Improved adhesion, growth and maturation of vascular smooth muscle cells on polyethylene grafted with bioactive molecules and carbon particles.
High-density polyethylene (PE) foils were modified by an Ar(+) plasma discharge and subsequent grafting with biomolecules, namely glycine (Gly), polyethylene glycol (PEG), bovine serum albumin (BSA), colloidal carbon particles (C) or BSA and C (BSA + C). As revealed by atomic force microscopy (AFM), goniometry and Rutherford Backscattering Spectroscopy (RBS), the surface chemical structure and surface morphology of PE changed dramatically after plasma treatment. The contact angle decreased for the samples treated by plasma, mainly in relation to the formation of oxygen structures during plasma irradiation. A further decrease in the contact angle was obvious after glycine and PEG grafting. The increase in oxygen concentration after glycine and PEG grafting proved that the two molecules were chemically linked to the plasma-activated surface. Plasma treatment led to ablation of the PE surface layer, thus the surface morphology was changed and the surface roughness was increased. The materials were then seeded with vascular smooth muscle cells (VSMC) derived from rat aorta and incubated in a DMEM medium with fetal bovine serum. Generally, the cells adhered and grew better on modified rather than on unmodified PE samples. Immunofluorescence showed that focal adhesion plaques containing talin, vinculin and paxillin were most apparent in cells on PE grafted with PEG or BSA + C, and the fibres containing alpha-actin, beta-actin or SM1 and SM2 myosins were thicker, more numerous and more brightly stained in the cells on all modified PE samples than on pristine PE. An enzyme-linked immunosorbent assay (ELISA) revealed increased concentrations of focal adhesion proteins talin and vinculin and also a cytoskeletal protein beta-actin in cells on PE modified with BSA + C. A contractile protein alpha-actin was increased in cells on PE grafted with PEG or Gly. These results showed that PE activated with plasma and subsequently grafted with bioactive molecules and colloidal C particles, especially with PEG and BSA + C, promotes the adhesion, proliferation and phenotypic maturation of VSMC.
Parizek
Martin
M
Centre for Cardiovascular Research, Institute of Physiology, Academy of Sciences of the Czech Republic / Videnska 1083, CZ-14220 Prague 4, Czech Republic.
Kasalkova
Nikola
N
Department of Solid State Engineering, Institute of Chemical Technology / Technicka 5, 16628 Prague 6, Czech Republic.
Bacakova
Lucie
L
Centre for Cardiovascular Research, Institute of Physiology, Academy of Sciences of the Czech Republic / Videnska 1083, CZ-14220 Prague 4, Czech Republic.
Slepicka
Petr
P
Department of Solid State Engineering, Institute of Chemical Technology / Technicka 5, 16628 Prague 6, Czech Republic.
Lisa
Vera
V
Centre for Cardiovascular Research, Institute of Physiology, Academy of Sciences of the Czech Republic / Videnska 1083, CZ-14220 Prague 4, Czech Republic.
Blazkova
Martina
M
Department of Biochemistry and Microbiology, Institute of Chemical Technology / Technicka 5, 16628 Prague 6, Czech Republic.
Svorcik
Vaclav
V
Department of Solid State Engineering, Institute of Chemical Technology / Technicka 5, 16628 Prague 6, Czech Republic.
D000199:Actins / Q000378:metabolism; D000818:Animals; D001011:Aorta / Q000166:cytology; D002244:Carbon / Q000737:chemistry*; D002417:Cattle; D002448:Cell Adhesion / Q000187:drug effects; D049109:Cell Proliferation / Q000187:drug effects; D002478:Cells, Cultured; D005998:Glycine / Q000494:pharmacology; D018625:Microscopy, Atomic Force; D009131:Muscle, Smooth, Vascular / Q000166:cytology / Q000187:drug effects* / Q000378:metabolism; D010100:Oxygen / Q000378:metabolism; D020959:Polyethylene / Q000737:chemistry / Q000494:pharmacology*; D011092:Polyethylene Glycols / Q000737:chemistry / Q000494:pharmacology; D051381:Rats; D012710:Serum Albumin, Bovine / Q000494:pharmacology
D000199:Actins; D012710:Serum Albumin, Bovine; D011092:Polyethylene Glycols; D002244:Carbon; D020959:Polyethylene; D010100:Oxygen; D005998:Glycine