Photoimmobilization of a Bioactive Laminin Fragment and Pattern-Guided Selective Neuronal Cell Attachment

Abstract

To attain light-dependent functionalization of biocompatible materials, a photolabel-derivatized, bioactive laminin fragment has been synthesized, chemically characterized, and photoimmobilized. Covalent high-resolution patterning of the laminin fragment CDPGYIGSR to hydroxylated fluorinated ethylene propylene (FEP-OH), poly(vinyl alcohol), and glycophase glass has been achieved. The synthetic peptide CDPGYIGSR was thermochemically coupled to either N-[m-[3-(trifluoromethyl)-diazirin-3-yl]phenyl]-4-maleimidobuty ramide or 4-maleimidobenzophenone. Photolabel-derivatized peptides were radiolabeled, and 20 and 300 microns-sized patterns were visualized by autoradiography. The biospecific interaction of photoimmobilized laminin fragments with cells was investigated by analyzing the selective attachment of NG 108-15 neuroblastoma x glioma cells which bear CDPGYIGSR-specific cell surface receptors. On photopatterned FEP-OH membranes NG 108-15 cells differentiated in serum-supplemented media within 1 day. Specific attachment to the immobilized oligopeptide CDPGYIGSR was assessed in serum-free media with competitive binding studies, showing an 82% decrease in cell adherence after the cell receptors were blocked with soluble CDPGYIGSR.

Surface modification of titanium, titanium alloys, and related materials for biomedical applications

“…The benzophenone derivatives could be linked covalently onto the solid surface by irradiation. Clemence et al [186] and Collioud et al [187] produced a photosensitive hetero-bifunctional crosslinker, which could couple petides containing the cell-adhesive motifs of RGD and YIGSR to maleimidyl groups using thiol/maleimide chemistry. The photosensitive diazirine group generates an intermediate carbene after irradiation with non-destructive UV light (!320 nm).…”

Three-dimensional extracellular matrix engineering in the nervous system

“…Photoimmobilization of BP-Mal-CDPGYIGSR BP-Mal-CDPGYIGSR was synthesized according to Clém-ence et al 22 One milliliter of 2% (w/v) agarose was mixed with 1 mL (2.1 mol) of BP-Mal-CDPGYIGSR. Irradiation of the BP-Mal-CDPGYIGSR agarose solution was carried out for 30 min at 15 mW/cm 2 with a mercury lamp.…”

Microfabricated elastomeric stencils for micropatterning cell cultures

“…Microfabrication techniques offer great potential to design, on a micrometer scale, the biochemical composition of the substrate, the medium surrounding a given cell, as well as the type of cell contacting each cell. Previous work has demonstrated the micron-scale control of cell-substrate contact area, 1-5 cell attachment and growth, [6][7][8][9][10][11][12][13] and cell-cell interactions. [14][15][16] Usually, selective cell attachment is achieved indirectly by microfabricating a template to which cells adhere preferentially.…”