Long-term cytotoxicity of orthodontic direct-bonding adhesives

The purpose of this study was to examine the potential toxic effects of several orthodontic adhesives immediately after polymerization and at various time intervals up to 2 years postopolymerization by means of an in vitro overlay assay. Adhesive samples were incubated on tissue cultures containing an agar overlay with a vital dye (neutral red). Viable cells were stained red; nonviable cells were unstained. The circular pattern of nonviable cells demonstrated a zone of inhibition that was measured and compared. All materials tested showed cytotoxic effects immediately after polymerization and the toxic effect decreased with time postpolymerization. However, 2 years after initial polymerization, significant zones of inhibition indicating continued in vitro toxicity were still evident in all but one of the adhesives.