Thermoresponsive Copolymer Nanofilms for Controlling Cell Adhesion, Growth, and Detachment

Dalian R&D Center for Stem Cell and Tissue Engineering, Dalian University of Technology, Dalian 116024, China.
Langmuir (Impact Factor: 4.46). 10/2010; 26(22):17304-14. DOI: 10.1021/la102411u
Source: PubMed


This study reports the development and use of a novel thermoresponsive polymeric nanofilm for controlling cell adhesion and growth at 37 °C, and then cell detachment for cell recovery by subsequent temperature drop to the ambient temperature, without enzymatic cleavage or mechanical scraping. A copolymer, poly(N-isopropylacrylamide-co-hydroxypropyl methacrylate-co-3-(trimethoxysilyl)propyl methacrylate) (abbreviated PNIPAAm copolymer), was synthesized by free radical polymerization. The thermoresponses of the copolymer in aqueous solution were demonstrated by dynamic light scattering (DLS) through detecting the sensitive changes of copolymer aggregation against temperature. The DLS measurements revealed the lower critical solution temperature (LCST) at approximately 30 °C. The PNIPAAm film stability and robustness was provided through silyl cross-linking within the film and with the hydroxyl groups on the substrate surface. Film thickness, stability, and reversibility with respect to temperature switches were examined by spectroscopic ellipsometry (SE), atomic force microscopy (AFM), and contact angle measurements. The results confirmed the high extent of thermosensitivity and structural restoration based on the alterations of film thickness and surface wettability. The effective control of adhesion, growth, and detachment of HeLa and HEK293 cells demonstrated the physical controllability and cellular compatibility of the copolymer nanofilms. These PNIPAAm copolymer nanofilms could open up a convenient interfacial mediation for cell film production and cell expansion by nonenzymatic and nonmechanical cell recovery.

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    ABSTRACT: In this article we review recent results on the applications of stimuli-responsive polymers in fields such as development of drug carriers and active microfluidics. A special emphasis has been put on poly(N-isopropylacrylamide) (PNIPAM) and the related water-insoluble copolymers poly(methyl methacrylate/N-isopropylacrylamide) [P(MMA/NIPAM)] and poly(methyl methacrylate/N-isopropylacrylamide/acrylic acid) [P(MMA/NIPAM/AAc)] for the ease of their tunability and “smart” behaviour triggered by the change of temperature and pH. In particular, we discuss formation of PNIPAM nanoparticles loaded with fluorescent dye (as a model drug), which change the release rate in response to the temperature variation. We also describe nanofibers with PNIPAM nano-“raisins” facilitating the release rate on demand. Then, we consider synthesis and operation of water-insoluble thermo- and pH-sensitive hydrogels, which can be used either as nanofibers for drug release, or as components of microscopic devices changing surface wettability and volumetric flow rate in membranes and microchannels, as well as can be potentially used for development of self-cleaning micro- and nanochannels and fibers which shake loose deposits on demand.
    No preview · Article · May 2011 · Journal of Materials Chemistry
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    ABSTRACT: Temperature-responsive platforms containing poly(N-isopropylacrylamide) (PNIPAAm) have been developed as an effective substitute for enzymatic treatment to recover adherent cells, but it remains unclear whether this alternative harvesting method tends to support stem cells preserving them being primitive. This study mainly investigated the biological properties of mesenchymal stem cells derived from rat bone marrow and human adipose tissue (BM-MSCs and AT-MSCs) after being cultured on PNIPAAm copolymer films and recovered by temperature drop, and compared the cells harvested from glass coverslips with trypsinization as controls. The experimental results demonstrated that after three serial passages, the released MSCs from thermal liftoff showed no significant differences in cell morphology, immunophenotype and osteogenesis for BM-MSCs or adipogenesis for AT-MSCs, but had higher viability, stronger proliferation and higher adipogenic differentiation for BM-MSCs or higher osteogenic differentiation for AT-MSCs compared with the trypsinization group. Besides, more proteins remained around or within the cell membranes upon temperature drop. It is concluded that cell detachment with more extracellular matrix proteins facilitates the maintenance of membrane proteins, and accordingly preserves MSC properties related to viability, proliferation and differentiation to some extent. This indicates that the PNIPAAm copolymer films and their matching cooling treatment can be used as effective alternatives to the existing culture substrates and traditional enzymatic digestion for MSCs.
    No preview · Article · Feb 2012 · Biomedical Materials
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    ABSTRACT: Two types of thermoresponsive microgels, poly(N-isopropylacrylamide) (PNIPAM) microgels and poly(N-isopropylacrylamide-co-acrylic acid) (PNIPAMAC) microgels were synthesized and used as templates for the mineralization of amorphous calcium carbonate (ACC) by diffusion of CO(2) vapor under ambient conditions. Thermosensitive PNIPAM/CaCO(3) hybrid macroscopic hydrogels and micrometer-sized PNIPAMAC/CaCO(3) hybrid microgels were controllably obtained and different mineralization mechanistic processes were proposed. The impact of the loaded CaCO(3) on the size, morphology, stability, and thermosensitivity of the microgels was also analyzed. PNIPAM/CaCO(3) hybrid macrogels had a slight decrease in thermoresponsive phase transition temperature, while PNIPAMAC/CaCO(3) hybrid microgels showed a clear increase in phase transition temperature. The difference reflected different amount and location of ACC in the gel network, causing different interactions with polymer chains. The PNIPAMAC/CaCO(3) microgels formed stable monolayer films on bare silica wafers and glass coverslips upon drying. The microgel films could facilitate the attachment and growth of 3T3 fibroblast cells and their subsequent detachment upon temperature drop from 37 °C to the ambient condition around 20 °C, thus, offering a convenient procedure for cell harvesting.
    Full-text · Article · Jun 2012 · Biomacromolecules
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