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The effect of capacitively coupled (CC) electrical stimulation on human disc nucleus pulposus cells and the relationship between CC and BMP-7

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Abstract

Purpose

An in vitro study using human intervertebral disc nucleus pulposus cells to evaluate the effects of CC stimulation on disc-matrix macromolecule production.

Methods

Nucleus pulposus cells were cultured in alginate beads and treated with CC stimulation. The effect of BMP on CC stimulation of the cells was evaluated by applying a BMP blocker (noggin) or by applying additional BMP-7 to the culture. The mRNA levels of the disc extracellular matrix genes (collagen I, II, aggrecan) and BMPs were measured by real-time PCR. The protein levels of aggrecan, collagen II, and BMPs were determined by ELISAs and Western blots. Sulfated glycosaminoglycan (sGAG) content was assayed using the DMMB method.

Results

(1) CC stimulation upregulates the production of the disc-matrix macromolecular components: sGAG, aggrecan and collagen II; (2) CC stimulation increases the ratio of mRNA expression levels of collagen II to collagen I; (3) CC stimulation induces the expression of endogenous BMP-4 and BMP-7; (4) inhibition of BMP activity (using noggin) reduces CC-mediated upregulation of aggrecan and collagen II; (5) CC and BMP-7 act in synergy to increase the upregulation of disc-matrix macromolecules.

Conclusion

CC stimulation upregulates the production of the intervertebral disc-matrix macromolecules aggrecan, collagen II, and sGAG by a mechanism involving BMPs. CC stimulation acts in synergy with BMP-7 to increase the upregulation of these disc-matrix macromolecules.

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Abbreviations

CC:

Capacitively coupled electrical stimulation

IVD:

Intervertebral disc

sGAG:

Sulfated glycosaminoglycan

NP:

Nucleus pulposus

DMMB:

Dimethylmethylene blue

ELISA:

Enzyme linked immunosorbent assay

BMP:

Bone morphogenic proteins

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Authors and Affiliations

  1. The Atlanta Department of Veterans Affairs Medical Center, The Emory Spine Center, Emory University School of Medicine, Atlanta, GA, USA

    Zili Wang, William C. Hutton & S. Tim Yoon

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  1. Zili Wang
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  2. William C. Hutton
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  3. S. Tim Yoon
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Correspondence to Zili Wang.

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Wang, Z., Hutton, W.C. & Yoon, S.T. The effect of capacitively coupled (CC) electrical stimulation on human disc nucleus pulposus cells and the relationship between CC and BMP-7. Eur Spine J 26, 240–247 (2017). https://doi.org/10.1007/s00586-016-4439-y

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  • DOI: https://doi.org/10.1007/s00586-016-4439-y

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