BMP-2, hypoxia, and COL1A1/HtrA1 siRNAs favor neo-cartilage hyaline matrix formation in chondrocytes.

Abstract

Osteoarthritis (OA) is an irreversible pathology that causes a decrease in articular cartilage thickness, leading finally to the complete degradation of the affected joint. The low spontaneous repair capacity of cartilage prevents any restoration of the joint surface, making OA a major public health issue. Here, we developed an innovative combination of treatment conditions to improve the human chondrocyte phenotype before autologous chondrocyte implantation. First, we seeded human dedifferentiated chondrocytes into a collagen sponge as a scaffold, cultured them in hypoxia in the presence of a bone morphogenetic protein (BMP), BMP-2, and transfected them with small interfering RNAs targeting two markers overexpressed in OA dedifferentiated chondrocytes, that is, type I collagen and/or HtrA1 serine protease. This strategy significantly decreased mRNA and protein expression of type I collagen and HtrA1, and led to an improvement in the chondrocyte phenotype index of differentiation. The effectiveness of our in vitro culture process was also demonstrated in the nude mouse model in vivo after subcutaneous implantation. We, thus, provide here a new protocol able to favor human hyaline chondrocyte phenotype in primarily dedifferentiated cells, both in vitro and in vivo. Our study also offers an innovative strategy for chondrocyte redifferentiation and opens new opportunities for developing therapeutic targets.

• Authors: David Ollitrault, Florence Legendre, Carole Drougard, Mélanie Briand, Hervé Benateau, Didier Goux, Hanane Chajra, Laurent Poulain, Daniel Hartmann, Denis Vivien, Vijayalakshmi Shridhar, Alfonso Baldi, Frédéric Mallein-Gerin, Karim Boumediene, Magali Demoor, Philippe Galera
• Journal: Tissue engineering. Part C, Methods (Tissue Eng Part C Methods) Vol. 21 Issue 2 Pg. 133-47 (Feb 2015) ISSN: 1937-3392 [Electronic] United States
• PMID: 24957638 (Publication Type: Journal Article, Research Support, Non-U.S. Gov't)

Chemical References

• Bone Morphogenetic Protein 2
• Collagen Type I
• Collagen Type I, alpha 1 Chain
• RNA, Messenger
• RNA, Small Interfering
• High-Temperature Requirement A Serine Peptidase 1
• HTRA1 protein, human
• Serine Endopeptidases

Topics

• Aged
• Aged, 80 and over
• Animals
• Bone Morphogenetic Protein 2 (pharmacology)
• Cartilage, Articular (cytology)
• Cattle
• Cell Hypoxia (drug effects)
• Cells, Cultured
• Chondrocytes
• Chondrogenesis (drug effects)
• Collagen Type I (metabolism)
• Collagen Type I, alpha 1 Chain
• Extracellular Matrix (drug effects, metabolism)
• High-Temperature Requirement A Serine Peptidase 1
• Humans
• Hyalin (metabolism)
• Hypertrophy
• Kinetics
• Mice, Nude
• Middle Aged
• Osteogenesis (drug effects)
• Phenotype
• RNA, Messenger (genetics, metabolism)
• RNA, Small Interfering (metabolism)
• Serine Endopeptidases (metabolism)