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Background/Aims: Mechanical overloading-induced nucleus pulposus (NP) apoptosis plays an important role in the pathogenesis of intervertebral disc degeneration. N-cadherin (N-CDH)-mediated signaling preserves normal NP cell phenotype. This study aims to investigate the effects of N-CDH on NP cell apoptosis under high-magnitude compression and the underlying mechanism behind this process. Methods: Rat NP cells seeded on scaffold were perfusion-cultured using a self-developed perfusion bioreactor for 5 days and experienced different magnitudes (2% and 20% compressive deformation, respectively) of compression at a frequency of 1.0 Hz for 4 hours once per day. The un-loaded NP cells were used as controls. Lentivirus-mediated N-CDH overexpression and inhibitor LY294002 were used to further investigate the role of N-CDH and PI3K/Akt pathway under highmagnitude compression, respectively. NP cell apoptosis was evaluated by caspase-3 activity measured using a commercial kit, flow cytometry, and expression of apoptosis-related molecules analyzed by real-time PCR and western blotting assays. Results: High-magnitude compression significantly increased apoptotic NP cells, caspase-3 activity and expression of pro-apoptotic molecules (Bax and caspase-3/cleaved caspase-3), but decreased expression of anti-apoptotic molecule (Bcl-2). High-magnitude compression decreased expression of N-CDH, p-Akt and p-GSK-3β. However, N-CDH overexpression attenuated NP cell apoptosis and increased expression of p-Akt and p-GSK-3β under high-magnitude compression. Further decreased expression of p-GSK-3β, but had no significant effects on N-CDH expression under high-magnitude compression. Conclusion: N-CDH can attenuate NP cell apoptosis through activating the PI3K/Akt-GSK-3β signaling under high-magnitude compression. [ABSTRACT FROM AUTHOR]

Mechanical stimuli participate in disc development and remodelling. However, the effects of mechanical load on the immature annulus fibrosus (AF) are largely unclear. This study aimed to investigate how the immature AF responded to dynamic compressive magnitude and duration. Immature porcine discs were bioreactor-cultured for 7 days and then dynamically compressed at various magnitudes (0.1, 0.2, 0.4, 0.8 and 1.3 MPa at a frequency of 1.0 Hz for 2 h/day) and durations (1, 2, 4 and 8 h/day at a magnitude of 0.4 MPa and a frequency of 1.0 Hz). Non-compressed discs were used as controls. The immature AF tissue was analysed for histology, gene expression (aggrecan, collagen I, ADAMTS-4, MMP-3, TIMP-1 and TIMP-3), biochemical content of glycosaminoglycans (GAG) and hydroxyproline (HYP) and aggrecan immunohistochemical staining. In the lower-compressivemagnitude groups (0.1, 0.2 and 0.4 MPa), the immature AF showed an up-regulation in the expression of matrix genes, GAG and HYP content and aggrecan deposition. In the compression duration groups, the GAG and HYP content and aggrecan deposition declined to a minimum in the 8-hour group, in which a catabolic gene expression profile was found. In conclusion, this study indicated that the effects of dynamic compression on the immature AF are magnitude and duration dependent and that catabolic remodelling within the immature AF can be induced by high compressive magnitudes and long compressive durations. [ABSTRACT FROM AUTHOR]

Background/Aims: Matrix homeostasis within the disc nucleus pulposus (NP) tissue is important for disc function. Increasing evidence indicates that sex hormone can influence the severity of disc degeneration. This study was aimed to study the role of 17β-estradiol (E2) in NP matrix synthesis and its underlying mechanism. Methods: Rat NP cells were cultured with (10-5, 10-7 and 10-9 M) or without (control) E2 for 48 hours. The estrogen receptor (ER)-βantagonist PHTPP and ERβ agonist ERB 041 were used to investigate the role mediated by ERβ. The p38 MAPK inhibitor SB203580 was used to investigate the role of p38 MAPK signaling pathway. Gene and protein expression of SOX9, aggrecan and collagen II, glycosaminoglycan (GAG) content, and immunostaining assay for aggrecan and collagen II were analyzed to evaluate matrix production in rat NP cells. Results: E2 enhanced NP matrix synthesis in a concentration-dependent manner regarding gene and proetin expression of SOX9, aggrecan and collagen II, protein deposition of aggrecan and collagen II, and GAG content. Moreover, activation of p38 MAPK signaling pathway was increased with elevating E2 concentration. Further analysis indicated that ERB 041 and PHTPP could respectively enhance and suppress effects of E2 on matrix synthesis in NP cells, as well as activation of p38 MAPK pathway. Additionally, inhibition of p38 MAPK signaling pathway significantly abolished the effects of E2 on matrix synthesis. Conclusion: E2 can enhance matrix synthesis of NP cells and the ERβ/p38 MAPK pathway is involved in this regulatory process. [ABSTRACT FROM AUTHOR]