Influence of particle size in the effect of polyethylene on human osteoblastic cells

Abstract

The influence of two different sizes of polyethylene particles (<30 and 20–200 μm) on osteoblastic function has been studied in primary human bone cell cultures. Cells were obtained from trabecular bone fragments of patients undergoing knee reconstructive surgery. On reaching confluency, cells were subcultured in three flasks: <30 μm polyethylene particles were added to the first flask, 20–200 μm particles to the second flask and none to the third flask, which was the control. The resulting subcultures were incubated until confluence. Osteoblastic function was evaluated by assaying the secretion of osteocalcin, alkaline phosphatase, and C-terminal type I procollagen (PICP), with or without 1,25(OH) 2D 3 stimulation in the cell-conditioned medium. Adding <30 μm polyethylene particles to these osteoblastic cell cultures increased the levels of osteocalcin secreted after 1,25(OH) 2D 3 stimulation. Treating stimulated or basal osteoblastic cultures with either polyethylene particle size did not affect alkaline phosphatase secretion. However, the addition of <30 μm polyethylene particles decreased PICP levels in the basal and stimulated cultures. A parallel series of osteoblastic cultures was treated with <30 μm polyethylene particles and stimulated or not with 1,25(OH) 2D 3 to determine the effect on osteocalcin mRNA expression using RT-PCR amplification. Polyethylene particle-treated cultures had higher osteocalcin mRNA expression regardless of whether they had been stimulated with 1,25(OH) 2D 3 or not. We conclude that particle size affects the influence of polyethylene on osteoblastic function markers. Particles with a diameter of less than 30 μm increase osteocalcin expression and secretion.