背景:钙化性主动脉瓣疾病(CAVD)是最常见的瓣膜病之一,致命心血管事件的风险增加了50%,仅在北美,每年就有超过15,000人死亡。治疗标准是瓣膜置换术作为早期诊断,缓解,药物策略仍然不发达。早期诊断和治疗策略的发展需要制造有效的体外瓣膜模拟模型来阐明早期CAVD机制。
方法:在本研究中,我们开发了一种多层生理相关的3D芯片瓣膜(VOC)系统,该系统结合了主动脉瓣模拟细胞外基质(ECM),猪主动脉瓣间质细胞(VIC)和内皮细胞(VEC)共培养和动态机械刺激。在双层中组装基于胶原和糖胺聚糖(GAG)的水凝胶以模拟天然纤维和海绵体的健康或患病组合物。对健康和患病的VOC进行多光子成像和蛋白质组学分析。
结果:基于胶原的双层水凝胶保持了VIC的表型。与细胞过程相关的蛋白质,如细胞周期进程,胆固醇生物合成,发现蛋白质稳态显着改变,并与患病VOC中细胞代谢的变化相关。这项研究表明,患病的VOC可能代表了早期,适应性疾病起始阶段,人类主动脉瓣蛋白质组学评估证实了这一点。
结论:在这项研究中,我们开发了一种基于胶原蛋白的双层水凝胶,以模拟天然纤维层和海绵体层的健康或患病成分。当凝胶与VEC和VIC组装在VOC中时,患病的VOC揭示了关于CAVD启动过程的关键见解。
结论:钙化性主动脉瓣疾病(CAVD)使心血管病理生理学导致的死亡风险增加50%,然而,缺乏预防和缓解策略,临床。开发评估早期疾病的工具将大大有助于疾病的预防和治疗方法的开发。以前,研究已经利用胶原和糖胺聚糖为基础的水凝胶用于瓣膜细胞共培养,动态环境中的瓣膜细胞共培养,和基于无机聚合物的多层水凝胶;然而,这些方法尚未结合起来建立CAVD研究的生理学相关模型.我们制造了一种双层水凝胶,其紧密模拟主动脉瓣,并将其用于动态平台中的瓣膜细胞共培养,以使用蛋白质组学和多光子成像评估获得对CAVD起始过程的机械见解。
BACKGROUND: Calcific aortic valve disease (CAVD) is one of the most common forms of valvulopathy, with a 50 % elevated risk of a fatal cardiovascular event, and greater than 15,000 annual deaths in North America alone. The treatment standard is valve replacement as early diagnostic, mitigation, and drug strategies remain underdeveloped. The development of early diagnostic and therapeutic strategies requires the fabrication of effective in vitro valve mimetic models to elucidate early CAVD mechanisms.
METHODS: In this study, we developed a multilayered physiologically relevant 3D valve-on-chip (VOC) system that incorporated aortic valve mimetic extracellular matrix (ECM), porcine aortic valve interstitial cell (VIC) and endothelial cell (VEC) co-culture and dynamic mechanical stimuli. Collagen and glycosaminoglycan (GAG) based hydrogels were assembled in a bilayer to mimic healthy or diseased compositions of the native fibrosa and spongiosa. Multiphoton imaging and proteomic analysis of healthy and diseased VOCs were performed.
RESULTS: Collagen-based bilayered hydrogel maintained the phenotype of the VICs. Proteins related to cellular processes like cell cycle progression, cholesterol biosynthesis, and protein homeostasis were found to be significantly altered and correlated with changes in cell metabolism in diseased VOCs. This study suggested that diseased VOCs may represent an early, adaptive disease initiation stage, which was corroborated by human aortic valve proteomic assessment.
CONCLUSIONS: In this study, we developed a collagen-based bilayered hydrogel to mimic healthy or diseased compositions of the native fibrosa and spongiosa layers. When the gels were assembled in a VOC with VECs and VICs, the diseased VOCs revealed key insights about the CAVD initiation process.
CONCLUSIONS: Calcific aortic valve disease (CAVD) elevates the risk of death due to cardiovascular pathophysiology by 50 %, however, prevention and mitigation strategies are lacking, clinically. Developing tools to assess early disease would significantly aid in the prevention of disease and in the development of therapeutics. Previously, studies have utilized collagen and glycosaminoglycan-based hydrogels for valve cell co-cultures, valve cell co-cultures in dynamic environments, and inorganic polymer-based multilayered hydrogels; however, these approaches have not been combined to make a physiologically relevant model for CAVD studies. We fabricated a bi-layered hydrogel that closely mimics the aortic valve and used it for valve cell co-culture in a dynamic platform to gain mechanistic insights into the CAVD initiation process using proteomic and multiphoton imaging assessment.