Effect of biphasic electrical current stimulation on IL-1β-stimulated annulus fibrosus cells using in vitro microcurrent generating chamber system

Spine (Phila Pa 1976). 2013 Oct 15;38(22):E1368-76. doi: 10.1097/BRS.0b013e3182a211e3.

Abstract

Study design: Human annulus fibrosus (AF) cells were stimulated in vitro with interleukin (IL)-1β and exposed to biphasic electrical currents.

Objective: To identify the effect of biphasic electrical currents on the production of the extracellular matrix-modifying enzymes and inflammatory mediators in IL-1β-stimulated AF cells.

Summary of background data: Symptomatic disc degeneration is an important cause of chronic intractable lumbar pain and is associated with macrophage-mediated inflammation in the AF. The inflammatory reaction relationship has not been studied in the AF.

Methods: Human AF cells were treated with 1 ng/mL IL-1β and cultured in a microcurrent generating chamber system. The levels of matrix metalloproteinase (MMP)-1, MMP-3, tissue inhibitor of metalloproteinase (TIMP)-1, TIMP-2, IL-6, IL-8, vascular endothelial growth factor (VEGF), insulin-like growth factor, and nitric oxide (NO) were measured. Expression of cyclooxygenase 2 and type I collagen mRNA was analyzed.

Results: Compared with unstimulated cells, IL-1β-stimulated AF cells produced significantly higher levels of MMP-1, MMP-3, IL-6, IL-8, NO, and VEGF, and lower levels of TIMP-1 and TIMP-2. Exposure to a 250-mV/mm field induced time-dependent increases in IL-6, NO, MMP-1, TIMP-1, VEGF, and insulin-like growth factor-1 production. The cells exposed to 500-mV/mm field produced significantly less MMP-1, TIMP-1, IL-6, and VEGF than unexposed cells (MMP-1, 17.2 ± 4.7 ng/mL vs. 27.3 ± 3.9 ng/mL, P< 0.05; TIMP-1, 12.4 ± 3.3 ng/mL vs. 22.3 ± 2.1 ng/mL, P< 0.02; IL-6, 2.5 ± 0.9 ng/mL vs. 6.39 ± 1.90 ng/mL, P< 0.05; and VEGF, 0.1 ± 0.04 ng/mL vs. 0.44 ± 0.15 ng/mL, P< 0.03). NO production was markedly increased at 500 mV/mm (P< 0.0001).

Conclusion: We showed that exposure of IL-1β-stimulated AF cells to a 500 mV/mm inhibited MMP-1, IL-6, VEGF, and TIMP-1 production. The results suggest that biphasic electrical current stimulation may have efficacy in diminishing symptomatic disc degeneration.

Level of evidence: N/A.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Cells, Cultured
  • Collagen Type I / genetics
  • Cyclooxygenase 2 / genetics
  • Electric Stimulation / instrumentation*
  • Electric Stimulation / methods*
  • Enzyme-Linked Immunosorbent Assay
  • Female
  • Gene Expression / drug effects
  • Humans
  • Interleukin-1beta / pharmacology*
  • Interleukin-6 / metabolism
  • Interleukin-8 / metabolism
  • Intervertebral Disc / drug effects*
  • Intervertebral Disc / metabolism
  • Intervertebral Disc / pathology
  • Intervertebral Disc Degeneration / genetics
  • Intervertebral Disc Degeneration / metabolism
  • Intervertebral Disc Degeneration / pathology
  • Male
  • Matrix Metalloproteinase 1 / metabolism
  • Matrix Metalloproteinase 3 / metabolism
  • Nitric Oxide / metabolism
  • Reverse Transcriptase Polymerase Chain Reaction
  • Tissue Inhibitor of Metalloproteinase-1 / metabolism
  • Tissue Inhibitor of Metalloproteinase-2 / metabolism
  • Vascular Endothelial Growth Factor A / metabolism

Substances

  • Collagen Type I
  • Interleukin-1beta
  • Interleukin-6
  • Interleukin-8
  • Tissue Inhibitor of Metalloproteinase-1
  • Vascular Endothelial Growth Factor A
  • Tissue Inhibitor of Metalloproteinase-2
  • Nitric Oxide
  • Cyclooxygenase 2
  • Matrix Metalloproteinase 3
  • Matrix Metalloproteinase 1