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Generation of Proinflammatory Mediator of Intervertebral Disc Cells by Nicotine Stimulation

  • Journal of Korean Society of Spine Surgery
  • Abbr : J Kor Spine Sur
  • 2014, 21(2), pp.84-89
  • Publisher : Korean Society Of Spine Surgery
  • Research Area : Medicine and Pharmacy > Orthopedic Surgery

서형연 1 김도연 2 윤주현 2

1전남대학교
2전남대학교 의과대학 정형외과학교실

Accredited

ABSTRACT

Study Design: Experimental investigation in vitro. Objectives: To evaluate the relationship between the degeneration of intervertebral disc cells, and low back pain induced bydegeneration of intervertebral disc cells and increases in use of proinflammatory mediators via nicotine stimulation. Summary of Literature Review: Smoking is a leading cause of degeneration of intervertebral disc cells and low back pain. Accordingto the existing literature, nicotine, one of the main ingredients in cigarettes, causes the degeneration of intervertebral disk cells includingdecrease of glycoprotein through generation of carboxy-hemoglobin, vasoconstriction, and disability of fibrinolysis and changes ofmetabolism of nucleus pulposus cells. Materials and Methods: Annulus fibrosus of intervertebral disc and knee joint cartilage were collected from pigs; these cells wereacquired by gradual enzyme decomposition. Using Trypan blue, concentration and survival rate of cells were examined; cells wereinserted on alginate beads for tertiary cultivation. Nicotine was then applied at 0, 50, 100, 200 and 300 nM, respectively, and the sampleswere cultivated for three, six and nine days, respectively. After collecting culture fluid, it was measured for interleukin(IL)-1β, IL-6 andIL-8 with the ELISA Test. DNA of cells used for cultivation was quantitated and the amount of the resulting proinflammatory mediatorwas normalized. The results were then compared with the result of same study on cartilage of porcine knee joints. Results: For changes of the inflammatory mediator based on the concentration of nicotine, in nicotine stimulation with low concentrationof 50 nM and the control group, there was no significant change, while transient increases of inflammatory mediator showed in nicotinestimulation with concentrations of 100, 200 nM, respectively. There was not a significant increase of IL-1β observed in all nicotinestimulation groups; these were the same results in porcine cartilage study. The level of IL-6 in 200, 300 nM nicotine concentrationshowed significant increases, respectively. The level of IL-8 in high dose nicotine stimulation groups also showed significant increasesof DNA on the sixth day. And in porcine cartilage study group, significant changes were observed in 200, 300 nM, but the absolute valuewas lower than that of annulus fibrous cells group. Conclusion: Inflammatory mediators such as IL-6 and IL-8 increased as the result of tertiary cultivation of annulus fibrosus cells ofporcine intervertebral disk and nicotine stimulation. It is believed that the cells of the disc annulus are more sensitive than articularchondrocytes to nicotine stimulation. This may be the focus of future long-term studies effects of nicotine other inflammatory cytokinesKey Words: Intervertebral disc, Annulus fibrosus, Proinflammatory mediator, Nicotine

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