Improved adhesion and growth of human osteoblast-like MG 63 cells on biomaterials modified with carbon nanoparticles

doi:10.1016/j.diamond.2007.07.015

Diamond and Related Materials

Volume 16, Issue 12, December 2007, Pages 2133-2140

https://doi.org/10.1016/j.diamond.2007.07.015 Get rights and content

Abstract

Three types of materials modified with carbon particles were prepared: (1) carbon fibre-reinforced carbon composites (CFRC), materials promising for hard tissue surgery, coated with a fullerene C₆₀ layer, (2) terpolymer of polytetrafluoroethylene, polyvinyldifluoride and polypropylene mixed with 4 wt.% of single- or multi-walled carbon nanotubes and (3) nanostructured or hierarchically micro- and nanostructured diamond layers deposited on silicon substrates. The materials were seeded with human osteoblast-like MG 63 cells (density from 8500 cells/cm² to 25,000 cells/cm²). On the fullerene layers, the cells (day 2 after seeding) adhered in numbers from 2.3 to 3.5 times lower than those on control non-coated CFRC or polystyrene dishes. However, their spreading area was larger by 68% to 145% than that on the control surfaces. These cells also assembled numerous dot-like vinculin-containing focal adhesion plaques and a rich fine mesh-like beta-actin cytoskeleton. Similar results were obtained on the terpolymers mixed with carbon nanotubes. The cells were well spread and contained distinct beta-actin filament bundles, whereas the cells on the pure terpolymer were often rounded and clustered into aggregates. An enzyme-linked immunosorbent assay revealed that the cells on the material with single-walled carbon nanotubes contained a higher concentration of vinculin and talin, i.e. components of focal adhesion plaques (by 56% and 35%, respectively, compared to the pure terpolymer). However, the concentration of osteocalcin, a marker of osteogenic differentiation, was lower in cells on the terpolymer containing multi-walled nanotubes, which was probably due to more active proliferation of these cells (on day 7, they reached a 4.5 times higher population density than cells on the unmodified terpolymer). Adding both single-and multi-walled nanotubes to the terpolymer did not increase the concentration of ICAM-1, a marker of immune activation, in MG 63 cells. On diamond layers, the number of initially adhered cells was higher on the nanostructured layers, whereas the subsequent proliferation was accelerated on the layers with a hierarchical micro-and nanostructure. Thus, all tested carbon nanoparticle-containing materials gave good support to adhesion and growth of bone-derived cells, and they can be considered as promising for construction of bone implants and bone tissue engineering.

Introduction

Carbon nanoparticles, such as fullerenes, nanotubes and nanodiamonds, are considered as promising building blocks for the construction of novel nanomaterials for emerging industrial technologies, such as molecular electronics, advanced optics or storage of hydrogen as a potential source of energy [1]. In addition, they are considered as promising materials for biomedical applications, such as photodynamic therapy against tumors and infectious agents, quenching oxygen radicals, biosensor technology, simulation of cellular components, such as membrane pores or ion channels, as well as controlled drug or gene delivery, particularly targeting the mineralized bone tissue [2], [3], [4], [5], [6]. Despite these exciting perspectives, relatively little is known about the influence of carbon nanoparticles present on the biomaterial surface on the adhesion and growth of cells.

In this paper, we present the influence of three types of carbon nanoparticle-modified materials on the adhesion and growth of cells. In the first set of experiments, a layer of fullerenes C₆₀ was deposited on carbon fibre-reinforced carbon composites, i.e. materials which are promising for hard tissue surgery [7]. The second set of experiments was carried out on a terpolymer of polytetrafluoroethylene, polyvinyldifluoride and polypropylene mixed with single- or multi-walled carbon nanotubes, and finally, diamond layers (nanostructured or hierarchically micro- and nanostructured) for potential biomaterial coating were prepared on a silicone substrate. These carbon nanoparticle-containing materials were seeded with human osteoblast-like MG 63 cells, and the adhesion and subsequent growth of these cells was investigated. We observed that all tested carbon nanoparticle-containing materials gave good support to adhesion and growth of bone-derived cells, and they can be considered as promising for construction of bone implants and bone tissue engineering.

Section snippets

Coating of carbon fibre-reinforced carbon composites (CFRC) with a fullerene layer

Two-dimensionally reinforced CFRC were prepared at the Institute of Rock Structure and Mechanics, Acad. Sci. CR, Prague [7]. Commercially available woven fabric (made of carbon fibres Toray T 800) was arranged in layers, infiltrated with a carbon matrix precursor (phenolic resin UMAFORM LE, Synpo Ltd., Pardubice, CR), pressed, cured, carbonised at 1000 °C, and finally graphitised at 2200 °C. In order to decrease the material surface micro-scale roughness, which has been shown to be

MG 63 cells on CFRC with a fullerene layer

Grinding with metallographic paper of 4000 grade lowered the surface roughness of CFRC about twice. As measured by a profilometer (Rank Taylor Hobson Ltd., England), the departures of the roughness profile from the mean line (i.e., R_a parameter) decreased from 6.5 ± 1.8 μm to 3.5 ± 0.6 μm, and the mean spacing of the adjacent local peaks (parameter S) lengthened from 38 ± 11 μm to 96 ± 49 μm. The fullerene coating did not significantly change this surface microroughness but created a nanostructured

Conclusion and further perspectives

The tested carbon nanoparticle-containing materials supported adhesion and growth of bone-derived cells. The carbon nanoparticle layers used in this study, such as fullerene layers and especially hard diamond coatings, could be used for surface modification of bone implants (e.g., bone-anchoring parts of joint prostheses or bone replacements) in order to improve their integration with the surrounding bone tissue. Polymers mixed with carbon nanoparticles could serve for the construction of

Acknowledgements

This study was supported by the Grant Agency of the Czech Republic (Grants No. 204/06/0225 and 202/05/2233). We also wish to express our thanks to Mrs. Ivana Zajanova (Inst. Physiol., Acad. Sci. CR) for her excellent technical assistance and Mr. Robin Healey (Czech Technical University, Prague) for the language revision of the manuscript.

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Improved adhesion and growth of human osteoblast-like MG 63 cells on biomaterials modified with carbon nanoparticles

Abstract

Introduction

Section snippets

Coating of carbon fibre-reinforced carbon composites (CFRC) with a fullerene layer

MG 63 cells on CFRC with a fullerene layer

Conclusion and further perspectives

Acknowledgements

Carbon

B. Lux: Diamond Relat. Mater.

Biomaterials

Biomaterials

Biomaterials

Biomaterials

Philos. Transact. A. Math. Phys. Eng. Sci.

Curr. Pharm. Biotechnol.

Anal. Bioanal. Chem.

J. Am. Chem. Soc.