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

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 (SiOx), which were oxygen-terminated (O-SiOx) 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-SiOx 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-SiOx 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 SiOx 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 SiOx meshes can therefore be considered as appropriate scaffolds for bone tissue engineering, particularly those with an O-terminated NCD coating.

Accelerated in vitro recellularization of decellularized porcine pericardium for cardiovascular grafts.

An ideal decellularized allogenic or xenogeneic cardiovascular graft should be capable of preventing thrombus formation after implantation. The antithrombogenicity of the graft is ensured by a confluent endothelial cell layer formed on its surface. Later repopulation and remodeling of the scaffold by the patient's cells should result in the formation of living autologous tissue. In the work presented here, decellularized porcine pericardium scaffolds were modified by growing a fibrin mesh on the surface and inside the scaffolds, and by attaching heparin and human vascular endothelial growth factor (VEGF) to this mesh. Then the scaffolds were seeded with human adipose tissue-derived stem cells (ASCs). While the ASCs grew only on the surface of the decellularized pericardium, the fibrin-modified scaffolds were entirely repopulated in 28 d, and the scaffolds modified with fibrin, heparin and VEGF were already repopulated within 6 d. Label free mass spectrometry revealed fibronectin, collagens, and other extracellular matrix proteins produced by ASCs during recellularization. Thin layers of human umbilical endothelial cells were formed within 4 d after the cells were seeded on the surfaces of the scaffold, which had previously been seeded with ASCs. The results indicate that an artificial tissue prepared by in vitro recellularization and remodeling of decellularized non-autologous pericardium with autologous ASCs seems to be a promising candidate for cardiovascular grafts capable of accelerating in situ endothelialization. ASCs resemble the valve interstitial cells present in heart valves. An advantage of this approach is that ASCs can easily be collected from the patient by liposuction.