Tandon Ishita, Woessner Alan E, Ferreira Laίs A, Shamblin Christine, Vaca-Diez Gustavo, Walls Amanda, Kuczwara Patrick, Applequist Alexis, Nascimento Denise F, Tandon Swastika, Kim Jin-Woo, Rausch Manuel, Timek Tomasz, Padala Muralidhar, Kinter Michael T, Province Dennis, Byrum Stephanie D, Quinn Kyle P, Balachandran Kartik
Department of Biomedical Engineering, University of Arkansas, Fayetteville, AR, USA.
Arkansas Integrative Metabolic Research Center, University of Arkansas, Fayetteville, AR, USA.
Acta Biomater. 2024 Sep 15;186:167-184. doi: 10.1016/j.actbio.2024.07.036. Epub 2024 Jul 30.
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.
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.
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.
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.
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.
钙化性主动脉瓣疾病（CAVD）是最常见的瓣膜病形式之一，发生致命心血管事件的风险升高50%，仅在北美每年就有超过15000人死亡。治疗标准是瓣膜置换，因为早期诊断、缓解和药物策略仍未充分发展。早期诊断和治疗策略的开发需要构建有效的体外瓣膜模拟模型，以阐明CAVD的早期机制。
在本研究中，我们开发了一种多层生理相关的片上瓣膜（VOC）系统，该系统整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）共培养以及动态机械刺激。基于胶原蛋白和糖胺聚糖（GAG）的水凝胶组装成双层，以模拟天然纤维层和海绵层的健康或病变组成。对健康和病变的VOC进行多光子成像和蛋白质组分析。
基于胶原蛋白的双层水凝胶维持了VIC的表型。发现与细胞周期进程、胆固醇生物合成和蛋白质稳态等细胞过程相关的蛋白质发生了显著变化，并且与病变VOC中细胞代谢的变化相关。本研究表明，病变的VOC可能代表疾病的早期适应性起始阶段，这一点得到了人类主动脉瓣蛋白质组评估的证实。
在本研究中，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层的健康或病变组成。当将这些凝胶与VEC和VIC组装到VOC中时，病变的VOC揭示了有关CAVD起始过程的关键见解。
钙化性主动脉瓣疾病（CAVD）因心血管病理生理学使死亡风险升高50%，然而临床上缺乏预防和缓解策略。开发评估早期疾病的工具将显著有助于疾病预防和治疗开发。此前，研究已将基于胶原蛋白和糖胺聚糖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以及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构建用于CAVD研究的生理相关模型。我们制造了一种紧密模拟主动脉瓣的双层水凝胶，并将其用于动态平台中的瓣膜细胞共培养，以通过蛋白质组学和多光子成像评估深入了解CAVD起始过程的机制。
