Radiation Induces Valvular Interstitial Cell Calcific Response in an in vitro Model of Calcific Aortic Valve Disease.
Radiation Induces Valvular Interstitial Cell Calcific Response in an in vitro Model of Calcific Aortic Valve Disease.
复制标题
辐射在钙化主动脉瓣疾病的体外模型中诱导瓣膜间质细胞钙化反应。
DOI：
10.3389/fcvm.2021.687885
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发表时间：
2021
影响因子：
3.6
通讯作者：
Bouten CVC
中科院分区：
文献类型：
--
作者：
Meerman M;Driessen R;van Engeland NCA;Bergsma I;Steenhuijsen JLG;Kozono D;Aikawa E;Hjortnaes J;Bouten CVC
背景：纵隔电离放疗与瓣膜疾病的风险增加相关，瓣膜疾病表现出与钙化主动脉瓣疾病（CAVD）相似的病理特征。尽管放疗技术取得了进步，但由于接受放疗的患者生存率提高，诸如辐射相关瓣膜病等合并症的发生率仍在增加。然而，与辐射相关的瓣膜疾病的机制在很大程度上是未知的。CAVD被认为是一种主动调节的疾病过程，主要由瓣膜间质细胞（VICs）控制。我们假设辐射暴露催化了vic的钙化反应，因此有助于辐射相关瓣膜疾病的发展。方法和结果：为了描述辐射与VIC行为（形态学、钙化和基质转换）之间的关系，建立了两种不同的体外模型：(1)VIC在对照培养基（CM）或成骨培养基（OM）的盖片上进行二维（2D）培养，并以0、2、4、8或16 Gy （Gy）照射；(2)设计三维（3D）水凝胶体系，加载vic，暴露于0、4或16 Gy的辐射下。在两种模型中，CM和OM的细胞活力和增殖均呈剂量依赖性下降。辐射暴露导致VICs的肌成纤维细胞样形态改变和分化，其特征是αSMA表达降低。钙化，根据碱性磷酸酶活性的增加来定义，主要存在于暴露于4 Gy的二维辐射vic中，而暴露于更高剂量后，vic获得了独特的巨大成纤维细胞样细胞形态。最后，在3D照射的vic中，基质转换受到辐射暴露的显著影响，表现为胶原染色降低，MMP-2和MMP-9活性升高。结论：目前的研究表明，辐射暴露增强了vic的钙化反应，这是CAVD的一个标志。此外，高辐射暴露诱导vic分化为终末分化的巨细胞成纤维细胞。进一步的研究是必要的，以阐明这些辐射引起的瓣膜改变的潜在机制。
Background: Mediastinal ionizing radiotherapy is associated with an increased risk of valvular disease, which demonstrates pathological hallmarks similar to calcific aortic valve disease (CAVD). Despite advances in radiotherapy techniques, the prevalence of comorbidities such as radiation-associated valvular disease is still increasing due to improved survival of patients receiving radiotherapy. However, the mechanisms of radiation-associated valvular disease are largely unknown. CAVD is considered to be an actively regulated disease process, mainly controlled by valvular interstitial cells (VICs). We hypothesize that radiation exposure catalyzes the calcific response of VICs and, therefore, contributes to the development of radiation-associated valvular disease. Methods and Results: To delineate the relationship between radiation and VIC behavior (morphology, calcification, and matrix turnover), two different in vitro models were established: (1) VICs were cultured two-dimensional (2D) on coverslips in control medium (CM) or osteogenic medium (OM) and irradiated with 0, 2, 4, 8, or 16 Gray (Gy); and (2) three-dimensional (3D) hydrogel system was designed, loaded with VICs and exposed to 0, 4, or 16 Gy of radiation. In both models, a dose-dependent decrease in cell viability and proliferation was observed in CM and OM. Radiation exposure caused myofibroblast-like morphological changes and differentiation of VICs, as characterized by decreased αSMA expression. Calcification, as defined by increased alkaline phosphatase activity, was mostly present in the 2D irradiated VICs exposed to 4 Gy, while after exposure to higher doses VICs acquired a unique giant fibroblast-like cell morphology. Finally, matrix turnover was significantly affected by radiation exposure in the 3D irradiated VICs, as shown by decreased collagen staining and increased MMP-2 and MMP-9 activity. Conclusions: The presented work demonstrates that radiation exposure enhances the calcific response in VICs, a hallmark of CAVD. In addition, high radiation exposure induces differentiation of VICs into a terminally differentiated giant-cell fibroblast. Further studies are essential to elucidate the underlying mechanisms of these radiation-induced valvular changes.