Histopathological characteristics of femoral head arteriovenous system induced by methylprednisolone and lipopolysaccharide in adult dogs

doi:10.3969/j.issn.2095-4344.0320

Abstract

Abstract:

BACKGROUND: Many mechanisms of action of glucocorticoids are closely related to the occurrence of osteonecrosis. However, the differences in pathomorphological changes between femoral head arteries and veins at different periods of steroid-induced osteonecrosis are rarely reported.
OBJECTIVE: To observe the histopathological changes of the arterial and venous system in adult dogs induced by high-dose corticosteroid combined with lipopolysaccharide, and to explore effects of these changes on the occurrence and development of osteonecrosis.
METHODS: Fifteen healthy adult dogs were randomly divided into, 4- and 8-week observation groups and control group (n=5 (10 hips) per group). The dogs in the observation groups underwent venous injection of 10 g/kg lipopolysaccharide, then intramuscular injection of 20 mg/kg methylprednisolone after 24 hours, for 3 times, with 24 hours in between. Afterwards, the dogs were sacrificed at 4 and 8 weeks respectively. The bilateral femoral heads were removed for pathological observation. The stenotic degree of arteries and veins in the bone and supporting band was measured.
RESULTS AND CONCLUSION: At 4 and 8 weeks after induction, the structure and morphology of normal intraosseous vein decreased significantly, while the proportion of moderate-stenotic and blocked-collapsed veins increased significantly. Compared with the 4-week observation and control groups, in the 8-week observation group, the thickness of arterial intima in the bone was significantly increased, and the stenotic degree was significantly increased (P < 0.05). These results indicate that there is difference in the occurrence of pathological changes between femoral artery and venous system in adult dogs under the induction of corticosteroids combined with lipopolysaccharide. The venous system presents the pathological changes at the early stage after induction (4 weeks), and the pathological changes occurred in the arterial system at 8 weeks.

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, Femur Head Necrosis, Lipopolysaccharides

CLC Number:

R318

Zhang Yao, Zhao De-wei, Wang Zi-hua, Yu Xiao-wei, Tian Feng-de, Yang Fan. Histopathological characteristics of femoral head arteriovenous system induced by methylprednisolone and lipopolysaccharide in adult dogs[J]. Chinese Journal of Tissue Engineering Research, 2018, 22(20): 3195-3200.

Figures/Tables 1

2.1 实验动物数量分析实验共纳入成年犬15只，均进入结果分析，无死亡及感染，无脱失值。 2.2 各组股骨头静脉系统病理改变利用连续切片方法，实验以血管穿出/穿入骨皮质的滋养孔为中心从骨内区域到骨外区域测量了穿经该区域的支持带静脉。在200倍显微镜下可以观察到，在骨内区域内，参与回流的静脉结构主要为薄壁窦状小管，其在股骨头的穿出部分逐渐变为典型的具有薄静脉壁的静脉结构，其外围包绕一层较厚的纤维间质。在邻近出骨点的骨外区域，可以观察到静脉结构在形态上存在差异，通过计算血管缩窄度(Stenotic Degree，SD=S2/(S1+S2)×100%)，可将静脉血管分为3种类型：第1种：正常型，静脉管壁基本正常，无明显管腔狭窄(SD<33%)；第2种：中度狭窄型，静脉管腔呈现中度狭窄(33%<SD<66%)；第3种：重度狭窄或阻塞塌陷型，静脉管腔呈现严重狭窄或阻塞(缩窄度为SD：66%-99%)甚至出现管壁塌陷或管腔消失(图2)。在3组中，观察到的静脉数量分别为51，57，47，平均5.2/股骨头，3组间所有观察到的3种类型静脉结构的分布见表1。从实验结果可以观察到，在经甲强龙及脂多糖联合诱导4周后，犬支持带静脉的结构形态发生了明显变化，表现为正常型静脉结构显著减少，而中度狭窄型、阻塞塌陷型静脉结构占比显著升高。在诱导8周后，中度狭窄型、阻塞塌陷型静脉在切片观察中仍然保持高比率，其相对于对照组仍然有显著差异(P < 0.05)，但对比4周观察组并无显著性意义(P > 0.05)。 2.3 各组股骨头动脉系统病理改变应用Motic Med 6.0 CMIAS数码医学图像分析系统，通过对小血管病理横切面面积的测量将不规则的血管切面近似转换为规则的圆形(图3)，通过血管直径，内膜厚度以进行血管缩窄都近似计算。3组支持带动脉血管狭窄度(Stenotic Degree， SD=S2/(S1+S2)×100%)在骨外和骨内区域测量计算数值如表2。将骨内区域和骨外区域3组血管缩窄度百分数测量数据进行比较：在骨内区域，3组总体比较差异有显著性意义(P =0.001)，但4周观察组与对照组比较无显著差异(P > 0.05)，4周观察组与8周观察组比较差异有显著性意义 (P < 0.05)，以及对照组与8周观察组之间差异有显著性意义(P=0.001)，即认为8周观察组骨内区域动脉血管内膜相对增厚，血管缩窄程度高(表2)；在骨外区域，3组的血管缩窄度差异无显著性意义(P > 0.05，表2)。"

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