Difference in osteogenesis- and angiogenesis-related protein expression in femoral head samples from patients with femoral head necrosis of different etiologies

doi:10.12307/2022.345

Abstract

Abstract: BACKGROUND: The etiology and pathogenesis of femoral head necrosis are still unclear, and in-depth research is urgently needed.
OBJECTIVE: To detect the protein expression of Hedgehog signaling pathway-, osteogenesis- and angiogenesis-related indicators in the necrotic femoral head samples after total hip arthroplasty, and attempt to explore the pathogenic mechanisms of different factors such as alcohol, hormones, and trauma, providing theoretical support for the subsequent clinical prevention and treatment of femoral head necrosis.
METHODS: The femoral head samples were collected from patients with femoral head necrosis who underwent total hip arthroplasty in the Affiliated Hospital of Shandong University of Traditional Chinese Medicine from December 2017 to December 2019. The femoral head samples meeting the inclusion criteria were selected and classified into control group, idiopathic group, alcohol group, hormone group and post-traumatic group according to the etiology. The expression of Hedgehog signaling pathway-, osteogenesis- and angiogenesis-related indicators in necrotic bone tissues of different etiologies were detected by western blot.
RESULTS AND CONCLUSION: By observing the general morphology and coronal plane of the femoral head samples in each group, it was found that the femoral head samples of patients with femoral head necrosis of different etiologies had similar pathological manifestations in general morphology and section, and had similar histological characteristics of necrosis and repair. Western blot results showed that the protein expression levels of osteogenesis-related indicators (alkaline phosphatase, type I collagen, osteopontin and RUNX2), angiogenesis-related indicators (CD31, vascular endothelial growth factor A, kinase insertion domain receptor, and von Willebrand factor), and Hedgehog signaling pathway indicators (hedgehog, zinc finger protein 1, and zinc finger protein 2) in the femoral head samples of patients with femoral head necrosis of different etiologies changed to different extents as compared with the control group. As the occurrence and development of femoral head necrosis may be closely related to osteogenesis, angiogenesis, and Hedgehog signaling pathway, further research is needed.

Key words: osteonecrosis of the femoral head, etiology, Hedgehog signaling pathway, osteogenesis, angiogenesis

CLC Number:

Luo Di, Liang Xuezhen, Liu Jinbao, Li Jiacheng, Yan Bozhao, Xu Bo, Li Gang. Difference in osteogenesis- and angiogenesis-related protein expression in femoral head samples from patients with femoral head necrosis of different etiologies[J]. Chinese Journal of Tissue Engineering Research, 2022, 26(11): 1641-1647.

Figures/Tables 6

(1)对照组股骨头样本形态未见明显异常，近似球形，软骨层完好且厚度正常，其冠状剖面骨小梁致密均匀，未见坏死与增生部分。 (2)特发组股骨头样本无明显塌陷与变形，但表面凹凸不光滑，负重区软骨有明显破坏与磨损特征，其颜色较正常软骨略显苍白，其冠状剖面未见明显软骨下骨折，但在负重区可见大范围不规则黄白色坏死区，其与正常区之间具有不连续纤维带增生表现。 (3)酒精组股骨头样本外形显著畸形，表面粗糙，负重区有明显塌陷下凹特征，表面软骨存在严重磨损与破坏，其冠状剖面可见负重区明显塌陷，软骨层变薄甚至消失，坏死区范围较广且呈散在分布且多由连续纤维组织带包裹，其中部分坏死区内呈现软组织囊性改变，周围则可见明显硬化带形成。 (4)激素组股骨头样本外观轮廓大致正常，但其表面软骨层存在明显断裂的特点，其剖面可见存在软骨下骨折线，负重区内侧有一明显内含无定型物质的囊性坏死病变，坏死区分布广泛且骨小梁稀疏，排布紊乱，坏死区、硬化区与正常区之间具有不连续纤维带。 (5)创伤术后组股骨头样本形态多呈不规则，股骨头表面凹凸不平，表面软骨层磨损破坏明显，其剖面可见大面积黄白色坏死区、硬化区伴有多处囊性坏死灶，负重软骨层破坏显著，坏死区周围可见不连续纤维带，其术后内固定螺钉隧道部分已由软组织填充。综合以上情况，各组样本的病理学与组织学特征存在一定程度的相似性，但不同组的坏死区与正常区分布以及病理分期尚存在差异，这提示激素组或具有更快的病理学变化。 2.3 Western blot分析结果为探究不同病因导致的股骨头坏死发生和发展的病理机制，对不同病因的典型股骨头坏死样本中成骨、成血管以及Hedgehog信号通路相关的指标蛋白表达水平进行分析。 2.3.1 不同病因坏死股骨头骨组织样本中成骨相关蛋白表达情况结果发现，在成骨方面，除创伤术后组外，各组骨样本中碱性磷酸酶的表达显著低于对照组(P < 0.05)；Ⅰ型胶原蛋白表达只有特发组与对照组差异无显著性意义(P > 0.05)，其余3组均明显低于对照组(P < 0.05)；激素组与创伤术后组的骨桥蛋白表达水平显著低于对照组(P < 0.05)，而特发组和酒精组与其相比差异无显著性意义(P > 0.05)；对照组中RUNX2的表达显著高于其余各组(P < 0.05)。通过实验结果发现，不同致病因素均可在不同程度上破坏骨的代谢平衡，抑制股骨头内正常的成骨过程。值得注意的是，创伤术后组样本内4个骨形成相关指标蛋白表达量均出现了显著下降，提示因股骨颈骨折行手术治疗却最终失败的患者，或因其术后股骨头内血供仍未得到有效改善，造成局部成骨能力明显下降，进而阻碍了骨修复过程导致最终发病，见图3。"

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