentration- dependent increased cytotoxicity, time-dependent increased intracellular ROS production were indicated on exposure to Cr6+. Pretreatment of osteoblasts with 1–5 mM NAC afforded dose-dependent cytoprotective effects against Cr6+-induced cytotoxicity in osteoblasts. NAC decreased the level of intracellular ROS induced by Cr6+, too. While Cr3+ and NAC did not have any significant effects on osteoblasts (5–20 μM). These results suggest that oxidative stress is involved in Cr6+-induced cytotoxicity in osteoblasts, and NAC can provide protection for osteoblasts against Cr6+-induced oxidative stress. Cr3+ (5–20 μM) have no significant cytotoxicity in osteoblasts based on the results of this study.
KeywordsReactive oxygen species (ROS) MTT Alkaline phosphatase (ALP) Antioxidant N-acetyl-cysteine (NAC) Ultrastructure
This study was supported by the Chinese National Science Foundation (Fund number 30271428) and the Distinguished Young Teacher Plan Fund (Education Bureau grant number 2002-40).
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