Novel pHEMA-gelatin SPHs as bone scaffolds in dynamic cultures.

The effectiveness of poly(2-hydroxyethyl methacrylate)-gelatin superporous hydrogels (pHEMA-gelatin SPHs) was investigated for bone tissue engineering. The cell culture studies were performed with preosteoblastic MC3T3-E1 cells. Dynamic culture conditions were provided using 100 ml spinner flask rotating at 50 rpm. According to the results of mitochondrial activity test (1-3-[4,5-dimethylthiazol-2-yl]-diphenyltetrazolium bromide), there is no significant difference between proliferation behavior of cells cultured under static and dynamic conditions during 28 days. Observations by scanning electron microscopy and confocal laser scanning microscopy showed that, cells attached well onto the scaffolds and spread through the pores for both culture conditions. However, it was found that, calcium deposition and alkaline phosphatase activity in the scaffolds cultured under dynamic conditions were higher than that of static conditions. The expression of osteogenic differentiation markers, i.e. collagen I and osteopontin, based on real-time reverse transcriptase-polymerase chain reaction demonstrated increased responses under the spinner flask culture conditions. The combination of dynamic culture conditions in spinner flask with the use of superporous pHEMA-gelatin scaffolds enhanced the outcomes related to bone tissue engineering.