Supramolecular Surface Coatings with Acetylated Polyrotaxane-Based Triblock Copolymers for Thermal Regulation of Cell Adhesion and Fabrication of Cell Sheets
Atsushi Tamura*, Tae Woong Kang, Asato Tonegawa, Yoshinori Arisaka, Hiroki Masuda, Ryo Mikami, Takanori Iwata, Tetsuya Yoda, and Nobuhiko Yui
Department of Organic Biomaterials, Institute of Biomaterials and Bioengineering, Tokyo Medical and Dental University (TMDU), 2-3-10 Kanda-Surugadai, Chiyoda, Tokyo 101-0062, Japan
Biomacromolecules 2022, XXXX, XXX, XXX-XXX
Publication Date: October 7, 2022
https://doi.org/10.1021/acs.biomac.2c01043

Abstract
Polyrotaxanes (PRXs) containing acetylated α-cyclodextrins exhibit a temperature-dependent phase transition in aqueous solutions across their lower critical solution temperature (LCST) of approximately 26.6 °C. To gain insights into the interactions of acetylated PRXs (Ac-PRXs) with biological components, thermoresponsive supramolecular surfaces were prepared by coating tissue culture polystyrene (TCPS) surfaces with Ac-PRX triblock copolymers, and their surface properties across the LCST were evaluated. The wettability and protein adsorption of Ac-PRX-coated surfaces changed significantly between 10 and 37 °C, whereas the uncoated TCPS and unmodified PRX-coated surfaces did not alter the wettability and protein adsorption at 10 and 37 °C. The adhesion, proliferation, morphology, and adhesion strength of NIH/3T3 cells on Ac-PRX-coated surfaces were found to be similar to those of the uncoated and unmodified PRX-coated surfaces. However, the adhesion strength of NIH/3T3 cells on Ac-PRX-coated surfaces decreased drastically at 10 °C. Consequently, the cells spontaneously detached from the Ac-PRX-coated surfaces without enzymatic treatment. Additionally, when incubating confluent cells at 10 °C, the cells detached from Ac-PRX-coated surfaces as cell sheets while retaining extracellular matrix proteins. The findings of this study provide new directions for the design of thermoresponsive supramolecular biointerfaces for applications in bioseparation and cell manipulation.
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摘要
含有乙酰化α-环糊精的聚轮烷(PRXs)在其约26.6°C的较低临界溶液温度(LCST)下,在水溶液中表现出温度依赖性相变。为了深入了解乙酰化PRX(Ac-PRX)与生物组分的相互作用,通过用Ac-PRX三嵌段共聚物涂覆组织培养聚苯乙烯(TCPS)表面来制备热响应超分子表面,并评估其在LCST上的表面性质。Ac-PRX涂层表面的润湿性和蛋白质吸附在10至37°C之间发生显著变化,而未涂层的TCPS和未改性的PRX涂层表面在10和37°C下不会改变润湿性及蛋白质吸附。发现Ac-PRX涂层表面上NIH/3T3细胞的粘附、增殖、形态和粘附强度与未涂层和未改性PRX涂层表面相似。然而,NIH/3T3细胞在Ac-PRX涂层表面上的粘附强度在10℃下急剧下降。因此,细胞在没有酶处理的情况下自发地从Ac-PRX涂层表面分离。此外,当在10°C下孵育融合细胞时,细胞从Ac-PRX涂层表面分离为细胞片,同时保留细胞外基质蛋白。本研究的发现为热响应超分子生物界面的设计提供了新的方向,用于生物分离和细胞操纵。