Abstract

Mechanical overloading is a risk factor of disc degeneration. Studies have demonstrated that resveratrol helps to maintain the disc cell’s healthy biology. The present study aims to investigate whether resveratrol can suppress mechanical overloading-induced nucleus pulposus (NP) cell senescence in vitro and the potential mechanism. The isolated rat NP cells were seeded in the decalcified bone matrix (DBM) and cultured under non-compression (control) and compression (20% deformation, 1.0 Hz, 6 h/day) for 5 days using the mechanically active bioreactor. The resveratrol (30 and 60 μM) was added into the culture medium of the compression group to investigate its protective effects against the NP cell senescence. NP cell senescence was evaluated by cell proliferation, cell cycle, senescence-associated β-galactosidase (SA-β-Gal) activity, telomerase (TE) activity, and gene expression of the senescence markers (p16 and p53). Additionally, the reactive oxygen species (ROS) content and activity of the NF-κB pathway were also analyzed. Compared with the non-compression group, the high-magnitude compression significantly promoted NP cell senescence, increased ROS generation and activity of the NF-κB pathway. However, resveratrol partly attenuated NP cell senescence, decreased ROS generation and activity of the NF-κB pathway in a concentration-dependent manner under mechanical compression. Resveratrol can alleviate mechanical overloading-induced NP cell senescence through regulating the ROS/NF-κB pathway. The present study provides that resveratrol may be a potential drug for retarding mechanical overloading-induced NP cell senescence.

Highlights

  • Intervertebral disc (IVD) degeneration (IDD) is a fundamental structure that interspaces and connects the adjacent vertebral bones [1]

  • Because subcultivation can lead to cellular senescence, passage 2 (P2) nucleus pulposus (NP) cells were first suspended in the collagen solution (1 mg/ml, Shengyou, China) and were seeded into the prepared bovine decalcified bone matrix (DBM, 10 × 10 × 5 mm, 1 × 107 cells per DBM) scaffold as previously described [25,26]

  • Results showed that the value of optical density value at 450 nm (OD450) and relative fluorescence units (RFU) (490/585 nm) in the compression group (20% deformation) was significantly decreased compared with the control group

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Summary

Introduction

Intervertebral disc (IVD) degeneration (IDD) is a fundamental structure that interspaces and connects the adjacent vertebral bones [1]. Degenerative changes first occur in the disc NP region, which leads to decrease in NP cellular density and increase in NP matrix degradation [5,6]. Amongst these degenerative changes, disc NP cell senescence is a classical feature during disc degeneration and is often identified to be positively and closely correlated with disc degeneration grade [7,8,9]

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