Effects of serum protein on ionic exchange between culture medium and microporous hydroxyapatite and silicate-substituted hydroxyapatite

Abstract

It has been proposed that one of the underlying mechanisms contributing to the bioactivity of osteoinductive or osteoconductive calcium phosphates involves the rapid dissolution and net release of calcium and phosphate ions from the matrix as alternatively a precursor to subsequent re-precipitation of a bone-like apatite at the surface and/or to facilitate ion exchange in biochemical processes. In order to confirm and evaluate ion release from sintered hydroxyapatite (HA) and to examine the effect of silicate substitution into the HA lattice on ion exchange under physiological conditions we monitored Ca(2+), PO(4)(3-) and SiO(4)(4-) levels in Earl's minimum essential medium (E-MEM) in the absence (serum-free medium, SFM) or presence (complete medium, C-MEM) of foetal calf serum (FCM), with both microporous HA or 2.6 wt% silicate-substituted HA (SA) sintered discs under both static and semi-dynamic (SD) conditions for up to 28 days. In SFM, variation in Ca(2+) ion concentration was not observed with either disc chemistry or culture conditions. In C-MEM, Ca(2+) ions were released from SA under static and SD conditions whereas with HA Ca(2+) was depleted under SD conditions. PO(4)(3-) depletion occurred in all cases, although it was greater in C-MEM, particularly under SD conditions. SiO(4)(4-) release occurred from SA irrespective of medium or culture conditions but a sustained release only occurred in C-MEM under SD conditions. In conclusion we showed that under physiological conditions the reservoir of exchangeable ions in both HA and SA in the absence of serum proteins is limited, but that the presence of serum proteins facilitated greater ionic exchange, particularly with SA. These observations support the hypothesis that silicate substitution into the HA lattice facilitates a number of ionic interactions between the material and the surrounding physiological environment, including but not limited to silicate ion release, which may play a key role in determining the overall bioactivity and osteoconductivity of the material. However, significant net release of Ca(2+) and PO(4)(3-) was not observed, thus rapid or significant net dissolution of the material is not necessarily a prerequisite for bioactivity in these materials.