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.

Objective:Viscum album L. has favorable cardiovascular effects including antihypertensive and vasorelaxant activity, and the nitric oxide (NO) pathway upregulation has been proposed to be the underlying mechanism. NO also plays an important role in the pathophysiology of heart failure. However, its effects on cardiac systolic function are unclear.Methods:A total of 30 male Wistar albino rats at 12 weeks of age were randomly divided into three groups: control, isoproterenol-induced heart failure group (ISO), and isoproterenol-induced heart failure + V. album treatment group (VA) groups (n=10 in each group). V. album was orally given at a dose of 250 mg/kg/day by gavage. Parameters of heart failure were compared among the groups. Tamhane’s T2 test, paired sample t-test, and Bonferroni methods were used for statistical analysis.Results:V. album resulted in an improvement in all parameters of heart failure including left ventricular diameters (6.34±0.23 mm, 6.98±0.35 mm, and 6.71±0.10 mm for left ventricular end-diastolic diameter in control, ISO, and VA groups, respectively, p<0.05), ejection fraction (73.3±3.1%, 56.7±2.6%, and 65.2±1.5% for control, ISO, and VA groups, respectively, p<0.05), serum NT-proBNP levels, and histopathological changes. V. album treatment resulted in a statistically significant attenuation of increased levels of NO and iNOS (p<0.0001). The levels of hs-CRP were also found to be lower in the VA group compared with the controls and ISO groups (p<0.01).Conclusion:V. album exerted favorable effects on left ventricular function in isoproterenol-induced heart failure rats. Upregulation of the NO pathway seems to be the possible pathophysiological mechanism. Favorable vascular outcomes can also be speculated considering the reduction in serum hs-CRP levels.

C-type natriuretic peptide (CNP) signaling has been implicated as an important regulator of chondrogenic differentiation during endochondral bone development. This preliminary study further investigated the putative effectors and/or targets of CNP signaling in transforming growth factor (TGF)-β induced in vitro chondrogenic differentiation of mesenchymal stem cells (MSCs). Previously characterized human trabecular bone derived MSCs were induced either with only TGF-β1 or with a combination of TGF-β1 and CNP in micromass culture for 10 or 20 days. Genome wide gene expression profile changes in between these two groups were analyzed on day-10 or day-20 of culture. Results revealed that there were only 7 genes, whose expression change was fourfolds or higher in TGF-β1 and CNP fed group in comparison to only TGF-β1 fed group. The up-regulated genes included matrilin-3 (MATN3), engulfment and cell motility 1 (ELMO1), CD24, and DCN1, defective in cullin neddylation 1, domain containing 1 (DCUN1D1). The down-regulated genes, on the other hand, included LIM domain kinase 2 (LIMK2), Ewing sarcoma breakpoint region 1, and guanine nucleotide binding protein (G protein), gamma 12 (GNG12). The up-regulation of MATN3 was confirmed on the basis of RT-PCR. The known literature on both CNP signaling and MATN3 function in chondrogenesis match with each other and suggest MATN3 as a putative effector and/or target of CNP signaling during this process.