Abstract
Introduction
Parathyroid hormone-related peptide (PTHrP) is an important inductive factor during chondrogenesis of mesenchymal stem cells (MSCs). PTHrP induces chondrogenesis and suppresses hypertrophy, yet the lack of an efficient delivery system limits its use for cartilage tissue engineering in clinical application.
Materials and Methods
In this study, a peptide of seven amino acids was first used to engineer PTHrP to construct a collagen-targeting system. This peptide functioned as a collagen-binding domain (CBD) to specially target the PTHrP to collagen. ELISA assay was used to determine the collagen-binding ability of CBD-PTHrP. The effect of CBD-PTHrP on chondrogenesis was measured by an in vitro pellet assay in bone marrow-derived MSCs (BM-MSCs).
Results
The CBD peptide promoted the binding of CBD-PTHrP to collagen when compared with NAT-PTHrP. Furthermore, the recombinant protein CBD-PTHrP induced the expression of COL2A1 and Sox-9, inhibited the expression of COL1A1 at the mRNA and protein levels as effectively as NAT-PTHrP. Safranin-O and immunohistochemistry for collagen types I, II, X and Sox-9 generally paralleled qRT-PCR and Western blotting findings with minor variations.
Conclusion
Our study demonstrated that CBD-PTHrP is a collagen-targeting system and promotes in vitro chondrogenesis in BM-MSCs. We suggest that this is an efficient delivery system for cartilage tissue engineering in clinical application.
None declared