Formation of composite polyacrylamide and silicone substrates for independent control of stiffness and strain.

Cells that line major tissues in the body such as blood vessels, lungs and gastrointestinal tract experience deformation from mechanical strain with our heartbeat, breathing, and other daily activities. Tissues also remodel in both development and disease, changing their mechanical properties. Taken together, cells can experience vastly different mechanical cues resulting from the combination of these interdependent stimuli. To date, most studies of cellular mechanotransduction have been limited to assays in which variations in substrate stiffness and strain were not combined. Here, we address this technological gap by implementing a method that can simultaneously tune both substrate stiffness and mechanical strain. Substrate stiffness is controlled with different monomer and crosslinker ratios during polyacrylamide gel polymerization, and strain is transferred from the underlying silicone platform when stretched. We demonstrate this platform with polyacrylamide gels with elastic moduli at 6 kPa and 20 kPa in combination with two different silicone formulations. The gels remain attached with up to 50% applied strains. To validate strain transfer through the gels into cells, we employ particle-tracking methods and observe strain transmission via cell morphological changes.

[1]  Andrew D McCulloch,et al.  Substrate stiffness affects the functional maturation of neonatal rat ventricular myocytes. , 2008, Biophysical journal.

[2]  L. Lanyon,et al.  Mechanical Strain and Bone Cell Function: A Review , 2002, Osteoporosis International.

[3]  S T Quek,et al.  Mechanical models for living cells--a review. , 2006, Journal of biomechanics.

[4]  D. Ingber,et al.  Reconstituting Organ-Level Lung Functions on a Chip , 2010, Science.

[5]  Jin Ho Kim,et al.  Surface modification of poly(dimethylsiloxane) microchannels , 2003, Electrophoresis.

[6]  M. Sudol,et al.  Modularity and functional plasticity of scaffold proteins as p(l)acemakers in cell signaling. , 2012, Cellular signalling.

[7]  Rajeev J Ram,et al.  Plastic-PDMS bonding for high pressure hydrolytically stable active microfluidics. , 2009, Lab on a chip.

[8]  Adam J Engler,et al.  Embryonic cardiomyocytes beat best on a matrix with heart-like elasticity: scar-like rigidity inhibits beating , 2008, Journal of Cell Science.

[9]  P. Janmey,et al.  Polyacrylamide hydrogels for cell mechanics: steps toward optimization and alternative uses. , 2007, Methods in cell biology.

[10]  Bruce K. Gale,et al.  Determining the optimal PDMS–PDMS bonding technique for microfluidic devices , 2008 .

[11]  Naveed Akbar,et al.  Biocompatibility of amorphous silica nanoparticles: Size and charge effect on vascular function, in vitro , 2011, Biotechnology and applied biochemistry.

[12]  H. Schnaper,et al.  Role of laminin in endothelial cell recognition and differentiation. , 1993, Kidney international.

[13]  Andrew K. Capulli,et al.  Combining Dynamic Stretch and Tunable Stiffness to Probe Cell Mechanobiology In Vitro , 2011, PloS one.

[14]  Tza-Huei Wang,et al.  PDMS-glass bonding using grafted polymeric adhesive--alternative process flow for compatibility with patterned biological molecules. , 2012, Lab on a chip.

[15]  Jay Lee,et al.  Hard top soft bottom microfluidic devices for cell culture and chemical analysis. , 2009, Analytical chemistry.

[16]  G. Pharr,et al.  An improved technique for determining hardness and elastic modulus using load and displacement sensing indentation experiments , 1992 .

[17]  R. Kramer,et al.  Integrin receptors on aortic smooth muscle cells mediate adhesion to fibronectin, laminin, and collagen. , 1990, Circulation research.

[18]  A. McCulloch,et al.  Cardiac myocyte force development during differentiation and maturation , 2010, Annals of the New York Academy of Sciences.

[19]  Cynthia A. Reinhart-King,et al.  Exogenous and endogenous force regulation of endothelial cell behavior. , 2010, Journal of biomechanics.

[20]  N. Kawazoe,et al.  Effects of Poly(L-lysine), Poly(acrylic acid) and Poly(ethylene glycol) on the Adhesion, Proliferation and Chondrogenic Differentiation of Human Mesenchymal Stem Cells , 2009, Journal of biomaterials science. Polymer edition.

[21]  Jennifer L. West,et al.  Synthetic Materials in the Study of Cell Response to Substrate Rigidity , 2009, Annals of Biomedical Engineering.

[22]  Joachim P. Spatz,et al.  Erratum: Extracellular-matrix tethering regulates stem-cell fate (Nature Materials (2012) 11 (642-649)) , 2012 .

[23]  J. Engel,et al.  Domain structure and organisation in extracellular matrix proteins. , 2002, Matrix biology : journal of the International Society for Matrix Biology.

[24]  R. Swerlick,et al.  HMEC-1: establishment of an immortalized human microvascular endothelial cell line. , 1992, The Journal of investigative dermatology.

[25]  S. Sen,et al.  Matrix Elasticity Directs Stem Cell Lineage Specification , 2006, Cell.

[26]  T D Brown,et al.  Techniques for mechanical stimulation of cells in vitro: a review. , 2000, Journal of biomechanics.

[27]  F. Guilak,et al.  Mechanical signals as regulators of stem cell fate. , 2004, Current topics in developmental biology.

[28]  P. Janmey,et al.  Tissue Cells Feel and Respond to the Stiffness of Their Substrate , 2005, Science.

[29]  G. Marshall,et al.  Nanoindentation of polydimethylsiloxane elastomers: Effect of crosslinking, work of adhesion, and fluid environment on elastic modulus , 2005 .

[30]  K. Rottner,et al.  Interplay between Rac and Rho in the control of substrate contact dynamics , 1999, Current Biology.

[31]  E. Jaffe,et al.  Synthesis of fibronectin by cultured human endothelial cells , 1978, Annals of the New York Academy of Sciences.

[32]  K. J. Grande-Allen,et al.  Effects of static and cyclic loading in regulating extracellular matrix synthesis by cardiovascular cells. , 2006, Cardiovascular research.

[33]  Mark Bachman,et al.  Covalent micropatterning of poly(dimethylsiloxane) by photografting through a mask. , 2005, Analytical chemistry.

[34]  Chelsey S Simmons,et al.  Microsystems for biomimetic stimulation of cardiac cells. , 2012, Lab on a chip.

[35]  Y. Wang,et al.  Cell locomotion and focal adhesions are regulated by substrate flexibility. , 1997, Proceedings of the National Academy of Sciences of the United States of America.