Acridine orange induces binucleation in chondrocytes

Although it is well known that binuclear cells commonly appear among the chondrocytes of normal cartilages as well as among neoplastic chondrocytes of chondrosarcomas, the mechanism of binucleation is still unclear. Therefore, this study was undertaken to clarify the mechanism of binucleation in chondrocytes, using primary culture cells of growth plate cartilage.

These chondrocytes were exposed to acridine orange (AO) which is a fluorescent dye for differentiating certain DNAs and RNAs in nuclei and cytoplasm, and which inhibits mitosis. After exposure to 0.5 microg/ml AO, for 0, 6, 24, 48, and 96 h, the following parameters were investigated: (1) cell growth rate (GR); (2) frequency of hyperdiploid cells (%HDC) by DNA cytofluorometry; (3) mitotic index (MI); (4) BrdU labeling index (LI); (5) frequency of binuclear cells (%BNC).

Compared with the control cells, which were cultured in AO-free medium, the GR was remarkably inhibited at 24 h. MI was also decreased from 6 to 24 h, and LI decreased at 48 h. However, these parameters were recovered at 96 h. The %HDC was increased from 6 to 96 h, and the %BNC was also increased to a maximum of six times that of the control cells at 96 h.

These results suggested that the binuclear cells observed among the cultured chondrocytes may be formed from G2 arrested cells by amitotic nuclear division, but not by mitosis without cytoplasmic division or cell fusion.