Does ischemia/reperfusion impact apoptosis of articular chondrocyte in the femoral head epiphyses

doi:10.3969/j.issn.2095-4344.2013.28.006

Abstract

Abstract:

BACKGROUND:Ischemia/reperfusion can induce degenerative alterations in articular cartilage. However, the precise mechanism remains poorly understood.

OBJECTIVE: To observe the morphological changes and the apoptosis of articular cartilage of femoral head epiphyses with ischemia/reperfusion.

METHODS: A total of 80 Sprague-Dawley rats were randomly assigned to two groups: ischemia/reperfusion (model of ischemia/reperfusion in hip joint) and sham-surgery (exposure of abdominal aorta for 5 minutes) groups, with 40 animals in each group. Articular cartilages of femoral head epiphysis were collected in 6, 12, 24, and 48 hours, 5 days, and 2 and 4 weeks after operation. Morphology of articular cartilage of femoral head epiphyses was examined by light microscope, and cell apoptosis was detected by TUNEL method.

RESULTS AND CONCLUSION: Light microscopy showed chondrocytes degeneration and reduction, as well as fibrosis in matrix of cartilage in the ischemia/reperfusion group. Chondrocyte apoptosis was observed in both groups by TUNEL. Several apoptotic cells, less than five, were observed in the sham-surgery, while 10-30 apoptotic cells were found in ischemia/reperfusion group at 48 hours. Results indicated that ischemia/reperfusion can induce degenerative changes in articular cartilage of femoral head epiphyses, and cell apoptosis in developing hip joint may participate in damage of articular cartilage. Inhibition of chondrocyte apoptosis in articular cartilage may be useful for the prevention and cure of early osteoarthritis.

Key words: tissue construction, cartilage tissue construction, ischemia/reperfusion, femoral head epiphyses, hip joint, articular cartilage, apoptosis, chondrocyte, animal model

CLC Number:

R318

Zhang Jing-dong, Yi Xian-hong, Li Yu-an. Does ischemia/reperfusion impact apoptosis of articular chondrocyte in the femoral head epiphyses[J]. Chinese Journal of Tissue Engineering Research, 2013, 17(28): 5133-5138.

Figures/Tables 2

2.1 实验动物数量分析实验选用SD大鼠80只，分为2组，无脱失，全部进入结果分析。 2.2 光镜观察关节软骨组织形态学变化软骨细胞的观察：24 h前软骨细胞无明显变化，24 h、48 h、5 d可见部分软骨细胞轻度肿胀，软骨细胞核膜模糊、核溶解，有核空泡化现象，提示少量软骨细胞变性，5 d、2周、4周可见局灶性软骨细胞减少，但未见明显软骨细胞坏死，深层软骨细胞排列不规则，部分软骨细胞有胞浆空泡化表现，局限性软骨细胞减少、消失，软骨陷窝未见软骨细胞，软骨细胞层次分界不清，4周偶见深层软骨细胞内钙化，深层柱状排列的软骨细胞数目明显减少，甚至仅残存1-3列，4周深层软骨细胞完全失去柱状排列，软骨细胞稀少。软骨基质的改变：光镜下48 h、5 d、2周、4周时点可观察到关节软骨浅层、中层基质局限性嗜酸性增强的红染表现，2周、4周时点深层软骨基质可有明显的嗜碱性，甚至可有嗜碱性染色增强，提示局部钙盐沉积即将钙化，但也有失去嗜碱性异染性，提示软骨基质中硫酸软骨素明显减少。软骨中各层有局灶性基质肿胀，部分切片可见胶原裸露，关节软骨各层基质可见局限性纤维化表现。关节软骨表面软骨下连接部的观察：4周可见关节软骨表面粗糙，表面有多条裂纹，裂纹处小梭形软骨细胞可有局限性增生。部分5 d、2周、4周软骨深层下方无成骨细胞、骨小梁样结构等骨样组织相连。见图1。 2.3 TUNEL法检测关节软骨细胞凋亡现象假手术组的观察：假手术组12 h，24 h，48 h，5 d，2周、4周各时点均未见集群的软骨细胞凋亡，但可偶见有散在分布5个以下的软骨细胞凋亡(48 h 1只，5 d时1只，2周时1只)，一般位于头骺关节软骨的浅层或中层。而在各切片的头骺关节软骨深层均未见到明显的细胞凋亡信号。缺血-再灌注组的观察：缺血-再灌注组48 h可见10- 30个软骨细胞发生凋亡(2只)，位于头骺关节软骨的浅层及中层，聚集存在。在其他各时点关节软骨的浅层或中层也偶见个别散在分布的软骨细胞凋亡(24 h 1只，5 d时1只，2周时1只)，但同样均未见5个以上的软骨细胞凋亡现象。同时也同样在各时点的头骺关节软骨深层肥大细胞也未见明显的阳性凋亡细胞。见图2-4。"

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