Micropattern Printing of Adhesion, Spreading, and Migration Peptides on Poly(tetrafluoroethylene) Films To Promote Endothelialization

Abstract

We report here the development of an original multistep micropatterning technique for printing peptides on surfaces, based on the ink-jet printer technology. Contrary to most micropatterning methods used nowadays, this technique is advantageous because it allows displaying 2D-arrays of multiple biomolecules. Moreover, this low cost procedure allies the advantages of computer-aided design with high flexibility and reproducibility. A Hewlett-Packard printer was modified to print peptide solutions, and Adobe Illustrator was used as the graphic-editing software to design high-resolution checkerboard-like micropatterns. In a first step, PTFE films were treated with ammonia plasma to introduce amino groups on the surface. These chemical functionalities were reacted with heterobifunctional cross-linker sulfo-succinimidyl 4-(N-maleimidomethyl)cycloexane-1-carboxylate (S-SMCC) to allow the subsequent surface covalent conjugation of various cysteine-modified peptides to the polymer substrate. These peptidic molecules containing RGD and WQPPRARI sequences were selected for their adhesive, spreading, and migrational properties toward endothelial cells. On one hand, our data demonstrated that the initial cell adhesion does not depend on the chemical structure and combination of the peptides covalently bonded either through conventional conjugation or micropatterning. On the other hand, spreading and migration of endothelial cells is clearly enhanced while coconjugating the GRGDS peptide in conjunction with WQPPRARI. This behavior is further improved by micropatterning these peptides on specific areas of the polymer surface.

Endothelial Cell Micropatterning: Methods, Effects, and Applications

“…60 Using a modified Hewlett-Packard inkjet printer, Gauvreau et al were able to print CRGD, GRGDS and WQPPRARI peptides as well as their combinations onto plasma-treated expanded polytetrafluoroethylene (ePTFE). 26 Similar results were obtain by Woodrow et al using a modified robotic microarray printer (Virtek ChipWriter Pro, BioRad, Hercules, CA) to print proteins singularly and in combination, specifically collagen I, collagen III, collagen IV, FN, laminin, and Matrigel (a solution of basement membrane proteins), onto electrospun PLA scaffolds or onto a HydroGel slide (a commercially available acrylamide surface on glass). 86 These researchers showed the preferential attachment of several cell types onto these protein-coated scaffolds for the purpose of tissue engineering.…”

Fabrication of paper-based microfluidic sensors by printing

“…Ink jet printing of biomolecules has been reported previously by many research groups [13,14]. Ink jet printing is a non-contact printing process, which offers a unique advantage of minimizing cross-sample contamination.…”

Low-cost bioanalysis on paper-based and its hybrid microfluidic platforms

“…This process provides paper devices with fine fluidic channels together with preserved folding and bending capability [73]. In addition, being a non-contact liquid deposition, it effectively minimizes cross-sample contamination and decreases risk of substrate damage, which is highly desirable for the printing of biomolecules [71, 74, 75]. …”