Abstract
Cartilage engineering is a new strategy for the treatment of cartilage damage due to osteoarthritis or trauma in humans. Racehorses are exposed to the same type of cartilage damage and the anatomical, cellular, and biochemical properties of their cartilage are comparable to those of human cartilage, making the horse an excellent model for the development of cartilage engineering. Human mesenchymal stem cells (MSCs) differentiated into chondrocytes with chondrogenic factors in a biomaterial appears to be a promising therapeutic approach for direct implantation and cartilage repair. Here, we characterized equine umbilical cord blood-derived MSCs (eUCB-MSCs) and evaluated their potential for chondrocyte differentiation for use in cartilage repair therapy. Our results show that isolated eUCB-MSCs had high proliferative capacity and differentiated easily into osteoblasts and chondrocytes, but not into adipocytes. A three-dimensional (3D) culture approach with the chondrogenic factors BMP-2 and TGF-β1 potentiated chondrogenic differentiation with a significant increase in cartilage-specific markers at the mRNA level (Col2a1, Acan, Snorc) and the protein level (type II and IIB collagen) without an increase in hypertrophic chondrocyte markers (Col10a1 and Mmp13) in normoxia and in hypoxia. However, these chondrogenic factors caused an increase in type I collagen, which can be reduced using small interfering RNA targeting Col1a2. This study provides robust data on MSCs characterization and demonstrates that eUCB-MSCs have a great potential for cartilage tissue engineering.
Highlights
Osteoarthritis is the most common musculoskeletal disease in horses and is characterized by varying degrees of lameness, the presence of joint effusion, the occurrence of osteophytes, an increase in bone density, and the reduction of joint space in more advanced cases [1]
Among the various biomaterials available, type I/III collagen sponges show efficient chondrogenic differentiation of human umbilical cord blood (UCB)-mesenchymal stem cells (MSCs) when used with bone morphogenetic proteins (BMPs)-2 and TGF-β1, leading to a strong increase in mature chondrocyte-specific markers (COL2A1, COL2A, ACAN) and cellular phenotype stability in the neo-tissue [30]
Equine MSCs were successfully isolated in 22 UCB samples; the two other UCB samples had been processed more than 40 h after collection, hindering the emergence of colonies
Summary
Osteoarthritis is the most common musculoskeletal disease in horses and is characterized by varying degrees of lameness, the presence of joint effusion, the occurrence of osteophytes, an increase in bone density, and the reduction of joint space in more advanced cases [1]. Supplementation with fibroblast growth factor-2 (FGF-2) during MSCs expansion maintains the differentiation potential and increases proliferative capacity [21,22] Owing to their abundance and the painless non-invasive harvesting procedure, UCB-derived MSCs (UCB-MSCs) are a promising alternative source to bone marrow. Among the various biomaterials available, type I/III collagen sponges show efficient chondrogenic differentiation of human UCB-MSCs when used with BMP-2 and TGF-β1, leading to a strong increase in mature chondrocyte-specific markers (COL2A1, COL2A, ACAN) and cellular phenotype stability in the neo-tissue [30]. We characterized equine UCB-MSCs and determined their potential for cartilage tissue engineering using type I/III collagen sponges as the three-dimensional (3D) scaffold and chondrogenic factors under two oxygen conditions (normoxia and hypoxia). To stabilize the phenotype of differentiated UCB-MSCs, the use of small interfering RNA (siRNA) targeting Col1a2 mRNA was tested
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