Thoracic spinal kinematics is affected by the grade of intervertebral disc degeneration, but not by the presence of the ribs: An in vitro study

Objectives. Aberrant transforming growth factor β (TGFβ) activation is detrimental to both nucleus pulposus (NP) cells and cartilage endplates (CEPs), which can lead to intervertebral disc degeneration (IDD). Ligustrazine (LIG) reduces the expression of inflammatory factors and TGFβ1 in hypertrophic CEP to prevent IDD. In this study, we investigate the effects of LIG on NP cells and the TGFβ signaling. Design. LIG was injected to the lumbar spinal instability (LSI) mouse model. The effect of LIG was evaluated by intervertebral disc (IVD) score in the LSI mouse model. The expression of activated TGFβ was examined using immunostaining with pSmad2/3 antibody. The upright posture (UP) rat model was also treated and evaluated in the same manner to assess the effect of LIG. In ex vivo study, IVDs from four-week old mice were isolated and treated with 10−5, 10−6, and 10−7 M of LIG. We used western blot to detect activated TGFβ expression. TGFβ-treated human nucleus pulposus cells (HNPCs) were cotreated with optimized dose of LIG in vitro. Immunofluorescence staining was performed to determine pSmad2/3, connective tissue growth factor (CCN2), and aggrecan (ACAN) expression levels. Results. IVD score and the percentage of pSmad2/3+ NP cells were low in LIG-treated LSI mice in comparison with LSI mice, but close to the levels in the Sham group. Similarly, LIG reduced the overexpression of TGFβ1 in NP cells. The inhibitory effect of LIG was dose dependent. A dose of 10−5 M LIG not only strongly attenuated Smad2/3 phosphorylation in TGFβ-treated IVD ex vivo but also suppressed pSmad2/3, CCN2, and ACAN expression in TGFβ-treated NP cells in vitro. Conclusions. LIG prevents IDD via suppression of TGFβ overactivation in NP cells.

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Testing the impact of discoplasty on the biomechanics of the intervertebral disc with simulated degeneration: An in vitro study

Medical Engineering & Physics ◽

10.1016/j.medengphy.2020.07.024 ◽

2020 ◽

Vol 84 ◽

pp. 51-59 ◽

Cited By ~ 1

Author(s):

Chloé Techens ◽

Marco Palanca ◽

Peter Endre Éltes ◽

Áron Lazáry ◽

Luca Cristofolini

Keyword(s):

Intervertebral Disc ◽

In Vitro Study ◽

Vitro Study ◽

The Impact

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The noncoding RNA linc-ADAMTS5 cooperates with RREB1 to protect from intervertebral disc degeneration through inhibiting ADAMTS5 expression

Clinical Science ◽

10.1042/cs20160918 ◽

2017 ◽

Vol 131 (10) ◽

pp. 965-979 ◽

Cited By ~ 15

Author(s):

Kun Wang ◽

Yu Song ◽

Wei Liu ◽

Xinghuo Wu ◽

Yukun Zhang ◽

...

Keyword(s):

Intervertebral Disc ◽

Disc Degeneration ◽

Histone Deacetylases ◽

Noncoding Rna ◽

Intervertebral Disc Degeneration ◽

Physical Interaction ◽

Loss Of Function ◽

Element Binding Protein ◽

Long Intergenic Noncoding Rna

Previous studies have indicated the important roles of ADAMTS5 in intervertebral disc degeneration (IDD). However, the mechanisms that regulate ADAMTS5 expression in nuclear pulposus (NP) cells remain largely unknown. Evidence suggests that intergenic transcription may be associated with genes that encode transcriptional regulators. Here, we identified a long intergenic noncoding RNA, linc-ADAMTS5, which was transcribed in the opposite direction to ADAMTS5. In the present study, through mining computational algorithm programs, and publicly available data sets, we identified Ras-responsive element-binding protein 1 (RREB1) as a crucial transcription factor regulating the expression of ADAMTS5 in NP cells. RNA pull-down, RNA immunoprecipitation (RIP), in vitro binding assays, and gain- and loss-of-function studies indicated that a physical interaction between linc-ADAMTS5 and splicing factor proline/glutamine-rich (SFPQ) facilitated the recruitment of RREB1 to binding sites within the ADAMTS5 promoter to induce chromatin remodeling. This resulted in subdued ADAMTS5 levels in cultured NP cells involving histone deacetylases (HDACs). In clinical NP tissues, linc-ADAMTS5 and RREB1 were correlated negatively with ADAMTS5 expression. Taken together, these results demonstrate that RREB1 cooperates with noncoding RNA linc-ADAMTS5 to inhibit ADAMTS5 expression, thereby affecting degeneration of the extracellular matrix (ECM) of the intervertebral disc (IVD).

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Do early stages of lumbar intervertebral disc degeneration really cause instability? Evaluation of an in vitro database. Kettler A, Rohlmann F, Ring C, Mack C, Wilke HJ. Eur Spine J 2011;20(4):578–84. Epub 2010 Dec 2

The Spine Journal ◽

10.1016/j.spinee.2011.08.424 ◽

2011 ◽

Vol 11 (8) ◽

pp. 793

Keyword(s):

Intervertebral Disc ◽

Disc Degeneration ◽

Intervertebral Disc Degeneration ◽

Lumbar Intervertebral Disc ◽

Early Stages ◽

F Ring

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In-vitro study of the cryopreserved intervertebral disc

10.5353/th_b4129038 ◽

2008 ◽

Author(s):

Chun-wai Chan

Keyword(s):

Intervertebral Disc ◽

In Vitro Study ◽

Vitro Study

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Measurement of disc bulge of the intervertebral disc: an in vitro study using human cadaver lumbar spine segments

ArgoSpine News & Journal ◽

10.1007/s12240-010-0016-0 ◽

2010 ◽

Vol 22 (2) ◽

pp. 73-80 ◽

Cited By ~ 2

Author(s):

M. Cuchanski ◽

D. Cook ◽

M. Jegapragasan ◽

D. M. Whiting ◽

Boyle C. Cheng

Keyword(s):

Lumbar Spine ◽

Intervertebral Disc ◽

In Vitro Study ◽

Vitro Study ◽

Human Cadaver ◽

Disc Bulge

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Follistatin-Like 1 Attenuation Suppresses Intervertebral Disc Degeneration in Mice through Interacting with TNF-α and Smad Signaling Pathway

Oxidative Medicine and Cellular Longevity ◽

10.1155/2021/6640751 ◽

2021 ◽

Vol 2021 ◽

pp. 1-13

Author(s):

Shaoyi Wang ◽

Jianlu Wei ◽

Jie Shi ◽

Qiting He ◽

Xiaocong Zhou ◽

...

Keyword(s):

Intervertebral Disc ◽

Disc Degeneration ◽

Intervertebral Disc Degeneration ◽

Inflammatory Diseases ◽

Morphological Changes ◽

Intervertebral Discs ◽

Inflammatory Factors ◽

Wild Type ◽

Tnf Α

Background. Inflammation plays an important role in intervertebral disc degeneration (IDD). The protein follistatin-like 1 (FSTL1) plays a proinflammatory role in a variety of inflammatory diseases. Objectives. The purpose of this study was to investigate whether IDD could be delayed by inhibiting FSTL-1 expression. Methods. We established a puncture-induced IDD model in wild-type and FSTL-1+/- mice and collected intervertebral discs (IVDs) from the mice. Safranin O staining was used to detect cartilage loss of IVD tissue, and HE staining was used to detect morphological changes of IVD tissue. We measured the expression of FSTL-1 and related inflammatory indicators in IVD tissues by immunohistochemical staining, real-time PCR, and Western blotting. Results. In the age-induced model of IDD, the level of FSTL-1 increased with the exacerbation of degeneration. In the puncture-induced IDD model, FSTL-1-knockdown mice showed a reduced degree of degeneration compared with that of wild-type mice. Further experiments showed that FSTL-1 knockdown also significantly reduced the level of related inflammatory factors in IVD. In vitro experiments showed that FSTL-1 knockdown significantly reduced TNF-α-induced inflammation. Specifically, the expression levels of the inflammatory factors COX-2, iNOS, MMP-13, and ADAMTS-5 were reduced. Knockdown of FSTL-1 attenuated inflammation by inhibiting the expression of P-Smad1/5/8, P-Erk1/2, and P-P65. Conclusion. Knockdown of FSTL-1 attenuated inflammation by inhibiting the TNF-α response and Smad pathway activity and ultimately delayed IDD.

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