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响应声音模式驱动的局部细胞密度增强的微毛细管网络的差异蛋白质组学图谱。

Differential proteomics profile of microcapillary networks in response to sound pattern-driven local cell density enhancement.

作者信息

Di Marzio N, Tognato R, Bella E Della, De Giorgis V, Manfredi M, Cochis A, Alini M, Serra T

机构信息

AO Research Institute Davos, 7270 Davos, Switzerland.

Department of Health Sciences, Center for Translational Research and Autoimmune and Allergic Diseases (CAAD), University of Piemonte Orientale, 28100 Novara, Italy.

出版信息

Biomater Biosyst. 2024 Mar 29;14:100094. doi: 10.1016/j.bbiosy.2024.100094. eCollection 2024 Jun.

Abstract

Spatial cell organization and biofabrication of microcapillary networks in vitro has a great potential in tissue engineering and regenerative medicine. This study explores the impact of local cell density enhancement achieved through an innovative sound-based patterning on microcapillary networks formation and their proteomic profile. Human umbilical vein endothelial cells (HUVEC) and human pericytes from placenta (hPC-PL) were mixed in a fibrin suspension. The mild effect of sound-induced hydrodynamic forces condensed cells into architected geometries showing good fidelity to the numerical simulation of the physical process. Local cell density increased significantly within the patterned areas and the capillary-like structures formed following the cell density gradient. Over five days, these patterns were well-maintained, resulting in concentric circles and honeycomb-like structures. Proteomic analysis of the pre-condensed cells cultured for 5 days, revealed over 900 differentially expressed proteins when cells were preassembled through mild-hydrodynamic forces. Gene ontology (GO) enrichment analysis identified cellular components, molecular functions, and biological processes that were up- and down-regulated, providing insights regarding molecular processes influenced by the local density enhancement. Furthermore, we employed Ingenuity Pathway Analysis (IPA) to identify altered pathways and predict upstream regulators. Notably, VEGF-A emerged as one of the most prominent upstream regulators. Accordingly, this study initiates the unraveling of the changes in microcapillary networks at both molecular and proteins level induced by cell condensation obtained through sound patterning. The findings provide valuable insights for further investigation into sound patterning as a biofabrication technique for creating more complex microcapillary networks and advancing in vitro models.

摘要

体外微毛细管网络的空间细胞组织和生物制造在组织工程和再生医学中具有巨大潜力。本研究探讨了通过创新的基于声音的图案化实现的局部细胞密度增强对微毛细管网络形成及其蛋白质组学特征的影响。将人脐静脉内皮细胞(HUVEC)和来自胎盘的人周细胞(hPC-PL)混合在纤维蛋白悬浮液中。声音诱导的流体动力的温和作用将细胞凝聚成结构化几何形状,与物理过程的数值模拟显示出良好的保真度。图案化区域内的局部细胞密度显著增加,并且沿着细胞密度梯度形成了毛细血管样结构。在五天的时间里,这些图案得到了很好的维持,形成了同心圆和蜂窝状结构。对预凝聚细胞培养5天进行蛋白质组学分析,结果显示当细胞通过温和流体动力进行预组装时,有900多种差异表达蛋白。基因本体(GO)富集分析确定了上调和下调的细胞成分、分子功能和生物学过程,为受局部密度增强影响的分子过程提供了见解。此外,我们采用 Ingenuity 通路分析(IPA)来识别改变的通路并预测上游调节因子。值得注意的是,VEGF-A 成为最突出的上游调节因子之一。因此,本研究开始揭示通过声音图案化获得的细胞凝聚在分子和蛋白质水平上对微毛细管网络的影响。这些发现为进一步研究声音图案化作为一种生物制造技术以创建更复杂的微毛细管网络和推进体外模型提供了有价值的见解。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/9963/11001772/8d70f25a6b1b/ga1.jpg

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