Redifferentiation of dedifferentiated bovine chondrocytes when cultured in vitro in a PLGA–collagen hybrid mesh

Bovine articular chondrocytes dedifferentiated and lost their ability to express articular cartilage‐specific extracellular matrices such as type II collagen and aggrecan when cultured in a culture flask during in vitro multiplication. A poly(DL‐lactic‐co‐glycolic acid) (PLGA)–collagen hybrid mesh was prepared and used to redifferentiate the dedifferentiated cells. The two passaged dedifferentiated chondrocytes were seeded in a PLGA–collagen hybrid mesh and cultured in vitro in Dulbecco's modified Eagle's medium containing 10% fetal bovine serum. The cells adhered to the hybrid mesh, distributed evenly, and proliferated to fill the spaces in the scaffold. The gene expression of type I collagen, type II collagen, and aggrecan was analyzed after the cells were cultured in the hybrid mesh for 2–12 weeks. The expression of the gene encoding type I collagen was downregulated, whereas those of type II collagen and aggrecan were upregulated. Histological examination by hematoxylin–eosin and safranin O/fast green staining indicates that the cells regained their original round morphology. In addition, a homogeneous distribution of articular cartilage extracellular matrices was detected around the cells. These results suggest redifferentiation of the differentiated chondrocytes in the hybrid mesh. The hybrid mesh, which facilitated the redifferentiation of the dedifferentiated multiplied chondrocytes, would be an effective scaffold for the assembly of cells to regenerate three‐dimensional cartilaginous tissue.

[1]  J. Vacanti,et al.  Tissue engineering : Frontiers in biotechnology , 1993 .

[2]  G. Schulze-Tanzil,et al.  Redifferentiation of dedifferentiated human chondrocytes in high-density cultures , 2002, Cell and Tissue Research.

[3]  Peter Müller,et al.  Relationship between cell shape and type of collagen synthesised as chondrocytes lose their cartilage phenotype in culture , 1977, Nature.

[4]  G. Lisignoli,et al.  Evidence for redifferentiation of human chondrocytes grown on a hyaluronan-based biomaterial (HYAff 11): molecular, immunohistochemical and ultrastructural analysis. , 2002, Biomaterials.

[5]  J. MacLeod,et al.  Phenotypic Stability of Articular Chondrocytes In Vitro: The Effects of Culture Models, Bone Morphogenetic Protein 2, and Serum Supplementation , 2000, Journal of bone and mineral research : the official journal of the American Society for Bone and Mineral Research.

[6]  Jeffrey A. Hubbell,et al.  Biomaterials in Tissue Engineering , 1995, Bio/Technology.

[7]  G. Vunjak‐Novakovic,et al.  Growth factors for sequential cellular de- and re-differentiation in tissue engineering. , 2002, Biochemical and biophysical research communications.

[8]  D. Heinegård,et al.  Chondrocyte-matrix interactions. Attachment to proteins isolated from cartilage. , 1989, Experimental cell research.

[9]  M. Heberer,et al.  Specific growth factors during the expansion and redifferentiation of adult human articular chondrocytes enhance chondrogenesis and cartilaginous tissue formation in vitro , 2001, Journal of cellular biochemistry.

[10]  R M Nerem,et al.  Tissue engineering: from biology to biological substitutes. , 1995, Tissue engineering.

[11]  J. Verhaar,et al.  Growth factors in cartilage tissue engineering. , 2002, Biorheology.

[12]  J. McPherson,et al.  Synergistic action of transforming growth factor-beta and insulin-like growth factor-I induces expression of type II collagen and aggrecan genes in adult human articular chondrocytes. , 1997, Experimental cell research.

[13]  J. Verhaar,et al.  Serum-free medium supplemented with high-concentration FGF2 for cell expansion culture of human ear chondrocytes promotes redifferentiation capacity. , 2002, Tissue engineering.

[14]  G. Vunjak‐Novakovic,et al.  Mammalian chondrocytes expanded in the presence of fibroblast growth factor 2 maintain the ability to differentiate and regenerate three-dimensional cartilaginous tissue. , 1999, Experimental cell research.

[15]  G. V. van Osch,et al.  In vitro redifferentiation of culture-expanded rabbit and human auricular chondrocytes for cartilage reconstruction. , 2001, Plastic and reconstructive surgery.

[16]  E. Vuorio,et al.  Expression of mRNAs for collagens and other matrix components in dedifferentiating and redifferentiating human chondrocytes in culture , 1989, FEBS letters.

[17]  P. Benya,et al.  Dedifferentiated chondrocytes reexpress the differentiated collagen phenotype when cultured in agarose gels , 1982, Cell.