Implication of Dickkopf-1 and cell apoptosis in steroid-induced avascular necrosis of the femoral head

doi:10.3969/j.issn.2095-4344.2017.28.002

Abstract

Abstract:

BACKGROUND: The roles of osteocyte and osteoblast apoptosis in steroid-induced avascular necrosis of the femoral head (ANFH) have arouse much attention, and its pathogenesis has been understood gradually. But there is a lack of knowledge about the mechanisms underlying osteocyte and osteoblast apoptosis. Meanwhile, hormones have been shown to enhance the Dickkopf-1 expression in Wnt signaling pathway.
OBJECTIVE: To investigate the roles of Dickkopf-1 and cell apoptosis in steroid-induced ANFH, and to understand their correlations with steroid-induced ANFH.
METHODS: Necrotic femoral head samples were removed from 14 patients with steroid-induced ANFH after total hip arthroplasty (experimental group), and normal femoral heads were from 8 patients with femoral neck fracture (control group). The cellular morphology was observed using transmission electron microscope; the number of empty lacunae was counted through hematoxylin-eosin staining; the cell apoptosis was detected by TUNEL assay; the expression level of Dickkopf-1 was detected by immunohistochemistry. Moreover, the correlation between the Dickkopf-1 positive rate and apoptosis index of osteocytes was analyzed by Spearman rank correlation analysis.
RESULTS AND CONCLUSION: The number of empty lacunae, expression level of Dickkopf-1, and apoptosis index of osteocytes in the experimental group were significantly higher than those in the control group (P < 0.05). In the experimental group, the Dickkopf-1 positive rate was positively correlated to the apoptosis index of osteocytes (r=0.623). These results indicate that Dickkopf-1 and osteocyte apoptosis both play key roles in steroid-induced ANFH, and increased Dickkopf-1 level may promote the development of ANFH.

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: Arthroplasty, Replacement, Hip, Femur Head Necrosis, Prosthesis Implantation, Tissue Engineering

CLC Number:

R318

Kong Ling-yue, Liu Wan-lin, Ren Yi-zhong. Implication of Dickkopf-1 and cell apoptosis in steroid-induced avascular necrosis of the femoral head[J]. Chinese Journal of Tissue Engineering Research, 2017, 21(28): 4436-4441.

Figures/Tables 1

2.1 大体形态学观察实验组和对照组股骨头大体形态无明显差异。 2.2 苏木精-伊红染色结果实验组：在实验组的病理切片上可以很清楚地看见非常具有特征性的骨坏死，光镜下可以看见骨小梁失去原有的形态，稀疏、变细，有些甚至出现断裂，结构紊乱，有碎片出现；造血细胞数量减少；成骨细胞排列不规则；空缺骨陷窝明显增加；骨髓内脂肪细胞体积增大，有的融合成泡状(图1A)。对照组：在对照组的病理切片上可以观察到形态完整的骨小梁，这些骨小梁排列非常整齐，致密而且饱满，骨小梁中的骨细胞清晰可见，核大并且位于中央；骨髓内造血细胞丰富，形态正常(图1B)。实验组空缺骨陷窝率均明显高于对照组，经统计分析，差异有显著性意义(t=28.18，P=0.00，P < 0.05)，见表1。 2.3 超薄切片透射电镜结果实验组：股骨头切片表现为胞膜完整，细胞核固缩，染色质边集，细胞器消失，凋亡小体出现(图2A)；对照组：骨细胞、骨髓细胞超微结构均正常(图2B)。 2.4 TUNEL骨细胞凋亡情况 TUNEL染色阳性在切片中的表现为：细胞核着色呈棕黄褐色，极为少数的细胞浆内可以出现阳性颗粒。实验组的切片上可见骨细胞的细胞核中有棕黄褐色的阳性颗粒，并且在骨髓组织中可见有大量的骨细胞凋亡(见图3)。实验组的骨细胞凋亡指数明显高于对照组，经统计分析，差异有显著性意义(Z=2.26，P=0.00， P < 0.05)，见表1。 2.5 Dickkopf-1表达率实验组Dickkopf-1表达率高于对照组，经统计分析，差异有显著性意义(t=24.77，P=0.00，P < 0.05)，见表1及图4。 2.6 实验组中Dickkopf-1表达率与骨细胞凋亡指数的线性相关分析实验组中Dickkopf-1表达率与骨细胞凋亡指数经统计分析，二者呈正相关性(r=0.623，P < 0.05)。"

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