Quantitation method for Cu2+ release from biomaterials for bone tissue engineering: development and validation

CATTALINI JUAN; MOURINO V.; LUCANGIOLI S.

Mendoza

Congreso; 7mo Congreso de Química Analítica; 2013

AACA

The new generation of biomaterials to develop scaffolds with application in bone tissue engineering (BTE) includes the enhancement of the interaction between the material and the tissue to be regenerated [1]. Lately, an interesting approach has been the incorporation of metallic ions as therapeutic agents into the biomaterials to develop scaffolds [1,2]. Particularly, vascularization of bone tissue plays an important role in the integration of scaffold with the tissue to be regenerated. In this regard, copper (Cu2+) ions are considered metallic angiogenic factors, because of their important role in blood vessels growth and the stimulation of the proliferation of endothelial cells [1,3,4]. In addition, Cu2+ ions also enhance the differentiation of stem cells towards the osteogenic linage [1]. Therefore, the study of the release of Cu2+ ions from these biomaterials is important to evaluate the effects that these ions could produce on bone tissue. As capillary electrophoresis (CE) methods are highly efficient for ions analysis in different matrices due to its high separation efficiency and resolution, versatility, low sample consumption, short analysis time and the possibility of automation, the aim of this work was to develop and validate an UV-direct method for Cu2+ release quantitation form biomaterials for BTE. Cu2+ cross-linked alginate films containing bioactive glass nanoparticles were similarly prepared as mentioned in a previous work [5]. The release study of Cu2+ from films was carried out in phosphate buffer and aliquots samples were withdrawn at regular time intervals for 30 days. The quantitation was made using a fused silica capillary (40 cm, 75 µm ID) and background electrolyte containing 8 mM EDTA, 5 mM TTAB (tetradecyltrimethylammonium bromide) and 50 mM acetate-acetic buffer at pH 5.5. Direct UV-detection mode at 225 nm was performed. The separations were achieved using reverse polarity at 15 kV, a cartridge temperature of 25 °C and 1 psi for 10 s for sample injection. Parameters of validation such as specificity, linearity, limit of detection and quantitation, accuracy, precision and robustness were evaluated according to the International Conference on Harmonization guidelines. The results indicated that this method is suitable for Cu2+ quantitation from biomaterials with application in bone tissue engineering.