PMID- 21168098
OWN - NLM
STAT- MEDLINE
DCOM- 20110414
LR  - 20131121
IS  - 1878-1632 (Electronic)
IS  - 1529-9430 (Linking)
VI  - 11
IP  - 1
DP  - 2011 Jan
TI  - Bupivacaine--the deadly friend of intervertebral disc cells?
PG  - 46-53
LID - 10.1016/j.spinee.2010.11.001 [doi]
AB  - BACKGROUND CONTEXT: Bupivacaine is commonly used as an adjunct during provocative 
      discography and is administered intradiscally in patients with discogenic back 
      pain. Recent studies demonstrated that bupivacaine is cytotoxic for articular 
      chondrocytes in vitro at clinically used concentrations (0.25%-0.5%). PURPOSE: To 
      analyze a concentration-dependent effect of bupivacaine on cell viability and 
      gene expression of human intervertebral disc (IVD) cells in an in vitro model. 
      STUDY DESIGN: In vitro cell culture study. PATIENT SAMPLE: Disc cells were 
      isolated from human disc biopsies from 11 patients undergoing surgery because of 
      degenerative disc disease or disc herniation. OUTCOME MEASURES: Cell viability 
      and gene expression after exposure to bupivacaine. METHODS: Human IVD cells were 
      treated with different concentrations of bupivacaine for 2 (n=5) or 18 hours 
      (n=5) and analyzed for cell viability and proliferation 
      (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide assay). 
      Additionally, cells were prestimulated with interleukin-1 beta (IL-1beta) (5 ng/mL) 
      to increase the levels of proinflammatory cytokines and matrix-degrading enzymes 
      and thereafter treated with 0.75 mmol bupivacaine (as determined in the cell 
      viability test) for 2 (n=5) or 18 hours (n=5). Prestimulated cells with or 
      without bupivacaine treatment were analyzed for gene expression of IL-1beta, IL-6, 
      IL-8, tumor necrosis factor-alpha (TNF-alpha), cyclooxygenase-2 (COX-2), matrix 
      metalloproteinase-3 (MMP3), MMP9, MMP13, and a disintegrin and metalloproteinase 
      with thrombospondin motifs 4 (ADAMTS4) using real-time reverse 
      transcription-polymerase chain reaction. Statistical analysis was performed by 
      using the Mann-Whitney U test with a significance level of p<.05. RESULTS: After 
      18 hours, bupivacaine exhibited either a cytotoxic or a proliferative effect on 
      human IVD cells, depending on the concentration. Similar but lower effects could 
      be observed already after 2 hours. With a concentration of 0.75 mmol 
      (proliferative effect), bupivacaine significantly decreased messenger RNA levels 
      of TNF-alpha, COX-2, MMP13, and ADAMTS4 after 18 hours. In contrast, expression of 
      IL-6, IL-8, and MMP9 did not differ; expression of IL-1beta and MMP3 was stimulated 
      with 0.75 mmol. After 2 hours, we observed a reduction in the expression of 
      COX-2, MMP3, MMP13, and ADAMTS4, without any effect regarding IL-1beta. CONCLUSIONS: 
      Application of bupivacaine in clinically relevant concentrations was toxic for 
      IVD cells in vitro. A low concentration stimulated cell proliferation and reduced 
      gene expression of certain matrix-degrading enzymes and proinflammatory 
      cytokines. If these results can be corroborated in tissue explant models or 
      animal studies, caution regarding provocative discography with bupivacaine is 
      prompted.
CI  - Copyright (c) 2011 Elsevier Inc. All rights reserved.
FAU - Quero, Lilian
AU  - Quero L
AD  - Spine Research Group, Competence Centre for Applied Biotechnology and Molecular 
      Medicine, University of Zurich, Winterthurerstrasse 190, Zurich, Switzerland.
FAU - Klawitter, Marina
AU  - Klawitter M
FAU - Nerlich, Andreas G
AU  - Nerlich AG
FAU - Leonardi, Massimo
AU  - Leonardi M
FAU - Boos, Norbert
AU  - Boos N
FAU - Wuertz, Karin
AU  - Wuertz K
LA  - eng
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
PL  - United States
TA  - Spine J
JT  - The spine journal : official journal of the North American Spine Society
JID - 101130732
RN  - 0 (Cytokines)
RN  - EC 3.4.24.- (Matrix Metalloproteinases)
RN  - Y8335394RO (Bupivacaine)
SB  - IM
MH  - Bupivacaine/*pharmacology
MH  - Cell Proliferation/*drug effects
MH  - Cell Survival/*drug effects
MH  - Cells, Cultured
MH  - Cytokines/genetics/metabolism
MH  - Dose-Response Relationship, Drug
MH  - Gene Expression/*drug effects
MH  - Humans
MH  - Intervertebral Disc/cytology/*drug effects/metabolism
MH  - Matrix Metalloproteinases/genetics/metabolism
MH  - Reverse Transcriptase Polymerase Chain Reaction
MH  - Statistics, Nonparametric
EDAT- 2010/12/21 06:00
MHDA- 2011/04/16 06:00
CRDT- 2010/12/21 06:00
PHST- 2010/01/13 00:00 [received]
PHST- 2010/10/20 00:00 [revised]
PHST- 2010/11/03 00:00 [accepted]
PHST- 2010/12/21 06:00 [entrez]
PHST- 2010/12/21 06:00 [pubmed]
PHST- 2011/04/16 06:00 [medline]
AID - S1529-9430(10)01336-7 [pii]
AID - 10.1016/j.spinee.2010.11.001 [doi]
PST - ppublish
SO  - Spine J. 2011 Jan;11(1):46-53. doi: 10.1016/j.spinee.2010.11.001.