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In Vitro Studies of Carbon Nanotubes Biocompatibility
In Vitro Studies of Carbon Nanotubes Biocompatibility
www.elsevier.com/locate/carbon
Abstract
Cellular tests have been applied to study the biocompatibility of high purity multiwalled carbon nanotubes (MWNTs). The viability
of broblasts, osteoblasts and osteocalcin concentrations in osteoblasts cultures in the presence of nanotubes has been examined, as well
as the degree of cells stimulation, based on the amount of released collagen type I, IL-6 and oxygen free radicals. The high level of via-
bility of the examined cells in contact with the nanotubes, the slight increase of collagen formation, the lack of pro-inammatory IL-6
cytokine as well as the induction of free radicals, conrm a good biocompatibility of nanotubes, which is similar to that of polysulfone
currently used in medicine. The collagen synthesis induced on nanotubes by both broblasts and osteoblasts may be signicant for future
medical applications of nanotubes, in particular as substrates for the regeneration of tissues.
2005 Elsevier Ltd. All rights reserved.
0008-6223/$ - see front matter 2005 Elsevier Ltd. All rights reserved.
doi:10.1016/j.carbon.2005.11.022
J. Chopek et al. / Carbon 44 (2006) 11061111 1107
Table 1
Examples of applications of carbon biomaterials [810]
Type of material Function Type of implant Area of medicine
Carboncarbon composites Bone xation Screws, plates, nails, stems of endoprosthesis Bone surgery
Braided carbon bers Tissue knitting, reconstruction of joint Surgical sutures, ligament and tendons prosthesis Orthopedics
ligaments and tendons
Unwoven carbon fabric Filling bone and cartilage losses Disks and rings Bone surgery
Coatings of diamond-like Coating of metal implantscorrosion Joint endoprosthesis, screws Bone surgery
carbon (DLC) protection
Glassy carbon Blood ow regulation Heart valves Cardiology
growth agents, and may also be used as a drug carrier [18]. MWNTs, free of any disordered carbon, that makes the
Very good mechanical properties, the possibility of surface purication very easy by simple dissolution of the catalyst
machining as well as the ability to form functional groups precursor in HCl. The elemental analysis on the puried
constitute good fundaments for the use of nanotubes in the material gives: C = 96 wt%, H = 0.85 wt%, Mg = 200 ppm
fabrication of composite materials. and Co = 2 wt%. The low magnesium and cobalt content
The possibility of forming composites with polymer shows that the treatment in hydrochloric acid is very
matrices, both biostable (polysulfone, PEEK), and biore- ecient.
sorbable (PLA, PGLA, co-polymers) is of particular The structure and texture of the puried carbon nano-
importance in the case of medical applications [1921]. tubes was controlled by scanning electron microscopy
This opens opportunities for the manufacture of multifunc- (SEM, Hitachi S 4200) and transmission electron micros-
tional implants useful in many areas of medicine. However, copy (TEM, Philips CM 20). For the TEM observations,
with resorbable polymers, the relation between the resorp- the samples were ground in ethanol and dropped over a
tion time and the time of tissue healing is of signicant copper holey grid covered by amorphous carbon. The
importance. A too high resorption rate may lead to a SEM picture presented in Fig. 1 shows a very dense and
release of nanotubes from the composite materials into entangled network of nanotubes, where graphitic particles,
the living body. disordered carbon and nanocapsules are completely absent.
In the case of the ceramic matrix composites, an The TEM observation (Fig. 2) demonstrates that this
improvement of the fracture toughness can be expected, material consists exclusively of multiwalled nanotubes.
which may be particularly important in the case of manu- The high-resolution images show that the central canal is
facturing reinforced nanostructure ceramics. The nano- quite well dened (from 2 to 5 nm in diameter) and the
composite system of nanotubes reinforced hydroxyapatite walls consist of an average number of 1015 continuous
[22] may be included in this category. layers oriented in parallel to the tube axis. Most of the car-
The use of the advantages of nanotubes in medical bon nanotubes have closed tips and for only few of them
applications, particularly their tubular morphology and cobalt particles are encapsulated near the tip or inside the
their excellent electrical and mechanical properties relies canal. The histogram of the outer diameters (Fig. 3) shows
heavily on their biocompatibility. Although both the nat- a good calibration of the carbon nanotubes, with outer
ure of carbon and positive experiences to date with various diameters mostly in the range from 10 to 15 nm. The histo-
forms of carbon would suggest also a good biocompatibil- gram of the inner diameters (diameters of the central
ity of nanotubes, basic cellular tests must be performed in canal), ranges mainly in a narrow range from 2 to 5 nm.
order to allow projects and application works to be opened
in medicine. In the present work, the viability of bro-
blasts, osteoblasts and osteocalcin concentrations in osteo-
blasts cultures in the presence of high purity multiwalled
carbon nanotubes (MWNTs) has been examined, as well
as the degree of cells stimulation, based on the amount of
released collagen type I, IL-6 and oxygen free radicals.
2. Experimental
50
40
30
pg/ml
20
10
0
Fb PSU PSU+nanotubes
Fig. 7. IL-6 release from broblasts cultured on PSU and PSU + nano-
Fig. 5. Prolograms of PSU (a) and PSU + nanotubes (b). (RaArith- tubes. The results represent the average SD of duplicates from 4
metic mean roughness value.) dierent experiments.
1110 J. Chopek et al. / Carbon 44 (2006) 11061111
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