Evaluation of boronate-containing polymer brushes and gels as substrates for carbohydrate-mediated adhesion and cultivation of animal cells.
Boronate-containing thin polyacrylamide gels (B-Gel), polymer brushes (B-Brush) and chemisorbed organosilane layers (B-COSL) were prepared on the surface of glass slides and studied as substrates for carbohydrate-mediated cell adhesion. B-COSL- and B-Brush-modified glass samples exhibited multiple submicron structures densely and irregularly distributed on the glass surface, as found by scanning electron microscopy and atomic force microscopy. B-Gel was ca. 0.1 mm thick and contained pores with effective size of 1-2 microm in the middle and of 5-20 microm on the edges of the gel sample as found by confocal laser scanning microscopy. Evidence for the presence of phenylboronic acid in the samples was given by time-of-flight secondary ion mass-spectrometry (ToF SIMS), contact angle measurements performed in the presence of fructose, and staining with Alizarin Red S dye capable of formation specific, fluorescent complexes with boronic acids. A comparative study of adhesion and cultivation of animal cells on the above substrates was carried out using murine hybridoma M2139 cell line as a model. M2139 cells adhered to the substrates in the culture medium without glucose or sodium pyruvate at pH 8.0, and then were cultivated in the same medium at pH 7.2 for 4 days. It was found that the substrates of B-Brush type were superior both regarding cell adhesion and viability of the adhered cells, among the substrates studied. MTT assay confirmed proliferation of M2139 cells on B-Brush substrates. Some cell adhesion was also registered in the macropores of B-Gel substrate. The effects of surface microstructure of the boronate-containing polymers on cell adhesion are discussed. Transparent glass substrates grafted with boronate-containing copolymers offer good prospects for cell adhesion studies and development of cell-based assays.
Ivanov
Alexander E
AE
Protista Biotechnology AB, IDEON, SE-22370, Lund, Sweden. alexander.ivanov@protista.se
Kumar
Ashok
A
Nilsang
Suthasinee
S
Aguilar
Maria-Rosa
MR
Mikhalovska
Lyubov I
LI
Savina
Irina N
IN
Nilsson
Lars
L
Scheblykin
Ivan G
IG
Kuzimenkova
Marina V
MV
Galaev
Igor Yu
IY
D000818:Animals; D001897:Boronic Acids / Q000494:pharmacology*; D002241:Carbohydrates / Q000494:pharmacology*; D002448:Cell Adhesion / Q000187:drug effects; D002460:Cell Line; D049109:Cell Proliferation / Q000187:drug effects; D002470:Cell Survival / Q000187:drug effects; D002478:Cells, Cultured; D005782:Gels / Q000378:metabolism*; D051379:Mice; D018625:Microscopy, Atomic Force; D018613:Microscopy, Confocal; D008855:Microscopy, Electron, Scanning; D011108:Polymers / Q000494:pharmacology*; D012821:Silanes / Q000494:pharmacology; D013050:Spectrometry, Fluorescence
D001897:Boronic Acids; D002241:Carbohydrates; D005782:Gels; D011108:Polymers; D012821:Silanes