[Determination of silver content in silver-loaded coral hydroxyapatite in relation to the biocompatibility]

To determine the amount of silver in silver-loaded coral hydroxyapatite (Ag(+)-CHA) bone substitute and its impact on the biocompatibility of this material with mouse embryonic osteoblast cells.

Ag(+)-CHA was prepared by immersing coral hydroxyapatite in a serial concentration of silver nitrate solutions. The amount of silver in the prepared Ag(+)-CHA was measured by inductively coupled plasma atomic emission spectrometry (ICP-AES). The viability of MC3T3-E1 cells incubated with Ag(+)-CHA was measured by MTT colorimetric assay, and the cell growth and morphological changes were observed by inverted phase-contrast microscope and confocal laser scanning microscope.

The amount of silver loading in the bone substitutes prepared by immersion in 1×10(-2), 1×10(-3), 5×10(-4), 10(-4), 8×10(-5), and 5×10(-5) mol/L silver nitrate solutions were 4127.67∓47.35, 167.90∓11.00, 83.42∓4.51, 30.20∓2.32, 22.39∓4.09, and 15.11∓0.55 µg/g, respectively. A low silver content in the material (prepared with silver nitrate solution of less than 8×10(-5) mol/L) showed no significant inhibitory effect on the growth of MC3T3-E1 cells or produced noticeable cytotoxic effect. On the materials prepared with 8×10(-5) and 10(-5) mol/L silver nitrate solution, the osteoblasts displayed active proliferation similar to those incubated on materials without silver loading. The confluent cells showed a normal fusiform morphology with tight arrangement.

Ag(+)-CHA with low silver content has a good biocompability and can promote the proliferation and growth of MC3T3-E1 cells in vitro, suggesting the clinical potential of this material as a anti-infection bone substitute.