Biologic response of degenerative living human nucleus pulposus cells to treatment with cytokines.

Abstract

PURPOSE:

To investigate the molecular responses of various genes and proteins related to disc degeneration upon treatment with cytokines that affect disc-cell proliferation and phenotype in living human intervertebral discs (IVDs). Responsiveness to these cytokines according to the degree of disc degeneration was also evaluated.

MATERIALS AND METHODS:

The disc specimens were classified into two groups: group 1 (6 patients) showed mild degeneration of IVDs and group 2 (6 patients) exhibited severe degeneration of IVDs. Gene expression was analyzed after treatment with four cytokines: recombinant human bone morphogenic protein (rhBMP-2), transforming growth factor-β (TGF-β), interleukin-1β (IL-1β), and tumor necrosis factor-α (TNF-α). Molecular responses were assessed after exposure of cells from the IVD specimens to these cytokines via real-time polymerase chain reaction and immunofluorescence staining.

RESULTS:

mRNA gene expression was significantly greater for aggrecan, type I collagen, type II collagen, alkaline phosphatase, osteocalcin, and Sox9 in group 1 than mRNA gene expression in group 2, when the samples were not treated with cytokines. Analysis of mRNA levels for these molecules after morphogen treatment revealed significant increases in both groups, which were much higher in group 1 than in group 2. The average number of IVD cells that were immunofluorescence stained positive for alkaline phosphatase increased after treatment with rhBMP-2 and TGF-β in group 1.

CONCLUSION:

The biologic responsiveness to treatment of rhBMP-2, TGF-β, TNF-α, and IL-1β in the degenerative living human IVD can be different according to the degree of degeneration of the IVD.

• Authors: Sang Hyun Kim, Sung Uk Kuh, Keung Nyun Kim, Jeong Yoon Park, Ki Hong Cho, Dong Kyu Chin, Keun Su Kim, Yong Eun Cho
• Journal: Yonsei medical journal (Yonsei Med J) Vol. 56 Issue 1 Pg. 277-86 (Jan 2015) ISSN: 1976-2437 [Electronic] Korea (South)
• PMID: 25510775 (Publication Type: Journal Article, Research Support, Non-U.S. Gov't)

Chemical References

• Aggrecans
• Biological Products
• Bone Morphogenetic Protein 2
• Collagen Type I
• Collagen Type II
• Cytokines
• Interleukin-1
• RNA, Messenger
• Recombinant Proteins
• SOX9 Transcription Factor
• Transforming Growth Factor beta
• Tumor Necrosis Factor-alpha
• recombinant human bone morphogenetic protein-2
• Osteocalcin
• Alkaline Phosphatase

Topics

• Adult
• Aggrecans (genetics, metabolism)
• Alkaline Phosphatase (genetics, metabolism)
• Biological Products (pharmacology, therapeutic use)
• Bone Morphogenetic Protein 2 (pharmacology, therapeutic use)
• Collagen Type I (genetics, metabolism)
• Collagen Type II (genetics, metabolism)
• Cytokines (pharmacology, therapeutic use)
• Female
• Fluorescent Antibody Technique
• Gene Expression Regulation (drug effects)
• Humans
• Interleukin-1 (pharmacology, therapeutic use)
• Intervertebral Disc (drug effects, pathology)
• Intervertebral Disc Degeneration (drug therapy, genetics, pathology)
• Male
• Middle Aged
• Osteocalcin (genetics, metabolism)
• RNA, Messenger (genetics, metabolism)
• Recombinant Proteins (pharmacology, therapeutic use)
• SOX9 Transcription Factor (genetics, metabolism)
• Transforming Growth Factor beta (pharmacology, therapeutic use)
• Tumor Necrosis Factor-alpha (pharmacology)