Toxic effects of Elaeagnus angustifolia fruit extract on chondrogenesis and osteogenesis in mouse limb buds.

Objectives: We determined the effect of Elaeagnus angustifolia extract on chondrogenesis and osteogenesis in mouse embryo limb buds in vitro and in vivo. Limb bud mesenchyme from day 12.5 embryos were used for high-density micromass cultures. Water/alcohol extract was added to culture media at 10, 100, 1000 and 10000 μg/L. Cytotoxicity was tested with neutral red. Chondogenesis was detected by alcian blue and osteogenesis was detected by alizarin red S and alkaline phosphatase activity. For in vivo experiments, 40 pregnant mice were given 0.5, 5.0 or 50.0 mg/kg of the extract between days 8 and 18 of gestation. Embryos were stained with alizarin red S and alcian blue to measure femur and ossified region lengths. Total bone mass volume was measured stereometrically. Data were compared with ANOVA and LSD.

Results: In limb bud cultures 10 μg/mL of extract reduced chondrogenesis but not osteogenesis. Higher concentrations had no effect on chondrogenesis or osteogenesis. In pregnant mice 50 mg/kg of the extract significantly increased fetal femur and ossified zone length, but significantly decreased bone and cartilage volumes.

Significance: The extract had no favorable effects on chodrification or ossification and appeared to reduce chondrogenesis. This is in apparent contradiction to its empirical effects in human adults.