Dose dependent effect of C-type natriuretic peptide signaling in glycosaminoglycan synthesis during TGF-β1 induced chondrogenic differentiation of mesenchymal stem cells

Abstract

Recent investigations credited important roles to C-type natriuretic peptide (CNP) signaling during chondrogenesis. This study investigated the putative role of CNP in transforming growth factor (TGF)-β1 induced in vitro chondrogenic differentiation of mesenchymal stem cells (MSCs) in pellet culture. MSCs were derived from human trabecular bone and were characterized on the basis of their cell surface antigens and adipogenic, osteogenic, and chondrogenic differentiation potential. TGF-β1 induced chondrogenic differentiation and glycosaminoglycan (GAG) synthesis was analyzed on the basis of basic histology, collagen type II, Sox 9 and aggrecan expressions, and Alcian blue staining. Results revealed that human trabecular bone-derived MSCs express CNP and NPR-B analyzed on the basis of RT-PCR and immunohistochemistry. In pellet cultures of MSCs TGF-β1 successfully induced chondrogenic differentiation and GAG synthesis. RT-PCR analyses of both CNP and NPR-B during this process revealed an activation of this signaling pathway in response to TGF-β1. Similar cultures induced with TGF-β1 and treated with different doses of CNP showed that CNP supplementation at 10(-8) and 10(-7) M concentrations significantly increased GAG synthesis in a dose dependent manner, whereas at 10(-6) M concentration this stimulatory effect was diminished. In conclusion, CNP/NPR-B signaling pathway is activated during TGF-β1 induced chondrogenic differentiation of human trabecular bone-derived MSCs and may strongly be involved in GAG synthesis during this process. This effect is likely to be a dose-dependent effect.

Role of C-type natriuretic peptide signalling in maintaining cartilage and bone function

“…Transforming growth factor b (TGFb) is known to be critical for the differentiation of chondrocytes. Indeed, the presence of TGFb upregulates CNP/Npr2 levels and subsequent cGMP production in bone marrow derived mesenchymal stem cells 45,46 . The combination of CNP and TGFb appears to be additive with effects dependent on peptide concentration 46,47 .…”

Design of a Novel 3D Printed Bioactive Nanocomposite Scaffold for Improved Osteochondral Regeneration

“…Organic TGF-β1 has been shown to direct early and late-stage MSC chondrogenic and osteogenic differentiation 63 from expedited proliferation to increased GAG production and hypertrophic maturation of seeded MSCs. Tezcan et al 64 examined TGF-β1 induced MSC chondrogenic differentiation and correlated their findings as a dose-dependent response wherein TGF-β1 was critical in the initiation of GAG synthesis and late stage tissue maturation. Although TGF-β1 was added only to the top cartilage layer, localized diffusion through the entire construct is facilitated by the inherent microporous nature of the PEG-DA hydrogel whose composition consisted of a 40% soluble fraction as characterized in our previous work.…”

Maintaining the Phenotype Stability of Chondrocytes Derived from MSCs by C-Type Natriuretic Peptide

“…In vitro studies have revealed that CNP can promote proliferation of primary chondrocytes (Waldman et al, 2008), as well as chondrocytes-derived from MSCs (Tezcan et al, 2010; Kocamaz et al, 2012). Woods et al revealed that CNP could regulate cellular condensation of mouse embryonic limbs bud cells during micromass culture by increasing the expression of N-cadherin (Woods et al, 2007).…”

“…By measuring the 35 SO 4 , Mericq V et al found that CNP increased the synthesis of glycosaminoglycan (GAG), one of the main cartilage matrix components (Mericq et al, 2000). Addition of CNP further increased the GAG synthesis in the human trabecular bone derived MSCs (Tezcan et al, 2010) and chicken bone marrow derived MSCs (Kocamaz et al, 2012). …”