Effect of preventive drugs on steroid-induced osteonecrosis of the femoral head / 中国组织工程研究

ABSTRACT

BACKGROUND: Due to glucocorticoid (GG) is used wildly on clinic, incidence rate of steroid-induced osteonecrosis of femoral head is increased.Apoptosis of bone cells plays a key role in steroid-induced osteonecrosis of femoral head during earlier period.OBJECTIVE: To induce models of femoral head necrosis of rabbits by steroid steroid so as to observe the effect of integrated Chinese and western medicine on preventing apoptosis of steroid-induced osteonecrosis of femoral head.DESIGN: Randomized controlled animal study.SETTING: Pharmacological Department and Anatomy Department of Medical College of Shaoguan University, Pathological Department of Yuebei People's Hospital of Guangdong Province.MATERIALS The experiment was carried out at the Central Laboratory of Medical College of Shaoguan University from April to July 2005. A total of 30 adult New Zealand white rabbits were selected and randomly divided into blank control group, steroid group and preventive medicine group with 10 in each group.Xuesai tongpian, main component of panax notoginseng saponin, was provided by Yuxi Weihe Pharmaceutical Company Limited (batch number 200410290; 25 mg/pill); zhibituo pian, main component of shanzha, baizhu, hongqu, was provided by Chengdu Di 'ao Jiuhong Pharmaceutical Factory (batch number 0410063, 350 mg/pill); alendronate, main component of 4-amido-1-hydroxy butylidene-1, was provided by Hainan Mankexing Pharmaceutical Factory (batch number 040501, 5 mg/pill); dexamethasone sodium phosphate injection was provided by Henan Tiankang Pharmaceutical Company Limited (batch number 040905, 5 g/L).METHODS: ① One week later, rabbits in steroid group and preventive medicine group were injected with 1 mL dexamethasone sodium phosphate solution twice a week to establish models of steroid-induced osteonecrosis of femoral head. Animals in blank control group did not model and were injected with the same volume of saline at the same time point. ② Rabbits in preventive medicine group were prefused with 25 mg xuesai tong, 350 mg zhibituo and 5 mg alendronate at the same time for 8 weeks. Animals in steroid group and blank control group did not treat with any medicine. Benzylpenicillin was injected into muscle of animals in each group for infection prevention with the dosage of 50 000 u each rabbit twice a week. ③ One week after discontinue, apoptosis of femoral head was detected with in situ end-labeling (ISEL) method. Ten areas were selected from each carry sheet glass to calculate numbers of apoptosis in every 100 cells. Pathological changes of ultrastructure were observed with double staining of uranium acetic acid and lead citromalic acid.MAIN OUTCOME MEASURES: ① Results of apoptosis of femoral head; ② Results of pathological changes of ultrastructure of femoral head under electron microscope.RESULTS: A total of 30 adult New Zealand white rabbits were selected in this study. Because one rabbit in preventive medicine group died during process, 29 rabbits were involved in the final analysis. ① Results of apoptosis of femoral head Buffy granulosum positive cells, i.e. apoptosis cells,were observed in nucleus in steroid group. Numbers of apoptosis every 100cells were more in steroid group than those in blank control group and preventive medicine group (24.14±5.46, 2.47±0.76, 8.12±3.23, q=8.76, q=13.45, P ＜ 0.01), and numbers in blank control group were similar to those in preventive medicine group (P ＞ 0.05). ② Results of pathological changes of ultrastructure of femoral head under electron microscope Volume of nucleus in steroid group was decreased; chromatin was aggregated at edge; part of chromatin was dark and thick; density of electron was increased; structure was unclear; characteristics of apoptosis were shown. Nuclear membrane in preventive medicine group was complete under electron microscope, chromatin was steady, and the nucleus was accounted for one third of cell.CONCLUSION: The integrated application of xuesai tong,, zhibituo and alendronate can inhibit hormonal-induced apoptosis of femoral head after necrosis of femoral head induced by steroid steroid.