Initial in vitro biocompatibility of a bone cement composite containing a poly-ε-caprolactone microspheres

The biocompatibility of a reinforced calcium phosphate injectable bone substitute (CPC-IBS) containing 30% poly-ε-caprolactone (PCL) microspheres was evaluated. The IBS consisted of a solution of chitosan and citric acid as the liquid phase and tetracalcium phosphate (TTCP) and dicalcium phosphate anhydrous (DCPA) powder as the solid phase with 30% PCL microspheres. The surface of the CPC-IBS was observed by SEM, and analyzed by EDX profiles. The initial setting of the sample was lower in the IBS containing 0% citric acid than in the IBS containing 10 or 20% citric acid. The compressive strength of the PCL-incorporated CPC-IBS was measured using a Universal Testing Machine. The 20% citric acid samples had the highest mechanical strength at day 12, which was dependent on both time and the citric acid concentration. The in vitro bioactivity experiments with simulated body fluid (SBF) confirmed the formation of apatite on the sample surfaces after 2, 7, and 14 days of incubation in SBF. Ca and P ion release profile by ICP method also confirmed apatite nucleation on the CPC-IBS surfaces. The in vitro biocompatibility of the CPC-IBS was evaluated by using MTT, cellular adhesion, and spreading studies. In vitro cytotoxicity tests by MTT assay showed that the 0 and 10% CPC-IBS was cytocompatible for fibroblast L-929 cells. The SEM micrograph confirmed that MG-63 cells maintained their phenotype on all of the CPC-IBS surfaces although cellular attachment was better in 0 and 10% CPC-IBS than 20% samples.