Institute of Bioengineering, University of London, Mile End Road, London E1 4NS, United Kingdom.
School of Engineering and Materials Science, Queen Mary, University of London, Mile End Road, London E1 4NS, United Kingdom.
Biomed Mater. 2023 Mar 17;18(3). doi: 10.1088/1748-605X/acbdda.
Bioemulsions are attractive platforms for the expansion of adherent cells in bioreactors. Their design relies on the self-assembly of protein nanosheets at liquid-liquid interfaces, displaying strong interfacial mechanical properties and promoting integrin-mediated cell adhesion. However, most systems developed to date have focused on fluorinated oils, which are unlikely to be accepted for direct implantation of resulting cell products for regenerative medicine, and protein nanosheets self-assembly at other interfaces has not been investigated. In this report, the composition of aliphatic pro-surfactants palmitoyl chloride and sebacoyl chloride, on the assembly kinetics of poly(L-lysine) at silicone oil interfaces and characterisation of ultimate interfacial shear mechanics and viscoelasticity is presented. The impact of the resulting nanosheets on the adhesion of mesenchymal stem cells (MSCs) is investigated via immunostaining and fluorescence microscopy, demonstrating the engagement of the classic focal adhesion-actin cytoskeleton machinery. The ability of MSCs to proliferate at the corresponding interfaces is quantified. In addition, expansion of MSCs at other non-fluorinated oil interfaces, based on mineral and plant-based oils is investigated. Finally, the proof-of-concept of such non-fluorinated oil systems for the formulation of bioemulsions supporting stem cell adhesion and expansion is demonstrated.
生物乳剂是在生物反应器中扩大贴壁细胞的有吸引力的平台。它们的设计依赖于蛋白质纳米片在液-液界面的自组装，显示出强大的界面力学性能，并促进整合素介导的细胞黏附。然而，迄今为止开发的大多数系统都集中在氟化油上，这些油不太可能被直接用于再生医学中细胞产物的植入，而其他界面上的蛋白质纳米片自组装尚未得到研究。在本报告中，介绍了脂肪族助表面活性剂棕榈酰氯和癸酰氯对聚（L-赖氨酸）在硅油界面上组装动力学的影响，以及最终界面剪切力学和粘弹性的特性。通过免疫染色和荧光显微镜研究了所得纳米片对间充质干细胞（MSCs）黏附的影响，证明了经典的黏附斑-肌动蛋白细胞骨架机制的参与。定量研究了 MSCs 在相应界面上的增殖能力。此外，还研究了基于矿物油和植物油的其他非氟化油界面上 MSCs 的扩展。最后，证明了这种非氟化油系统在支持干细胞黏附和扩展的生物乳剂配方中的概念验证。
