Experimental design
Gel fabrication
Gel functionalization with fibronectin
Gel functionalization with TAMRA-RGD peptide
Mechanical characterization
Interfacial rheology measurements
Contact angle measurements
Steel ball indentation measurements
Gel preparation with Span85 surfactant treatment for cellular assays
Cell lines and culture
Antibodies and reagents
Cell morphology and spreading assays
Cell proliferation assays
Gene expression analysis
Isolation and culture of PBMCs
Preparation of a collagen master mix for 3D collagen I gels
PBMC-silicone interaction assay
PBMC—silicone droplets interaction assay
Immunofluorescence imaging
Hertz contact model with surface tension
Statistics
Silicone implants were gifts from Mentor Worldwide LLC. The fourth generation (catalog numbers 6580031 and 7333875) and fifth generation (catalog number CPG323) of implants were tested. Implants were cut open with razor blades. The interior materials were pinched with tweezers and spread onto glass-bottom dishes. The gels were functionalized by adsorption of fibronectin at 10 μg/ml in PBS in a 4°C refrigerator overnight. Cells were seeded at 5000 cells/cm2 and analyzed 24 hours after seeding.