Influence of DL-β-hydroxybutyric acid on cell proliferation and calcium influx

Poly(hydroxybutyrate-co-hydroxyhexanoate) (PHBHHx), a member of the polyhydroxyalkanoate family of biopolyesters, has superior mechanical properties and biocompatibilities that enable it to meet diverse biomedical requirements. The main component of PHBHHx is DL-β-hydroxybutyric acid (HB), a ketone body that is also produced in vivo. The effects of HB treatment on murine fibroblast L929 cells, human umbilical vein endothelial cells, and rabbit articular cartilages were investigated. HB (0.005-0.10 g/L) promoted cell proliferation for each cell line. Cell cycle analysis indicated that HB had a stimulatory effect on DNA synthesis. Flow cytometric analysis of L929 cells revealed changes in the [Ca²⁺]_i for different stages of the cell cycle. In L929 cells, HB (0.02 g/L) stimulated a rapid increase in the concentration of cytosolic calcium that was blocked by verapamil and diltiazem, inhibitors of L-type Ca²⁺ channels. Finally, verapamil inhibited HB-induced L929 cell proliferation. Collectively, these results indicated that HB had a stimulatory effect on cell cycle progression that is mediated by a signaling pathway dependent upon increases in [Ca²⁺]_i. This trophic effect may underlie the good biocompatibility observed for PHBHHx.