Osteoblast-like MG Research Articles

TiO2 doped chitosan/poly (vinyl alcohol) nanocomposite film with enhanced mechanical properties for application in bone tissue regeneration

Here, TiO2 nanoparticles have been doped into the polymer film-construct of Chitosan/poly (vinyl alcohol)/Nano-hydroxyapatite (CPHT I – III) to enhance the mechanical and biological properties of the film so as to mimic the human bone extracellular matrix for application in human bone regeneration. The synthesized films are highly porous in nature along with the presence of macrovoids. Significantly enhanced mechanical properties were obtained upon the addition of TiO2 in comparison to previous literature. Increasing content of n-HAP-TiO2 increased the elasticity, tensile strength of the films and the antibacterial efficacy against both Gram-Positive and Gram-Negative Bacteria. The pH of CPHT I-III films in saline remained in the low alkalinity range of (7.48–7.53) on day 14. CPHT I-III films were compatible with the human erythrocytes as their hemolysis was well below the limit of acute hemolysis. The in-vitro studies revealed the highly cytocompatible nature of CPHT III (15% n-HAP-TiO2) for osteoblast-like MG – 63 cell attachment and proliferation. The study has revealed that CPHT III has the potential to be used for bone tissue regeneration, our future studies will be focused on the in-vivo investigations to establish its use in clinical settings.

Biodegradable dendrimer functionalized carbon nanotube-hybrids for biomedical applications

In the present study, biodegradable poly(amidoamine) dendrimers (PAMAM G1/G2/G3) was covalently functionalized with carbon nanotubes (CNTs) and nanostructured hydroxyapatite was immobilized onto CNTs-PAMAM matrix through coordination bond between the –NH2 groups of dendrimer and Ca2+ of hydroxyapatite. The structural and morphological behaviors of the nanohybrids were established through spectroscopic and microscopic analyses. In vitro cytotoxicity and cell proliferation was assessed through osteoblast-like MG 63 cell line using 3–4, 5-dimethylthiazole-2-yl, 2,5-diphenyl tetrazolium bromide assay for 3 days. The more abundant of –NH2 group exist in PAMAM(G3) dendrimer attracts more number of HAp molecules onto their surface which exhibits enhanced activity in cell proliferations even at higher concentrations.

Heat-treated carbon coatings on poly (l-lactide) foils for tissue engineering

Carbon-based materials have emerged as promising candidates for a wide variety of biomedical applications, including tissue engineering. We have developed a simple but unique technique for patterning carbon-based substrates in order to control cell adhesion, growth and phenotypic maturation. Carbon films were deposited on PLLA foils from distances of 3 to 7 cm. Subsequent heat-treatment (60 °C, 1 h) created lamellar structures with dimensions decreasing from micro- to nanoscale with increasing deposition distance. All carbon films improved the spreading and proliferation of human osteoblast-like MG 63 cells, and promoted the alignment of these cells along the lamellar structures. Similar alignment was observed in human osteoblast-like Saos-2 cells and in human dermal fibroblasts. Type I collagen fibers produced by Saos-2 cells and fibroblasts were also oriented along the lamellar structures. These structures increased the activity of alkaline phosphatase in Saos-2 cells. Carbon coatings also supported adhesion and growth of vascular endothelial and smooth muscle cells, particularly flatter non-heated carbon films. On these films, the continuity of the endothelial cell layer was better than on heat-treated lamellar surfaces. Heat-treated carbon-coated PLLA is therefore more suitable for bone and skin tissue engineering, while carbon-coated PLLA without heating is more appropriate for vascular tissue engineering.

Hydrogen-Terminated Diamond Sensors for Electrical Monitoring of Cells

In this paper we introduce fully optically transparent impedance biosensors based on intrinsic nanocrystalline diamond (NCD) films deposited on glass substrate. Prepared sensors have an interdigital electrode (IDE) structures realized by local hydrogen and oxygen termination of diamond surface, which mean in-plane configuration of active sensor area. Sensors were tested by real time monitoring of human osteoblast-like MG 63 cells in wide frequency range from 10 Hz to 100 kHz for several days. Two different measurement setups were used and compared regarding to their advantages and disadvantages. Proof of concept of diamond-based impedance sensor is showed, i.e. time dependence and frequency dependence (Nyquist plots) of absolute impedance.

Polylactide nanofibers with hydroxyapatite as growth substrates for osteoblast-like cells

Various types of nanofibers are increasingly used in tissue engineering, mainly for their ability to mimic the architecture of tissue at the nanoscale. We evaluated the adhesion, growth, viability, and differentiation of human osteoblast-like MG 63 cells on polylactide (PLA) nanofibers prepared by needle-less electrospinning and loaded with 5 or 15 wt % of hydroxyapatite (HA) nanoparticles. On day 7 after seeding, the cell number was the highest on samples with 15 wt % of HA. This result was confirmed by the XTT test, especially after dynamic cultivation, when the number of metabolically active cells on these samples was even higher than on control polystyrene. Staining with a live/dead kit showed that the viability of cells on all nanofibrous scaffolds was very high and comparable to that on control polystyrene dishes. An enzyme-linked immunosorbent assay revealed that the concentration of osteocalcin was also higher in cells on samples with 15 wt % of HA. There was no immune activation of cells (measured by production of TNF-alpha), associated with the incorporation of HA. Moreover, the addition of HA suppressed the creep behavior of the scaffolds in their dry state. Thus, nanofibrous PLA scaffolds have potential for bone tissue engineering, particularly those with 15 wt % of HA.

Human osteoblast-like MG 63 cells on polysulfone modified with carbon nanotubes or carbon nanohorns

Composites of polysulfone (PSU) and 0.5, 1 or 2wt.% of single-wall carbon nanohorns (SWCNHs) or multi-wall carbon nanotubes (MWCNTs) were prepared. Reinforcement with SWCNHs, and especially with MWCNTs, increased the tensile strength and the Young modulus of the material. The composites provided good support for the initial adhesion of human osteoblast-like MG 63 cells, and also for their further viability and metabolic activity. From day 1 to 3, the cell population doubling time was longer on composites with higher nanoparticle concentrations, and the DNA synthesis in cells on the composites was often lower than on unmodified PSU or on polystyrene culture dishes (PS). Nevertheless, from day 3 to 5, the proliferation activity of cells on the composites increased. On day 5, the concentrations of osteocalcin were lower in the cells on both pure PSU and PSU/nanocarbon composites than in the cells on polystyrene dishes, but on day 11, these differences equalized. The composites did not promote a significant increase in the concentration of ICAM-1, i.e., a marker of cell immune activation. Therefore, after some improvements, PSU/carbon nanotubule composites may be considered for application as bone tissue replacements.

Influence of surface roughness of carbon materials on human osteoblast‐like cell growth

This article presents a study of the dependence of the biocompatibility of a carbon-based material, namely a 2D C/C composite, on mechanical and chemical surface modifications. The mechanical modifications were surface grinding and polishing, and chemical modifications were made by depositing thin layers of pyrolytic carbon, titanium-carbon and DLC layers. Human osteoblast-like MG 63 cells were cultivated on these materials. The densities of the cells after one-day cultivation and after four-day cultivation, and the average cell spreading area after one-day cultivation, were evaluated in dependence on particular surface roughness parameters. The minima of the cell density on pyrolytic carbon and titanium-carbon layers were found; they were connected with the maxima of the average cell area. For DLC, the cell area decreased as the roughness parameter Ra increased in the range 0.1-10 µm, although the minimum appeared for the density of the cells. Using a multivariate test, the dependences of the biocompatibility parameters on the layer material and on surface grinding were statistically significant. The results suggest that the optimal roughness parameters for MG 63 cell on carbon based surface were Ra ∼ 3.5 µm, RSm ∼0.03-0.08 mm, Rsk ∼0 or negative and Rku ∼ 20, DLC being the best material choice. These values of roughness were obtained by simple mechanical grinding of substrate and coating by DLC layer.

On the role of Nb-related sites of an oxidized β-TiNb alloy surface in its interaction with osteoblast-like MG-63 cells

β-Stabilized titanium (Ti) alloys containing non-toxic elements, particularly niobium (Nb), are promising materials for the construction of bone implants. Their biocompatibility can be further increased by oxidation of their surface. Therefore, in this study, the adhesion, growth and viability of human osteoblast-like MG 63 cells in cultures on oxidized surfaces of a β-TiNb alloy were investigated and compared with the cell behavior on thermally oxidized Ti, i.e. a metal commonly used for constructing bone implants. Four experimental groups of samples were prepared: Ti or TiNb samples annealed to 600°C for 60min in a stream of dry air, and Ti and TiNb samples treated in Piranha solution prior to annealing. We found that on all TiNb-based samples, the cell population densities on days 1, 3 and 7 after seeding were higher than on the corresponding Ti-based samples. As revealed by XPS and Raman spectroscopy, and also by isoelectric point measurements, these results can be attributed to the presence of T-Nb2O5 oxide phase in the surface of the alloy sample, which decreased its negative zeta (ζ)-potential in comparison with zeta (ζ)-potential of the Ti sample at physiological pH. This effect was tentatively explained by the presence of positively charged defects acting as Lewis sites of the surface Nb2O5 phase. Piranha treatment slightly decreases the biocompatibility of the samples, which for the alloy samples may be explained by a decrease in the number of defective sites with this treatment. Thus, the presence of Nb and thermal oxidation of β-stabilized Ti alloys play a significant role in the increased biocompatibility of TiNb alloys.

The Study of Human Osteoblast-Like MG 63 Cells Proliferation on Resorbable Polymer-Based Nanocomposites Modified with Ceramic and Carbon Nanoparticles

Polymer-based nanocomposites containing biocompatible and bioactive nanocomponents seem to be excellent materials that could be used in many biomedical applications. The aim of this study was biological evaluation of resorbable polymer-based nanocomposites (PLA, PCL) and their modifications with ceramic nanoparticles (silica — SiO2, montmorylonit — MMT) or carbon nanotubes. The nanocomposites were seeded with the human osteoblast-like MG 63 cells. After 1, 3 and 7 days of incubation, Trypan blue exclusion test was used to determine the viability and number of cells. The cell population density depending on incubation time and cell population doubling time was calculated. The cell proliferation abilities on the all applied nanocomposites and on control material (polystyrene cell culture plate) were also compared. The number of cells growing on the nanocomposite surfaces increased with the incubation time. The cell viability was not decreased for all applied materials during the entire study (97‐100%). The ceramic nanoparticles and carbon nanotubes modified the bone cell growth and proliferation rate. Results of this study confirm that all types of the nanocomposites are appropriate to the growing and proliferation of human osteoblast-like cells.

Surface treatment by electric discharge machining of Ti–6Al–4V alloy for potential application in orthopaedics

This study investigated the properties of Ti–6Al–4V alloy after surface treatment by the electric discharge machining (EDM) process. The EDM process with high peak currents proved to induce surface macro-roughness and to cause chemical changes to the surface. Evaluations were made of the mechanical properties by means of tensile tests, and of surface roughness for different peak currents of the EDM process. The EDM process with peak current of 29 A was found to induce sufficient surface roughness, and to have a low adverse effect on tensile properties. The chemical changes were studied by scanning electron microscopy equipped with an energy dispersive X-ray analyser (EDX). The surface of the benchmark samples was obtained by plasma-spraying a titanium dioxide coating. An investigation of the biocompatibility of the surface-treated Ti–6Al–4V samples in cultures of human osteoblast-like MG 63 cells revealed that the samples modified by EDM provided better substrates for the adhesion, growth and viability of MG 63 cells than the TiO2 coated surface. Thus, EDM treatment can be considered as a promising surface modification to orthopaedic implants, in which good integration with the surrounding bone tissue is required.

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.

Adhesion and Growth of Human Osteoblast-Like Cell in Cultures on Nanocomposite Carbon-Based Materials

This paper summarizes our recent research on carbon nanoparticles (fullerenes, nanotubes, nanodiamond) as substrates for the adhesion, growth and phenotypic maturation of osteogenic cells. Fullerenes C60 were deposited on microscopic glass coverslips in the form of continuous layers or layers micropatterned with bulge-like prominences of 128 ± 8 nm, 238 ± 3 nm, 326 ± 5n m and 1043 ± 57 nm in height. On continuous layers and on layers with prominences up to 326 ± 5 nm, the adhesion and proliferation of human osteoblast-like MG 63 cells was similar as in control cells on polystyrene dishes. On layers with prominences 1043 ± 57 nm in height, the cells grew preferentially in grooves among the prominences. Similar cell responses were found in MG 63 cells cultured on continuous and micropatterned films made of binary C60/Ti composites. In the second set of experiments, single wall carbon nanohorns and multi-wall carbon nanotubes were mixed with a terpolymer of polytetrafluoroethylene, polyvinyldifluoride and polypropylene (in concentrations from 2 to 8 wt%) or with polysulfone (concentrations from 0.5 to 2 wt%). Adding carbon nanotubes to the terpolymer markedly improved the cell adhesion, spreading and subsequent growth of MG 63 cells, while the adhesion and growth of these cells on the pure and modified polysulfone were similar, which was probably due to a more hydrophilic character of polysulfone compared to the terpolymer. Nanocrystalline diamond (NCD) was deposited on silicon substrates and provided an excellent substrate for the adhesion, growth and osteogenic differentiation of MG 63 cells, measured by the concentration of osteocalcin. These beneficial effects of NCD films were further enhanced by boron doping, which can be attributed to increased electroactivity (i.e., electrical potential and conductivity) of these films.

Fullerene (C 60)–transitional metal (Ti) composites: Structural and biological properties of the thin films

Thin films of the binary C 60/Ti composites (with various phase ratios) were deposited on the Si(001) wafers and microscopic glass coverslips in continuous and micropatterned forms. The composites acquired a nanogranular structure with granules of about 50 nm in size. The RBS inspection confirmed homogeneous distribution of the phases and also the presence of oxygen. The Raman study suggested polymerization of the C 60/Ti composites into polymeric structures. The hybrid substrates were tested on biocompatibility—the films were seeded with human osteoblast-like MG 63 cells, and their proliferation was analyzed for 7 days. It has been found that the C 60/Ti composites can be counted as good supports for the adhesion of the selected MG 63 cells. The composites exhibited similar biocompatibility as the mix of amorphous carbon and titanium, but different than fullerene (C 60) solids.

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/SiO 2 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.

Nanodiamond as Promising Material for Bone Tissue Engineering

The adhesion, growth and differentiation of human osteoblast-like MG 63 cells were investigated in cultures grown on nanostructured nanocrystalline diamond (NCD) films with either low surface roughness (rms of 8.2 nm) or hierarchically organized surfaces made of low roughness NCD films deposited on Si surfaces with the original microroughness (rms of 301.0 nm and 7.6 nm, respectively). The NCD films were grown using a microwave plasma-enhanced CVD method in an ellipsoidal cavity reactor. The films were treated in oxygen plasma to enhance the hydrophilic character of the diamond surface (water drop contact angle approx. 20 degrees). The samples were then sterilized by 70% ethanol, inserted into 12-well polystyrene multidishes (diameter 2.2 cm), seeded with human osteoblast-like MG 63 cells (40,000 cells/dish, 10,530 cells/cm2) and incubated in 2 ml of DMEM medium with 10% of fetal bovine serum. On day 3 after seeding, the cell numbers were significantly higher on the nanostructured NCD films (72,020 +/- 6540 cells/cm2) and also on the hierarchically micro- and nanostructured films (60200 +/- 6420 cells/cm2) than on the control polystyrene culture dish (40750 +/- 2,530 cells/cm2). The cells on hierarchically micro- and nanostructured diamond substrates also adhered over a significantly larger area (3730 +/- 180 microm2 compared to 2740 +/- 130 microm2 on polystyrene). The cell viability, measured by a LIVE/DEAD viability/cytotoxicity kit, reached 98% to 100% on both types of NCD films. The XTT test showed that the cells on both nanodiamond layers had significantly higher metabolic activity than those on the control polystyrene dish (approx. 2 to 3 times). Immunofluorescence staining of the cells on both NCD films revealed talin-containing focal adhesion plaques and beta-actin filaments, well apparent particularly at the cell periphery, as well as the presence of considerable amounts of osteocalcin, i.e., a marker of osteogenic cell differentiation. These results suggest that nanocrystalline diamond films give good support for adhesion, growth and differentiation of osteogenic cells and could be used for surface modification of bone implants in order to improve their integration with the surrounding bone tissue.

Molecular markers of adhesion, maturation and immune activation of human osteoblast-like MG 63 cells on nanocrystalline diamond films

A semi-quantitative evaluation was made of the concentration of α v- and β 1-integrins, integrin-associated proteins talin and vinculin, cytoskeletal protein β-actin, differentiation markers osteocalcin and osteopontin, and immunoglobulin adhesion molecule ICAM-1 (CD 54), using an enzyme-linked immunosorbent assay (ELISA), in human osteoblast-like MG 63 cells in 5-day-old cultures on hydrophilic oxygen-terminated nanocrystalline diamond (NCD) films with either nanoscale surface roughness (nano-NCD, RMS of 8.2 nm) or hierarchically organized submicron- and nanoscale roughness ( RMS of 301.0 nm and 7.6 nm, respectively). The concentration of talin was significantly higher in cells on submicron-nano-NCD (by 36.0 ± 6.2% compared to the value on the polystyrene dishes). In addition, the concentration of vinculin increased on both nano-NCD and submicron-nano-NCD (by 19.8 ± 3.4% and 26.0 ± 6.1%, respectively). The concentration of integrins α v and β 1, β-actin, osteocalcin, osteopontin and ICAM-1 were similar in the cells on both the NCD films and the control polystyrene surfaces. These results suggest that nanocrystalline diamond films give good support for colonization with osteogenic cells and could be used for surface modification of bone implants in order to improve their integration with the surrounding bone tissue, or for biosensor technology, where firm cell–substrate adhesion is also required.

Fullerene C 60 films of continuous and micropatterned morphology as substrates for adhesion and growth of bone cells

Fullerenes C 60 were deposited on microscopic glass coverslips in the form of continuous layers (thickness 505 ± 43 nm or 1090 ± 8 nm) or micropatterned layers using a metallic mask with rectangular openings (128 × 98 µm, spacing 50 µm). Below the openings, the fullerenes formed bulges 484 ± 5 nm or 1043 ± 57 nm in thickness. Very thin fullerene films were also found below the metallic bars of the grid. The adhesion and proliferation activities of human osteoblast-like MG 63 cells in cultures on the fullerene layers were similar as that on the control polystyrene dishes and glass coverslips. On the thick patterned layers, the cells grew preferentially in the grooves among the fullerene bulges. Although these grooves occupied only approximately 41% of the surface, they contained from 80% to 98% of the cells. Immunocytochemistry showed that the cells on all tested surfaces formed β 1 integrin- and talin-containing focal adhesion plaques, a β-actin cytoskeleton, and contained osteocalcin, a marker of osteogenic cell differentiation. These results suggest that fullerene C 60 layers can act as good substrates for cell colonization, and could also serve for the construction of micropatterned surfaces for guided cell adhesion and growth.

Resorbable polymeric scaffolds for bone tissue engineering: The influence of their microstructure on the growth of human osteoblast‐like MG 63 cells

Degradable three-dimensional porous scaffolds applicable as cell carriers for bone tissue engineering were developed by an innovative solvent casting/particulate leaching technique from poly(L-lactide-co-glycolide) (PLG). Three types of PLG scaffolds were prepared, and these had the same high porosity (83%) but increasing diameter of the pores (180-200 microm, 250-320 microm, and 400-600 microm) and increasing pore interconnectivity. The colonization of the scaffolds with human osteoblast-like MG 63 cells was then studied in vitro in a conventional static cell culture system. The number of cells growing on the scaffolds on days 1 and 7 after seeding was highest in the material with the largest pore diameter, but on day 15, the differences among the scaffolds disappeared. Confocal microscopy revealed that on day 1 after seeding, the cells penetrated to a depth of 490 +/- 100 microm, 720 +/- 170 microm, and 720 +/- 120 microm into the scaffolds of small, medium, and large pore size, respectively. Incorporation of bromodeoxyuridine into newly synthesized DNA and the concentration of vinculin, beta-actin, osteopontin, and osteocalcin in cells on the scaffolds of all pore sizes were similar to the values obtained on standard tissue culture polystyrene, which indicated good biocompatibility of the scaffolds. These results suggest that all scaffolds could serve as good carriers for bone cells, although the quickest colonization with cells was found in the scaffolds with the largest pore diameter from 400 to 600 microm.

Nanocomposite Ti/hydrocarbon plasma polymer films from reactive magnetron sputtering as growth support for osteoblast‐like and endothelial cells

Nanocomposite Ti/hydrocarbon plasma polymer (Ti/ppCH) films were deposited by DC magnetron sputtering of titanium target in n-hexane, argon, or a mixture of these two gases. The resultant films were heterogeneous, with inorganic regions of nanometer scale distributed within a plasma polymer matrix. The titanium content was controlled by adjusting the argon/n-hexane ratio in the working gas. In the pure n-hexane atmosphere, the Ti concentration was found to be below 1 at %, whereas in pure argon it reached 20 at %, as measured by Rutherford backscattering spectroscopy and elastic recoil detection analysis (RBS/ERDA). A high level of titanium oxidation is detected with TiO(2), substoichiometric titania, and titanium carbide, composing an inorganic phase of the composite films. In addition, high hydrogen content is detected in films rich with titanium. Ti-deficient and Ti-rich films proved equally good substrates for adhesion and growth of cultured human osteoblast-like MG 63 cells. In these cells, the population densities on days 1, 3, and 7 after seeding, spreading area on day 1, formation of talin-containing focal adhesion plaques as well as concentrations of talin and osteocalcin (per mg of protein) were comparable to the values obtained in cells on the reference cell culture materials, represented by microscopic glass coverslips or a polystyrene dish. An interesting finding was made when the Ti/ppCH films were seeded with calf pulmonary artery endothelial cells of the line CPAE. The cell population densities, the spreading area and also the concentration of von Willebrand factor, a marker of endothelial cell maturation, were significantly higher on Ti-rich than on Ti-deficient films. On Ti-rich films, these parameters were also higher or similar in comparison with the reference cell culture materials. Thus, both types of films could be used for coating bone implants, of which the Ti-rich film remains effective in enhancing the endothelialization of blood contacting artificial materials.

Bone and vascular endothelial cells in cultures on nanocrystalline diamond films

The adhesion, growth and viability of human osteoblast-like MG 63 cells and bovine pulmonary artery endothelial CPAE cells were investigated in cultures grown on nanocrystalline diamond (NCD) films with either low surface roughness (RMS of 8.2 nm) or hierarchically organized surfaces made of nano-dimensional roughness NCD films deposited on Si surfaces with the original microroughness (RMS of 301.1 nm and 7.6 nm, respectively). The number and viability of both cell types on NCD films was similar to the values obtained on control tissue culture polystyrene dishes, or even better. It can be concluded that nanocrystalline diamond films give good support for adhesion and growth of osteogenic and endothelial cells, and could be used in tissue engineering applications.