Chondrogenic differentiation of canine myoblasts induced by cartilage-derived morphogenetic protein-2 and transforming growth factor-β1 in vitro

Abstract

Abstract. The meniscus has limited ability to repair itself following injury. However, tissue engineering provides new means of meniscus repair. Myoblasts, which possess the potential of multi-directional differentiation, may be ideal seed cells in meniscus tissue engineering. Myoblasts from different animals showed slight differences in morphology and in the potential to differentiate. In the present study, we isolated myoblasts from canines and induced chondrogenesis in order to establish a new experimental model of seed cells. Myoblasts were isolated and harvested from Beagle canines. To induce chondrogenesis, cartilage-derived morphogenetic protein-2 (CDMP-2) at different concentrations (10, 20, 50 and 100 ng/ml), transforming growth factor-β1 (TGF-β1; 10, 20, 30 and 50 ng/ml), and different concentrations of CDMP-2 (10, 20, 50 and 100 ng/ml) together with TGF-β1 (20 ng/ml) were added to the cultured pellets. After 21 days of in vitro culture, chondrogenic differentiation was evaluated by histological and immunohistochemical techniques. The degree of gene expression was measured by quantitative RT-PCR. Based on the histological staining of glycosaminoglycan, using the toluidine blue dye-binding method, we found that CDMP-2 initiated chondrogenic differentiation of myoblasts, as did TGF-β1. Furthermore, CDMP-2 conferred a stronger stimulatory effect than TGF-β1. The combination of CDMP-2 and TGF-β1 synergistically induced chondrogenesis of myoblasts. This synergistic chondrogenic effect of CDMP-2 together with TGF-β1 was further confirmed by quantification of glycosaminoglycan using dimethylmethylene blue dye-binding assay and immunohistochemical analysis of the expression of cartilage-specific proteins collagen I and II. Canine myoblasts can be induced into chondrocytes by CDMP-2 and TGF-β1 in vitro, suggesting that myoblasts are suitable as seed cells for meniscus tissue engineering.

Nucleus pulposus cell apoptosis is attenuated by CDMP-2 through regulating oxidative damage under the hyperosmotic environment

“…Cartilage-derived morphogenetic protein-2 (CDMP-2) is an endogenous growth factor belonging to the BMP family and participates in regulating chondrocyte’s biology [ 25 , 26 ]. Previous studies have demonstrated that CDMP-2 is down-regulated in the osteoarthritic human articular chondrocytes and is able to enhance cartilaginous matrix synthesis [ 27 , 28 ].…”

Meniscal allograft transplants and new scaffolding techniques

“…152 Nanotechnology also permits modification of the surfaces of biomaterials. 153 Biologics are biologically active natural components that can activate cells and enhance tissue healing, including specific growth factors 154,155 and platelet rich plasma (PRP). 114 PRP is an autologous source of a ‘cocktail’ of several autologous growth factors (including platelet-derived growth factor, endothelial growth factor, and transforming growth factor) as well as anti- and pro-inflammatory cytokines (including interleukin-4, -8, -13, -17, tumor necrosis factor-α, and interferon-α) which might influence the process of tissue healing.…”

An Overview of Intervertebral Disc Degeneration Therapies and an Evaluation of the Chondrogenic and Chemotactic Potential of CDMP-2

“…More recently, an in vitro study of cultured canine myoblasts, which have potential for multilineage differentiation, observed that direct injection of CDMP-2 resulted in chondrogenic differentiation and significantly increased glycosaminoglycan synthesis (p<0.05, n=4) on immunehistological staining, and the molecule had stronger chondrogenic effects than TGF-β1 [96]. The study also confirmed the results with RT-PCR, which showed increased expression of collagen I and II in groups treated with CDMP-2, suggesting its role in chondrogenic differentiation of precursor cells.…”