Analysis Of Cellular Adhesion On Superhydrophobic And Superhydrophilic Vertically Aligned Carbon Nanotube Scaffolds

We analyzed GFP cells after 24 h cultivated on superhydrophilic vertically aligned carbon nanotube scaffolds. We produced two different densities of VACNT scaffolds on Ti using Ni or Fe catalysts. A simple and fast oxygen plasma treatment promoted the superhydrophilicity of them. We used five different substrates, such as: as-grown VACNT produced using Ni as catalyst (Ni), as-grown VACNT produced using Fe as catalyst (Fe), VACNT-O produced using Ni as catalyst (NiO), VACNT-O produced using Fe as catalyst (FeO) and Ti (control). The 4′,6-diamidino-2-phenylindole reagent nuclei stained the adherent cells cultivated on five different analyzed scaffolds. We used fluorescence microscopy for image collect, ImageJ® to count adhered cell and GraphPad Prism 5® for statistical analysis. We demonstrated in crescent order: Fe, Ni, NiO, FeO and Ti scaffolds that had an improved cellular adhesion. Oxygen treatment associated to high VACNT density (group FeO) presented significantly superior cell adhesion up to 24 h. However, they do not show significant differences compared with Ti substrates (control). We demonstrated that all the analyzed substrates were nontoxic. Also, we proposed that the density and hydrophilicity influenced the cell adhesion behavior.48365371Haubold, A.D., Shim, H.S., Bokros, J.C., (1979) Biocompatibility of Clinical Implants Materials, 2, pp. 520-532. , D.F. WilliamsCenni, E., Granchi, D., Arciola, C.R., Ciapetti, G., Savarinol, S.S., Cavedagna, D., Di Leo, A., Pizzoferrato, A., (1995) Biomaterials, 16 (16), pp. 1223-1227Salvetat, J.P., Bonard, J.M., Thomson, N.H., Kulik, A.J., Forró, L., Benoit, W., Zuppiroli, L., (1999) Appl. Phys. A Mater. Sci. Process., 69 (3), pp. 255-260Tran, P.A., Zhang, L., Webster, T.J., (2009) Adv. Drug Deliv. Rev., 61 (12), pp. 1097-1114Lim, J.Y., Shaughnessy, M.C., Zhou, Z.Y., Noh, H., Vogler, E.A., Donahue, H.J., (2008) Biomaterials, 29 (12), pp. 1776-1784Smart, S.K., Cassady, A.I., Lu, G.Q., Martin, D.J., (2006) Carbon, 44 (6), pp. 1034-1047Lin, Y., Taylor, S., Li, H., Fernando, K.A., Qu, L., Wang, W., Gu, L., Sun, Y.P., (2004) J. Mater. Chem., 14 (4), pp. 527-541Tasis, D., Tagmatarchis, N., Bianco, A., Prato, M., (2006) Chem. Rev., 106 (3), pp. 1105-1136Miller, D.C., Vance, R.J., Thapa, A., Haberstroh, M., Webster, T.J., (2004) Appl. Bionics Biomech., 2 (1), pp. 1754-2103Thomas, V., Dean, D.R., Vohra, Y.K., (2006) Curr. Nanosci., 2 (3), pp. 155-177Stevens, M.M., George, J.H., (2005) Science, 310, pp. 1135-1138Monteiro-Riviere, N.A., Nemanich, R.J., Inman, A.O., Wang, Y.Y., Riviere, J.E., (2005) Toxicol. Lett., 155 (3), pp. 377-384Ramos, S.C., Vasconcelos, G., Antunes, E.F., Lobo, A.O., Trava-Airoldi, V.J., Corat, E.J., (2010) Surf. Coat. Technol., 204, pp. 3073-3077Lobo, A.O., Marciano, F.R., Ramos, S.C., Machado, M.M., Corat, E.J., Corat, M.A.F., (2011) Mater. Sci. Eng. C, 31 (7), pp. 1505-1511Antunes, E.F., Lobo, A.O., Corat, E.J., Trava-Airoldi, V.J., Martin, A.A., Veríssimo, C., (2006) Carbon, 44 (11), pp. 2202-2211Hogan, B., Beddington, R., Costantini, F., Lacy, E., (1994) Manipulating the Mouse Embryo - A Laboratory Manual, p. 260. , 2nd ed. EUAOwens, D.K., Wendt, R.C., (1969) J. Appl. Polym. Sci., 13, pp. 1741-1747Van Oss, C.J., Good, R.J., Chaundhury, M.K., (1986) J. Colloid Interface Sci., 111, pp. 378-390Schneider, R.P., (1996) J. Colloid Interface Sci., 182, pp. 204-213Bokara, K.K., Kim, J.Y., Lee, Y.I., Yun, K., Webster, T.J., Lee, J.E., (2013) Anat. Cell Biol., 46 (2), pp. 85-92Harrison, B.S., Atala, A., (2007) Biomaterials, 28 (2), pp. 344-353Vardharajula, S., Ali, S.Z., Tiwari, P.M., Eroʇlu, E., Vig, K., Dennis, V.A., Singh, S.R., (2012) Int. J. Nanomedicine, 7, pp. 5361-5374Van Hooijdonk, E., Bittencourt, C., Snyders, R., Colomer, J.F., Beilstein, (2013) J. Nanotechnol., 4, pp. 129-152Okabe, M., Ikawa, M., Kominami, K., Nakanishi, T., Nishimune, Y., (1997) FEBS Lett., 407 (3), pp. 313-319Marsi, T.C.O., Santos, T.G., Pacheco-Soares, C., Corat, E.J., Marciano, F.R., Lobo, A.O., (2012) Langmuir, 28 (9), pp. 4413-4424Ruiz, O.N., Fernando, K.A., Wang, B., Brown, N.A., Luo, P.G., McNamara, N.D., Vangsness, M., Bunker, C.E., (2011) ACS Nano, 5 (10), pp. 8100-810

In Vitro And In Vivo Studies Of A Novel Nanohydroxyapatite/superhydrophilic Vertically Aligned Carbon Nanotube Nanocomposites

An association between in vitro and in vivo studies has been demonstrated for the first time, using a novel nanohydroxyapatite/superhydrophilic vertically aligned multiwalled carbon nanotube (nHAp/VAMWCNT-O2) nanocomposites. Human osteoblast cell culture and bone defects were used to evaluate the in vitro extracellular matrix (ECM) calcification process and bone regeneration, respectively. The in vitro ECM calcification process of nHAp/VAMWCNT-O2 nanocomposites were investigated using alkaline phosphatase assay. The in vivo biomineralization studies were carried out on bone defects of C57BL/6/JUnib mice. Scanning electron microscopy, micro-energy dispersive spectroscopy, X-ray photoelectron spectroscopy, and X-ray difractometry analyses confirmed the presence of the nHAp crystals. nHAp/VAMWCNT-O2 nanocomposites induced in vitro calcification of the ECM of human osteoblast cells in culture after only 24 h. Bone regeneration with lamellar bone formation after 9 weeks was found in the in vivo studies. Our findings make these new nanocomposites very attractive for application in bone tissue regeneration. 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An evaluation of cell proliferation and adhesion on vertically-aligned multi-walled carbon nanotube films

FAPESP - FUNDAÇÃO DE AMPARO À PESQUISA DO ESTADO DE SÃO PAULOBiocompatibility tests were performed on vertically-aligned multi-walled carbon nanotube (MWCNT) films produced by microwave plasma chemical vapor deposition on titanium substrates with iron (Fe) and nickel (Ni) as catalysts. The cell adhesion and morphology of L-929 mouse fibroblast cells were studied by high resolution scanning electron microscopy, after up to 7 days incubation periods. Cell viability and proliferation were evaluated by two "in vitro" tests: (1) 2-(4,5-dimethyl-2-thiazolyl)-3,5-diphenyl-2H-tetrazolium bromide (MTT), and (2) lactate dehydrogenase (LDH) assays. Low level of bioavailable Fe and Ni was determined by inductively coupled plasma optical emission spectrometry. Neither functionalization nor purification of MWCNT films was necessary to obtain good response to the biocompatibility tests. Efficient cell growth and non-toxicity suggest the use MWCNTs in tissue regeneration. The MWCNT films stimulated the cell growth, showing a proliferation 20% higher than on Ti481245254FAPESP - FUNDAÇÃO DE AMPARO À PESQUISA DO ESTADO DE SÃO PAULOFAPESP - FUNDAÇÃO DE AMPARO À PESQUISA DO ESTADO DE SÃO PAULOFAPESP [08/116425, 07/00013-4]08/116425; 07/00013-

Biocompatibility Differences Between Dispersed And Vertically-aligned Carbon Nanotubes: An In Vitro Assays Review

An overview about carbon nanotube (CNT) production and quality parameters will be presented, as well a review of current literature about "in vitro"assays commonly used to evaluate the biocompatibility of CNT. The limits of colorimetric assays for CNTs evaluation will be discussed, using comparisons between dispersed CNT and CNT arrays. The influence of nanotopography and wettability of CNT scaffolds for cell adhesion will be shown. Studies carried out in our laboratories with vertically-aligned carbon nanotubes (VACNT) will also be presented. We have shown the interaction among CNT (VACNT) and four cell lines: mouse fibroblasts (L-929), mouse embryo fibroblast (C57/BL6) with or without green fluorescent protein (GFP) and human osteoblast (SaOS-2). The biocompatibility tests were performed with in vitro tests on raw-VACNT and after superficial modification by O2 plasma, which changes its hydrophobic character. The non-toxicity, cell viability, proliferation and cell adhesion were evaluated by: (i) 2-(4,5-dimethyl-2-thioazoly)-3,5-diphenyl-2H-tetrazolium bromide (MTT) assay; (ii) Lactate dehydrogenase (LDH) assay; (iii) neutral red (NR) assay; (iv) Scanning electron microscopy (SEM); and fluorescence microscopy. The influence of catalyst type, VACNT density and superficial modification were evaluated by morphological, structural and superficial techniques: SEM, Transmission electron microscopy (TEM), Raman spectroscopy, contact angle (CA) and X-Ray Photoelectron Spectroscopy (XPS). High cell viability, exceptional cell adhesion and preference were achieved. © 2009 by Nova Science Publishers, Inc. 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Columnar Cvd Diamond Growth Structure On Irregular Surface Substrates

Columnar grain structure is always observed in CVD-diamond growth and is an important parameter to identify the morphology of thin and thick films. Structure defects, aspects of onset nucleation and film growth mechanisms can also be related to columnar growth. In this work we focused our attention on the columnar structure of CVD-diamond grown on irregular surfaces. We observed that there is a relationship among curvature radius of the substrate surface, the spread of the column volume and the growth rate of the diamond film. Growth rates on spherical surfaces of around 0.5 mm curvature radius have been observed to be up to three times bigger than the growth rates on planar surfaces. Also, the grain size distribution on planar and on the corner surfaces as a function of the growth rate has been studied. Characterization with scanning electron microscopy (SEM) and Raman scattering spectroscopy (RSS) has been performed. © 1995.41112551259Bachmann, van Enckevort, (1992) Diamond Relat. Mater., 1, p. 1021Koidl, Klages, (1992) Diamond Relat. Mater., 1, p. 1065Buckley-Golder, Collins, (1992) Diamond Relat. Mater., 1, p. 1083Lux, Haubner, Renard, (1992) Diamond Relat. Mater., 1, p. 1035Lux, Haubner, (1991) Diamond and Diamond-like Films and Coatings, NATO-ASI Ser. B, Physics, 266, p. 579. , 5th edn., Plenum, New YorkClausing, Heatherly, Specht, More, (1991) Int. Conf. Proc. on New Diamond Science and Technology, p. 575. , MRS, Pittsburgh, PABruckner, Mantyla, (1993) Diamond Relat. Mater., 2, p. 373Jin, Graebner, Tiefl, Kammlott, (1993) Diamond Relat. Mater., 2, p. 1038Wild, Heres, Koidl, (1990) J. Appl. Phys., 68, p. 973Trava-Airoldi, Rodrigues, Fukui, Baranauskas, <title>Characterization of diamond films deposited by hot-filament CVD using CF4 as doping gas by Raman spectroscopy, FTIR spectroscopy, and atomic force microscopy</title> (1992) SPIE Proc. Diamond Optics V, 1759, p. 87. , 5th ednVan der Drift, (1967) Philips Res. Rep., 2, p. 26

Graphene And Carbon Nanotube Nanocomposite For Gene Transfection

Graphene and carbon nanotube nanocomposite (GCN) was synthesised and applied in gene transfection of pIRES plasmid conjugated with green fluorescent protein (GFP) in NIH-3T3 and NG97 cell lines. The tips of the multi-walled carbon nanotubes (MWCNTs) were exfoliated by oxygen plasma etching, which is also known to attach oxygen content groups on the MWCNT surfaces, changing their hydrophobicity. The nanocomposite was characterised by high resolution scanning electron microscopy; energy-dispersive X-ray, Fourier transform infrared and Raman spectroscopies, as well as zeta potential and particle size analyses using dynamic light scattering. BET adsorption isotherms showed the GCN to have an effective surface area of 38.5 m2/g. The GCN and pIRES plasmid conjugated with the GFP gene, forming π-stacking when dispersed in water by magnetic stirring, resulting in a helical wrap. The measured zeta potential confirmed that the plasmid was connected to the nanocomposite. The NIH-3T3 and NG97 cell lines could phagocytize this wrap. The gene transfection was characterised by fluorescent protein produced in the cells and pictured by fluorescent microscopy. Before application, we studied GCN cell viability in NIH-3T3 and NG97 line cells using both MTT and Neutral Red uptake assays. Our results suggest that GCN has moderate stability behaviour as colloid solution and has great potential as a gene carrier agent in non-viral based therapy, with low cytotoxicity and good transfection efficiency. © 2014 Elsevier B.V.391288298Vardharajula, S., Ali, S.Z., Tiwari, P.M., Eroglu, E., Vig, K., Dennis, V.A., Functionalized carbon nanotubes: Biomedical applications (2012) Int. J. 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Friction Coefficient Measurements By Lfm On Dlc Films As Function Of Sputtering Deposition Parameters

Diamond-like carbon (DLC) coatings are widely used as protective overcoats on space junctions, magnetic media and magnetic head sliders in hard disk-drive systems, etc. In the present work, the friction coefficient of DLC films was investigated by lateral force microscopy (LFM) as a function of sputtering deposition parameters. The lateral force acts on the pyramidal tip attached to the end of the cantilever, due to friction or viscous forces, resulting in cantilever measurable torsion and deflection, related to the friction's magnitude. The relationship among data from friction coefficient distribution on DLC films surface with flow of precursor gases, surface composition, morphology structure, hardness and elastic module parameters was evaluated. Characterization techniques such as Raman scattering spectroscopy (RSS), X-ray photoelectron spectroscopy (XPS) and nanoindentation were used. The results have showed good agreement with the literature for DLC tribological parameters. © 2002 Elsevier Science B.V. All rights reserved.113-611351138Cunningham, J.M., Marirrodriga, C.G., (1995) Proceedings of the Sixth European Space Mechanisms and Tribology Symposium, p. 374. , Technopark, Zurich, Switzerland, ESA SPBriscoe, H.M., (1990) Tribol. Int, 23 (2), p. 67Spalvins, T., (1973) ASLE Trans, 1, p. 17Lifshitz, Y., (1999) Diamond Relat. Mater, 8, p. 1659Santos, L.V., Trava-Airoldi, V.J., Iha, K., Corat, E.J., Salvador, M.C., (2001) Diamond Relat. Mater, 10, p. 1049Maillat, M., Hintermann, H.E., (1990) Proceedings of the Fourth European on Space Mechanisms and Tribology Symposium, p. 299. , Canes, France (20-22 Sep 1989), ESA SPGrill, A., Patel, V., (1993) Diamond Relat. Mater, 2, p. 597Mogne, T.L., Donnet, C., Martin, J.M., Tonck, A., Millard, J.N., (1994) Vacuum Sci. Technol, 12 A (4), p. 1998Liu, E., Blanpain, B., Shia, X., Celis, J.-P., Tan, H.-S., Tay, B.-K., Cheah, L.-K., Ross, J.R., (1998) Surf. Coat. Technol, 106, p. 72Donnet, C., Belin, M., Auge, J.C., Martin, J.M., Grill, A., Patel, V., (1994) Surf. Coat. Technol, 68-69, p. 626Grill, A., (1999) J. Res. Dev. IBM-plasma Process, 1-2, p. 43Erlandsson, R., Hadziioannou, G., Mate, C.M., McClelland, G.M., Chiangs, S.J., (1988) Chem. Phys, 89 (8), p. 5190Prioli, R., Reigada, D.C., Freire, F.L., (1999) Appl. Phys. Lett, 75 (9), p. 1317Miyake, S., Kaneko, R., (1992) Thin Solids Films, 212, p. 256Mar-tines, E., Andujur, J.L., Polo, M.C., Esteve, J., Robertson, J., Milne, W.I., (2001) Diamond Relat. Mater, 10, pp. 145-152Marti, O., Colchero, J., Mlynek, J., (1990) Nanotechnol, 1, p. 141Meyer, E., Overney, R., Brodbeck, D., Howald, L., Ltithi, R., Frommer, J., Gtintherodt, H.J., (1992) Phys. Rev. Lett, 69 (12), p. 1777Neumeister, J.M., Ducker, W.A., (1994) Sci. Instrum, 68 (8), p. 2527Hu, J., Xiao, X.-D., Ogletree, D.F., Salmerom, M., (1995) Surf. Sci, 344, pp. 221-23

Determination Of Gas Phase Reactive Species By Mass Spectrometry In Chlorine Assisted Hot-filament Chemical Vapor Deposition Of Diamond

In this paper we studied the CVD diamond growth from CCl4 mixtures. We observed by mass spectrometry that CCl4 completely dissociates and converts into hydrocarbons and HCl. The quartz microprobe experiments confirm dissociation and shows that the gas phase is completely reacted to its equilibrium for distances larger than 4mm from the gas inlet. The observation of no other chloro-carbon species indicates a fast and short range conversion that probably prevents the participation of chloro-carbon radicals in the growth process. The observation of other chlorinated species in the range from 2 to 4mm may explain the annular regions observed in the growth experiments with gas inlet close to the substrate. Growth experiments have been analysed by Scanning Electron Microscopy (SEM) and Raman Spectroscopy.274142145Spear, K.E., Dismuskes, J.P., (1994) Synthetic Diamond: Emerging CVD Science and Technology, , John Wiley, New YorkCorat, E.J., Trava-Airoldi, V.J., Leite, N.F., Nono, M.C.A., Baranauskas, V., (1997) J. Mater. Sci., 32, p. 941Bai, B.J., Chu, C.J., Patterson, D.E., Hauge, R.H., Margrave, J.L., (1993) J. Mater. Res., 8, p. 233Pan, C., Margrave, J.L., Hauge, R.H., (1994) Mat. Res. Soc. Symp. Proc. Vol. 349, Novel Forms of Carbon II, 349, p. 427. , Ed. C.L. Renschler, D.M. Cox, J.J. Pouch and Y. Achiba, Materials Research Society, San Francisco, CAFox, C., McMaster, M.C., Hsu, W.L., Kelly, M.A., Hagstrom, S.B., (1995) Appl. Phys. Lett., 67, p. 2379Mendes De Barros, R.C., Corat, E.J., Trava-Airoldi, V.J., Ferreira, N.G., Leite, N.F., Iha, K., (1997) Diamond Relat. Mater., 6, p. 49