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Osteoarthritis Cartilage. 2007 Oct;15(10):1207-16. Epub 2007 May 23.

Structure of pericellular matrix around agarose-embedded chondrocytes.

Author information

  • 1Center for Biomedical Engineering and Biological Engineering Division, Massachusetts Institute of Technology, Cambridge, MA, USA. michael.dimarco@genzyme.com

Abstract

OBJECTIVE:

Determine whether the structure of the type VI collagen component of the chondrocyte pericellular matrix (PCM) generated by agarose-embedded chondrocytes in culture is similar to that found in native articular cartilage.

METHODS:

Confocal microscopy, quick-freeze deep-etch electron microscopy, and real-time polymerase chain reaction (PCR) were used to investigate temporal and spatial patterns of type VI collagen protein deposition and gene expression by bovine chondrocytes during 4 weeks of culture within a 2% agarose hydrogel. Similar analyses were performed on chondrocytes within samples of intact cartilage obtained from the same joint surfaces as those used for cell isolation for comparison.

RESULTS:

Type VI collagen accumulated uniformly around cells embedded in agarose, with the rate of deposition slowing after the second week. After 1 week, PCM fibrils were observed to be oriented perpendicular to the cell surface, in contrast with the primarily tangential fibrillar arrangement observed in native articular cartilage. Expression of col6 in agarose-embedded cells was initially much higher ( approximately 400%) than that in chondrocytes within cartilage. Expression of col6 in the cultured chondrocytes declined by approximately 60% after 1 week, and remained stable thereafter.

CONCLUSIONS:

PCM structure and composition around cells in a hydrogel scaffold may be different than that in native cartilage, with potential implications for mass transport, mechanotransduction, and ultimately, the success of tissue engineering approaches.

PMID:
17524677
[PubMed - indexed for MEDLINE]
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