Microfluidic 3D platform to evaluate endothelial progenitor cell recruitment by bioactive materials.
Microfluidic 3D platform to evaluate endothelial progenitor cell recruitment by bioactive materials.
复制标题
微流体 3D 平台用于评估生物活性材料对内皮祖细胞的募集。
DOI：
10.1016/j.actbio.2022.08.019
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发表时间：
2022
影响因子：
9.7
通讯作者：
Castaño,Oscar
中科院分区：
文献类型：
--
作者：
López-Canosa,Adrián;Pérez-Amodio,Soledad;Engel,Elisabeth;Castaño,Oscar
大多数用于测试生物材料驱动的血管形成的常规体外模型过于简单，无法概括实际细胞微环境中发生的复杂相互作用，这导致对材料存活性能的预测很差。然而，在过去的十年里，基于微流控技术的细胞培养模型已经使组织仿生达到了前所未有的水平。在这项工作中，我们提出了一个基于微流体的3D模型来评估能够释放信号线索(如离子或蛋白质)的生物活性生物材料在内源性内皮祖细胞招募中的效果，这是血管形成过程中的关键步骤。通过实验验证的有限元模型以及对大鼠内皮祖细胞和骨髓间充质基质细胞的迁移和增殖研究，验证了该平台的可用性。为了验证生物材料评价的概念，在与BM-RMSC共培养的微环境中，比较了rEPC对聚乳酸和磷酸钙纳米颗粒(PLA+CAP)电纺复合材料的响应。我们的结果显示，在解放军+CAP的情况下，rEPC的迁移显著增加，几种促炎和促血管生成蛋白的表达上调。利用该平台还研究了骨桥蛋白(OPN)对rEPC迁移反应的影响，表明OPN在调节rEPC重新聚集到富钙微环境中具有重要作用。这一新工具可用于筛选各种生物活性支架诱导血管形成的能力，并加快生物材料的临床前测试。多年来，研究人员使用新生血管模型在体外评估生物活性生物材料，但由于其较差的生物仿真性和对细胞线索(如时空生物分子信号)和活体模型的最小控制，预测结果较低，存在成本高、耗时、人类外推能力差和伦理争议等缺点。我们描述了一个紧凑的微生理学平台，旨在通过量化模拟内皮细胞中能够对基于纤维蛋白的细胞外基质中生物材料释放的信号的梯度做出反应的萌芽水平来评估生物材料中的前血管生成。该模型对于组织再生的临床前可信性研究和更好地理解复杂的血管形成过程中涉及的不同因素是一个有用的工具。
Most of the conventionalin vitromodels to test biomaterial-driven vascularization are too simplistic to recapitulate the complex interactions taking place in the actual cell microenvironment, which results in a poor prediction of thein vivoperformance of the material. However, during the last decade, cell culture models based on microfluidic technology have allowed attaining unprecedented levels of tissue biomimicry. In this work, we propose a microfluidic-based 3D model to evaluate the effect of bioactive biomaterials capable of releasing signaling cues (such as ions or proteins) in the recruitment of endogenous endothelial progenitor cells, a key step in the vascularization process. The usability of the platform is demonstrated using experimentally-validated finite element models and migration and proliferation studies with rat endothelial progenitor cells (rEPCs) and bone marrow-derived rat mesenchymal stromal cells (BM-rMSCs). As a proof of concept of biomaterial evaluation, the response of rEPCs to an electrospun composite made of polylactic acid with calcium phosphates nanoparticles (PLA+CaP) was compared in a co-culture microenvironment with BM-rMSC to a regular PLA control. Our results show a significantly higher rEPCs migration and the upregulation of several pro-inflammatory and proangiogenic proteins in the case of the PLA+CaP. The effects of osteopontin (OPN) on the rEPCs migratory response were also studied using this platform, suggesting its important role in mediating their recruitment to a calcium-rich microenvironment. This new tool could be applied to screen the capacity of a variety of bioactive scaffolds to induce vascularization and accelerate the preclinical testing of biomaterials.Statement of significanceFor many years researchers have used neovascularization models to evaluate bioactive biomaterials bothin vitro, with low predictive results due to their poor biomimicry and minimal control over cell cues such as spatiotemporal biomolecule signaling, andin vivomodels, presenting drawbacks such as being highly costly, time-consuming, poor human extrapolation, and ethically controversial. We describe a compact microphysiological platform designed for the evaluation of proangiogenesis in biomaterials through the quantification of the level of sprouting in a mimicked endothelium able to react to gradients of biomaterial-released signals in a fibrin-based extracellular matrix. This model is a useful tool to perform preclinical trustworthy studies in tissue regeneration and to better understand the different elements involved in the complex process of vascularization.