柚皮苷抑制PMMA颗粒诱导的破骨细胞形成及骨溶解

目的从体外和体内实验2个方面评价柚皮苷对聚甲基丙烯酸甲酯（PMMA）诱发的破骨细胞性骨溶解的作用。方法培养破骨细胞前体细胞RAW264.7,使用PMMA颗粒刺激细胞,观察柚皮苷的治疗作用。检测抗酒石酸酸性磷酸酶（TRAP）染色、Ca2＋释放实验以及TRAP、组织蛋白酶（CPK）、NF-κB的基因表达。采用小鼠膝关节置钉模型评价柚皮苷对PMMA诱发骨吸收的作用,对实验标本采用生物力学拔出试验进行评价。结果柚皮苷可以有效地抑制破骨细胞的形成并下调骨吸收基因标记物的表达,剂量为10μg/ml时表现出对TRAP活性的最大抑制作用。在颗粒刺激培养液中柚皮苷降低了钙的释放。柚皮苷缓解了膝关节置钉模型中PMMA颗粒刺激所造成的长期的骨吸收,表现为骨体积分数的增加和最大钉拔出力的增加。300 mg/kg的灌胃剂量可达到最好的疗效。结论柚皮苷抑制PMMA诱导的破骨细胞形成,降低PMMA颗粒刺激的钙释放。在观察慢性骨吸收的长期模型中,柚皮苷也有效抑制PMMA诱发的骨吸收,并保留了置入物的稳定性。

Polymethylmethacrylate-induced osteoclastogenesis and osteolysis in mouse model and the therapeutic influence of naringin

Objective To evaluate the influence of naringin on polymethylmethacrylate（PMMA）-induced osteoclastic bone resorption both in vitro and in vivo. Methods Osteoclast precursor cell RAW 264.7 were cultured and stimulated with PMMA particles followed by naringin treatment. Tartrate resistant acid phosphatase（TRAP）, Ca2＋release and gene expression copies of TRAP, cathepsin K（CPK） and nuclear factor NF-κB（RANK） were sequentially evaluated. Mouse knee-pin implantation model was adopted to evaluate the effects of naringin in treating PMMA-induced bone resorptions. Biomechanical pin-pullout tests was performed. Results Naringin was effective in inhibiting osteoclastogenesis and down-regulating bone resorption gene marker expressions in vitro. To suppress TRAP activities, the dose of 10 μg/ml exhibited the most significant influence.Naringin decreased calcium release in the stimulated cell culture medium. Naringin alleviated long term bone resorption in knee-pin model with PMMA stimulation by increasing BV/TV ratio and pin pullout peak strength. Gavage dosage of 300 mg/kg appeared to be an optimal dosage to deliver the therapeutic effects. Conclusion Naringin inhibits PMMA-induced osteoclastogenesis and reduced Ca2 ＋release. In a long-term experimental osteolysis model, naringin appeared effective in protecting against PMMA-induced osteolysis and reserved the implant stability. The study suggests that naringin may serve as a potent therapeutic means to treat osteolysis.