The formation of a functionally-confluent endothelial cell (EC) monolayer affords
proliferation of EC, which only happens in case of appropriate migratory activity.
AIM OF THE STUDY:
The migratory pathway of human umbilical endothelial cells (HUVEC) was investigated on
different polymeric substrates.
MATERIAL AND METHODS:
Surface characterization of the polymers was performed by contact angle measurements
and atomic force microscopy under wet conditions. 30,000 HUVEC per well were seeded on
polytetrafluoroethylene (PTFE) (θadv = 119°±2°), on
low-attachment plate LAP (θadv = 28°±2°) and on polystyrene
based tissue culture plates (TCP, θadv = 22°±1°). HUVEC
tracks (trajectories) were recorded by time lapse microscopy and the euclidean distance
(straight line between starting and end point), the total distance and the velocities of
HUVEC not leaving the vision field were determined.
RESULTS:
On PTFE, 42 HUVEC were in the vision field directly after seeding. The mean length of
single migration steps (SML) was 6.1±5.2 μm, the mean velocity (MV) 0.40±0.3
μm·min-1 and the complete length of the trajectory (LT) was 710±440 μm. On
TCP 82 HUVEC were in the vision field subsequent to seeding. The LT was 840±550 μm, the
SML 6.1±5.2 μm and the MV 0.44±0.3 μm·min-1. The trajectories on LAP differed
significantly in respect to SML (2.4±3.9 μm, p < 0.05), the MV
(0.16±0.3 μm·min-1, p < 0.05) and the LT (410±300 μm,
p < 0.05), compared to PTFE and TCP. Solely on TCP a nearly
confluent EC monolayer developed after three days. While on TCP diffuse signals of
vinculin were found over the whole basal cell surface organizing the binding of the
cells by focal adhesions, on PTFE vinculin was merely arranged at the cell rims, and on
the hydrophilic material (LAP) no focal adhesions were found.
CONCLUSION:
The study revealed that the wettability of polymers affected not only the initial
adherence but also the migration of EC, which is of importance for the proliferation and
ultimately the endothelialization of polymer-based biomaterials.
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