The Attachment Affinity of Hemoglobin toward Silver-Containing Bioactive Glass Functionalized with Glutaraldehyde

Abstract

Bioactive glasses belonging to the 56SiO(2)center dot(40 - x)CaO center dot 4P(2)O(5)center dot xAg(2)O system, with x = 0, 2, and 8 mol %, were surface functionalized with the protein coupling agent glutaraldehyde (GA) and further evaluated in terms of hemoglobin affinity. The bare and GA-functionalized samples were investigated before and after protein attachment, by electron paramagnetic resonance (EPR) spectroscopy combined with spin-labeling procedure. Methanethiosulfonate spin label was used to explore the local environment of beta-93 cysteine in horse hemoglobin, in terms of spin label side chain mobility. The EPR simulation methods were employed to quantify the rotational correlational times and fraction of the immobilized spin labels. The EPR absorption spectrum was further exploited to estimate the amount of hemoglobin loaded on the substrates. The surface elemental composition obtained by X-ray photoelectron spectroscopy revealed similar tendency in terms of surface coverage. Changes in surface architecture, that is, changes in surface morphology after protein coverage, were observed by scanning electron microscopy. It was concluded that GA improves the stability of protein attachment and induces polymerization of hemoglobin molecules.