Ribavirin, but Not Interferon α-2b, is Associated with Impaired Osteoblast Proliferation and Differentiation In Vitro

Abstract

Hepatitis C treatment with interferon alpha-2b (IFN-alpha) and ribavirin has been related to decreased bone mineral density. The aim of this study was to investigate the in vitro effects of different concentrations of ribavirin and IFN-alpha on osteoblast-like cells. Human osteoblast-like cells obtained by the outgrowth of cells from bone chips were exposed to ribavirin (0.1-10 microg/mL) or IFN-alpha (0.1-1000 UI/mL). At regular time-points, cultures were harvested for posterior analysis. Alkaline phosphatase (ALP) activity was determined on days 7 and 14, and cell growth was accessed by C3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT) and cell count on days 1, 3, 5, and 7. Flow cytometry analysis was used for investigating cell death on days 1, 3, 5, and 7. IFN-alpha affected ALP expression only at the higher concentration (1000 UI/mL) after 7 days (P < 0.05). No effects were detected in cell growth. In ribavirin treated cultures, concentrations higher than 2.5 microg/mL were associated with a decrease in ALP activity within 7 and 14 days (P < 0.01 and P < 0.001, respectively). Furthermore, the reduction in cell growth was dose-dependent and was detected after the fifth day. This decrease can be explained by an increase in the number of dead cells and a decrease in cell proliferation. In conclusion, our experiments demonstrated that ribavirin reduced, in a time- and dose-dependent manner, the number of metabolically active cells through a decrease in proliferation and an increase in cell death, and induced an impairment in osteoblast differentiation. These negative effects of ribavirin on osteblast-like cells might contribute to the bone loss reported in vivo.

Bone mineral density and cytokine levels during interferon therapy in children with chronic hepatitis B: does interferon therapy prevent from osteoporosis?

“…Trombetti et al [47], on the other hand, did not find any effect of ribavirin in bone metabolism. The impact, therefore, of IFN-alpha and ribavirin in bone remains unclear [48]. In our study, there was no statistically significant difference between patients with hepatitis B received IFN-alpha alone and combination with lamivudine.…”

Long-Term Cytotoxicity, pH and Dissolution Rate of AH Plus and MTA Fillapex

“…It was also reported that hOB cells provide a useful tool able to help predicting the effects of biomaterials on regenerative capability of periapical tissues (14). It is important to mention that the used cell line has been previously characterized (15).…”

Classical and Emerging Roles of Vitamin D in Hepatitis C Virus Infection

“…When human osteoblasts were incubated with ribavirin, a dose-dependent decrease in cell proliferation occurred and there was a significant increase in osteoblast cell death. 88 In contrast, interferon had no effect on osteoblast cell proliferation or death. Other in vitro studies demonstrated that ribavirin directly enhances osteoclast formation, 89 decreases intestinal calcium absorption by directly acting on intestinal mucosa cells, and interferes with 1,25(OH) 2 D production.…”