Ovariectomy (OVX) Murine Model Establishment

We examined BA’s effects on bone loss in mice by constructing an osteoporosis model (OVX mice). All experimental procedures involving animals had approval from the Animal Care Committee of Guangxi Medical University. Female C57/BL6 mice (10 weeks old) were randomized into four groups (n = eight per group): sham, OVX + PBS, OVX + 5 mg/kg BA, and OVX + 10 mg/kg BA groups. Extraction of bilateral ovaries and partial fallopian tube resection were performed in all mice except the sham group; intraperitoneal injection of penicillin (once daily) was given 3 days after the operation for preventing wound infection. After 1 week, the OVX + 5 mg/kg BA and OVX + 10 mg/kg BA groups were intraperitoneally injected BA (5 and 10 mg/kg, respectively) for treatment every other day for six weeks. The mice were weighed before each treatment, the corresponding drug dose of mice were calculated according to the high and low concentration groups, diluted with PBS to make a volume of 0.5 ml liquid medicine injected into each mouse. The remaining two groups were given the same amount of PBS as vehicle controls. Upon treatment, euthanasia was performed by excessive anesthesia. The tibias were removed and submitted to fixation with 4% formalin, and prepared for micro-CT and histological analyses.