REGULATION OF GELATINASE-A (MMP-2) PRODUCTION BY OVINE INTERVERTEBRAL DISC NUCLEUS PULPOSUS CELLS GROWN IN ALGINATE BEAD CULTURE BY TRANSFORMING GROWTH FACTOR-β1AND INSULIN LIKE GROWTH FACTOR-I
| Autor: | Peter Ghosh, Thomas K. F. Taylor, Scott T. Pattison, James Melrose |
|---|---|
| Rok vydání: | 2001 |
| Předmět: |
Matrix Metalloproteinases
Membrane-Associated Alginates Angiogenesis medicine.medical_treatment Cell Gelatinase A Cell Culture Techniques Matrix metalloproteinase Transforming Growth Factor beta1 Insulin-like growth factor Glucuronic Acid Transforming Growth Factor beta medicine Animals Insulin-Like Growth Factor I Intervertebral Disc Cells Cultured Enzyme Precursors Tissue Inhibitor of Metalloproteinase-2 Wound Healing Sheep Chemistry Hexuronic Acids Metalloendopeptidases Intervertebral disc Cell Biology General Medicine Anatomy Tissue inhibitor of metalloproteinase Microspheres Cell biology medicine.anatomical_structure Gelatinases Matrix Metalloproteinase 2 Transforming growth factor |
| Zdroj: | Cell Biology International. 25:679-689 |
| ISSN: | 1065-6995 |
| DOI: | 10.1006/cbir.2000.0718 |
| Popis: | The aim of this study was to gain information relevant to disc repair processes. Limited degradation of the collagen matrix by matrix metalloproteases (MMPs) may facilitate the loosening of cell-cell and cell-matrix interactions within the injured intervertebral disc (IVD) to favour the penetration of blood vessels and migration of fibroblasts into the defect to promote repair processes. Gelatinase A (MMP-2) has a particularly important role to play in angiogenesis, in the present study we investigated the in vitro regulation of MMP-2 by Transforming Growth Factor-beta 1 (TGF-beta 1) and Insulin-like Growth Factor-1 (beta IGF-I) in cells from the nucleus pulposus (NP) of the ovine IVD. Ovine NP cells were grown in alginate bead cultures in complete medium (10% foetal calf serum) for 7 days, established in serum-free conditions for 24 h, then stimulated with TGF-beta 1 (0.1 or 10 ng/ml) or IGF-I (2 or 50 ng/ml) +/-Concanavalin A (20 microg/ml) for an additional 48 h. Conditioned medium was examined for matrix metalloproteases using gelatin zymography, Tissue Inhibitor of Metalloproteinase 2 (TIMP-2) and Membrane Type 1 Matrix Metalloproteinase (MT1-MMP) were immunolocalised in beads. Pro (72 kDa) and active (59 kDa) MMP-2 were the major gelatinolytic MMPs detected in control cultures, the TGF-beta 1 and IGF-I treatments significantly decreased levels of the active MMP-2, inclusion of Concanavalin A resulted in a complete reversal of this trend with IGF-I, and to a lesser extent with TGF-beta 1. Cell surface levels of TIMP-2 and MT1-MMP were decreased by the TGF-beta 1 treatment while IGF-I only appeared to decrease TIMP-2 expression. The findings of this study provide some insight as to why dense avascular connective tissues such as the intervertebral disc have such a poor healing potential. |
| Databáze: | OpenAIRE |
| Externí odkaz: |
|
Plný text