Enhanced Growth and Osteogenic Differentiation of Human Osteoblast-Like Cells on Boron-Doped Nanocrystalline Diamond Thin Films

Intrinsic nanocrystalline diamond (NCD) films have been proven to be promising substrates for the adhesion, growth and osteogenic differentiation of bone-derived cells. To understand the role of various degrees of doping (semiconducting to metallic-like), the NCD films were deposited on silicon substrates by a microwave plasma-enhanced CVD process and their boron doping was achieved by adding trimethylboron to the CH4:H2 gas mixture, the B∶C ratio was 133, 1000 and 6700 ppm. The room temperature electrical resistivity of the films decreased from >10 MΩ (undoped films) to 55 kΩ, 0.6 kΩ, and 0.3 kΩ (doped films with 133, 1000 and 6700 ppm of B, respectively). The increase in the number of human osteoblast-like MG 63 cells in 7-day-old cultures on NCD films was most apparent on the NCD films doped with 133 and 1000 ppm of B (153,000±14,000 and 152,000±10,000 cells/cm2, respectively, compared to 113,000±10,000 cells/cm2 on undoped NCD films). As measured by ELISA per mg of total protein, the cells on NCD with 133 and 1000 ppm of B also contained the highest concentrations of collagen I and alkaline phosphatase, respectively. On the NCD films with 6700 ppm of B, the cells contained the highest concentration of focal adhesion protein vinculin, and the highest amount of collagen I was adsorbed. The concentration of osteocalcin also increased with increasing level of B doping. The cell viability on all tested NCD films was almost 100%. Measurements of the concentration of ICAM-1, i.e. an immunoglobuline adhesion molecule binding inflammatory cells, suggested that the cells on the NCD films did not undergo significant immune activation. Thus, the potential of NCD films for bone tissue regeneration can be further enhanced and tailored by B doping and that B doping up to metallic-like levels is not detrimental for cells

Semiconducting to metallic-like boron doping of nanocrystalline diamond films and its effect on osteoblastic cells

The impact of boron doping level of nanocrystalline diamond (NCD) films on the character of cell growth (i.e., adhesion, proliferation and differentiation) is presented. Intrinsic and boron-doped NCD films were grown on Si/SiO2 substrates by microwave plasma CVD process. The boron-doped samples were grown by adding trimethylboron (TMB) to the gas mixture of methane and hydrogen. Highly resistive (0 ppm), semiconducting (133 or 1000 ppm), and metallic-like (6700 ppm) NCD films were tested as the artificial substrates for the cultivation of osteoblast-like MG 63 cells. The conductivity and surface charge increased monotonically with the increasing boron content. All NCD substrates showed good biocompatibility and stimulated the adhesion and growth of MG 63 cells. Higher osteocalcin concentration (by more than 30%) for the cells growing on 1000 and 6700 ppm boron-doped NCD films was found which indicates an enhancement in the cell growth biochemistry

V.: Regionally-selective adhesion and growth of human osteoblast-like MG 63 cells on micropatterned fullerene C60 layers

Fullerenes C60 were deposited on to microscopic glass coverslips using the Leybold Univex-300 vacuum system through metallic masks with rectangle openings. Below the openings, the C60 formed prominences 128 ± 8 nm (A), 238 ± 3 nm (B), 326 ± 5 nm (C) or 1043 ± 57 nm (D) in height. Human osteoblast-like MG 63 cells in the cultures on samples A, B and C were distributed almost homogeneously, while on samples D, they were localized almost exclusively in the grooves among the prominences. Thus, fullerene C60 films can act as substrates for guided cell adhesion and growth

Micro- and nanopatterned surfaces for guided adhesion, growth and phenotypic maturation of cells

Micropatterned surfaces were created by UV light-irradiation of polytetrafluoroethylene through a metallic mask, by successive plasma polymerization of acrylic acid and 1,7-octadiene, or by creation of prominences and grooves by deposition of fullerenes C60 through a metallic mask. All these surface types were capable of inducing regionally-selective adhesion, proliferation and phenotypic maturation of vascular endothelial cells, vascular smooth muscle cells or human bone-derived MG 63 cells. Nanopatterned surfaces created by tethering GRGDSG oligopeptides through polyethylene oxide chains on a polymeric surface promoted spreading, formation of focal adhesion plaques and DNA synthesis in vascular smooth muscle cells. Surfaces nanopatterned with nanocrystalline diamond gave good support for the adhesion, growth and metabolic activity of osteoblast-like MG 63 cells

Staphylococcal bacteremia in cancer patients: Risk factors and outcome in 134 episodes prior to and after introduction of quinolones into infection prevention in neutropenia

A total of 134 episodes of staphylococcal bacteremia (SBE) appearing among 9987 admissions, and 979 episodes of bacteremia in cancer patients within 5 years, were analyzed for risk factors, clinical course and outcome; 64 were monomicrobial and 70 polymicrobial. The most frequent risk factors were acute leukemia, catheter insertion, long-lasting neutropenia, and prior prophylaxis with quinolones. There was no significant difference between polymicrobial and monomicrobial SBE in risk factors. The two groups differed only in the source of bacteremia (gastrointestinal and respiratory-tract infections were more common in monomicrobial SBE) and etiology - Staphylococcus aureus appeared more frequently in monomicrobial than in polymicrobial bacteremia (20.3% compared to 4.3%, P<0.05). More complications (14.3%) such as abscesses, endocarditis, etc. appeared in the group of polymicrobial SBE (P<0.05). No difference was observed in clinical course and outcome between monomicrobial and polymicrobial SBE. The incidence of SBE has increased since 1991, when quinolones were first used in prophylaxis in afebrile neutropenia at our center; however, the infection-associated mortality in monomicrobial SBE was low (4.3%)