Pamidronate Down-regulates Tumor Necrosis Factor-alpha Induced Matrix Metalloproteinases Expression in Human Intervertebral Disc Cells.
BACKGROUND
N-containing bisphosphonates (BPs), such as pamidronate and risedronate, can inhibit osteoclastic function and reduce osteoclast number by inducing apoptotic cell death in osteoclasts. The aim of this study is to demonstrate the effect of pamidronate, second generation nitrogen-containing BPs and to elucidate matrix metallo-proteinases (MMPs) mRNA expression under serum starvation and/or tumor necrosis factor alpha (TNF-α) stimulation on metabolism of intervertebral disc (IVD) cells in vitro.


METHODS
Firstly, to test the effect of pamidronate on IVD cells in vitro, various concentrations (10(-12), 10(-10), 10(-8), and 10(-6) M) of pamidronate were administered to IVD cells. Then DNA and proteoglycan synthesis were measured and messenger RNA (mRNA) expressions of type I collagen, type II collagen, and aggrecan were analyzed. Secondly, to elucidate the expression of MMPs mRNA in human IVD cells under the lower serum status, IVD cells were cultivated in full serum or 1% serum. Thirdly, to elucidate the expression of MMPs mRNA in IVD cells under the stimulation of 1% serum and TNF-α (10 ng/mL) In this study, IVD cells were cultivated in three dimensional alginate bead.


RESULTS
Under the lower serum culture, IVD cells in alginate beads showed upregulation of MMP 2, 3, 9, 13 mRNA. The cells in lower serum and TNF-α also demonstrated upregulation of MMP-2, 3, 9, and 13 mRNA. The cells with various doses of pamidronate and lower serum and TNF-α were reveled partial down-regulation of MMPs.


CONCLUSIONS
Pamidronate, N-containing second generation BPs, was safe in metabolism of IVD in vitro maintaining chondrogenic phenotype and matrix synthesis, and down-regulated TNF-α induced MMPs expression.
The Brain Korea 21 PLUS Project for Medical Science, Yonsei University College of Medicine, Seoul, Korea.
Young-Mi
YM
Kang
Department of Orthopaedic Surgery, Yonsei University College of Medicine, Seoul, Korea.
Seong-Hwan
SH
Hong
Department of Orthopaedic Surgery, Yonsei University College of Medicine, Seoul, Korea.
Jae-Ho
JH
Yang
Department of Orthopaedic Surgery, Yonsei University College of Medicine, Seoul, Korea.
Jin-Cheol
JC
Oh
Department of Orthopaedic Surgery, Yonsei University College of Medicine, Seoul, Korea.
Jin-Oh
JO
Park
Department of Orthopedic Surgery, Catholic Kwandong University College of Medicine, Incheon, Korea.
Byung Ho
BH
Lee
Department of Orthopaedic Surgery, Yonsei University College of Medicine, Seoul, Korea.
Sang-Yoon
SY
Lee
Department of Orthopaedic Surgery, Yonsei University College of Medicine, Seoul, Korea.
Hak-Sun
HS
Kim
Department of Orthopaedic Surgery, Yonsei University College of Medicine, Seoul, Korea.
Hwan-Mo
HM
Lee
The Brain Korea 21 PLUS Project for Medical Science, Yonsei University College of Medicine, Seoul, Korea.; Department of Orthopaedic Surgery, Yonsei University College of Medicine, Seoul, Korea.
Seong-Hwan
SH
Moon
Collagen; Intervertebral disc; Matrix metalloproteinases; Pamidronate; Tumor necrosis factor-alpha