The advance of mitral valve repair techniques through tissue engineering is impeded by the lack of information regarding the cellular and extracellular components of the mitral valve. The present study aims to expand our understanding of the mitral valve structure by analysing the synthesis of extracellular matrix (ECM) proteins and the expression of nitric oxide synthase (NOS). Valvular endothelial cells (VECs) and valvular interstitial cells (VICs) were isolated from porcine mitral valves. Immunochemical staining of ECM components, including type I, II, III, IV and V collagen, laminin, fibronectin, elastin and chondroitin sulphate (CS), was performed on both mitral valve tissue and cell cultures. Reverse transcription polymerase chain reaction and immunochemistry were used to analyse NOS expression in native valve and in culture. Both VECs and VICs synthesised the basement membrane components, laminin and type IV collagen both in vivo and in vitro, amongst other fibrous ECM proteins. Synthesis of type I collagen and CS was absent in VEC cultures. Each cell type had a characteristic profile of NOS expression. VECs synthesised endothelial NOS both in vivo and in vitro, with a minority of VICs expressing neuronal NOS in vitro. The present study reports newly recognised aspects of the mitral valve structure and the in vitro behaviour of mitral valve cell populations based on ECM synthesis and NOS expression. The presented profiles can be used as base tools for the generation of data necessary for the selection of ideal cell sources and for the design of appropriate scaffolds for the development of effective tissue-engineered mitral valves.

译文

缺乏有关二尖瓣细胞和细胞外成分的信息,阻碍了通过组织工程进行二尖瓣修复技术的发展。本研究旨在通过分析细胞外基质 (ECM) 蛋白的合成和一氧化氮合酶 (NOS) 的表达来扩大我们对二尖瓣结构的理解。从猪二尖瓣分离瓣膜内皮细胞 (VECs) 和瓣膜间质细胞 (VICs)。对二尖瓣组织和细胞培养物进行了ECM成分的免疫化学染色,包括I,II,III,IV和V型胶原蛋白,层粘连蛋白,纤连蛋白,弹性蛋白和硫酸软骨素 (CS)。逆转录聚合酶链反应和免疫化学用于分析天然瓣膜和培养物中NOS的表达。VECs和VICs均在体内和体外合成了基底膜成分,层粘连蛋白和IV型胶原蛋白以及其他纤维ECM蛋白。在VEC培养物中没有I型胶原蛋白和CS的合成。每种细胞类型都有NOS表达的特征。VECs在体内和体外合成了内皮NOS,少数VICs在体外表达神经元NOS。本研究报告了基于ECM合成和NOS表达的二尖瓣结构和二尖瓣细胞群体的体外行为的新认识。所呈现的配置文件可用作基础工具,用于生成选择理想细胞源所需的数据以及设计用于开发有效组织工程二尖瓣的适当支架。

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