Poly(L-lactide) nanofibrous meshes loaded with detonation diamond particles as substrates for the adhesion, growth and osteogenic differentiation of human osteoblast-like cells
At a Glance
Section titled āAt a Glanceā| Metadata | Details |
|---|---|
| Publication Date | 2016-01-01 |
| Journal | Frontiers in Bioengineering and Biotechnology |
| Authors | Bacakova Lucie, Lubica StaÅkovĆ”, Jana MusıĢlkovĆ”, Alexander Kromka, Å tÄpĆ”n PotockĆ½ |
Abstract
Section titled āAbstractāEvent Abstract Back to Event Poly(L-lactide) nanofibrous meshes loaded with detonation diamond particles as substrates for the adhesion, growth and osteogenic differentiation of human osteoblast-like cells Lucie Bacakova1*, Lubica Stankova1*, Jana Musilkova1*, Alexander Kromka2*, Stepan Potocky2* and Denisa Stranska3 1 Institute of Physiology, Academy of Sciences of the Czech Republic, Dept. of Biomaterials and Tissue Engineering, Czechia 2 Institute of Physics, Academy of Sciences of the Czech Republic, Czechia 3 InStar Technologies, Joint-Stock Co., Czechia Introduction: Diamond nanoparticles (DNPs) are considered as a highly biocompatible material for biomedical applications. In our earlier studies, nanocrystalline diamond films proved as excellent substrates for the adhesion, growth and osteogenic differentiation of human-bone derived cells, particularly after their doping with boron[1] or termination with oxygen[2]. In addition, diamond nanoparticles could be added into nanofibrous polymeric scaffolds in order to improve their mechanical properties and bioactivity for bone tissue engineering. Thus, composite poly(L-lactide) (PLLA) nanofibrous membranes with six different concentrations of DNPs were prepared, and the adhesion, growth and osteogenic differentiation of human osteoblast-like MG-63 and Saos-2 cells were studied on these matrices. Materials and Methods: Five grams of PLLA were dissolved in 100 ml of chloroform, and DNPs (NanoAmando, Nanocarbon Research Institute Co., Ltd., Japan) were added in 6 concentrations ranging from 0.021875 to 0.7 g/100 ml of PLLA solution, i.e. from 0.38 to 12.28 wt.% in dry PLLA. Nanofibrous membranes were then prepared by needle-less electrospinning. The cell growth was estimated using XTT test, measuring the activity of mitochondrial enzymes. The concentrations of focal adhesion proteins talin and vinculin, and also of osteogenic proteins collagen I, alkaline phosphatase, osteopontin and osteocalcin, were measured in cell homogenates by an enzyme linked-immunosorbent assay (ELISA) on day 7 after seeding. The expression of mRNA for these proteins was measured by real-time PCR on day 14 after seeding. Results and Discussion: The activity of mitochondrial enzymes decreased with increasing nanoparticle concentration in both MG-63 and Saos-2 cells (Fig. 1). The concentration of focal adhesion and osteogenic proteins, measured on day 7 at the protein level, was similar in cells on pure PLLA scaffolds and scaffolds with all concentrations with DNPs, except of osteocalcin in MG-63 cells, which decreased with increasing nanoparticle concentration. On day 14 after seeding, the mRNA expression of talin and vinculin, as well as of collagen I, alkaline phosphatase and osteopontin showed a general tendency to decrease with increasing DNP concentration (Fig. 2). On the other hand, the addition of DNPs to PLGA nanofibers in our earlier studies supported the growth of MG-63 cells[3] and human bone marrow mesenchymal stem cells[4], even in a higher concentration (23 wt.%). This disproportion could be explained by different origin and physicochemical properties of DNPs used for PLLA (detonation diamond) and for PLGA (DNPs prepared by a RF-PACVD method[5]). The DNPs used for PLLA scaffolds were hydrogen-terminated and hydrophobic, which markedly increased the hydrophobicity of the PLLA-DNP scaffolds, making them less appropriate for the colonization with cells. A direct cytotoxicity of DNPs cannot also be excluded. DNPs can damage cells by oxidative mechanisms and by an increased influx of Na+ ions into cells (for a review, see[4]). Conclusion: Addition of diamond nanoparticles produced by detonation and terminated with H into nanofibrous scaffolds had negative effects on the adhesion, growth and osteogenic differentiation of human bone-derived cells, while the effects of diamond nanoparticles produced by RF-PACVD method were beneficial. Thus, the mode of preparation and surface properties of diamond nanoparticles could be important for their biocompatibility and for their use as components of biomaterials. Supported by the Grant Agency of the Czech Republic (grants No. P108/12/1168 and 14-04790S) and the Ministry of Health of the Czech Republic (grant No. 15-32497A).References:[1] Grausova L et al.: PLoS One 6(6):e20943, 2011[2] Liskova J et al.: Int J Nanomedicine 10:869-84, 2015[3] Parizek M et al.: Int J Nanomedicine 7:1931-51, 2012[4] Brady MA et al.: J Nanosci Nanotechnol 15:1060-9, 2015[5] Mitura S et al.: J AMME 16:9-16, 2006 Keywords: Bone Regeneration, Cell Differentiation, Cell Proliferation, Bioactivity Conference: 10th World Biomaterials Congress, MontrĆ©al, Canada, 17 May - 22 May, 2016. Presentation Type: Poster Topic: Biomaterials in constructing tissue substitutes Citation: Bacakova L, Stankova L, Musilkova J, Kromka A, Potocky S and Stranska D (2016). Poly(L-lactide) nanofibrous meshes loaded with detonation diamond particles as substrates for the adhesion, growth and osteogenic differentiation of human osteoblast-like cells. Front. Bioeng. Biotechnol. Conference Abstract: 10th World Biomaterials Congress. doi: 10.3389/conf.FBIOE.2016.01.02086 Copyright: The abstracts in this collection have not been subject to any Frontiers peer review or checks, and are not endorsed by Frontiers. They are made available through the Frontiers publishing platform as a service to conference organizers and presenters. The copyright in the individual abstracts is owned by the author of each abstract or his/her employer unless otherwise stated. Each abstract, as well as the collection of abstracts, are published under a Creative Commons CC-BY 4.0 (attribution) licence (https://creativecommons.org/licenses/by/4.0/) and may thus be reproduced, translated, adapted and be the subject of derivative works provided the authors and Frontiers are attributed. For Frontiersā terms and conditions please see https://www.frontiersin.org/legal/terms-and-conditions. Received: 27 Mar 2016; Published Online: 30 Mar 2016. * Correspondence: Dr. Lucie Bacakova, Institute of Physiology, Academy of Sciences of the Czech Republic, Dept. of Biomaterials and Tissue Engineering, Prague, Czechia, Email1 Dr. Lubica Stankova, Institute of Physiology, Academy of Sciences of the Czech Republic, Dept. of Biomaterials and Tissue Engineering, Prague, Czechia, [email protected] Dr. Jana Musilkova, Institute of Physiology, Academy of Sciences of the Czech Republic, Dept. of Biomaterials and Tissue Engineering, Prague, Czechia, [email protected] Dr. Alexander Kromka, Institute of Physics, Academy of Sciences of the Czech Republic, Prague, Czechia, [email protected] Dr. Stepan Potocky, Institute of Physics, Academy of Sciences of the Czech Republic, Prague, Czechia, [email protected] Login Required This action requires you to be registered with Frontiers and logged in. To register or login click here. Abstract Info Abstract The Authors in Frontiers Lucie Bacakova Lubica Stankova Jana Musilkova Alexander Kromka Stepan Potocky Denisa Stranska Google Lucie Bacakova Lubica Stankova Jana Musilkova Alexander Kromka Stepan Potocky Denisa Stranska Google Scholar Lucie Bacakova Lubica Stankova Jana Musilkova Alexander Kromka Stepan Potocky Denisa Stranska PubMed Lucie Bacakova Lubica Stankova Jana Musilkova Alexander Kromka Stepan Potocky Denisa Stranska Related Article in Frontiers Google Scholar PubMed Abstract Close Back to top Javascript is disabled. Please enable Javascript in your browser settings in order to see all the content on this page.