BACKGROUND: Nanofibrous scaffolds loaded with bioactive nanoparticles are promising materials for bone tissue engineering. METHODS: In this study, composite nanofibrous membranes containing a copolymer of L-lactide and glycolide (PLGA) and diamond nanoparticles were fabricated by an electrospinning technique. PLGA was dissolved in a mixture of methylene chloride and dimethyl formamide (2:3) at a concentration of 2.3 wt%, and nanodiamond (ND) powder was added at a concentration of 0.7 wt% (about 23 wt% in dry PLGA). RESULTS: In the composite scaffolds, the ND particles were either arranged like beads in the central part of the fibers or formed clusters protruding from the fibers. In the PLGA-ND membranes, the fibers were thicker (diameter 270 ± 9 nm) than in pure PLGA meshes (diameter 218 ± 4 nm), but the areas of pores among these fibers were smaller than in pure PLGA samples (0.46 ± 0.02 μm(2) versus 1.28 ± 0.09 μm(2) in pure PLGA samples). The PLGA-ND membranes showed higher mechanical resistance, as demonstrated by rupture tests of load and deflection of rupture probe at failure. Both types of membranes enabled the attachment, spreading, and subsequent proliferation of human osteoblast-like MG-63 cells to a similar extent, although these values were usually lower than on polystyrene dishes. Nevertheless, the cells on both types of membranes were polygonal or spindle-like in shape, and were distributed homogeneously on the samples. From days 1-7 after seeding, their number rose continuously, and at the end of the experiment, these cells were able to create a confluent layer. At the same time, the cell viability, evaluated by a LIVE/DEAD viability/cytotoxicity kit, ranged from 92% to 97% on both types of membranes. In addition, on PLGA-ND membranes, the cells formed well developed talin-containing focal adhesion plaques. As estimated by the determination of tumor necrosis factor-alpha levels in the culture medium and concentration of intercellular adhesion molecule-1, MG-63 cells, and RAW 264.7 macrophages on these membranes did not show considerable inflammatory activity. CONCLUSION: This study shows that nanofibrous PLGA membranes loaded with diamond nanoparticles have interesting potential for use in bone tissue engineering.

Department of Biomaterials and Tissue Engineering Institute of Physiology Academy of Sciences of the Czech Republic Prague Czech Republic

Erratum v

Int J Nanomedicine. 2012;7:5873 PubMed   Warnke, Patrick H [added]

Zobrazit více v PubMed

Webster TJ, Ergun C, Doremus RH, Siegel RW, Bizios R. Specific proteins mediate enhanced osteoblast adhesion on nanophase ceramics. J Biomed Mater Res. 2000;51:475–483. PubMed

Price RL, Waid MC, Haberstroh KM, Webster TJ. Selective bone cell adhesion on formulations containing carbon nanofibers. Biomaterials. 2003;24:1877–1887. PubMed

Boland ED, Matthews JA, Pawlowski KJ, Simpson DG, Wnek GE, Bowlin GL. Electrospinning collagen and elastin: preliminary vascular tissue engineering. Front Biosci. 2004;9:1422–1432. PubMed

Meinel AJ, Kubow KE, Klotzsch E, et al. Optimization strategies for electrospun silk fibroin tissue engineering scaffolds. Biomaterials. 2009;30:3058–3067. PubMed PMC

Park YJ, Kim KH, Lee JY, et al. Immobilization of bone morphogenetic protein-2 on a nanofibrous chitosan membrane for enhanced guided bone regeneration. Biotechnol Appl Biochem. 2006;43:17–24. PubMed

Sargeant TD, Rao MS, Koh CY, Stupp SI. Covalent functionalization of NiTi surfaces with bioactive peptide amphiphile nanofibers. Biomaterials. 2008;29:1085–1098. PubMed PMC

Stylianopoulos T, Bashur CA, Goldstein AS, Guelcher SA, Barocas VH. Computational predictions of the tensile properties of electrospun fiber meshes: effect of fiber diameter and fiber orientation. J Mech Behav Biomed Mater. 2008;1:326–335. PubMed PMC

Pektok E, Nottelet B, Tille JC, et al. Degradation and healing characteristics of small-diameter poly(epsilon-caprolactone) vascular grafts in the rat systemic arterial circulation. Circulation. 2008;118:2563–2570. PubMed

Jeong SI, Ko EK, Yum J, Jung CH, Lee YM, Shin H. Nanofibrous poly(lactic acid)/hydroxyapatite composite scaffolds for guided tissue regeneration. Macromol Biosci. 2008;8:328–338. PubMed

Bashur CA, Dahlgren LA, Goldstein AS. Effect of fiber diameter and orientation on fibroblast morphology and proliferation on electrospun poly(D,L-lactic-co-glycolic acid) meshes. Biomaterials. 2006;27:5681–5688. PubMed

Kumbar SG, Nukavarapu SP, James R, Nair LS, Laurencin CT. Electrospun poly(lactic acid-co-glycolic acid) scaffolds for skin tissue engineering. Biomaterials. 2008;29:4100–4107. PubMed PMC

Nie H, Soh BW, Fu YC, Wang CH. Three-dimensional fibrous PLGA/ HAp composite scaffold for BMP-2 delivery. Biotechnol Bioeng. 2008;99:223–34. PubMed

Schneider OD, Loher S, Brunner TJ, et al. Cotton wool-like nanocomposite biomaterials prepared by electrospinning: in vitro bioactivity and osteogenic differentiation of human mesenchymal stem cells. J Biomed Mater Res B Appl Biomater. 2008;84:350–362. PubMed

Jose MV, Thomas V, Johnson KT, Dean DR, Nyairo E. Aligned PLGA/ HA nanofibrous nanocomposite scaffolds for bone tissue engineering. Acta Biomater. 2009;5:305–315. PubMed

Lee JH, Rim NG, Jung HS, Shin H. Control of osteogenic differentiation and mineralization of human mesenchymal stem cells on composite nanofibers containing poly[lactic-co-(glycolic acid)] and hydroxyapatite. Macromol Biosci. 2010;10:173–182. PubMed

Pamula E, Filova E, Bacakova L, Lisa V, Adamczyk D. Resorbable polymeric scaffolds for bone tissue engineering: the influence of their microstructure on the growth of human osteoblast-like MG 63 cells. J Biomed Mater Res A. 2009;89:432–443. PubMed

Huang Z-M, Zhang Y-Z, Kotaki M, Ramakrishna S. A review on polymer nanofibers by electrospinning and their applications in nanocomposites. Compos Sci Technol. 2003;63:2223–2253.

Vasita R, Katti DS. Nanofibers and their applications in tissue engineering. Int J Nanomedicine. 2006;1:15–30. PubMed PMC

Tan J, Saltzman WM. Biomaterials with hierarchically defined micro-and nanoscale structure. Biomaterials. 2004;25:3593–3601. PubMed

Gizdavic-Nikolaidis M, Ray S, Bennett JR, Easteal AJ, Cooney RP. Electrospun functionalized polyaniline copolymer-based nanofibers with potential application in tissue engineering. Macromol Biosci. 2010;10:1424–1431. PubMed

Gizdavic-Nikolaidis MR, Bennett JR, Swift S, Ray S, Bowmaker G. Electrospun poly(aniline-co-ethyl 3-aminobenzoate)/poly(lactic acid) nanofibers and their potential in biomedical application. J Polym Sci A Polym Chem. 2011;49:4902–4910.

Kim HW, Lee HH, Chun GS. Bioactivity and osteoblast responses of novel biomedical nanocomposites of bioactive glass nanofiber filled poly(lactic acid) J Biomed Mater Res A. 2008;85:651–663. PubMed

McCullen SD, Stevens DR, Roberts WA, et al. Characterization of electrospun nanocomposite scaffolds and biocompatibility with adipose-derived human mesenchymal stem cells. Int J Nanomedicine. 2007;2:253–263. PubMed PMC

Sargent LM, Reynolds SH, Castranova V. Potential pulmonary effects of engineered carbon nanotubes: in vitro genotoxic effects. Nanotoxicology. 2010;4:396–408. PubMed

Wang L, Luanpitpong S, Castranova V, et al. Carbon nanotubes induce malignant transformation and tumorigenesis of human lung epithelial cells. Nano Lett. 2011;11:2796–2803. PubMed PMC

Schrand AM, Huang H, Carlson C, et al. Are diamond nanoparticles cytotoxic? J Phys Chem B. 2007;111:2–7. PubMed

Amaral M, Dias AG, Gomes PS, et al. Nanocrystalline diamond: in vitro biocompatibility assessment by MG 63 and human bone marrow cells cultures. J Biomater Res A. 2008;87:91–99. PubMed

Grausova L, Kromka A, Burdikova Z, et al. Enhanced growth and osteogenic differentiation of human osteoblast-like cells on boron-doped nanocrystalline diamond thin films. PLoS One. 2011;6:e20943. PubMed PMC

Liu KK, Wang CC, Cheng CL, Chao JI. Endocytic carboxylated nanodiamond for the labeling and tracking of cell division and differentiation in cancer and stem cells. Biomaterials. 2009;30:4249–4259. PubMed

McDevitt MR, Chattopadhyay D, Jaggi JS, et al. PET imaging of soluble yttrium-86-labeled carbon nanotubes in mice. PLoS One. 2007;2:e907. PubMed PMC

Ruggiero A, Villa CH, Bander E, et al. Paradoxical glomerular filtration of carbon nanotubes. Proc Natl Acad Sci U S A. 2010;107:12369–12374. PubMed PMC

Behler KD, Stravato A, Mochalin V, Korneva G, Yushin G, Gogotsi Y. Nanodiamond-polymer composite fibers and coatings. ACS Nano. 2009;3:363–369. PubMed

Zhang Q, Mochalin VN, Neitzel I, et al. Fluorescent PLLA-nanodiamond composites for bone tissue engineering. Biomaterials. 2011;32:87–94. PubMed

Dos Santos AM, Dierck J, Troch M, Podevijn M, Schacht E. Production of continuous electrospun mats with improved mechanical properties. Macromol Mater Eng. 2011;296:637–644.

Huang L, Nagapudi K, Apkarian RP, Chaikof EL. Engineered collagen-PEO nanofibers and fabrics. J Biomater Sci Polym Ed. 2001;12:979–993. PubMed

Jo JH, Lee EJ, Shin DS, et al. In vitro/in vivo biocompatibility and mechanical properties of bioactive glass nanofiber and poly(epsilon-caprolactone) composite materials. J Biomed Mater Res B Appl Biomater. 2009;91:213–220. PubMed

Jang TS, Lee EJ, Jo JH, et al. Fibrous membrane of nano-hybrid poly-L-lactic acid/silica xerogel for guided bone regeneration. J Biomed Mater Res B Appl Biomater. 2011 Nov; [Epub ahead of print.] PubMed

Mitura S, Mitura K, Niedzielski P, Louda P, Danilenko V. Nanocrystalline diamond, its synthesis, properties and applications. J AMME. 2006;16:9–16.

Kromka A, Rezek B, Remes Z, et al. Formation of continuous nano-crystalline diamond layer on glass and silicon at low temperatures. Chem Vapor Depos. 2008;14:181–186.

Kromka A, Rezek B, Kalbacova M, et al. Diamond seeding and growth of hierarchically structured films for tissue engineering. Adv Eng Mater. 2009;11:B71–76.

Kromka A, Babchenko O, Kozak H, Rezek B, Vanecek M. Role of polymers in CVD growth of nanocrystalline diamond films on foreign substrates. Phys Stat Sol B. 2009;246:2654–2657.

Janicki P, Boeuf S, Steck E, Egermann M, Kasten P, Richter W. Prediction of in vivo bone forming potency of bone marrow-derived human mesenchymal stem cells. Eur Cell Mater. 2011;21:488–507. PubMed

Maheshwari G, Brown G, Lauffenburger DA, Wells A, Griffith LG. Cell adhesion and motility depend on nanoscale RGD clustering. J Cell Sci. 2000;113:1677–1686. PubMed

Yim ES, Zhao B, Myung D, et al. Biocompatibility of poly(ethylene glycol)/poly(acrylic acid) interpenetrating polymer network hydrogel particles in RAW 264.7 macrophage and MG-63 osteoblast cell lines. J Biomed Mater Res A. 2009;91:894–902. PubMed

Delpino MV, Fossati CA, Baldi PC. Proinflammatory response of human osteoblastic cell lines and osteoblast-monocyte interaction upon infection with Brucella spp. Infect Immun. 2009;77:984–895. PubMed PMC

Zhao Y-Q, Lau K-T, Kim J-K, Xu C-L, Zhao D-D, Li HL. Nanodiamond/ poly (lactic acid) nanocomposites: Effect of nanodiamond on structure and properties of poly (lactic acid) Composites: Part B. 2010;41:646–653.

Jose MV, Thomas V, Dean DR, Nyairo E. Fabrication and characterization of aligned nanofibrous PLGA/collagen blends as bone tissue scaffolds. Polymer. 2009;50:3778–3785.

Deitzel JM, Kleinmeyer J, Harris D, Beck Tan NC. A review on polymer nanofibers by electrospinning and their applications in nanocomposites. J Nanosci Nanotechnol. 2009;9:3535–3545. PubMed

Henriques C, Vidinha R, Botequim D, Borges JP, Silva JA. A systematic study of solution and processing parameters on nanofiber morphology using a new electrospinning apparatus. J Nanosci Nanotechnol. 2009;9:3535–3545. PubMed

Liang D, Luu YK, Kim K, Hsiao BS, Hadjiargyrou M, Chu B. In vitro non-viral gene delivery with nanofibrous scaffolds. Nucleic Acids Res. 2005;33:e170. PubMed PMC

Beachley V, Wen X. Effect of electrospinning parameters on the nanofiber diameter and length. Mater Sci Eng C Mater Biol Appl. 2009;29:663–668. PubMed PMC

Bisht GS, Canton G, Mirsepassi A, et al. Controlled continuous patterning of polymeric nanofibers on three-dimensional substrates using low-voltage near-field electrospinning. Nano Lett. 2011;11:1831–1837. PubMed

Hu J, Liu X, Ma PX. Induction of osteoblast differentiation phenotype on poly(L-lactic acid) nanofibrous matrix. Biomaterials. 2008;29:3815–3821. PubMed PMC

Engler A, Bacakova L, Newman C, Hategan A, Griffin M, Discher D. Substrate compliance versus ligand density in cell on gel responses. Biophys J. 2004;86:617–628. PubMed PMC

Prasad KE, Das B, Maitra U, Ramamurty U, Rao CN. Extraordinary synergy in the mechanical properties of polymer matrix composites reinforced with 2 nanocarbons. Proc Natl Acad Sci U S A. 2009;106:13186–13189. PubMed PMC

Bacakova L, Stary V, Kofronova O, Lisa V. Polishing and coating carbon fiber-reinforced carbon composites with a carbon-titanium layer enhances adhesion and growth of osteoblast-like MG63 cells and vascular smooth muscle cells in vitro. J Biomed Mater Res. 2001;54:567–578. PubMed

Huang S, Chen CS, Ingber DE. Control of cyclin D1, p27(Kip1), and cell cycle progression in human capillary endothelial cells by cell shape and cytoskeletal tension. Mol Biol Cell. 1998;9:3179–3193. PubMed PMC

Puzyr AP, Neshumaev DA, Tarskikh SV, Makarskaia GV, Dolmatov VI, Bondar VS. Destruction of human blood cells upon interaction with detonation nanodiamonds in experiments in vitro. Biofizika. 2005;50:101–106. Russian. PubMed

Karpukhin AV, Avkhacheva NV, Yakovlev RY, et al. Effect of detonation nanodiamonds on phagocyte activity. Cell Biol Int. 2011;35:727–733. PubMed

Krueger A. New carbon materials: biological applications of functionalized nanodiamond materials. Chemistry. 2008;14:1382–1390. PubMed

Fucikova A, Valenta J, Pelant I, Brezina V. Novel use of silicon nanocrystals and nanodiamonds in biology. Chem Pap. 2009;63:704–708.

Bacakova L, Mares V, Lisa V, Bottone MG, Pellicciari C, Kocourek F. Sex-related differences in the migration and proliferation of rat aortic smooth muscle cells in short and long term culture. In Vitro Cell Dev Biol. 1997;33:410–413. PubMed

Huang TH, Lu YC, Kao CT. Low-level diode laser therapy reduces lipopolysaccharide (LPS)-induced bone cell inflammation. Lasers Med Sci. 2011 Oct 16; [Epub ahead of print.] PubMed

Takaoka Y, Matsuura S, Boda K, Nagai H. The effect of mesoporphyrin on the production of cytokines by inflammatory cells in vitro. Jpn J Pharmacol. 1999;80:33–40. PubMed

Kurokouchi K, Kambe F, Kikumori T, et al. Effects of glucocorticoids on tumor necrosis factor alpha-dependent activation of nuclear factor kappaB and expression of the intercellular adhesion molecule 1 gene in osteoblast-like ROS17/2.8 cells. J Bone Miner Res. 2000;15:1707–1715. PubMed

Shi Q, Benderdour M, Lavigne P, Ranger P, Fernandes JC. Evidence for two distinct pathways in TNFalpha-induced membrane and soluble forms of ICAM-1 in human osteoblast-like cells isolated from osteoarthritic patients. Osteoarthritis Cartilage. 2007;15:300–308. PubMed

Tanaka Y, Maruo A, Fujii K, et al. Intercellular adhesion molecule 1 discriminates functionally different populations of human osteoblasts: characteristic involvement of cell cycle regulators. J Bone Miner Res. 2000;15:1912–1923. PubMed

Whitehead GS, Grasman KA, Kimmel EC. Lung function and airway inflammation in rats following exposure to combustion products of carbon-graphite/epoxy composite material: comparison to a rodent model of acute lung injury. Toxicology. 2003;183:175–197. PubMed

Li Q, Xia YY, Tang JC, Wang RY, Bei CY, Zeng Y. In vitro and in vivo biocompatibility investigation of diamond-like carbon coated nickel-titanium shape memory alloy. Artif Cells Blood Substit Immobil Biotechnol. 2011;39:137–142. PubMed

Dimitrievska S, Petit A, Ajji A, Bureau MN, Yahia L. Biocompatibility of novel polymer-apatite nanocomposite fibers. J Biomed Mater Res A. 2008;84:44–53. PubMed

Influence of Biomimetically Mineralized Collagen Scaffolds on Bone Cell Proliferation and Immune Activation

Morphology of a fibrin nanocoating influences dermal fibroblast behavior

Enhanced Mitogenic Activity of Recombinant Human Vascular Endothelial Growth Factor VEGF121 Expressed in E. coli Origami B (DE3) with Molecular Chaperones

Interaction of human osteoblast-like Saos-2 and MG-63 cells with thermally oxidized surfaces of a titanium-niobium alloy