Collagen and fibronectin immobilization on PHEMA microcarriers for hepatocyte attachment.
Polyhydroxyethylmethacrylate (PHEMA) microcarriers in a size range of 150-250 microns were prepared by a suspension polymerization in an aqueous phase containing magnesium oxide. The hydroxyl groups on the PHEMA microcarriers were activated by cyanogen bromide. In order to improve cell attachment, cell-adhesive proteins, namely, collagen and fibronectin were immobilized onto PHEMA microcarriers. The nonspecific adsorption values for collagen and fibronectin were 0.10 mg collagen/g PHEMA and 0.044 mg fibronectin/g PHEMA, respectively. Collagen and fibronectin immobilization on PHEMA microcarriers were studied at different pH by using single protein solutions containing different amounts of proteins, at a constant temperature of 20 degrees C. The maximum immobilizations were 0.85 mg collagen/g PHEMA (at pH: 9.5) and 0.52 mg fibronectin/g PHEMA (at pH: 7.4). Hepatocyte attachment onto these biologically modified PHEMA microcarriers was studied. Hydrophilic PHEMA microcarriers did not support cell attachment. High hepatocyte attachment yields (up to 75% surface coverage) were observed on collagen and fibronectin immobilized PHEMA microcarriers.
Denizli
A
A
Hacettepe University, Chemical Engineering Department, Ankara, Turkey.
Pişkin
E
E
Dixit
V
V
Arthur
M
M
Gitnick
G
G
D000818:Animals; D002448:Cell Adhesion / Q000502:physiology; D003094:Collagen / Q000737:chemistry / Q000378:metabolism*; D003432:Cross-Linking Reagents; D003488:Cyanogen Bromide / Q000737:chemistry; D004337:Drug Carriers; D005353:Fibronectins / Q000737:chemistry / Q000378:metabolism*; D006863:Hydrogen-Ion Concentration; D008099:Liver / Q000166:cytology*; D008297:Male; D008863:Microspheres; D011102:Polyhydroxyethyl Methacrylate / Q000008:administration & dosage / Q000737:chemistry*; D051381:Rats; D017208:Rats, Wistar; D013696:Temperature
D003432:Cross-Linking Reagents; D004337:Drug Carriers; D005353:Fibronectins; D011102:Polyhydroxyethyl Methacrylate; D003094:Collagen; D003488:Cyanogen Bromide