Effect of honeycomb film on protein adsorption, cell adhesion and proliferation

Abstract This article describes novel methods for controlling of cell adhesion by using micro porous polymer films. Recently we found the highly ordered micro porous films were formed when poly(ɛ-caprolactone) (PCL) solution was cast on substrates at high atmospheric humidity. The micro porous film has regular honeycomb morphology with a size of 5 μm per cell (honeycomb film). Endothelial cells grew rapidly on the honeycomb film. After 24 h cell culture, the cell number on honeycomb films was lager than that on PCL flat films. In order to elucidate the effect of honeycomb films as a scaffold for cell culture, the adsorbed proteins on honeycomb films under cell culture condition were observed. After conditioning of the honeycomb film and the flat film in DMEM containing 10% foetal bovine serum (FBS) for 72 h at 37 °C in 5% CO2 atmosphere, the adsorbed fibronectin-FITC and albumin-Texasred on the honeycomb films was observed by using confocal laser scanning microscope (CLSM). The observation revealed that fibronectin showed site-selective adsorption behavior on the honeycomb film. Albumin adsorbed on the honeycomb film non site-selectively, while fibronectin mainly adsorbed on inside of honeycomb pores. On the flat film, fibronectin was hardly observed. Since the honeycomb film accelerate the adsorption of fibronectin which is a typical protein as a cell adhesion molecule, the film could be a scaffold with excellent cell adhesion properties.

[1]  S. P. Fodor,et al.  Light-directed, spatially addressable parallel chemical synthesis. , 1991, Science.

[2]  E. Delamarche,et al.  Patterned delivery of immunoglobulins to surfaces using microfluidic networks. , 1997, Science.

[3]  G. G. Stokes "J." , 1890, The New Yale Book of Quotations.

[4]  F. Ligler,et al.  New approach to producing patterned biomolecular assemblies , 1992 .

[5]  T. Matsuda,et al.  Photochemical Protein Fixation on Polymer Surfaces via Derivatized Phenyl Azido Group , 1995 .

[6]  Emmanuel Delamarche,et al.  Microcontact Printing of Proteins , 2000 .

[7]  Masaru Tanaka,et al.  Preparation of the Honeycomb Patterned Porous Film of Biodegradable Polymer for Tissue Engineering Scaffolds , 2002 .

[8]  G M Whitesides,et al.  Using self-assembled monolayers to understand the interactions of man-made surfaces with proteins and cells. , 1996, Annual review of biophysics and biomolecular structure.

[9]  S. Nishimura,et al.  Preparation of Self-Organized Micro-Patterned Polymer Films Having Cell Adhesive Ligands , 2002 .

[10]  Barbara Lom,et al.  A versatile technique for patterning biomolecules onto glass coverslips , 1993, Journal of Neuroscience Methods.

[11]  S. Nishimura,et al.  Mesoscopic patterning of cell adhesive substrates as novel biofunctional interfaces , 1999 .

[12]  Daniel I. C. Wang,et al.  Engineering cell shape and function. , 1994, Science.

[13]  Lance C. Kam,et al.  Patterning Hybrid Surfaces of Proteins and Supported Lipid Bilayers , 2000 .

[14]  G M Whitesides,et al.  Patterning cells and their environments using multiple laminar fluid flows in capillary networks. , 1999, Proceedings of the National Academy of Sciences of the United States of America.

[15]  Masatsugu Shimomura,et al.  Design of novel biointerfaces (II). Fabrication of self-organized porous polymer film with highly uniform pores. , 2004, Bio-medical materials and engineering.

[16]  M L Yarmush,et al.  Controlling cell interactions by micropatterning in co-cultures: hepatocytes and 3T3 fibroblasts. , 1997, Journal of biomedical materials research.

[17]  J M Calvert,et al.  Deep UV photochemistry of chemisorbed monolayers: patterned coplanar molecular assemblies. , 1991, Science.

[18]  K. Sugioka,et al.  Micropatterning of neurons using organic substrates in culture , 1997 .

[19]  G. Whitesides,et al.  Patterning ligands on reactive SAMs by microcontact printing , 1999 .

[20]  M. Shimomura,et al.  Morphological changes in neurons by self-organized patterned films , 2005 .

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

[22]  D. Nicolau,et al.  Negative and Positive Tone Protein Patterning on E-Beam/Deep-UV Resists , 1999 .

[23]  Masatsugu Shimomura,et al.  Mesoscopic patterns of molecular aggregates on solid substrates , 1998 .

[24]  Shin-Ichiro Nishimura,et al.  Honeycomb-patterned thin films of amphiphilic polymers as cell culture substrates , 1999 .

[25]  M. Shimomura,et al.  Novel Cell Culture Substrates based on Micro-Porous Films of Amphiphilic Polymers , 2001 .

[26]  C. S. Chen,et al.  Geometric control of cell life and death. , 1997, Science.