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Quantification of changes in morphology, mechanotransduction, and gene expression in bovine articular chondrocytes in response to 2-dimensional culture indicates the existence of a novel phenotype

Quantification of changes in morphology, mechanotransduction, and gene expression in bovine articular chondrocytes in response to 2-dimensional culture indicates the existence of a novel phenotype

Qusous, Ala and Kerrigan, Mark J.P. (2012) Quantification of changes in morphology, mechanotransduction, and gene expression in bovine articular chondrocytes in response to 2-dimensional culture indicates the existence of a novel phenotype. Cartilage, 3 (3). pp. 222-234. ISSN 1947-6035 (Print), 1947-6043 (Online) (doi:10.1177/1947603511427556)

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Abstract

Objective: Matrix-induced autologous chondrocyte implantation (ACI) offers a potential solution for cartilage repair but is currently hindered by loss of the chondrocyte differentiated phenotype. To further our understanding of the mechanism of dedifferentiation, changes in the phenotype in relation to mechanotransduction were recorded in response to monolayer culture.

Methods: Bovine cartilage explants were excised and chondrocytes cultured for 9 days (P1), 14 days (P2), and 21 (P3) days. Changes in morphology and regulatory volume increase (RVI; a mechanotransduction response) were determined by the expression of key genes by RT-PCR and confocal microscopy, respectively.

Results: A loss of a differentiated phenotype was observed in P1 with a reduction in sphericity and an overall increase in cell volume from 474.7 ± 32.1 µm3 to 725.2 ± 35.6 µm3. Furthermore, the effect of 2-dimensional (2-D) culture-induced dedifferentiation on mechanotransduction was investigated, whereby RVI and Gd3+-sensitive REV5901-induced calcium rise were only observed in 2-D cultured chondrocytes. A significant up-regulation of types I and II collagens and Sox9 was observed in P1 chondrocytes and no further significant change in type I collagen but a return to baseline levels of type II collagen and Sox9 upon further culture.

Conclusion: These data indicated the presence of an intermediate, mesodifferentiated phenotype and highlight the importance of mechanotransduction as a marker of the chondrocytic cell type.

Item Type: Article
Additional Information: [1] Originally published online 17 February 2012, more recent version published July 17, 2012. [2] Supplementary material for this article is available on the Cartilage website at http://cart.sagepub.com/supplemental. [3] Cartilage is published on behalf of the International Cartilage Repair Society.
Uncontrolled Keywords: phenotype, regulatory volume increase, collagen, dedifferentiation, calcium, REV5901
Subjects: Q Science > QR Microbiology
R Medicine > RM Therapeutics. Pharmacology
Pre-2014 Departments: School of Science
Related URLs:
Last Modified: 14 Oct 2016 09:23
Selected for GREAT 2016: None
Selected for GREAT 2017: None
Selected for GREAT 2018: None
URI: http://gala.gre.ac.uk/id/eprint/9302

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