Effect of reduction of mechanical loading on subchondral bone and articular cartilage of early osteoarthritis in mice

doi:10.3969/j.issn.2095-4344.1432

Abstract

Abstract:

BACKGROUND: Osteoarthritis has a high incidence in the world, but its underlying pathogenesis is still not thoroughly studied.
OBJECTIVE: To investigate the effect of reduction of mechanical loading on subchondral bone and articular cartilage of early osteoarthritis in mice.
METHODS: The study was approved by the Laboratory Animal Ethics Committee of the First Affiliated Hospital of Soochow University. Forty-eight male adult C57BL/6 mice (8 weeks old) were randomly divided into normal group, osteoarthritis model group, motor nerve transection+osteoarthritis model group, and motor nerve transection+sham group, 12 mice in each group. We simulated the reduction of mechanical loading in knee joint via tibial nerve combined with common peroneal nerve transection, which was performed at 1 week before the osteoarthritis induction. Six mice in each group were sacrificed at 4 and 8 weeks after modeling. The right knee joints of each group were collected for micro-CT scanning, histological staining, footprint analysis and muscle atrophy detection, respectively.
RESULTS AND CONCLUSION: (1) The bone volume fraction in the motor nerve transection+osteoarthritis model group was lower than that in the osteoarthritis model group at 4 and 8 weeks after modeling (P < 0.05). There was no significant difference in the bone volume fraction between motor nerve transection+osteoarthritis model and motor nerve transection+sham groups (P > 0.05). The bone volume fraction in the motor nerve transection+osteoarthritis model group was increased significantly at 8 weeks compared with that at 4 weeks after modeling (P < 0.05). (2) The OARSI score in the motor nerve transection+osteoarthritis model group was lower than that in the osteoarthritis model group at 4 and 8 weeks (P < 0.05). OARSI score in the motor nerve transection+osteoarthritis model group at 8 weeks was higher than that at 4 weeks after modeling (P < 0.05). (3) Compared with the osteoarthritis model group, the hindfoot base widths in the motor nerve transection+osteoarthritis model group at 4 and 8 weeks were decreased significantly (P < 0.05), and the wet weights of the right tibial anterior muscle and the calf triceps muscle were decreased (P < 0.05). (4) These results indicate that reduction of mechanical loading in knee joint induced by peripheral motor nerve transection leads to bone loss of subchondral bone and reduction thickness of articular cartilage layer, suggesting that the appropriate joint mechanical loading plays an important role in maintaining the homeostasis of articular cartilage and subchondral bone.

Key words: osteoarthritis, mechanical loading, articular cartilage, subchondral bone, micro-CT, footprint analysis, bone volume fraction, the National Natural Science Foundation of China

CLC Number:

Shao Yijie1, Jiang Huaye1, Gao Chao1, Luo Zongping2, Yang Huilin1. Effect of reduction of mechanical loading on subchondral bone and articular cartilage of early osteoarthritis in mice [J]. Chinese Journal of Tissue Engineering Research, 2019, 23(35): 5611-5618.

Figures/Tables 2

2.1 实验动物数量分析所有48只小鼠耐受手术良好，无小鼠出现意外死亡，未出现感染迹象。 2.2 Micro-CT分析结果造模后4周及8周，典型的Micro-CT冠状面图像见图3A，无论4周还是8周，关节炎模型组的内侧胫骨软骨下骨的骨体积分数增加，表现为骨硬化。相反，运动神经(胫神经联合腓总神经)切断后，小鼠膝关节内侧胫骨软骨下骨的骨体积分数急剧减少，表现为骨丢失。与关节炎模型组比较，造模后4周及8周运动神经切断+模型组中骨体积分数降低(P < 0.001)，见图3B，C。运动神经切断+模型组与运动神经切断+假手术组之间差异无显著性意义。另外，运动神经切断+模型组8周时骨体积分数较4周有所增加(P < 0.05)。 2.3 组织学染色结果如图4A所示，造模后4周，关节炎模型组软骨表面蛋白聚糖丢失(局部番红O失染)，软骨表面粗糙，软骨细胞死亡。运动神经切断+模型组软骨表面轻微蛋白聚糖丢失，软骨表面较为光滑。造模后8周，骨关节炎模型组软骨表面磨损较为严重，软骨细胞大量死亡。运动神经切断+模型组软骨表面蛋白聚糖丢失范围较4周大。运动神经切断+假手术组4周及8周并未见到明显的蛋白聚糖丢失。图4B为OARSI半定量评分，随着时间的进展，关节炎模型组及运动神经切断+模型组软骨退变逐渐加重(P < 0.001)，与关节炎模型组比较，造模后4周及8周运动神经切断+模型组软骨退变减轻(P < 0.001)。运动神经切断+假手术组造模后4周及8周相比于正常组软骨OARSI评分无明显差异。图4C，D示，造模后4周及8周时运动神经切断+假手术组软骨厚度较正常组降低，且8周时降低更为明显。 2.4 足印分析结果运动神经(胫神经联合腓总神经)切断后，小鼠右足行五趾并拢、足轻度外翻。图5A为小鼠足印示意图，图5B为各组小鼠的足印图，从中可以发现运动神经切断组造模后4周及8周右前足和右后足足印重叠或接近。造模后4周及8周时，相比于关节炎模型组，运动神经切断+模型组后足足宽降低(P < 0.001)，见图5C，D。此外，运动神经切断+模型组和运动神经切断+假手术组后足足宽在4周时与在8周时无明显差别。 2.5 肌肉湿质量造模后4周及8周时，相比于关节炎模型组，运动神经切断+模型组右侧小腿三头肌肌肉湿质量明显降低(P < 0.001)，见图6A，B。运动神经切断+模型组右侧小腿三头肌肌肉湿质量在造模后8周时要比4周稍降低，但差异无显著性意义。造模后4周及8周时，相比于关节炎模型组，运动神经切断+模型组右侧胫前肌肌肉湿质量明显降低(P < 0.001)，见图6C，D。运动神经切断+模型组右侧胫前肌肌肉湿质量在造模后4周与8周无明显差异。 "

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