PMID- 31101043 OWN - NLM STAT- MEDLINE DCOM- 20191230 LR - 20200225 IS - 1471-2474 (Electronic) IS - 1471-2474 (Linking) VI - 20 IP - 1 DP - 2019 May 17 TI - Expression of the RANK/RANKL/OPG system in the human intervertebral disc: implication for the pathogenesis of intervertebral disc degeneration. PG - 225 LID - 10.1186/s12891-019-2609-x [doi] LID - 225 AB - BACKGROUND: The expression of the receptor activator of nuclear factor kappa B (RANK) /RANK ligand (RANKL) /osteoprotegerin (OPG) system and its association with the progression of intervertebral disc (IVD) degeneration has recently been reported in a human IVD. However, the effect of the RANK/RANKL/OPG system on the matrix metabolism of human IVD cells, especially on the expression of catabolic factors relevant to IVD degeneration, remains unknown. The purpose of this study was to examine the expression of the RANK/RANKL/OPG system, and then to evaluate the effect of this system on the expression of catabolic factors by human IVD cells. METHODS: Annulus fibrosus (AF) and nucleus pulposus (NP) cells isolated by sequential enzyme digestion from human IVD tissues obtained during spine surgeries were monolayer cultured. The expression of the RANK/RANKL/OPG system was determined using immunohistochemical methods and real-time polymerase chain reaction (PCR). To evaluate the influence of interleukin-1 beta (IL-1beta) stimulation on the mRNA expression of RANK, RANKL, and OPG, recombinant human IL-1beta (rhIL-1beta) was administered in the culture media of IVD cells. To examine the influence of RANKL signaling on the expression of matrix metalloprotease-3 (MMP-3), MMP-13, and IL-1beta, the cells were cultured with exogenous recombinant human RANKL (rhRANKL), recombinant human OPG (rhOPG) or anti-human RANKL mouse monoclonal antibody (ahRANKL-mAB) with or without rhIL-1beta. RESULTS: Immunoreactivity to RANK/RANKL/OPG and the mRNA expression of the three genes were obviously identified in both AF and NP cells. rhIL-1beta stimulation significantly upregulated the mRNA expression level of RANK/RANKL/OPG. The mRNA expression of catabolic factors was significantly upregulated by stimulation of rhRANKL in the presence of rhIL-1beta. On the other hand, the administration of either rhOPG or ahRANKL-mAB significantly suppressed the mRNA expression of catabolic factors that had been upregulated by rhIL-1beta stimulation. The suppressive effect of ahRANKL-mAB against rhIL-1beta stimulation was also confirmed by the protein expression of MMP-3. CONCLUSIONS: The present study showed that the RANK/RANKL/OPG system may be involved in the progression of IVD degeneration. This study also suggested the potential use of anti-RANKL monoclonal antibody and OPG as therapeutic agents to suppress the progression of IVD degeneration. FAU - Sano, Tomohiko AU - Sano T AD - Department of Orthopaedic Surgery, Mie University Graduate School of Medicine, 2-174 Edobashi, Tsu City, Mie, 514-8507, Japan. FAU - Akeda, Koji AU - Akeda K AUID- ORCID: 0000-0001-9468-9387 AD - Department of Orthopaedic Surgery, Mie University Graduate School of Medicine, 2-174 Edobashi, Tsu City, Mie, 514-8507, Japan. k_akeda@clin.medic.mie-u.ac.jp. FAU - Yamada, Junichi AU - Yamada J AD - Department of Orthopaedic Surgery, Mie University Graduate School of Medicine, 2-174 Edobashi, Tsu City, Mie, 514-8507, Japan. FAU - Takegami, Norihiko AU - Takegami N AD - Department of Orthopaedic Surgery, Mie University Graduate School of Medicine, 2-174 Edobashi, Tsu City, Mie, 514-8507, Japan. FAU - Sudo, Takao AU - Sudo T AD - Department of Orthopaedic Surgery, Mie University Graduate School of Medicine, 2-174 Edobashi, Tsu City, Mie, 514-8507, Japan. FAU - Sudo, Akihiro AU - Sudo A AD - Department of Orthopaedic Surgery, Mie University Graduate School of Medicine, 2-174 Edobashi, Tsu City, Mie, 514-8507, Japan. LA - eng GR - 26861184/Grant-Aid for Scientific Research grants from the Japan Society for the Promotion of Science/ PT - Journal Article DEP - 20190517 PL - England TA - BMC Musculoskelet Disord JT - BMC musculoskeletal disorders JID - 100968565 RN - 0 (Osteoprotegerin) RN - 0 (RANK Ligand) RN - 0 (Receptor Activator of Nuclear Factor-kappa B) RN - 0 (Recombinant Proteins) RN - 0 (TNFRSF11A protein, human) RN - 0 (TNFRSF11B protein, human) RN - 0 (TNFSF11 protein, human) SB - IM MH - Aged MH - Aged, 80 and over MH - Animals MH - Cells, Cultured MH - Chondrocytes MH - Disease Progression MH - Female MH - Fibroblasts MH - Humans MH - Intervertebral Disc/cytology/*pathology MH - Intervertebral Disc Degeneration/*pathology/surgery MH - Male MH - Mice MH - Middle Aged MH - Osteoprotegerin/*metabolism MH - Primary Cell Culture MH - RANK Ligand/antagonists & inhibitors/*metabolism MH - Receptor Activator of Nuclear Factor-kappa B/*metabolism MH - Recombinant Proteins/metabolism PMC - PMC6525349 OTO - NOTNLM OT - Anti-receptor activator of nuclear factor kappa B ligand antibody OT - Degeneration OT - Intervertebral disc OT - Matrix-degrading enzyme OT - Osteoprotegerin OT - Proinflammatory cytokine OT - Receptor activator of nuclear factor kappa B OT - Receptor activator of nuclear factor kappa B ligand COIS- The authors declare that they have no competing interests. EDAT- 2019/05/19 06:00 MHDA- 2019/12/31 06:00 PMCR- 2019/05/17 CRDT- 2019/05/19 06:00 PHST- 2018/09/05 00:00 [received] PHST- 2019/05/02 00:00 [accepted] PHST- 2019/05/19 06:00 [entrez] PHST- 2019/05/19 06:00 [pubmed] PHST- 2019/12/31 06:00 [medline] PHST- 2019/05/17 00:00 [pmc-release] AID - 10.1186/s12891-019-2609-x [pii] AID - 2609 [pii] AID - 10.1186/s12891-019-2609-x [doi] PST - epublish SO - BMC Musculoskelet Disord. 2019 May 17;20(1):225. doi: 10.1186/s12891-019-2609-x.