Human monocyte response to cementum extracts from periodontally diseased teeth: Effect of conditioning with tetracycline

Monocyte inflammatory cytokines, such as TNFα and IL-Iβ, have been implicated in the pathogenesis of periodontal destruction. The present study was designed to test the ability of extracts of cementum from periodontally diseased teeth to induce the secretion of these mediators by monocytes, to evaluate the role of adsorbed endotoxin in this process, and to test the effect of cementum conditioning with tetracycline on the monocyte response. Human monocytes were incubated with varying concentrations of cementum extracts, and TNFα and IL-Iβ levels in the media were measured. The results showed that while extracts of healthy cementum had no effect on monocyte secretion, concentration as low as 0.5 mg/ml of cementum from diseased sites raised the levels of TNFα and IL-1β secretion 10-fold. This response was dose-dependent. Diseased cementum were found to contain 1.5 ng/mg endotoxin, while endotoxin was not detectable in the extracts of the healthy cementum. However, neutralization of the endotoxin by polymyxin B only partially reduced the monocyte secretory response by 50 to 70%, suggesting that other factors in the extracts are also involved in monocyte stimulation. To simulate the effect of root conditioning, cementum was first agitated in a tetracycline or control solution prior to its extraction in media. Pretreatment of diseased cementum with tetracycline (50 mg/ml) was found to block the secretion of TNFα from cementum-stimulated monocytes. Pretreatment of the diseased cementum with 10 mg/ml tetracycline was not more effective than saline and HCl controls, with all treatments reducing cytokine secretion by ≈80%. The direct addition of tetracycline to cementum-stimulated monocyte culture was found to block TNFα secretion in a dose dependent manner. The results suggest that extracts from diseased cementum are potent stimulators of monocyte secretion, and that endotoxin as well as other factor(s) appear to be involved. These factors are partially extracted by washing and a 10 mg/ml tetracycline solution is not more effective than saline in achieving this goal. In addition, tetracycline was found to be a potent inhibitor of TNFα secretion by cementum-stimulated monocytes, suggesting a novel mechanism for this drug in periodontal therapy.