[Research on inhibition of areca nut extract on human buccal keratinocytes].

OBJECTIVE

To culture human oral mucosa epithelial cells in vitro and study the mechanism of changes in the epithelia of oral submucous fibrosis (OSF).

METHODS

The keratinocytes were obtained from normal buccal mucosa (NM-KC) and OSF buccal mucosa (OSF-KC) with dispase trypsine and cultured in vitro. Then the cell proliferation of keratinocytes incubated with or without areca nut extract in the presence of 10% fetal calf serum for 48 hours at 37 degrees C in 5% CO2 and air were monitored by use of thiazolyl blue (MTT) assay.

RESULTS

Primary culture grew in small islands that coalesced at confluency. Ultrastructurally the cells contained a large of tonofilament and microvilli. Immunohistochemistry demonstrated uniform staining of the cells with antibodies to keratins. There was no difference in cell proliferation between OSF-KC and NM-KC, and areca nut extract inhibited keratinocyte proliferation in a concentration-dependent manner.

CONCLUSION

The results suggested that utilizing dispase II can establish fibroblast-free culture for growing of human oral mucosa keratinocytes and areca nut extract is a cytotoxic agent to human buccal keratinocytes.