Calcific aortic valve disease (CAVD) is a progressive disorder that increases in prevalence with age. An important role in aortic valve calcification is played by valvular interstitial cells (VIC), that with age or in pathological conditions acquire an osteoblast-like phenotype that advances the disease. Therefore, pharmacological interventions aiming to stop or reverse the osteoblastic transition of VIC may represent a therapeutic option for CAVD. In this study, we aimed at developing a nanotherapeutic strategy able to prevent the phenotypic switch of human aortic VIC into osteoblast-like cells. We hypothesize that nanocarriers designed for silencing the Runt-related transcription factor 2 (Runx2) will stop the progress or reverse the osteodifferentiation of human VIC, induced by high glucose concentrations and pro-osteogenic factors. We report here the potential of fullerene (C60)-polyethyleneimine (PEI)/short hairpin (sh)RNA-Runx2 nano-polyplexes to efficiently down-regulate Runx2 mRNA and protein expression leading subsequently to a significant reduction in the expression of osteogenic proteins (i.e. ALP, BSP, OSP and BMP4) in osteoblast-committed VIC. The data suggest that the silencing of Runx2 could represent a novel strategy to impede the osteoblastic phenotypic shift of VIC and the ensuing progress of CAVD.

中文翻译：

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纳米多聚体介导的Runx2-shRNA转染可减轻人类瓣膜间质细胞的骨分化。

钙化性主动脉瓣疾病（CAVD）是一种进行性疾病，其患病率随着年龄的增长而增加。瓣膜间质细胞（VIC）在主动脉瓣钙化中起重要作用，随着年龄的增长或在病理条件下，它们会获得成骨样表型，从而使疾病进展。因此，旨在阻止或逆转VIC成骨细胞转变的药理干预措施可能代表了CAVD的治疗选择。在这项研究中，我们旨在开发一种能够防止人类主动脉VIC向成骨细胞样细胞表型转换的纳米治疗策略。我们假设设计用于沉默Runt相关转录因子2（Runx2）的纳米载体将阻止由高葡萄糖浓度和促成骨因子引起的人类VIC的骨分化或逆转。我们在这里报告富勒烯（C60）-聚乙烯亚胺（PEI）/短发夹（sh）RNA-Runx2纳米复合物潜在地有效下调Runx2 mRNA和蛋白质表达的潜力，从而导致成骨蛋白的表达显着降低（即成骨细胞承诺的VIC中的ALP，BSP，OSP和BMP4）。数据表明，Runx2沉默可能代表了一种新的策略，可以阻止VIC的成骨细胞表型转移和随后的CAVD进展。