Effect of periodic mechanical stress on inflammatory responses and apoptosis of chondrocytes

doi:10.12307/2021.157

Abstract

Abstract: BACKGROUND: As chondrocyte is the only cell component of cartilage, chondrocyte phenotype loss is a characteristic change of osteoarthritis, while periodic mechanical stress stimulation has great influence on the biological characteristics of chondrocytes. Therefore, clarification of the mechanism of periodic mechanical stress on chondrocytes can provide reference for treating osteoarthritis.
OBJECTIVE: To observe the effect of periodic mechanical stress on inflammatory responses and apoptosis of chondrocytes in vitro.
METHODS: Primary chondrocytes from rats were inoculated into a Bioflex plate coated with type I collagen and were cultured for 1 day. Then, the cells were treated with Flexcell-5000T Tension System. Under the stretching frequency of 0.5 Hz, the chondrocytes were stretched for 24 hours. According to the stretching strain rate, the chondrocytes were divided into control group (0% stretching), 2% stretching group, 6% stretching group, 8% stretching group, 10% stretching group and 14% stretching group. Inflammatory responses of chondrocytes (interleukin-1β, tumor necrosis factor alpha) were detected by ELISA, cell apoptosis was detected by TUNEL staining and Annexin V-FITC/PI method, and the expression of endoplasmic reticulum stress signal molecules [Caspase-12, PKR-like endoplasmic reticulum kinase (PERK), glucose regulated protein 78 (GRP78)] was detected using RT-PCR.
RESULTS AND CONCLUSION: Under normal light microscope, normal cells (control group) were polygonal and irregular in shape. With the increase of mechanical stretching strain rate, the cells were exfoliated to varying degrees, with the edge being blurred and rounded. Moreover, cell apoptosis appeared. Compared with the control group, interleukin-1β and tumor necrosis factor alpha levels were higher in the force groups, especially in the 10% and 14% stretching groups (P < 0.05). Under the action of periodic mechanical stress, the apoptosis rates of the 2% stretching group and 6% stretching group were (12.45±1.64)% and (12.48±1.75)%, respectively, which did not significantly differ from that of the control group [(12.14±1.43)%; P > 0.05]. Compared with the control group, the apoptosis rates of 8%, 10% and 14% stretching groups were significantly higher [(29.18±3.04)%, (34.12±3.57)% and (44.32±45.08)%; P < 0.05]. The results of RT-PCR detection showed that the mRNA levels of Caspase-12, PERK and GRP78 in the 8%, 10% and 14% stretching groups were significantly higher than that in the control group (P < 0.05). The above findings indicate that periodic mechanical stress can accelerate the apoptosis of chondrocytes and increase the inflammatory responses of chondrocytes, which may be related to the initiation of endoplasmic reticulum stress pathway.

Key words: periodic mechanical stress, in vitro study, chondrocyte, inflammatory response, apoptosis

CLC Number:

Wu Weiting, Li Runguang, Cao Shenglu, Liang Shuangwu. Effect of periodic mechanical stress on inflammatory responses and apoptosis of chondrocytes[J]. Chinese Journal of Tissue Engineering Research, 2021, 25(29): 4608-4613.

Figures/Tables 9

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