Implication of C-type natriuretic peptide-3 signaling in glycosaminoglycan synthesis and chondrocyte hypertrophy during TGF-beta 1 induced chondrogenic differentiation of chicken bone marrow-derived mesenchymal stem cells


Kocamaz E., Gok D. , Cetinkaya A., TUFAN A. Ç.

JOURNAL OF MOLECULAR HISTOLOGY, vol.43, no.5, pp.497-508, 2012 (Journal Indexed in SCI) identifier identifier identifier

  • Publication Type: Article / Article
  • Volume: 43 Issue: 5
  • Publication Date: 2012
  • Doi Number: 10.1007/s10735-012-9430-2
  • Title of Journal : JOURNAL OF MOLECULAR HISTOLOGY
  • Page Numbers: pp.497-508
  • Keywords: CNP, NPR-B, Chondrogenesis, Endochondral ossification, Glycosaminoglycan synthesis, Chondrogenic hypertrophy, HUMAN TRABECULAR BONE, ALTERED N-CADHERIN, RECEPTOR-B, MUTATIONS, COLLAGEN, GROWTH

Abstract

This study investigated the involvement of CNP-3, chick homologue for human C-type natriuretic peptide (CNP), in TGF-beta 1 induced chondrogenic differentiation of chicken bone marrow-derived mesenchymal stem cells (MSCs). Chondrogenic differentiation of MSCs in pellet cultures was induced by TGF-beta 1. Chondrogenic differentiation and glycosaminoglycan synthesis were analyzed on the basis of basic histology, collagen type II expression, and Alcian blue staining. Antibodies against CNP and NPR-B were used to block their function during these processes. Results revealed that expression of CNP-3 and NPR-B in MSCs were regulated by TGF-beta 1 in monolayer cultures at mRNA level. In pellet cultures of MSCs, TGF-beta 1 successfully induced chondrogenic differentiation and glycosaminoglycan synthesis. Addition of CNP into the TGF-beta 1 supplemented chondrogenic differentiation medium further induced the glycosaminoglycan synthesis and hypertrophy of differentiated chondrocytes in these pellets. Pellets induced with TGF-beta 1 and treated with antibodies against CNP and NPR-B, did show collagen type II expression, however, Alcian blue staining showing glycosaminoglycan synthesis was significantly suppressed. In conclusion, CNP-3/NPR-B signaling may strongly be involved in synthesis of glycosaminoglycans of the chondrogenic matrix and hypertrophy of differentiated chondrocytes during TGF-beta 1 induced chondrogenic differentiation of MSCs.