Construction of a rabbit model of steroid-induced osteonecrosis of the femoral head by combining different concentrations of glucocorticoids with horse serum

doi:10.3969/j.issn.2095-4344.2017.08.014

Abstract

Abstract:

BACKGROUND: Glucocorticoid has been shown to be a major factor of osteonecrosis of the femoral head
(ONFH), so constructing a reliable, effective and low mortality ONFH model will be helpful for searching for a better treatment strategy of ONFH.
OBJECTIVE: To construct a rabbit model of early ONFH by intravenous injection of different concentrations of glucocorticoids and horse serum.
METHODS: Thirty healthy male New Zealand rabbits were randomly allotted to six groups, followed by given the injection of 10 mg/kg horse serum combined with 5 mg/kg dexamethasone (group A), 10 mg/kg horse serum combined with 10 mg/kg dexamethasone (group B), 20 mg/kg horse serum combined with 5 mg/kg dexamethasone (group C), 20 mg/kg horse serum combined with 10 mg/kg dexamethasone (group D), 10 mg/kg dexamethasone (group E), and 2 mL/kg normal saline (control group) via ear veins, respectively.
RESULTS AND CONCLUSION: Abnormal MRI signal of the femoral head appeared in the group D at postoperative 2 weeks, while abnormal signal was seen at postoperative 4 weeks in the other groups except the controls. Six weeks postoperatively, 80% rabbits in the group D showed abnormal signals, which were significantly more than those in the groups C (50%), B (40%), A (25%), and E (20%) (P < 0.05). The serum levels of triglyceride and total cholesterol in the groups A, B, C, D were significantly higher than those in the control group at 3, 7, 14 and 30 days after injection (P < 0.05). Compared with the control group, the ratio of empty lacuna sigmificantly increased in the group D (P < 0.05). These results indicate that the injection of high concentration of horse serum combined with the high concentration of dexamethasone is successful and safe to make an animal model of early ONFH.

中国组织工程研究杂志出版内容重点：组织构建；骨细胞；软骨细胞；细胞培养；成纤维细胞；血管内皮细胞；骨质疏松；组织工程

Key words: Tissue Engineering, Dexamethasone, Models, Animal

CLC Number:

R318

Cao Liang-quan1, Du Bin2, Sun Guang-quan2, Liu Xin2, Chen Guo-qing3, Gu Lei1, Liu Bing4. Construction of a rabbit model of steroid-induced osteonecrosis of the femoral head by combining different concentrations of glucocorticoids with horse serum[J]. Chinese Journal of Tissue Engineering Research, 2017, 21(8): 1229-1235.

Figures/Tables 4

2.1 实验动物数量分析实验共纳入30只新西兰白兔， 2只白兔在使用马血清和地塞米松后死亡，5只白兔在实验过程中患病但仍存活，其中4只是因皮肤软组织感染，1只因肛门直肠感染。最终进入结果分析28只，分别为高剂量地塞米松联合低剂量组5只，高剂量地塞米松联合高剂量马血清组5只，低剂量地塞米松联合高剂量马血清组4只，低剂量地塞米松联合低剂量马血清组5只，地塞米松组4只，对照组5只。 2.2 兔一般情况在实验过程中，高剂量地塞米松联合低剂量马血清组、高剂量地塞米松联合高剂量马血清组、低剂量地塞米松联合高剂量马血清组、低剂量地塞米松联合低剂量马血清组在静脉注射地塞米松后自主性活动减少，体质量显著增加，尤其在注射马血清之后尤为明显。2只白兔在使用马血清和地塞米松后死亡(低剂量地塞米松联合高剂量马血清组组1只，地塞米松组1只)，5只白兔在实验过程中患病但仍存活(高剂量地塞米松联合低剂量马血清组组2只，高剂量地塞米松联合高剂量马血清组组1只，低剂量地塞米松联合低剂量马血清组组1只，对照组1只)，其中4只是因皮肤软组织感染，1只因肛门直肠感染。存活的白兔精神及饮食均正常。 2.3 各组兔健康率、患病率、死亡率变化与其他组相比，高剂量地塞米松联合高剂量马血清不会增加患病率和死亡率，模型的安全性是可靠的。见图1。 2.4 各组核磁共振成像(MRI)检测结果如图2所示，在对照组，通过MRI成像研究发现所有股骨头均正常。高剂量地塞米松联合高剂量马血清组在干预2周后在MRI成像发现第一个异常信号。高剂量地塞米松联合低剂量马血清组、低剂量地塞米松联合高剂量马血清组、低剂量地塞米松联合低剂量马血清组、地塞米松组在干预第4周出现异常，都表现为在MRI T1WI表现为小结节低信号影，在T2WI相上表现为斑片状或是小结节状高信号影。干预第6周，高剂量地塞米松联合高剂量马血清组中80%(4/5)的白兔于MRI上表现出骨坏死信号，同时，低剂量地塞米松联合高剂量马血清组中50%(2/4)白兔出现骨坏死信号，在高剂量地塞米松联合低剂量马血清组为40%(2/5)，地塞米松组为50%(2/5)，低剂量地塞米松联合低剂量马血清组为20%(1/5)。 2.5 各组兔血脂谱变化在进行第1次注射地塞米松后的第3，7，14，30天检测模型组的兔子的血清胆固醇和三酰甘油水平明显高于对照组，在高剂量地塞米松联合低剂量马血清组、高剂量地塞米松联合高剂量马血清组、地塞米松组尤为明显(P < 0.05)，各地塞米松组联合马血清组与总胆固醇之间差异无显著性意义(表2，3)。然而，高剂量地塞米松联合高剂量马血清组与低剂量地塞米松联合高剂量马血清组、低剂量地塞米松联合低剂量马血清组相比，其血清胆固醇水平明显升高(P < 0.05)。见表2，3。 2.6 各组兔骨组织变化在6组实验标本中，未发现肉眼可见的骨坏死或是塌陷的股骨头。与对照组相比，空骨陷窝程度在高剂量地塞米松联合高剂量马血清组逐渐增加，一些骨小梁已破损不完整。在低剂量地塞米松组或低剂量马血清组或地塞米松联合生理盐水组，空骨陷窝程度中度增加(P < 0.05)，骨小梁部分破碎，脂肪细胞明显增加，见表4。"

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