Induction of Re-Differentiation of Passaged Rat Chondrocytes Using a Naturally Obtained Extracellular Matrix Microenvironment
- Induction of Re-Differentiation of Passaged Rat Chondrocytes Using a Naturally Obtained Extracellular Matrix Microenvironment
- 차명화; 도선희; 박가람; 두 핑; 한기철; 한동근; 박귀덕
- Issue Date
- Tissue engineering. Part A.
- VOL 19, NO 7-8, 978-988
- Dedifferentiated human chondrocytes severely limit successful hyaline cartilage repair in clinical practice. The
primary interest of this study is to evaluate the naturally obtained cell-derived matrix (CDM) as a physical
microenvironment for chondrocyte re-differentiation. Once different cell types were cultured for 6 days and
decellularized using detergents and enzymes, the fibroblast-derived matrix (FDM), preosteoblast-derived matrix
(PDM), and chondrocyte-derived matrix (CHDM) were obtained. From scanning electron microscope observation,
each CDM was found to resemble a fibrous mesh with self-assembled fibrils. Both the FDM and PDM
showed a more compact matrix structure compared to the CHDM. For compositional analysis, sodium dodecyl
sulfate–polyacrylamide gel electrophoresis displayed numerous matrix proteins, which were quite different
from each CDM in quantity and type. Specific matrix components, such as fibronectin, type I collagen (Col I),
and laminin, were detected using immunofluorescent staining. In addition, the water contact angle suggests that
the FDM is more hydrophilic than the PDM or CHDM. The proliferation of rat primary chondrocytes growing
on CDMs was better than those growing on a plastic coverslip (control) or gelatin. Meanwhile, synthesis of
glycosaminoglycan (GAG) was more effective for passaged chondrocytes (P4) cultivated on CDMs, and the
difference was significant compared to cells grown on the control or on gelatin. As for the gene expression of
cartilage-specific markers, CDMs exhibited good chondrocyte re-differentiation with time: the dedifferentiating
marker, Col I was restrained, whereas the ratio between Col II and Col I, and between aggrecan and Col I, as an
indicator of re-differentiation, was greatly improved. In addition, immunofluorescence of Col II showed a very
positive signal in chondrocytes cultivated for 2 weeks on the CDMs.
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