Characterization and Inhibition of Fibrin Hydrogel-Degrading Enzymes During Development of Tissue Engineering Scaffolds

The goal of articular cartilage tissue engineering is to provide cartilaginous constructs to replace abnormal cartilage. We have evaluated the chondroprogenitor clonal cell line RCJ3.1C5.18 (C5.18) as a model to guide the development of appropriate scaffolds for tissue engineering. Rapid degradation...

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Bibliographic Details
Published in:Tissue engineering 2007-07, Vol.13 (7), p.1469-1477
Main Authors: Ahmed, Tamer A.E., Griffith, May, Hincke, Max
Format: Article
Language:English
Subjects:
Animals
Biocompatible Materials - metabolism
Cartilage
Cartilage, Articular - cytology
Cartilage, Articular - enzymology
Cell Line
Enzyme Inhibitors
Enzymes
Enzymes - chemistry
Fibrin - metabolism
Hydrogels - metabolism
Proteins
Rats
TECHNOLOGY
TEKNIKVETENSKAP
Tissue Engineering
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Summary:The goal of articular cartilage tissue engineering is to provide cartilaginous constructs to replace abnormal cartilage. We have evaluated the chondroprogenitor clonal cell line RCJ3.1C5.18 (C5.18) as a model to guide the development of appropriate scaffolds for tissue engineering. Rapid degradation of fibrin hydrogels was observed after encapsulation of C5.18 cells. The enzymes responsible for this fibrin gel breakdown were characterized to control their activity and regulate gel stability. Western blotting, confirming zymography, revealed bands due to matrix metalloproteinases (MMP-2, MMP-3) that are secreted concomitantly with fibrin hydrogels breakdown. High plasmin activity was detected in conditioned media during hydrogel breakdown but not in the confluent cells before encapsulation. Reverse transcriptase polymerase chain reaction indicated the expression of MMP-2, -3, and -9 and plasminogen in the cells. MMP-9 was 100 times higher at day 1, whereas MMP-2 started to increase and reached its maximum level by day 7. Aprotinin, a known serine protease inhibitor, and galardin (GM6001), a potent MMP inhibitor, in combination or separately, prevented the breakdown of fibrin-C5.18 hydrogels, whereas only the combination of both promoted the accumulation of extracellular matrix. These findings suggest that plasmin and MMPs contribute independently to fibrin hydrogel breakdown, but that either enzyme can achieve extracellular matrix breakdown.
ISSN:1076-3279
1557-8690
1557-8690
DOI:10.1089/ten.2006.0354