Measuring cell adhesion on RGD-modified, self-assembled PEG monolayers using the quartz crystal microbalance technique.
In this study, the suitability of a flow-through quartz crystal microbalance system for the detection of the adhesion of rMSCs and 3T3-L1 fibroblasts on different surfaces is demonstrated. Frequency shifts for rMSCs of -6.7 mHz x cell(-1) and -2.0 mHz x cell(-1) for 3T3-L1 cells could be detected on non-modified gold sensors, revealing that the frequency shift per cell is comparable to that of a static setup. Modifying the sensor surface with SAMs of thioalkylated omega-amine-terminated PEG derivatives led to cell-adhesion-resistant surfaces. Total frequency shifts of only -20 +/- 7 Hz showed that protein adsorption was also significantly reduced. Attaching 35 pmol x mm(-2) of the GRGDS cell adhesion motif to the SAMs induced specific cell adhesion due to RGD-integrin interactions; the resonance frequency dropped by 3.4 mHz x cell(-1). Furthermore, the kinetics of cell detachment could be determined. The corresponding processes were completed after 10 min for trypsin, and not before 90 min with GRGDS. Moreover, the detectability of cell adhesion was shown to increase after the addition of manganese cations. The total decrease in the resonance frequency was almost 80 Hz in the presence of Mn(2+) (6.4 mHz x cell(-1)). [image: see text] Staining the cytoskeleton of the rMSCs shows that the GRGDS-modified surfaces are almost completely covered with well-spread cells.
Knerr
Robert
R
Department of Pharmaceutical Technology, University of Regensburg, Universitätsstrasse 31, 93040 Regensburg, Germany.
Weiser
Barbara
B
Drotleff
Sigrid
S
Steinem
Claudia
C
Göpferich
Achim
A
D000818:Animals; D015374:Biosensing Techniques / Q000379:methods*; D001854:Bone Marrow Cells / Q000502:physiology*; D002448:Cell Adhesion; D002460:Cell Line; D051379:Mice; D019946:Propylene Glycol / Q000737:chemistry*; D011791:Quartz / Q000737:chemistry*; D051381:Rats; D013499:Surface Properties
D011791:Quartz; D019946:Propylene Glycol