Abstract
Microgels are cross-linked soft particles with a three-dimensional network structure that are swollen in a good solvent. Poly(N-isopropylacrylamide) (pNIPAAm)-based microgels have attracted great attentions for their temperature responsive property, particularly in recent years, pNIPAAm-based microgel films were utilized as a new kind of thermoresponsive surface to tune cell attachment/detachment behavior via temperature stimuli. However, some results are not consistent, for example, different polymerization conditions may bring out different results even for pure pNIPAAm microgel. This work aims to find out which factor plays the critical role for successful cell detachment on the pNIPAAm-based microgel films. The results unraveled that the structure and swelling ratio of the microgel rather than the film thickness plays a key role on the successful cells detachment, unlike linear pNIPAAm films in which the cells’ attach/detach property is only determined by the film thickness. For poly(N-isopropylacrylamide–styrene) microgel film, NIH3T3 cells could only detach when the microgel has a uniform structure and the volume dilatation of the microgel (20/38 °C) is larger than 4.
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Elloumi-Hannachi I, Yamato M, Okano T (2010) Cell sheet engineering: a unique nanotechnology for scaffold-free tissue reconstruction with clinical applications in regenerative medicine. Biomaterials 31:8974–8979
Takahashi H, Nakayama M, Itoga K, Yamato M, Okano T (2011) Micropatterned thermoresponsive polymer brush surfaces for fabricating cell sheets with well-controlled orientational structures. Biomacromolecules 12:1414–1418
Ohtsuki K, Miyai S, Yamaguchi A, Morikawa K, Okano T (2012) Biochemical characterization of novel lignans isolated from the wood of Taxus yunnanensis as effective stimulators for glycogen synthase kinase-3beta and the phosphorylation of basic brain proteins by the kinase in vitro. Biol Pharm Bull 35:385–393
Patel NG, Cavicchia JP, Zhang G, Zhang NB (2012) Rapid cell sheet detachment using spin-coated pNIPAAm films retained on surfaces by an aminopropyltriethoxysilane network. Acta Biomater 8:2559–2567
Tang Z, Akiyama Y, Yamato M, Okano T (2010) Comb-type grafted poly(N-isopropylacrylamide) gel modified surfaces for rapid detachment of cell sheet. Biomaterials 31:7435–7443
Chen YH, Chung YC, Wang IJ, Young TH (2012) Control of cell attachment on pH-responsive chitosan surface by precise adjustment of medium pH. Biomaterials 33:1336–1342
Zahn R, Thomasson E, Guillaume-Gentil O, Voros J, Zambelli T (2012) Ion-induced cell sheet detachment from standard cell culture surfaces coated with polyelectrolytes. Biomaterials 33:3421–3427
Hong Y, Yu M, Weng W, Cheng K, Wang H, Lin J (2013) Light-induced cell detachment for cell sheet technology. Biomaterials 34:11–18
Nandkumar MA, Yamato M, Kushida A, Konno C, Hirose M, Kikuchi A, Okano T (2002) Two-dimensional cell sheet manipulation of heterotypically co-cultured lung cells utilizing temperature-responsive culture dishes results in long-term maintenance of differentiated epithelial cell functions. Biomaterials 23:1121–1130
Kwon OH, Kikuchi A, Yamato M, Sakurai Y, Okano T (2000) Rapid cell sheet detachment from poly(N-isopropylacrylamide)-grafted porous cell culture membranes. J Biomed Mater Res 50:82–89
Halperin A, Kröger M (2012) Thermoresponsive cell culture substrates based on PNIPAM brushes functionalized with adhesion peptides: theoretical considerations of mechanism and design. Langmuir 28:16623–16637
Vaquette C, Fan W, Xiao Y, Hamlet S, Hutmacher DW, Ivanovski S (2012) A biphasic scaffold design combined with cell sheet technology for simultaneous regeneration of alveolar bone/periodontal ligament complex. Biomaterials 33:5560–5573
Halperin A, Kroger M (2012) Theoretical considerations on mechanisms of harvesting cells cultured on thermoresponsive polymer brushes. Biomaterials 33:4975–4987
Takahashi H, Nakayama M, Yamato M, Okano T (2010) Controlled chain length and graft density of thermoresponsive polymer brushes for optimizing cell sheet harvest. Biomacromolecules 11:1991–1999
Schmaljohann D, Oswald J, Jorgensen B, Nitschke M, Beyerlein D, Werner C (2003) Thermo-responsive PNiPAAm-g-PEG films for controlled cell detachment. Biomacromolecules 4:1733–1739
Matsuda TSN, Yamato M, Okano T (2007) Tissue engineering based on cell sheet technology. Adv Mater 19:3089–3099
Akiyama Y, Kikuchi A, Yamato M, Okano T (2004) Ultrathin poly(N-isopropylacrylamide) grafted layer on polystyrene surfaces for cell adhesion/detachment control. Langmuir 20:5506–5511
Fukumori K, Akiyama Y, Yamato M, Kobayashi J, Sakai K, Okano T (2009) Temperature-responsive glass coverslips with an ultrathin poly(N-isopropylacrylamide) layer. Acta Biomater 5:470–476
Li L, Zhu Y, Li B, Gao C (2008) Fabrication of thermoresponsive polymer gradients for study of cell adhesion and detachment. Langmuir 24:13632–13639
Nagase K, Watanabe M, Kikuchi A, Yamato M, Okano T (2011) Thermo-responsive polymer brushes as intelligent biointerfaces: preparation via ATRP and characterization. Macromol Biosci 11:400–409
Schmidt S, Zeiser M, Hellweg T, Duschl C, Fery A, Möhwald H (2010) Adhesion and mechanical properties of PNIPAM microgel films and their potential use as switchable cell culture substrates. Adv Funct Mater 20:3235–3243
Pan G, Guo Q, Ma Y, Yang H, Li B (2013) Thermo-responsive hydrogel layers imprinted with RGDS peptide: a system for harvesting cell sheets. Angew Chem Int Ed 52:6907–6911
Heinen S, Cuéllarcamacho JL, Weinhart M (2017) Thermoresponsive poly(glycidyl ether) brushes on gold: surface engineering parameters and their implication for cell sheet fabrication. Acta Biomater 59:117–128
Xia Y, He X, Cao M, Chen C, Xu H, Pan F, Lu JR (2013) Thermoresponsive microgel films for harvesting cells and cell sheets. Biomacromolecules 14:3615–3625
Xia Y, He X, Cao M, Wang X, Sun Y, He H, Xu H, Lu JR (2014) Self-assembled two-dimensional thermoresponsive microgel arrays for cell growth/detachment control. Biomacromolecules 15:4021–4031
Uhlig K, Wegener T, He J, Zeiser M, Bookhold J, Dewald I, Godino N, Jaeger M, Hellweg T, Fery A, Dusch C (2016) Patterned thermoresponsive microgel coatings for noninvasive processing of adherent cells. Biomacromolecules 17:1110–1116
Wei J, Cai J, Li Y, Wu B, Gong X, Ngai T (2015) Investigation of cell behaviors on thermo-responsive PNIPAM microgel films. Colloid Surf B 132:202–207
Nash ME, Carroll WM, Nikoloskya N, Yang R, O’Connell C, Gorelov AV, Dockery P, Liptrot C, Lyng FM, Garcia A, Rochev YA (2011) Straightforward, one-step fabrication of ultrathin thermoresponsive films from commercially available pNIPAm for cell culture and recovery. ACS Appl Mater Interfaces 3:1980–1990
Tsai H-Y, Vats K, Yates MZ, Benoit DSW (2013) Two-dimensional patterns of poly(N-isopropylacrylamide) microgels to spatially control fibroblast adhesion and temperature-responsive detachment. Langmuir 29:12183–12193
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The authors gratefully acknowledge the financial support from the Natural Science Fund of Shandong Province (ZR2015BM013) and the National Natural Science Foundation of China (21773310).
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Xia, Y., Tang, D., Wu, H. et al. Cell attachment/detachment behavior on poly(N-isopropylacrylamide)-based microgel films: the effect of microgel structure and swelling ratio. J Mater Sci 53, 8795–8806 (2018). https://doi.org/10.1007/s10853-018-2217-4
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DOI: https://doi.org/10.1007/s10853-018-2217-4