Protein coating of
Ti discs was carried out with 20 μg/mL fibronectin isolated
from human plasma (R&D Systems Europe Ltd.). For atomic force
microscopy (AFM) assays, the coating was performed for a time of 10
min; then, samples were washed once with PBS and three times with
ultrapure water.19 (link) For all other experiments,
titanium samples were UV-sterilized for 20 min and coated with 20
μg/mL FN solution in DPBS (Gibco, Life Technologies) for 1 h.7 (link),19 (link)After that, the samples were washed with DPBS and then coated with
25 ng/mL or 100 ng/mL BMP-7 solutions in DPBS (human recombinant BMP-7
cat. no. 354-BP, R&D Systems Europe Ltd.) for 1 h. This was followed
by a single wash with DPBS; GF-negative samples were treated with
DBPS. The volume of coating solution and subsequent washes was 200
μL per sample in all cases. After the coating, samples were
preserved in DPBS until cell seeding.
Ti discs was carried out with 20 μg/mL fibronectin isolated
from human plasma (R&D Systems Europe Ltd.). For atomic force
microscopy (AFM) assays, the coating was performed for a time of 10
min; then, samples were washed once with PBS and three times with
ultrapure water.19 (link) For all other experiments,
titanium samples were UV-sterilized for 20 min and coated with 20
μg/mL FN solution in DPBS (Gibco, Life Technologies) for 1 h.7 (link),19 (link)After that, the samples were washed with DPBS and then coated with
25 ng/mL or 100 ng/mL BMP-7 solutions in DPBS (human recombinant BMP-7
cat. no. 354-BP, R&D Systems Europe Ltd.) for 1 h. This was followed
by a single wash with DPBS; GF-negative samples were treated with
DBPS. The volume of coating solution and subsequent washes was 200
μL per sample in all cases. After the coating, samples were
preserved in DPBS until cell seeding.
