Redifferentiation of aged human articular chondrocytes by combining bone morphogenetic protein-2 and melanoma inhibitory activity protein in 3D-culture

Stephan Payr; Brigitte Tichy; Clemens Atteneder; Marc Michel; Thomas Tiefenboeck; Nikolaus Lang; Sylvia Nuernberger; Stefan Hajdu; Elizabeth Rosado-Balmayor; Stefan Marlovits; Christian Albrecht

doi:10.1371/journal.pone.0179729

Redifferentiation of aged human articular chondrocytes by combining bone morphogenetic protein-2 and melanoma inhibitory activity protein in 3D-culture

PLoS One. 2017 Jul 13;12(7):e0179729. doi: 10.1371/journal.pone.0179729. eCollection 2017.

Authors

Stephan Payr^{1

2}, Brigitte Tichy^{1

2}, Clemens Atteneder^{1

2}, Marc Michel^{1

2}, Thomas Tiefenboeck^{1

2}, Nikolaus Lang^{1

2}, Sylvia Nuernberger^{1

2}, Stefan Hajdu^{1

2}, Elizabeth Rosado-Balmayor³, Stefan Marlovits^{1

2}, Christian Albrecht^{1

2}

Affiliations

¹ Department of Trauma Surgery, General Hospital, Medical University of Vienna, Austria.
² Austrian Cluster for Tissue Regeneration, Vienna, Austria.
³ Department of Experimental Trauma Surgery, Klinikum rechts der Isar, Technical University Munich, Germany.

Abstract

Melanoma inhibitory activity (MIA) affects the differentiation to hyaline cartilage and can inhibit the osteogenic potential of bone morphogenetic protein (BMP)-2 in mesenchymal stem cells (MSC). The aim of this study was to investigate if MIA also inhibits the osteogenic potential of BMP-2 in human articular chondrocytes during redifferentiation, which may lead to a higher grade of differentiation without calcification. HAC of four female patients (mean age: 73.75 ±6.98) were seeded into 3D culture for 28 days; after adding the recombinant proteins, four groups were formed (Control, BMP-2, MIA, BMP-2+MIA). Samples were analysed for gene expression, glycosaminoglycan (GAG) content and histology on day 0, 14 and 28. Collagen type 2 (COL2A1) was significantly increased in the BMP-2 containing groups on day 28; BMP-2 (100-fold, p = 0.001), BMP-2+MIA (65-fold, p = 0.009) and similar to the level of native cartilage. Higher aggrecan (Agg) levels were present in the BMP-2 (3-fold, p = 0.007) and BMP-2+MIA (4-fold, p = 0.002) group after 14 days and in the BMP-2 (9-fold, p = 0.001) group after 28 days. Collagen type 10 (COL10A1) was increased in the BMP-2 containing groups (6-fold, p = 0.006) but these levels were significantly below native cartilage. Alkaline phosphatase (ALP), collagen type 1 (COL1A1) and the glycosaminoglycan (GAG) content did not reveal any relevant differences between groups. BMP-2 is a potent inducer for differentiation of HAC. A significant enhancement of this effect in combination with MIA could not be observed. Furthermore no significant reduction of osteogenic markers during re-differentiation of chondrocytes was present combining BMP-2 and MIA.

MeSH terms

Aged
Aged, 80 and over
Bone Morphogenetic Protein 2 / pharmacology*
Cell Culture Techniques / methods*
Cell Differentiation
Cells, Cultured
Chondrocytes / cytology*
Chondrocytes / drug effects
Chondrocytes / metabolism
Collagen Type II / genetics*
Extracellular Matrix Proteins / pharmacology*
Female
Gene Expression Profiling
Gene Expression Regulation / drug effects
Glycosaminoglycans / metabolism
Humans

Substances

Bone Morphogenetic Protein 2
Collagen Type II
Extracellular Matrix Proteins
Glycosaminoglycans

Grants and funding

Study was funded by the medical scientific fund of the mayor of the city of Vienna (C.A.); project number 15002. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.