Highly porous PHB-based bioactive scaffolds for bone tissue engineering by in situ synthesis of hydroxyapatite

In this study bioactive and bioresorbable porous scaffolds for bone tissue regeneration, based on poly(3-hydroxybutyrate) (PHB), are presented. The porous structure is obtained by thermally induced phase separation (TIPS) technique, whereas the osteoinductivity and osteoconductivity are enhanced through the incorporation of hydroxyapatite (HA). The HA particles are generated in PHB using an innovative filler in situ synthesis, and the properties of the composite scaffolds are then compared to scaffolds obtained by conventional mechanical dispersion of ex situ synthesized HA particles. The in situ synthesis leads to composite materials with improved porosity, even at high filler content, without any degradation of the polymeric matrix as confirmed by GPC and DSC measurements. On the contrary, the samples prepared by ex situ method show a suppressed porosity by increasing the inorganic filler content, therefore limiting the amount of HA that can be loaded in PHB and the resulting bioactivity. The possibility to use PHB/HA porous composites as scaffolds for bone tissue regeneration, is assessed by preliminary cell viability in vitro studies. In particular, it is observed that the composites are fully cytocompatible and able to sustain MC3T3-E1 mouse pre-osteoblast cells adhesion and proliferation. Investigations on cell morphology reveal, for all PHB/HA scaffolds, the presence of differentiated cells with a predominance of osteocyte-like morphology, which are not observed for neat PHB scaffolds. Moreover, the MC3T3-E1 cells differentiation towards osteoblastic phenotype is further supported by the evaluation of the early osteogenic markers. In particular, samples loaded with HA in situ synthesized showed the highest ALP production and typical morphology of the terminal differentiation stages of osteoblasts.