Human osteoblast-like SAOS-2 cells on submicron-scale fibers coated with nanocrystalline diamond films

Abstract

A unique composite nanodiamond-based porous material with a hierarchically-organized submicron-nano-structure was constructed for potential bone tissue engineering. This material consisted of submicron fibers prepared by electrospinning of silicon oxide (SiO_x), which were oxygen-terminated (O-SiO_x) and were hermetically coated with nanocrystalline diamond (NCD) films. The NCD films were then terminated with hydrogen (H-NCD) or oxygen (O-NCD). The materials were tested as substrates for the adhesion, growth and osteogenic differentiation of human osteoblast-like Saos-2 cells. The number and the spreading area of the initially adhered cells, their growth rate during 7 days after seeding and the activity of alkaline phosphatase (ALP) were significantly higher on the NCD-coated samples than on the uncoated O-SiO_x samples. In addition, the concentration of type I collagen was significantly higher in the cells on the O-NCD-coated samples than on the bare O-SiO_x samples. The observed differences could be attributed to the tunable wettability of NCD and to the more appropriate surface morphology of the NCD-coated samples in contrast to the less stable, rapidly eroding bare SiO_x surface. The H-NCD coatings and the O-NCD coatings both promoted similar initial adhesion of Saos-2 cells, but the subsequent cell proliferation activity was higher on the O-NCD-coated samples. The concentration of beta-actin, vinculin, type I collagen and alkaline phosphatase (ALP), the ALP activity, and also the calcium deposition tended to be higher in the cells on the O-NCD-coated samples than on the H-NCD-coated samples, although these differences did not reach statistical significance. The improved cell performance on the O-NCD-coated samples could be attributed to higher wettability of these samples (water drop contact angle less than 10°), while the H-NCD-coated samples were hydrophobic (contact angle >70°). NCD-coated porous SiO_x meshes can therefore be considered as appropriate scaffolds for bone tissue engineering, particularly those with an O-terminated NCD coating.

Keywords: Alkaline phosphatase; Collagen; Hierarchical organization; Nanocrystalline diamond; Osteoblast proliferation; Osteocalcin; Osteogenic differentiation; Silica nanostructures; Submicron fibers.