Calcium modulates interactions between bacteria and hydroxyapatite.

Bacterial adhesion onto hydroxyapatite is known to depend on the surface properties of both the biomaterial and the bacterial strain, but less is known about the influence of the composition of the aqueous medium. Here, the adhesion of Streptococcus mutans and 3 different Lactobacilli on powdered hydroxyapatite was shown to change with Ca2+ concentration. The effect depends on the surface properties of each strain. Adhesion of Lactobacillus fermentum and salivarius (and of Streptococcus mutans at low Ca2+) was enhanced with increasing Ca2+ concentration. Lactobacillus casei was efficiently removed by adhesion on hydroxyapatite, even without Ca2+ addition, and the effect of this ion was only marginal. The results are interpreted in terms of Ca2+-mediated adhesion, and relative to the hydrophobic properties of each strain and the electrical properties of the bacterial and solid surfaces (electrophoretic mobility).

Surface properties of various powdered hydroxyapatites.

Electrophoretic mobilities of various synthetic and semisynthetic hydroxyapatites (Ca10(PO4)6(OH)2, HAP) suspended in aqueous solutions have been measured as a function of pH and calcium concentration. The studied powders differ in particle size, crystallinity degree and surface contamination (carbonate). When equilibrated in mineral acids or bases, a large plateau of negative mobility is observed in the pH range 5-8, with increasing negative values at higher pH. Only in the case of the sample composed of nanoparticles, positive mobility obtains at pH < 8.9. When Ca2+ is added, positive mobility values are observed for all samples, and a bell-shaped profile results as a function of pH. Two possible models are explored to describe the results: the Nernstian approach, which assumes solubility equilibrium and surface potentials determined by the three potential-determining ions (Ca2+, PO3-4, and OH-), and the surface complexation approach, based on the idea of negligible phase transfer of structural phosphate. The Nernstian model is inadequate, whereas a very simple surface complexation model based on the equations Ca5(PO4)+3 = Ca4(PO4)-3 + Ca2+,Ca4(PO4)-3 + H+ = Ca4(PO4)2(PO4H),Ca5(PO4)+3 + OH- = Ca5(PO4)3(OH),coupled with a very simple electrical double layer, model suffices to reproduce the bell-shaped profile of the mobility as a function of pH in the presence of added calcium salts. The results also show that the sample composed of nanoparticles exchanges ions more easily with the solution, without reaching the solubility equilibrium in the explored timespans. In the presence of soluble phosphate salts, it is postulated that the same surface ensembles define the surface charge, with participation of phosphate as described by the equation Ca5(PO4)+3 + PO3-4 = Ca4(PO4)-3.HAP is just one member of a family of calcium phosphates with different (Ca)/(P) ratios. Electrophoretic mobilities of another member, tricalcium diphosphate, Ca3(PO4)2, were also measured and shown to be described by the same basic model. Comparison with previous literature data shows that the negative plateau in the mobility is a general feature of many HAP samples at low Ca2+, again in agreement with the surface complexation model. FTIR data demonstrates that surface phosphate indeed undergoes protonation, as postulated in the model.

Acidity constants of benzidine in aqueous solutions.

The acidity constants of benzidine (Bz) in aqueous solutions determined potentiometrically at 25 degrees were K(a1) = (1.11 +/- 0.08) x 10(-5), K(a2) = (1.45 +/- 0.12) x 10(-4). The apparent mixed constants in 0.1M sodium nitrate are K(a1) = (5.37 +/- 0.28) x 10(-6) and K(a2) = (1.14 +/- 0.09) x 10(-4). The ultraviolet spectra were recorded as a function of pH and analysed with these constants to obtain the absorption spectra of H(2)Bz(2+), HBz(+) and Bz; the corresponding wavelengths of maximal absorption are 247, 273 and 278 nm, and molar absorptivities 1.63 x 10(4), 1.76 x 10(4) and 2.26 x 10(4) 1.mole(-1).cm(-1).