Lack of Teratogenicity of 3‐Chloro‐4‐(Dichloromethyl)‐5‐Hydroxy‐2 (5H)‐Furanone (MX) in Embryonic Mouse Palate Culture in vitro

Abstract

ABSTRACT3‐Chloro‐4‐(dichloromethyl)‐5‐hydroxy‐2(5H)‐furanone (MX) is known as a by‐product of wood pulp manufacture and a contaminant of chlorinated drinking water. Since our previous studies (Teramoto et al., 1998, 1999) demonstrated in a micromass in vitro test a strong inhibitory effect of MX on rat embryo cell differentiation, the potential teratogenicity was investigated in this study by using a suspension organ culture system. Twelve‐day mouse embryo palatal explants were cultivated for 72 hr in the MX‐containing medium at a concentration of 0, 1, 10, 100 or 300 μg/ml and examined for closure of the palatal shelves. All control explants showed almost complete closure of the palatal shelves. Similar results were also obtained in the MX‐treated explants at concentrations up to and including 100 μg/ml. Immunohistochemistry revealed no difference between the control and MX‐treated explants in distribution of PCNA‐and TUNEL‐positive cells in the palatal mesenchyme and medial edge epithelium, respectively. When the MX concentration was raised to 300 μg/ml, palatal shelves remained wide open. However, histopathology revealed extensive pyknosis of the mesenchymal cells and loss of the epithelium. These results may indicate that MX is cytotoxic against the mouse palate at a high concentration, and that it has no cleft‐palate inducing effects in mice.