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; Kerrigan, Mark J.P.

doi:10.1177/1947603511427556

Cited by 3 publications

(2 citation statements)

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“…This phenomenon is termed chondrocyte dedifferentiation. Chondrocytes, which are normally embedded in a 3‐dimensional cartilage matrix, lose their intrinsic phenotypes and exhibit a shift to fibroblast‐like cells in 2D culture (Qusous and Kerrigan, ). This marked change in cellular morphology is accompanied by changes in gene and protein expression, during which chondrocyte‐specific genes, such as collagen type II, aggrecan, and SOX9, are suppressed and the expression of a representative fibroblastic marker, collagen type I, is upregulated (Baghaban Eslaminejad et al, ; Ma et al, ).…”

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confidence: 99%

Focal Adhesion Assembly Induces Phenotypic Changes and Dedifferentiation in Chondrocytes

et al. 2016

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The expansion of autologous chondrocytes in vitro is used to generate sufficient populations for cell-based therapies. However, during monolayer culture, chondrocytes lose inherent characteristics and shift to fibroblast-like cells as passage number increase. Here, we investigated passage-dependent changes in cellular physiology, including cellular morphology, motility, and gene and protein expression, as well as the role of focal adhesion and cytoskeletal regulation in the dedifferentiation process. We found that the gene and protein expression levels of both the focal adhesion complex and small Rho GTPases are upregulated with increasing passage number and are closely linked to chondrocyte dedifferentiation. The inhibition of focal adhesion kinase (FAK) but not small Rho GTPases induced the loss of fibroblastic traits and the recovery of collagen type II, aggrecan, and SOX9 expression levels in dedifferentiated chondrocytes. Based on these findings, we propose a strategy to suppress chondrogenic dedifferentiation by inhibiting the identified FAK or Src pathways while maintaining the expansion capability of chondrocytes in a 2D environment. These results highlight a potential therapeutic target for the treatment of skeletal diseases and the generation of cartilage in tissue-engineering approaches. J. Cell. Physiol. 231: 1822-1831, 2016. © 2015 Wiley Periodicals, Inc.

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confidence: 99%

Focal Adhesion Assembly Induces Phenotypic Changes and Dedifferentiation in Chondrocytes

et al. 2016

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“…Moreover, LB treatment induced a significant increase in the number of cells expressing “fast” RVD (Kerrigan and Hall,2005), thereby implying that the actin cytoskeleton plays a role in the regulation of membrane transporters, possibly the NKCC (Kerrigan et al,2006; Ong et al,2010), and thus the RVI/RVD mechanisms. Conversely, both 2D cultured primary bovine articular chondrocytes and the chondrocytic cell line C‐20/A4 have been shown to exhibit NKCC‐dependent volume regulation (Qusous et al,2011; Qusous and Kerrigan,2011), possibly regulated by monolayer culture‐induced changes in actin organization (Idowu et al,2000).…”

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confidence: 99%

Novel methods for the quantification of changes in actin organization in chondrocytes using fluorescent imaging and linear profiling

Qusous

Parker

Geewan

et al. 2012

Microscopy Res & Technique

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We present three novel reproducible methodologies for the quantification of changes in actin organization from microscope images. Striation and integrative analysis were devised for the investigation of trans-cellular filaments and F-actin localization, respectively, in response to physiological or mechanical actin-modulatory conditions. Additionally, the Parker-Qusous (PQ) formula was developed as a measure of total quantity of F-actin, independent of cell volume changes, whereby fluorescence intensity was divided by the cube root of cell volume, squared. Values obtained were quantified in Mauricean Units (Mu; pixel/μm(3)). Upon isolation, there was a 49% decrease in total F-actin fluorescence from 1.91 ± 0.16 pixel/μm(3) (Mu) to 0.95 ± 0.55 Mu, whereas upon culture, an apparent increase in total fluorescence was deemed insignificant due to an increase in average cell volume, with a rise, however, in striation units (StU) from 1 ± 1 to 5 ± 1 StU/cell, and a decrease in percentage cortical fluorescence to 30.45% ± 1.52% (P = 7.8 × 10(-5)). Freshly isolated chondrocytes exhibited a decrease in total F-actin fluorescence to 0.61 ± 0.05 Mu and 0.32 ± 0.02 Mu, 10 min posthypertonic and hypotonic challenges, respectively. Regulatory volume decrease was inhibited in the presence of REV5901 with maintenance of actin levels at 1.15 Mu. Following mechanical impact in situ, there was a reduction in total F-actin fluorescence to 0.95 ± 0.08 Mu and 0.74 ± 0.06 Mu under isotonic and hypotonic conditions, respectively, but not under hypertonic conditions. We report simple methodologies for quantification of changes in actin organization, which will further our understanding of the role of actin in various cellular stress responses. These techniques can be applied to better quantify changes in localization of various proteins using fluorescent labeling.

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Adult Mesenchymal Stem Cell-Based Approaches for Osteoarthritis: Current Perspectives and Challenges

Hsiao

Chang

et al. 2021

J. Musculoskelet. Res.

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Osteoarthritis (OA) is a degenerative joint disease that may cause joint inflammation, stiffness, and pain. Current therapy for OA involves symptomatic treatment, mainly pain management. Therefore, it does not repair degenerated cartilage or attenuate joint inflammation. Because articular cartilage cannot heal or regenerate these tissues, tissue regeneration remains one of the most important objectives of new and potential OA therapeutics. The main features of adult mesenchymal stem cells (MSCs) are simple acquisition from adult tissues, rapid proliferation in vitro, immunomodulation in vivo, and lasting existence in the host, which are beneficial for OA treatment. In the past 15 years, adult MSCs, including bone marrow-, adipose-, and synovial membrane-derived MSCs, and their secretome have been successfully used in different animal (preclinical) models or in genetic manipulation for regenerating degenerated cartilage, reducing inflammation, or relieving pain. Furthermore, the implantation of adult MSCs showed pain reduction, anti-inflammation, and cartilage protection or healing in early-phase clinical trials. Adult MSCs are the most extensively explored as potential regenerative medicine for OA because of their efficacy in chondrocyte differentiation and their immunomodulatory properties. In this review paper, we highlighted current knowledge and future perspectives regarding preclinical tests, clinical application, and MSC-based/related products for curing OA.

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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

Cited by 3 publications

References 54 publications

Focal Adhesion Assembly Induces Phenotypic Changes and Dedifferentiation in Chondrocytes

Focal Adhesion Assembly Induces Phenotypic Changes and Dedifferentiation in Chondrocytes

Novel methods for the quantification of changes in actin organization in chondrocytes using fluorescent imaging and linear profiling

Adult Mesenchymal Stem Cell-Based Approaches for Osteoarthritis: Current Perspectives and Challenges

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