Establishment and identification of the rat models of chondrocyte apoptosis induced by tumor necrosis factor-alpha

doi:10.3969/j.issn.2095-4344.2017.04.006

Abstract

Abstract:

Abstract
BACKGROUND: Tumor necrosis factor-α (TNF-α), a main cytokine inducing chondrocyte apoptosis of osteoarthritis, plays a regulatory role in the process of osteoarthritis.
OBJECTIVE: To compare the rat models of chondrocyte apoptosis induced by different mass concentrations of TNF-α, thus providing theoretical basis for further study on the regulation of drugs on chondrocyte apoptosis.
METHODS: Chondrocytes were isolated from the knee cartilage of 4-week-old Sprague-Dawley rats of clean grade by mechanical ll collagenase digestion and were then incubated with different mass concentrations of TNF-α to induce apoptosis. The morphology of chondrocytes was observed under inverted phase contrast microscope, cells were identified using immunohistochemical staining of type II collagen, as well as the cell viability and apoptosis were detected by MTT and DAPI, respectively.
RESULTS AND CONCLUSION: (1) In vitro, the cytoplasm of chondrocytes was stained brown-yellow by using immunohistochemical staining of type II collagen. (2) At 48 hours, the apoptosis rate of chondrocytes induced by 10, 20 and 30 μg/L TNF-α was significantly higher than that of the 0 μg/L TNF-α (P < 0.01), and the apoptosis rate of chondrocytes induced by 40 μg/L TNF-α was significantly higher than that of the 10 μg/L TNF-α (P < 0.01). (3) The viability of chondrocytes induced by 10, 20 and 40 μg/L TNF-α was significantly lower than that of the 0 μg/L TNF-α (P < 0.01). In detail, the viability of chondrocytes induced by 20 μg/L TNF-α was lower than that of the 10 μg/L TNF-α (P < 0.05); the viability of chondrocytes induced by 40 μg/L TNF-α was significantly lower than that of the 10 and 20 μg/L TNF-α (P < 0.01, P < 0.05). (4) These results suggest that 20 μg/L TNF-α can successfully induce the chondrocyte apoptosis model.

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: Chondrocytes, Tumor Necrosis Factor-alpha, Apoptosis, Inflammation, Tissue Engineering

CLC Number:

R318

Chen Hou-huang1, Shao Xiang1, Li Li2, Wu Ming-xia2, Li Xi-hai1. Establishment and identification of the rat models of chondrocyte apoptosis induced by tumor necrosis factor-alpha[J]. Chinese Journal of Tissue Engineering Research, 2017, 21(4): 527-531.

Figures/Tables 3

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