Shi Lijun, Mao Tianli, Luo Pan, Li Tengqi, Gao Fuqiang, Sun Wei, Li Zirong
Department of Orthopedics, Graduate School of Peking Union Medical College, China-Japan Friendship Hospital, Beijing, 100029, P.R.China.
Department of Orthopedics, Peking University Third Hospital, Beijing, 100191, P.R.China.
Zhongguo Xiu Fu Chong Jian Wai Ke Za Zhi. 2020 Feb 15;34(2):206-212. doi: 10.7507/1002-1892.201905112.
To explore the effect of icariin on early steroid-induced osteonecrosis of the femoral head in rabbits.
Fifty mature New Zealand rabbits (weighing, 2.5-3.0 kg) were randomly divided into control group ( =10), model group ( =20), and experimental group ( =20). The rabbits of model and experimental groups were injected with lipopolysaccharide and methylprednisolone to establish the animal model of early steroid-induced osteonecrosis of the femoral head. The rabbits of experimental group were feeded with icariin solution once a day for 6 weeks since the first injection of methylprednisolone, whereas the rabbits of control and model groups were given normal saline at the same time points. The left femoral heads were removed after 6 weeks and gross morphological features were evaluated. Micro-CT scan was performed to analyze the trabecular microstructure with the following parameters: trabecular bone volume to total volume (BV/TV), trabecular number (Tb.N), trabecular thickness (Tb.Tn), and trabecular separation (Tb.Sp). The Micro-CT scan was also converted to three-dimensional reconstruction images for observation. HE staining was applied to observe the trabecular structure and morphological changes of osteocytes and marrow adipocytes. It was also used to determine whether the samples of femoral heads occurred osteonecrosis based on the criteria for pathological diagnosis, and calculate the rate of empty lacunae.
Seven rabbits died during the study, and 9, 16, and 18 rabbits in the control, model, and experimental groups, respectively, enrolled the final analysis. Compared with control group, the femoral head collapse and trabecular breaks were more obvious, and the trabeculae were sparse with irregular arrangement in the model group according to the results of gross observation, Micro-CT scan, and three-dimensional reconstruction images. But in the experimental group, the surface of femoral head was slight shrinking without obvious collapse, and the degeneration of trabecular structure was mild. According to bone microstructures analysis, the Tb.N, Tb.Tn, and BV/TV of femoral head in model and experimental groups were lower than those in control group, while the Tb.Sp in the model and experimental groups were significantly higher. The Tb.N, Tb.Tn, and BV/TV of femoral head in experimental group were higher than those in model group, while the Tb.Sp in the experimental group was significantly lower. The differences between groups were all significant ( <0.05). In the model group, HE staining showed that the number of osteocytes reduced, the number of empty lacunae increased, and the marrow adipocytes piled up in the space between femoral trabeculae, some even mashed together like a cyst. In the experimental group, the trabecular structure was still relatively complete compared with model group, no obvious apoptosis of osteocytes was observed, the size and number of adipocytes were basically normal. None of the animals in control group occurred osteonecrosis of the femoral head based on the criteria for pathological diagnosis, and the incidence of osteonecrosis were 81.3% (13/16) in the model group and 66.7% (12/18) in the experimental group, and the difference was not significant ( =0.448). The rate of empty lacunae of osteonecrotic femoral heads in the model group was 33.1%±1.4%, which was higher than that in experimental group (18.9%±0.8%) and in control group (12.7%±1.5%), and the differences between groups were significant ( <0.05).
The icariin has a protective effect on the early steroid-induced osteonecrosis of the femoral head in rabbits, which can decrease osteocytes apoptosis, improve the bone microstructure, and delay such disease processes.
探讨淫羊藿苷对兔早期激素性股骨头坏死的影响。
将50只成年新西兰兔(体重2.5 - 3.0 kg)随机分为对照组(n = 10)、模型组(n = 20)和实验组(n = 20)。模型组和实验组兔注射脂多糖和甲基泼尼松龙以建立早期激素性股骨头坏死动物模型。自首次注射甲基泼尼松龙起,实验组兔每日给予淫羊藿苷溶液,持续6周,而对照组和模型组兔在相同时间点给予生理盐水。6周后取出左侧股骨头,评估大体形态特征。进行Micro-CT扫描以分析骨小梁微观结构,参数如下:骨小梁体积与总体积之比(BV/TV)、骨小梁数量(Tb.N)、骨小梁厚度(Tb.Tn)和骨小梁间距(Tb.Sp)。Micro-CT扫描图像还转换为三维重建图像进行观察。应用HE染色观察骨小梁结构以及骨细胞和骨髓脂肪细胞的形态变化。根据病理诊断标准判断股骨头样本是否发生骨坏死,并计算空骨陷窝率。
研究期间7只兔死亡,最终纳入分析的对照组、模型组和实验组兔分别为9只、16只和18只。大体观察、Micro-CT扫描及三维重建图像结果显示,与对照组相比,模型组股骨头塌陷、骨小梁断裂更明显,骨小梁稀疏且排列不规则。但实验组股骨头表面轻度萎缩,无明显塌陷,骨小梁结构退变较轻。根据骨微观结构分析,模型组和实验组股骨头的Tb.N、Tb.Tn及BV/TV均低于对照组,而模型组和实验组的Tb.Sp显著高于对照组。实验组股骨头的Tb.N、Tb.Tn及BV/TV高于模型组,而实验组的Tb.Sp显著低于模型组。组间差异均有统计学意义(P < 0.05)。模型组HE染色显示骨细胞数量减少,空骨陷窝数量增加,骨髓脂肪细胞在股骨小梁间隙堆积,部分甚至像囊肿一样融合在一起。与模型组相比,实验组骨小梁结构仍相对完整,未观察到明显的骨细胞凋亡,脂肪细胞大小和数量基本正常。根据病理诊断标准,对照组动物均未发生股骨头坏死,模型组骨坏死发生率为81.3%(13/16),实验组为66.7%(12/18),差异无统计学意义(P = 0.448)。模型组骨坏死股骨头的空骨陷窝率为33.1%±1.4%,高于实验组(18.9%±0.8%)和对照组(12.7%±1.5%),组间差异有统计学意义(P < 0.05)。
淫羊藿苷对兔早期激素性股骨头坏死具有保护作用,可减少骨细胞凋亡,改善骨微观结构,延缓疾病进程。