Human intervertebral disc engineering with atelocollagen scaffold and by adenovirus-mediated gene transfer

Abstract

[한글]

[영문]

Tissue engineering is regarded as a new experimental technique for the biological treatment about degenerative intervertebral disc (IVD) diseases. Among artificial scaffolds, atelocollagen has an advantage in safety over others. And growth factors were known to stimulate matrix synthesis of IVD cells, among of them, transforming growth factor-β1 and bone morphogenetic protein-2 are the best candidates for IVD regeneration. Gene transfer to IVD cells provided sustained mechanism for matrix synthesis. Hence, the object of this study was to examine the cellular proliferation, proteoglycan (PG) synthesis, and phenotypical expression of IVD cells transduced by adenovirus transforming growth factor beta-1(Ad/TGF-β1) construct and adenovirus bone morphogenetic protein-2(Ad/BMP-2) construct and seeded on type I atelocollagen scaffolds.The IVD cells were isolated by sequential enzymatic digestion. Atelocollagen scaffold were prepared by lypophilization and cross-linking method. Ad/TGF-β1 or Ad/BMP-2 treated IVD cells were cultured for 7days. At 3 and 7 days, cellular proliferation, newly synthesized proteoglycan, mRNA expressions and the inner morphology of cell seeded scaffolds was determined by scanning electron microscopy (SEM).As result, transduced IVD cultures in atelocollagen type I with Ad/TGF-β1 and mixture group demonstrated increase in proteoglycan synthesis, but did not show significant change in mRNA expressions, compared to control. On the other hand, the IVD of transduced with Ad/ TGF-β1 in types I atelocollagen scaffolds decresed in cellular proliferation compared with others. And there were not showed osteocalcin mRNA expression in any groups.Human IVD cells were viable in type I atelocollagen scaffolds. Type I atelocollagen scaffold was suitable for cell proliferation, extracellular matrix synthesis. And Ad/TGF-β1 and Ad/BMP-2 combination was potential for IVD regeneration. Taken together, IVD cells in atelocollagen scaffolds, with therapeutic gene, provide a mechanism for tissue engineering of IVD.