Heinen Silke, Rackow Simon, Cuellar-Camacho Jose Luis, Donskyi Ievgen S, Unger Wolfgang E S, Weinhart Marie
Institute of Chemistry and Biochemistry, Freie Universitaet Berlin, Takustr. 3, 14195 Berlin, Germany.
J Mater Chem B. 2018 Mar 14;6(10):1489-1500. doi: 10.1039/c7tb03263c. Epub 2018 Feb 21.
Thermoresponsive polymer coatings can facilitate cell sheet fabrication under mild conditions by promoting cell adhesion and proliferation at 37 °C. At lower temperatures the detachment of confluent cell sheets is triggered without enzymatic treatment. Thus, confluent cell sheets with intact extracellular matrix for regenerative medicine or tissue engineering applications become available. Herein, we applied the previously identified structural design parameters of functional, thermoresponsive poly(glycidyl ether) brushes on gold to the more application-relevant substrate glass via the self-assembly of a corresponding block copolymer (PGE-AA) with a short surface-reactive, amine-presenting anchor block. Both, physical and covalent immobilization on glass via either multivalent ionic interactions of the anchor block with bare glass or the coupling of the anchor block to a polydopamine (PDA) adhesion layer on glass resulted in stable coatings. Atomic force microscopy revealed a high degree of roughness of covalently attached coatings on the PDA adhesion layer, while physically attached coatings on bare glass were smooth and in the brush-like regime. Cell sheets of primary human dermal fibroblasts detached reliably (86%) and within 20 ± 10 min from physically tethered PGE-AA coatings on glass when prepared under cloud point grafting conditions. The presence of the laterally inhomogeneous PDA adhesion layer, however, hindered the spontaneous temperature-triggered cell detachment from covalently grafted PGE-AA, decreasing both detachment rate and reliability. Despite being only physically attached, self-assembled monolayer brushes of PGE-AA block copolymers on glass are functional and stable thermoresponsive coatings for application in cell sheet fabrication of human fibroblasts as determined by X-ray photoelectron spectroscopy.
热响应性聚合物涂层可以在温和条件下通过促进细胞在37°C时的粘附和增殖来促进细胞片的制备。在较低温度下,无需酶处理即可触发汇合细胞片的脱离。因此,可获得具有完整细胞外基质的汇合细胞片,用于再生医学或组织工程应用。在此,我们通过具有短表面反应性、含胺锚定嵌段的相应嵌段共聚物(PGE-AA)的自组装,将先前确定的功能性热响应性聚(缩水甘油醚)刷在金上的结构设计参数应用于更具应用相关性的基底玻璃上。通过锚定嵌段与裸玻璃的多价离子相互作用或锚定嵌段与玻璃上的聚多巴胺(PDA)粘附层的偶联,在玻璃上进行物理和共价固定均产生了稳定的涂层。原子力显微镜显示,PDA粘附层上共价连接的涂层具有高度粗糙度,而裸玻璃上物理连接的涂层则是光滑的且处于刷状状态。当在浊点接枝条件下制备时,原代人真皮成纤维细胞片能可靠地(86%)在20±10分钟内从玻璃上物理连接的PGE-AA涂层上脱离。然而,横向不均匀的PDA粘附层的存在阻碍了共价接枝的PGE-AA上细胞的自发温度触发脱离,降低了脱离速率和可靠性。尽管只是物理连接,但通过X射线光电子能谱测定,玻璃上PGE-AA嵌段共聚物的自组装单分子层刷是用于人成纤维细胞片制备的功能性且稳定的热响应性涂层。