Shenzhen Key Laboratory of Biomimetic Materials and Cellular Immunomodulation, Shenzhen Institute of Advanced Technology, Chinese Academy of Sciences, Shenzhen, Guangdong 518055, China.
University of Chinese Academy of Sciences, Beijing 100049, China.
ACS Appl Mater Interfaces. 2021 Apr 28;13(16):18563-18580. doi: 10.1021/acsami.1c02969. Epub 2021 Apr 16.
Hybrid surfaces with tunable topography, chemistry, and stiffness have potential to rebuild native extracellular matrix (ECM) and manipulate cell behavior in vitro. However, the fabrication of controllable hybrid surfaces is still challenging. In this study, colloidal self-assembly technology was used to program particles into highly ordered structures with hybrid chemistry and stiffness at biointerfaces. These colloidal self-assembled patterns (cSAPs), including unary, binary, and ternary cSAPs, composed of silicon (Si), polystyrene (PS), and/or poly(-isopropylacrylamide) (pNIPAM) nanogels (PNGs), were fabricated using either coassembly or layer-by-layer (LBL) methods. The selected binary cSAPs (i.e., PS/PNG and PNG/PS) have a tunable surface topography and wettability between 25 and 37 °C; thus, they can be used as dynamic cell culture substrates. Human adipose-derived mesenchymal stem cells (hASCs), bone marrow-derived mesenchymal stem cells (hBMSCs), and macrophages (THP-1) were investigated on these hybrid cSAPs under a static or dynamic system. The results showed that hybrid cSAPs significantly influenced the focal adhesions, cell morphology, cell migration, and gene expressions of stem cells. In general, stem cells had more vinculin puncta, smaller spreading size, and faster migration speed than the TCPS control. Hybrid cSAPs up-regulated gene expressions of focal adhesion kinase (FAK) and chondrocytes (AGG and SOX9) under static culture, while they also up-regulated osteocytes (COL1 and RUNX2) under dynamic culture. THP-1 macrophages were at M0 state on all cSAPs under static culture. However, cells became sensitive under dynamic culture. For example, some M1 genes (i.e., IL6, CD68, and TNFα) and M2 genes (i.e., IL10 and CD206) were down-regulated, while other M1 genes (i.e., IL1β) and M2 genes (i.e., TGF-β and IL1ra) were up-regulated, depending on the particle combinations. In conclusion, new hybrid cSAPs with thermoresponsive surface properties are versatile materials for stem cells and macrophages manipulation.
具有可调形貌、化学性质和力学性质的杂化表面有潜力在体外重建天然细胞外基质 (ECM) 并调控细胞行为。然而，可控杂化表面的制备仍然具有挑战性。在这项研究中，胶体自组装技术被用于在生物界面上构建具有杂化化学性质和力学性质的高度有序结构。这些胶体自组装图案（cSAPs），包括单一组分、双组分和三组分 cSAPs，由硅（Si）、聚苯乙烯（PS）和/或聚（异丙基丙烯酰胺）（pNIPAM）纳米凝胶（PNGs）组成，通过共组装或层层（LBL）方法制备。选择的双组分 cSAPs（即 PS/PNG 和 PNG/PS）在 25-37°C 之间具有可调的表面形貌和润湿性；因此，它们可用作动态细胞培养的基底。在静态或动态系统下，将人脂肪间充质干细胞（hASCs）、骨髓间充质干细胞（hBMSCs）和巨噬细胞（THP-1）接种到这些杂化 cSAPs 上进行研究。结果表明，杂化 cSAPs 显著影响了干细胞的焦点附着、细胞形态、细胞迁移和基因表达。一般来说，与 TCPS 对照相比，干细胞具有更多的 vinculin 斑点、更小的扩展面积和更快的迁移速度。在静态培养下，杂化 cSAPs 上调了粘着斑激酶（FAK）和软骨细胞（AGG 和 SOX9）的基因表达，而在动态培养下，也上调了成骨细胞（COL1 和 RUNX2）的基因表达。在静态培养下，THP-1 巨噬细胞在所有 cSAPs 上都处于 M0 状态。然而，在动态培养下，细胞变得敏感。例如，一些 M1 基因（如 IL6、CD68 和 TNFα）和 M2 基因（如 IL10 和 CD206）下调，而其他 M1 基因（如 IL1β）和 M2 基因（如 TGF-β 和 IL1ra）上调，这取决于颗粒的组合。总之，具有温度响应表面性质的新型杂化 cSAPs 是一种用于调控干细胞和巨噬细胞的多功能材料。
