Glycophenotyping of Osteoarthritic Cartilage and Chondrocytes by RT-qPCR, Mass Spectrometry, Histochemistry with Plant/human Lectins and Lectin Localization with a Glycoprotein

Overview

Journal Arthritis Res Ther

Publisher Biomed Central

Specialty Rheumatology

Date 2013 Dec 3

PMID 24289744

Citations 25

Authors

Stefan Toegel

Daniela Bieder

Sabine Andre

Friedrich Altmann

Sonja M Walzer

Herbert Kaltner

Jochen G Hofstaetter

Reinhard Windhager

Hans-Joachim Gabius

Affiliations

Soon will be listed here.

Abstract

Introduction: This study aimed to characterize the glycophenotype of osteoarthritic cartilage and human chondrocytes.

Methods: Articular knee cartilage was obtained from nine osteoarthritis (OA) patients. mRNA levels for 27 glycosyltransferases were analyzed in OA chondrocytes using RT-qPCR. Additionally, N- and O-glycans were quantified using mass-spectrometry. Histologically, two cartilage areas with Mankin scores (MS) either ≤ 4 or ≥ 9 were selected from each patient representing areas of mild and severe OA, respectively. Tissue sections were stained with (1) a selected panel of plant lectins for probing into the OA glycophenotype, (2) the human lectins galectins-1 and -3, and (3) the glycoprotein asialofetuin (ASF) for visualizing β-galactoside-specific endogenous lectins.

Results: We found that OA chondrocytes expressed oligomannosidic structures as well as non-, mono- and disialylated complex-type N-glycans, and core 2 O-glycans. Reflecting B4GALNT3 mRNA presence in OA chondrocytes, LacdiNAc-terminated structures were detected. Staining profiles for plant and human lectins were dependent on the grade of cartilage degeneration, and ASF-positive cells were observed in significantly higher rates in areas of severe degeneration.

Conclusions: In summary, distinct aspects of the glycome in OA cartilage are altered with progressing degeneration. In particular, the alterations measured by galectin-3 and the pan-galectin sensor ASF encourage detailed studies of galectin functionality in OA.

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