Oxygenator assisted dynamic microphysiological culture elucidates the impact of hypoxia on valvular interstitial cell calcification.

IF 5.7 3区生物学 Q1 BIOCHEMICAL RESEARCH METHODS

Journal of Biological Engineering Pub Date : 2024-08-23 DOI:10.1186/s13036-024-00441-4

Claudia Dittfeld, Florian Schmieder, Stephan Behrens, Anett Jannasch, Klaus Matschke, Frank Sonntag, Sems-Malte Tugtekin

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引用次数: 0

Abstract

Introduction: Microphysiological systems (MPS) offer simulation of (patho)physiological parameters. Investigation includes items which lead to fibrosis and calcification in development and progress of calcific aortic valve disease, based e.g. on culturing of isolated valvular interstitial cells (VICs). Hypoxia regulated by hypoxia inducible factors impacts pathological differentiation in aortic valve (AV) disease. This is mimicked via an MPS implemented oxygenator in combination with calcification inducing medium supplementation.

Methods: Human valvular interstitial cells were isolated and dynamically cultured in MPS at hypoxic, normoxic, arterial blood oxygen concentration and cell incubator condition. Expression profile of fibrosis and calcification markers was monitored and calcification was quantified in induction and control media with and without hypoxia and in comparison to statically cultured counterparts.

Results: Hypoxic 24-hour culture of human VICs leads to HIF1α nuclear localization and induction of EGLN1, EGLN3 and LDHA mRNA expression but does not directly impact expression of fibrosis and calcification markers. Dependent on medium formulation, induction medium induces monolayer calcification and elevates RUNX2, ACTA2 and FN1 but reduces SOX9 mRNA expression in dynamic and static MPS culture. But combining hypoxic oxygen concentration leads to higher calcification potential of human VICs in calcification and standard medium formulation dynamically cultured for 96 h.

Conclusion: In hypoxic oxygen concentration an increased human VIC calcification in 2D VIC culture in an oxygenator assisted MPS was detected. Oxygen regulation therefore can be combined with calcification induction media to monitor additional effects of pathological marker expression. Validation of oxygenator dependent VIC behavior envisions future advancement and transfer to long term aortic valve tissue culture MPS.

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充氧器辅助动态微生理学培养阐明了缺氧对瓣膜间质细胞钙化的影响。

引言微生理学系统（MPS）可模拟（病理）生理参数。研究包括在钙化性主动脉瓣疾病的发生和发展过程中导致纤维化和钙化的项目，例如基于分离的瓣膜间质细胞（VICs）的培养。由缺氧诱导因子调节的缺氧会影响主动脉瓣（AV）疾病的病理分化。方法：分离人瓣膜间质细胞，并在缺氧、常氧、动脉血氧浓度和细胞培养箱条件下在 MPS 中进行动态培养。监测纤维化和钙化标志物的表达情况，并在有缺氧和无缺氧的诱导培养基和对照培养基中量化钙化情况，并与静态培养的对应细胞进行比较：结果：24小时缺氧培养人VICs会导致HIF1α核定位并诱导EGLN1、EGLN3和LDHA mRNA的表达，但不会直接影响纤维化和钙化标志物的表达。根据培养基配方的不同，在动态和静态 MPS 培养中，诱导培养基可诱导单层钙化并提高 RUNX2、ACTA2 和 FN1 的表达，但会降低 SOX9 mRNA 的表达。但在钙化培养基和标准培养基配方中，结合低氧浓度动态培养 96 小时后，人 VICs 的钙化潜能更高：结论：在缺氧氧气浓度下，氧气辅助 MPS 中的二维 VIC 培养可增加人 VIC 的钙化。因此，氧气调节可与钙化诱导培养基相结合，以监测病理标记表达的其他影响。氧合器依赖性 VIC 行为的验证预示着未来的进步，并将转移到长期主动脉瓣组织培养 MPS 中。

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来源期刊

Journal of Biological Engineering BIOCHEMICAL RESEARCH METHODS-BIOTECHNOLOGY & APPLIED MICROBIOLOGY

CiteScore

7.10

自引率

1.80%

发文量

审稿时长

17 weeks

期刊介绍： Biological engineering is an emerging discipline that encompasses engineering theory and practice connected to and derived from the science of biology, just as mechanical engineering and electrical engineering are rooted in physics and chemical engineering in chemistry. Topical areas include, but are not limited to: Synthetic biology and cellular design Biomolecular, cellular and tissue engineering Bioproduction and metabolic engineering Biosensors Ecological and environmental engineering Biological engineering education and the biodesign process As the official journal of the Institute of Biological Engineering, Journal of Biological Engineering provides a home for the continuum from biological information science, molecules and cells, product formation, wastes and remediation, and educational advances in curriculum content and pedagogy at the undergraduate and graduate-levels. Manuscripts should explore commonalities with other fields of application by providing some discussion of the broader context of the work and how it connects to other areas within the field.