title: The covalent attachment of adhesion molecules to silicone membranes for cell stretching applications.
abstract: Strain devices with expandable polydimethylsiloxane (PDMS) culture membranes are frequently used to stretch cells in vitro, mimicking mechanically dynamic tissue environments. To immobilize cell-adhesive molecules to the otherwise non-adhesive PDMS substrate, hydrophobic, electrostatic and covalent surface coating procedures have been developed. The efficacy of different coating strategies to transmit stretches to cells however is poorly documented and has not been compared. We describe a novel and simple procedure to covalently bind extracellular matrix proteins to the surface of stretchable PDMS membranes. The method comprises PDMS oxygenation, silanization, and covalent protein cross-linking to the silane. We demonstrate improved attachment ( approximately 2-fold), spreading ( approximately 2.5-fold) and proliferation ( approximately 1.2-fold) of fibroblasts to our new coating over established coating procedures. Further, we compared the efficiency of different PDMS coating techniques to transmit stretches. After 15% stretch, the number of maximally (15 +/- 5%) stretched cells on our PDMS surface coating was approximately 7-fold higher compared with alternative coating protocols. Hence, covalent linkage of adhesive molecules is superior to non-covalent methods in providing a coating that resists large deformations and that fully transmit this stretch to cultured cells.
authors.affiliation: Ecole Polytechnique Fédérale de Lausanne, Switzerland.
authors.forename: Pierre-Jean
authors.initials: PJ
authors.lastname: Wipff
authors.forename: Hicham
authors.initials: H
authors.lastname: Majd
authors.forename: Chitrangada
authors.initials: C
authors.lastname: Acharya
authors.forename: Lara
authors.initials: L
authors.lastname: Buscemi
authors.forename: Jean-Jacques
authors.initials: JJ
authors.lastname: Meister
authors.forename: Boris
authors.initials: B
authors.lastname: Hinz
mesh_terms: D000818:Animals; D002448:Cell Adhesion / Q000187:drug effects; D015815:Cell Adhesion Molecules / Q000378:metabolism*; D049109:Cell Proliferation / Q000187:drug effects; D048430:Cell Shape / Q000187:drug effects; D020099:Coated Materials, Biocompatible; D003094:Collagen / Q000378:metabolism; D003432:Cross-Linking Reagents / Q000494:pharmacology; D004129:Dimethylpolysiloxanes / Q000378:metabolism; D005109:Extracellular Matrix / Q000187:drug effects / Q000378:metabolism; D005347:Fibroblasts / Q000166:cytology* / Q000187:drug effects / Q000648:ultrastructure; D005976:Glutaral / Q000494:pharmacology; D008567:Membranes, Artificial; D018625:Microscopy, Atomic Force; D011437:Propylamines; D051381:Rats; D012821:Silanes / Q000494:pharmacology; D012828:Silicones / Q000378:metabolism*; D013499:Surface Properties / Q000187:drug effects
chemical_list: D015815:Cell Adhesion Molecules; D020099:Coated Materials, Biocompatible; D003432:Cross-Linking Reagents; D004129:Dimethylpolysiloxanes; D008567:Membranes, Artificial; D011437:Propylamines; D012821:Silanes; D012828:Silicones; D003094:Collagen; C477625:amino-propyl-triethoxysilane; D005976:Glutaral