Neural cell pattern formation on glass and oxidized silicon surfaces modified with poly(N-isopropylacrylamide).

Control over the adsorption of proteins and over the adsorption and spatial orientation of mammalian cells onto surfaces has been achieved by modification of glass and other silicon oxide substrates with poly(N-isopropylacrylamide) (PNIPAM). The functionalization of the substrates was achieved either by a polymer-analogous reaction of aminosilanes with reactive N-(isopropylacrylamide) (NIPAM)-copolymers and by copolymerization of NIPAM with surface-bound methacrylsilane. The obtained coatings were characterized by FT-1R, ellipsometry, and surface plasmon resonance measurements. The adsorption of two proteins-fibrinogen and ribonuclease A-on these surfaces was studied in situ by real time surface plasmon resonance measurements. The PNIPAM-grafted surfaces prepared by either chemical procedure inhibited the adsorption of both proteins. More importantly they prevented the adhesion of neuroblastomaXglioma hybrid cells cultured either in serum-free medium or in a medium containing serum proteins. Deep-UV irradiation was used to perform ablation processes and to create patterns permitting the examination of spatially controlled adhesion and growth of cells. This study showed that patterned ultrathin polymer films on glass are suitable substrates for controlling the interactions of cells with surfaces and are capable of directing the attachment and spreading of cells.

[1]  G Blume,et al.  Molecular mechanism of the lipid vesicle longevity in vivo. , 1993, Biochimica et biophysica acta.

[2]  H. Sasabe,et al.  Micropatterning of cultured cells on polystyrene surface by using an excimer laser , 1994 .

[3]  Jack L. Koenig,et al.  An investigation of the coupling agent/matrix interface of fiberglass reinforced plastics by fourier transform infrared spectroscopy , 1979 .

[4]  M. Heskins,et al.  Solution Properties of Poly(N-isopropylacrylamide) , 1968 .

[5]  A. Hoffman,et al.  Preparation of a thermally phase‐separating copolymer, poly(N‐isopropylacrylamide‐co‐N‐acryloxysuccinimide), with a controlled number of active esters per polymer chain , 1990 .

[6]  E. Yavin,et al.  ATTACHMENT AND CULTURE OF DISSOCIATED CELLS FROM RAT EMBRYO CEREBRAL HEMISPHERES ON POLYLYSINE-COATED SURFACE , 1974, The Journal of cell biology.

[7]  H. Ringsdorf,et al.  Wechselwirkung von hydrophob modifizierten Poly‐N‐isopropylacrylamiden mit Modellmembranen ‐ oder das Spielen einer molekularen Ziehharmonika , 1991 .

[8]  W. Knoll,et al.  Patterning molecularly thin films of polymers—new methods for photolithographic structuring of surfaces , 1995 .

[9]  D Needham,et al.  Repulsive interactions and mechanical stability of polymer-grafted lipid membranes. , 1992, Biochimica et biophysica acta.

[10]  Charles S. Dulcey,et al.  Coplanar molecular assemblies of amino- and perfluorinated alkylsilanes : characterization and geometric definition of mammalian cell adhesion and growth , 1992 .

[11]  Joseph A. Gardella,et al.  Monolayer chemical lithography and characterization of fluoropolymer films , 1992 .

[12]  Joseph D. Andrade,et al.  Protein—surface interactions in the presence of polyethylene oxide , 1991 .

[13]  George M. Whitesides,et al.  Convenient methods for patterning the adhesion of mammalian cells to surfaces using self-assembled monolayers of alkanethiolates on gold , 1993 .

[14]  P C Letourneau,et al.  Guidance of neurite outgrowth by pathways of substratum‐adsorbed laminin , 1985, Journal of neuroscience research.

[15]  J. Hubbell,et al.  Covalent surface immobilization of Arg-Gly-Asp- and Tyr-Ile-Gly-Ser-Arg-containing peptides to obtain well-defined cell-adhesive substrates. , 1990, Analytical biochemistry.

[16]  B D Ratner,et al.  New ideas in biomaterials science--a path to engineered biomaterials. , 1993, Journal of biomedical materials research.

[17]  A Cooper,et al.  The growth of mouse neuroblastoma cells in controlled orientations on thin films of silicon monoxide. , 1976, Experimental cell research.

[18]  E. Kretschmann.  Die Bestimmung optischer Konstanten von Metallen durch Anregung von Oberflächenplasmaschwingungen , 1971 .

[19]  Wolfgang Knoll,et al.  Optical Characterization of Organic Thin Films and Interfaces with Evanescent Waves , 1991 .

[20]  N. Busis,et al.  Modulation of synapse formation by cyclic adenosine monophosphate. , 1983, Science.

[21]  H. G. Schild.  Poly(N-isopropylacrylamide): experiment, theory and application , 1992 .

[22]  A. Wlodawer,et al.  Structure of phosphate-free ribonuclease A refined at 1.26 A. , 1988, Biochemistry.

[23]  T. Okano,et al.  Dynamic contact angle measurement of temperature-responsive surface properties for poly(N-isopropylacrylamide) grafted surfaces , 1994 .

[24]  G. Whitesides,et al.  Self-assembled organic monolayers: model systems for studying adsorption of proteins at surfaces , 1991, Science.

[25]  Ingemar Lundström,et al.  Structure of 3-aminopropyl triethoxy silane on silicon oxide , 1991 .

[26]  M. Woodle,et al.  Sterically stabilized liposomes. , 1992, Biochimica et biophysica acta.

[27]  S. Carter,et al.  Principles of Cell Motility: The Direction of Cell Movement and Cancer Invasion , 1965, Nature.

[28]  F. Winnik.  Fluorescence studies of aqueous solutions of poly(N-isopropylacrylamide) below and above their LCST , 1990 .

[29]  K. Ulbrich,et al.  Synthetic polymer derivatives as substrata for neuronal adhesion and growth , 1993, Brain Research Bulletin.

[30]  P. Fromherz,et al.  A neuron-silicon junction: a Retzius cell of the leech on an insulated-gate field-effect transistor. , 1991, Science.

[31]  D Kleinfeld,et al.  Controlled outgrowth of dissociated neurons on patterned substrates , 1988, The Journal of neuroscience : the official journal of the Society for Neuroscience.

[32]  H. Ringsdorf,et al.  Fluorescence studies of hydrophobically modified poly(N-isopropylacrylamides) , 1991 .

[33]  V. Torchilin,et al.  Activity of amphipathic poly(ethylene glycol) 5000 to prolong the circulation time of liposomes depends on the liposome size and is unfavorable for immunoliposome binding to target. , 1991, Biochimica et biophysica acta.

[34]  A. Ulman,et al.  Ultrathin organic films: From Langmuir-Blodgett to self assembly , 1991 .

[35]  E Ruoslahti,et al.  New perspectives in cell adhesion: RGD and integrins. , 1987, Science.

[36]  T. Okano,et al.  Thermo‐responsive polymeric surfaces; control of attachment and detachment of cultured cells , 1990 .

[37]  E. Sackmann,et al.  Wetting and dewetting of Si/SiO2-wafers by free and lipidmonolayer covered aqueous solutions under controlled humidity , 1994 .