Asporin Reduces Adult Aortic Valve Interstitial Cell Mineralization Induced by Osteogenic Media and Wnt Signaling Manipulation In Vitro

Figure 7

Human recombinant Asporin (rhAsp) treatment also decreases matrix mineralization in Xav939-treated osteogenic (OS) media-induced avian adult AVICs in culture. (a–c) Bright-field microscopic images (×10) of alizarin stained OS-induced AVICs show unchanged matrix mineralization in Xav939-treated OS-induced AVICs compared to OS-induced AVICs only. Moreover, decreased matrix mineralization is detected in Xav939+rhAsp-treated OS-induced AVICs (arrowheads). (d) Quantitative assay of Alizarin Red S shows that the absorbance of Xav939-treated OS-induced AVICs is similar (0.089) compared to only OS-induced AVICs (0.088), and the absorbance of rhWnt3A+rhAsp-treated OS-induced AVICs decreased (0.068) compared to Xav939-treated OS-induced AVICs (, ). (e) (I and VI): qRT-PCR data show that Asporin is unchanged in both OS-induced AVICs and Xav939-treated AVICs, whereas it increased by 1.71 fold in Xav939+rhAsp OS-induced AVICs (I). Osteopontin and Runx2 mRNA expressions are unchanged in both OS-induced AVICs and Xav939-treated AVICs, whereas these decreased by 0.28 fold and 0.35 fold in Xav939+rhAsp OS-induced AVICs, respectively (II and III). β-Catenin decreased by 0.78 fold and 0.27 fold in Xav939-treated and in Xav939+rhAsp-treated OS-induced AVICs, respectively (IV). Similarly, Wnt3A decreased by 0.05 fold and 0.65 fold in Xav939-treated and inXav939+rhAsp-treated OS-induced AVICs, respectively (V). Also, Dkk1 increased by 0.18 fold and 3.09 fold inXav939-treated and in Xav939+rhAsp-treated OS-induced AVICs, respectively. Statistical significance was determined by Student’s -test, where denotes and denotes and .

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