High-content profiling of cell responsiveness to graded substrates based on combinyatorially variant polymers.
We have developed a novel approach combining high information and high throughput analysis to characterize cell adhesive responses to biomaterial substrates possessing gradients in surface topography. These gradients were fabricated by subjecting thin film blends of tyrosine-derived polycarbonates, i.e. poly(DTE carbonate) and poly(DTO carbonate) to a gradient temperature annealing protocol. Saos-2 cells engineered with a green fluorescent protein (GFP) reporter for farnesylation (GFP-f) were cultured on the gradient substrates to assess the effects of nanoscale surface topology and roughness that arise during the phase separation process on cell attachment and adhesion strength. The high throughput imaging approach allowed us to rapidly identify the "global" and "high content" structure-property relationships between cell adhesion and biomaterial properties such as polymer chemistry and topography. This study found that cell attachment and spreading increased monotonically with DTE content and were significantly elevated at the position with intermediate regions corresponding to the highest "gradient" of surface roughness, while GFP-f farnesylation intensity descriptors were sensitively altered by surface roughness, even in cells with comparable levels of spreading.
Liu
Er
E
Department of Biomedical Engineering, Rutgers University, Piscataway, NJ 08854, USA.
Treiser
Matthew D
MD
Patel
Hiral
H
Sung
Hak-Joon
HJ
Roskov
Kristen E
KE
Kohn
Joachim
J
Becker
Matthew L
ML
Moghe
Prabhas V
PV
D001681:Biological Assay / Q000379:methods*; D002448:Cell Adhesion / Q000502:physiology; D002460:Cell Line; D020650:Combinatorial Chemistry Techniques / Q000379:methods*; D049452:Green Fluorescent Proteins / Q000737:chemistry; D006801:Humans; D018625:Microscopy, Atomic Force; D009757:Nylons / Q000737:chemistry*; D014443:Tyrosine / Q000737:chemistry
D009757:Nylons; C114440:poly(DTE carbonate); D049452:Green Fluorescent Proteins; D014443:Tyrosine