Multiparametric In Vitro Evaluation of Cytocompatibility of 1% Strontium-Containing Nanostructured Hydroxyapatite

E-ISSN： 1662-9795|2014|631|345-350

ISSN： 1013-9826

Source： Key Engineering Materials, Vol.2014, Iss.631, 2015-01, pp. : 345-350

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Abstract

Hydroxiapatite (HA), one of the most widely employed bioceramic bone substitutes, when applied on its nanostructured form (nHA) may contribute to achieve a crystalline structure which is closer to the size and morphology of biological apatite. Furthermore, HA might also be doped with several different cations with biological effects including Sr2+. Therefore, a biomaterial based on nanostructured HA containing 1% Strontium (nSrHA) could present interesting biological properties, as strontium is described as a modulator of both osteoblast and osteoclast activities, presenting an important regulatory role on bone resorption. However, such modifications may also affect the biocompatibility of this material, which should be accessed initially by in vitro methods. Therefore, the present work aimed to evaluate the in vitro biocompatibility of 1% nSrHA discs with human primary osteoblasts through a multiparametric assay which assesses simultaneously metabolic activity (XTT assay), membrane integrity (NR test) and cell density (CVDE). Extracts of nSrHA, latex fragments (positive control), polystyrene beads (negative control) and nHA (for comparison) were prepared and exposed to 104 cells for 24h at 37°C/5% CO₂ on test plates, according to ISO 10993-5:2009, on quintuplicates. Cells exposed to unconditioned media were used as experimental control. After exposure, cells were tested for viability with a commercial multiparametric kit (In Cytotox, Xenometrix, Germany). The positive and negative controls presented the expected results, validating the assay. Both nHA and SrnHA were considered biocompatible, since the presented a cell viability after exposure statistically similar to the experimental control. In conclusion, the synthesized nSrHA discs are cytocompatible and, consequently, adequate for further in vitro tests on cell adhesion, proliferation and differentiation.