Abstract:In experiments in vivo using diffusion chambers, the morphology and composition of calcium-containing deposits on natural and artificial biomaterials that had no direct contact with cells were studied using scanning electron microscopy with energy dispersion X-ray microanalysis. It was revealed that the formation of a protein layer containing protein-calcium complexes is the key event in biomaterial calcification. A mechanism of formation of a calciumcontaining protein matrix that creates the conditions for supersaturation of the crystal-forming medium over critical value has been proposed. The formation of nuclei of insoluble calcium phosphate starts predominantly deep in an adsorbed protein layer enriched by calcium ions.
The results of these in vivo experiments show that bovine pericardium can undergo calcification without direct contact with tissue. It is clear that the direct interaction of cells with implanted samples both promotes and accelerates the process of calcification. Moreover, dietary calcium supplements, calcium chloride and vitamin D can intensify the rate and extent of this process.
In experiments in vivo using diffusion chambers, the morphology and composition of calcium-containing deposits on natural and artificial biomaterials that had no direct contact with cells were studied using scanning electron microscopy with energy dispersion X-ray microanalysis. It was revealed that the formation of a protein layer containing protein-calcium complexes is the key event in biomaterial calcification. A mechanism of formation of a calcium-containing protein matrix that creates the conditions for supersaturation of the crystal-forming medium over critical value has been proposed. The formation of nuclei of insoluble calcium phosphate starts predominantly deep in an adsorbed protein layer enriched by calcium ions.
The nature and mechanism of calcium complexation with human serum components in the presence of silicone rubber were assessed in in vitro model experiments. Silicone rubber samples were incubated for 67 h in serum of both physiologic calcium concentrations and induced hypercalcemic levels. Physiologic calcium incubation decreased the intrinsic and nth binding site association constants by factors of 2 and 4, respectively, from initial serum values. Incubation with silicone rubber did not change the association constants. Hypercalcemia incubation lowered the intrinsic and nth binding site association constants by 39 and 61%, respectively. Polymer incubation increased the intrinsic association constant and decreased the number of binding sites by a factor of 2. With initial binding site association constants unperturbed by incubation and Hill coefficients equivalent to 1, complexation was non-cooperative and binding sites were considered to be identical and independent. Both Scatchard and Hill plots show nonlinearity for the polymer incubation case giving two distinct linear regions. This is indicative of cooperative binding for incubation with polymer; the Hill coefficient for the physiologic calcium concentration range was 0.97, while the coefficient for the hypercalcemic calcium range was 6.
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