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A Novel in Vitro Model to Investigate Behavior of Articular Chondrocytes in Osteoarthritis

Lookup NU author(s): Dr Kenneth RankinORCiD, Dr Rachel Lakey, Craig Gerrand, Andrew Sprowson, Professor Andrew McCaskie, Dr Mark Birch

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Abstract

Objective. To investigate in vivo simulation of the microenvironment in which osteoarthritis (OA) chondrocytes are cultured in vitro. Methods. Human articular chondrocytes were cultured Under normoxic and hypoxic conditions cells were cultured on standard culture plastic or a porous polyHEMA surface that closely resembles the in vivo cartilage microarchitecture. Morphological changes to the cells were demonstrated by fluorescent staining with DAPI and vinculin. Proteoglycan and type II collagen protein levels were assessed using established techniques. Matrix metalloproteinase-1 (MMP-1) production was assessed by ELISA. The gene expression of type II collagen and SOX9 was measured Using real-time polymerase chain reaction. Results. Cells grown on culture plastic were seen to be flat and hexagonal. Cells cultured on the porous polyHEMA surface exhibited morphology in keeping with the in vivo microenvironment. Glycosaminoglycan release in hypoxia was from cells cultured on standard culture plastic. Transcriptional expression of type II collagen was upregulated in hypoxia and by culture on the polyHEMA surface. Transcriptional expression of SOX9 in hypoxia was upregulated compared to normoxia: no significant effect was seen by varying the culture surface. Translational expression of type II collagen was upregulated at 20% oxygen on the polyHEMA surface compared to culture plastic and this was related to MMP-1 expression. Conclusion. Culture of chondrocytes in hypoxia and on a porous surface simulates the in vivo microenvironment and illustrates the molecular mechanisms of OA. (First Release Dec 23 2009: J Rheumatol 2010:37:426-31: doi: 10.3899/jrheum.080080)

Publication metadata

Author(s): Rankin KS, Lakey RL, Gerrand CH, Sprowson AP, McCaskie AW, Birch MA

Publication type: Article

Publication status: Published

Journal: Journal of Rheumatology

Year: 2010

Volume: 37

Issue: 2

Pages: 426-431

ISSN (print): 0315-162X

ISSN (electronic): 1499-2752

Publisher: Journal of Rheumatology Publishing Co. Ltd.

URL: http://dx.doi.org/10.3899/jrheum.080080

DOI: 10.3899/jrheum.080080

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