Micropattern array with gradient size (µPAGS) plastic surfaces fabricated by PDMS (polydimethylsiloxane) mold-based hot embossing technique for investigation of cell–surface interaction

Recently, it was found that the variations of physical environment significantly affect cell behaviors including cell proliferation, migration and differentiation. Through a plastic surface with controlled mechanical properties such as stiffness, one can change the orientation and migration of cells in a particular direction, thereby determining cell behaviors. In this study, we demonstrate a polydimethylsiloxane (PDMS) mold-based hot embossing technique for rapid, simple and low-cost replication of polystyrene (PS) surfaces having micropatterns. The PDMS mold was fabricated by UV-photolithography followed by PDMS casting; the elastomeric properties of PDMS enabled us to obtain conformal contact of the PDMS mold to a PS surface and to create high transcription quality of micropatterns on the PS surface. Two different types of circular micropillar and microwell arrays were successfully replicated on the PS surfaces based on the suggested technique. The micropatterns were designed to have various diameters (2-150 µm), spacings (2-160 µm) and heights (1.4, 2.4, 8.2 and 14.9 µm), so as to generate the gradient of physical properties on the surface. Experimental parametric studies indicated that (1) the embossing temperature became a critical processing parameter as the aspect ratio of micropattern increased and (2) the PDMS mold-based hot embossing could successfully replicate micropatterns, even having an aspect ratio of 2.7 for micropattern diameter of 6 µm, with an optimal processing condition (embossing pressure and temperature of 0.4 MPa and 130 °C, respectively) in this study. We carried out cell experiments with adipose-derived stem cells on the replicated PS surface with the height of 1.4 µm to investigate cellular behaviors in response to the micropattern array with gradient size. Cellular experiment results showed that the micropillar-arrayed surface improved cell proliferation as compared with the microwell-arrayed surface. We could also estimate the ranges of pattern sizes having the desired effects on the cellular behaviors.

[1]  Daniel T Chiu,et al.  Fabrication of thermoset polyester microfluidic devices and embossing masters using rapid prototyped polydimethylsiloxane molds. , 2003, Lab on a chip.

[2]  Jian Yu,et al.  Femtosecond laser ablation enhances cell infiltration into three-dimensional electrospun scaffolds. , 2012, Acta biomaterialia.

[3]  Won Mook Choi,et al.  The fabrication of submicron patterns on curved substrates using a polydimethylsiloxane film mould , 2004 .

[4]  I. Weissman,et al.  Self-renewal, differentiation or death: regulation and manipulation of hematopoietic stem cell fate. , 1999, Molecular medicine today.

[5]  Joyce Y. Wong,et al.  Directed Movement of Vascular Smooth Muscle Cells on Gradient-Compliant Hydrogels† , 2003 .

[6]  Maxence Bigerelle,et al.  Effect of grooved titanium substratum on human osteoblastic cell growth. , 2002, Journal of biomedical materials research.

[7]  B. Cappella,et al.  Solvent-assisted nanolithography on polystyrene surfaces using the atomic force microscope , 2007 .

[8]  C. Lohmann,et al.  Response of MG63 osteoblast-like cells to titanium and titanium alloy is dependent on surface roughness and composition. , 1998, Biomaterials.

[9]  Andre Levchenko,et al.  Biomimetic Nanopatterns as Enabling Tools for Analysis and Control of Live Cells , 2010, Advanced materials.

[10]  J. Jansen,et al.  The influence of nanoscale topographical cues on initial osteoblast morphology and migration. , 2010, European cells & materials.

[11]  Andrew J Ewald,et al.  Collective epithelial migration and cell rearrangements drive mammary branching morphogenesis. , 2008, Developmental cell.

[12]  W. Su,et al.  Micropillar substrate influences the cellular attachment and laminin expression. , 2009, Journal of biomedical materials research. Part A.

[13]  C. Fotakis,et al.  Tuning cell adhesion by controlling the roughness and wettability of 3D micro/nano silicon structures. , 2010, Acta biomaterialia.

[14]  Ian Papautsky,et al.  Institute of Physics Publishing Journal of Micromechanics and Microengineering Polymer Embossing Tools for Rapid Prototyping of Plastic Microfluidic Devices , 2022 .

[15]  C J Murphy,et al.  Effects of synthetic micro- and nano-structured surfaces on cell behavior. , 1999, Biomaterials.

[16]  T. V. van Kooten,et al.  Influence of silicone (PDMS) surface texture on human skin fibroblast proliferation as determined by cell cycle analysis. , 1998, Journal of biomedical materials research.

[17]  Dong Sung Kim,et al.  Effect of replicated polymeric substrate with lotus surface structure on adipose-derived stem cell behaviors. , 2011, Macromolecular bioscience.

[18]  Kedar G. Shah,et al.  MICROSCALE CONTROL OF MICROPOST STIFFNESS TO INDUCE CELLULAR DUROTAXIS , 2008 .

[19]  T. Mitchison,et al.  A high-throughput cell migration assay using scratch wound healing, a comparison of image-based readout methods , 2004, BMC biotechnology.

[20]  William P King,et al.  Hot embossing for micropatterned cell substrates. , 2004, Biomaterials.

[21]  C. Fotakis,et al.  Controlling cell adhesion via replication of laser micro/nano-textured surfaces on polymers , 2011, Biofabrication.

[22]  David Beebe,et al.  Engineers are from PDMS-land, Biologists are from Polystyrenia. , 2012, Lab on a chip.

[23]  M. Toner,et al.  Microfabrication of an analog of the basal lamina: biocompatible membranes with complex topographies , 2000, FASEB journal : official publication of the Federation of American Societies for Experimental Biology.

[24]  Luke P. Lee,et al.  Dynamic single cell culture array. , 2006, Lab on a chip.

[25]  M. Dembo,et al.  Cell movement is guided by the rigidity of the substrate. , 2000, Biophysical journal.

[26]  Sangwon Chung,et al.  Design concepts and strategies for tissue engineering scaffolds , 2011, Biotechnology and applied biochemistry.

[27]  Claire McCague,et al.  Spatially controlled cell adhesion via micropatterned surface modification of poly(dimethylsiloxane). , 2007, Langmuir : the ACS journal of surfaces and colloids.

[28]  John P. Puccinelli,et al.  Thermal aging and reduced hydrophobic recovery of polydimethylsiloxane , 2006 .

[29]  Teruo Fujii,et al.  Cell Culture in 3-Dimensional Microfluidic Structure of PDMS (polydimethylsiloxane) , 2003 .

[30]  Marcus Textor,et al.  An inverted microcontact printing method on topographically structured polystyrene chips for arrayed micro-3-D culturing of single cells. , 2005, Biomaterials.

[31]  Po Ki Yuen,et al.  Hot embossing of plastic microfluidic devices using poly(dimethylsiloxane) molds , 2010 .