Cell behavior depends strongly on the physical and chemical properties of the material surface, for example, its chemistry and topography. The authors have therefore assessed the influence of materials of different chemical composition (i.e., glass substrates with and without TiO(2) films in anatase form) and different surface roughness (R(a) = 0, 40, 100, or 170 nm) on the adhesion, proliferation, and osteogenic differentiation of human osteoblast-like MG63 cells. On day 1 after seeding, the largest cell spreading area was found on flat TiO(2) films (R(a) = 0 nm). On TiO(2) films with R(a) = 170 nm, the cell spreading area was larger and the number of initially adhering cells was higher than the values on the corresponding uncoated glass. On day 3 after seeding, the cell number was higher on the TiO(2) films (R(a) = 0 and 40 nm) than on the corresponding glass substrates and the standard polystyrene dishes. On day 7, all TiO(2) films contained higher cell numbers than the corresponding glass substrates, and the cells on the TiO(2) films with R(a) = 40 and 100 nm also contained a higher concentration of β-actin. These results indicate that TiO(2) coating had a positive influence on the adhesion and subsequent proliferation of MG63 cells. In addition, on all investigated materials, the cell population density achieved on day 7 decreased with increasing surface roughness. The concentration of osteocalcin, measured per mg of protein, was significantly lower in the cells on rougher TiO(2) films (R(a) = 100 and 170 nm) than in the cells on the polystyrene dishes. Thus, it can be concluded that the adhesion, growth, and phenotypic maturation of MG63 cells were controlled by the interplay between the material chemistry and surface topography, and were usually better on smoother and TiO(2)-coated surfaces than on rougher and uncoated glass substrates.
- MeSH
- Cell Adhesion MeSH
- Cell Division MeSH
- Cell Line MeSH
- Enzyme-Linked Immunosorbent Assay MeSH
- Immunohistochemistry MeSH
- Humans MeSH
- Nanotechnology MeSH
- Osteoblasts cytology MeSH
- Surface Properties MeSH
- Titanium chemistry MeSH
- Check Tag
- Humans MeSH
- Publication type
- Journal Article MeSH
- Research Support, Non-U.S. Gov't MeSH
- Publication type
- Meeting Abstract MeSH
Úvod a cíl: Oxidické vrstvy na povrchu titanové slitiny mají vliv na odolnost proti korozi a biokompatibilitu. Vynikající kompatibilita mezi tkání, kostí a slitinou titanu je převážně řízena vlastnostmi jeho stabilní povrchové vrstvy složené z oxidu titaničitého. Oxidové vrstvy mohou být připraveny mnoha různými metodami; oxidační proces a jeho podmínky vedou k různým vrstvám: rozdíly jsou v chemickém složení, mechanických vlastnostech, struktuře atd. Odchylka ve struktuře vrstvy může ovlivnit stabilitu, přilnavost nebo biokompatibilitu vrstvy. Anodická oxidace titanových slitin ve vhodném médiu (a za určitého napětí a proudové hustoty) může vytvořit nejen oxidickou vrstvu, ale také strukturu na povrchu oxidické vrstvy. Tato struktura je obvykle charakterizována póry o velikosti od desítek do stovek nanometrů. Strukturovaný povrch radikálně mění interakci mezi povrchem titanu a buňkami, a tím i chování tohoto materiálu uvnitř těla. Chování buněk na strukturovaném povrchu různých slitin titanu není dosud řádně popsáno. Tato experimentální práce povede k lepšímu pochopení těchto strukturovaných oxidových vrstev. Metody: Anodická oxidace byla provedena na vyleštěných vzorcích z Ti6Al4V ELI. Oxidační proces probíhal v elektrolytu 1M H2SO4 s napětím kolem 100 V a proudovou hustotou 50 mA/cm2. Výsledná oxidická vrstva byla hodnocena a dokumentována pomocí řádkovací elektronové mikroskopie (SEM), kterou byla kontrolována tloušťka oxidické vrstvy a povrchová morfologie. Pozorována byla také změna zbarvení a drsnosti povrchové vrstvy po oxidaci související s růstem oxidů. Cytokompatibilita povrchu materiálu je vyjádřena stanovením plochy povrchu, kterou obsadí buňky po třídenní kultivaci. Jedná se o metodu, která je běžně užívána a akreditována Českým institutem pro akreditaci. K pokusu byly užity buňky MG63 a bylo stanoveno procento buňkami kolonizované plochy povrchu. Hodnocení bylo provedeno na leštěných a anodizovaných površích vzorků z Ti6Al4V ELI. Výsledky byly navzájem porovnány. Výsledky: Byly připraveny vzorky s povrchem nanostrukturovaným pomocí anodické oxidace, přičemž struktura povrchu byla tvořena póry o velikostech v řádech desítek až stovek nanometrů. Výsledky pokusů ukázaly větší vůli buněk kolonizovat anodizovaný povrch. Neanodizovaný povrch byl kolonizován v 56,9 %, kdežto anodizovaný byl při stejných podmínkách kolonizován v 63,5 %. Všechny výběrové soubory byly gaussovsky distribuovány. Závěr: Anodickou oxidací byla připravena nanostrukturovaná oxidická vrstva na vzorcích z Ti6Al4V ELI. Cytokompatibilita vytvořené vrstvy byla porovnávána vůči neoxidovaným vzorkům. Bylo ukázáno, že buňky kolonizují větší plochu povrchu vzorku v případě oxidovaných vzorků.
Introduction, aim: The oxide layers on surface of titanium alloy are infl uencing corrosion resistance and biocompatibility. The compatibility between the bony tissue and titanium alloy is prevalently dependent on properties of a stable titanium dioxide layer. These layers can be prepared by various methods. The oxidation process (and its conditions) is resulting in diff erent types of oxide layer: diff erence in chemical composition, mechanical properties, inner structure etc. The deviation inside of the layers structure may infl uence the stability of the layer, its adhesion or biocompatibility. The anodic oxidation of titanium alloys in appropriate electrolyte (under certain conditions) can lead not only to creation of the oxide layer, but to creation of an oxide layer with structured surface. This kind of structure is usually characterized by pores in nanometer scale. The structured surface radically changes the interaction between the titanium alloys surface and cells; and thus influencing its behavior inside a body. The cell interaction with the structured surfaces is not properly described yet. This work aims for better understanding of such structured layers. Methods: The anodic oxidation was carried out on Ti6Al4V ELI polished samples. The oxidation process was realized in 1M H2SO4 electrolyte with the voltage 100 V and current density 50 mA/cm2. The thickness and surface morphology of the resulting oxide layer were evaluated and documented using a scanning electron microscope (SEM). The changes of color and roughness of the surface after the oxidation were observed as well. The cytocompatibility of the materials surface is expressed by a surface area colonized by cells after the three days of cultivation. This method is standardly used and accredited by ČIA. The MG63 cells were used for the experiment and the percentage of colonized surface area was evaluated. The evaluation was done on polished and oxidized Ti6Al4V ELI samples and the results were compared. Result: The structure of samples prepared using the anodic oxidation consisted of pores with size ranging from tens to hundreds of nanometers. The cytocompatibility testings showed that the cells colonized larger area on the oxidized samples. The cells covered 56.9% of the surface area of the polished samples, while 63.5% of the surface area of the anodized samples. Results of all samples exhibited Gaussian distribution. Conclusion: The anodic oxidation lead to a creation of nanostructured oxide layer on Ti6Al4V ELI samples. The cytocompatibility of this layer was compared to polished samples. It was shown that cells are colonizing the larger surface area on the oxidized samples.
- Keywords
- oxidická vrstva, cytokompatibilita, MG63, anodická oxidace,
- MeSH
- Biocompatible Materials * MeSH
- Cells MeSH
- Humans MeSH
- Titanium MeSH
- Research MeSH
- Check Tag
- Humans MeSH
Metals and their alloys show a variety of applications in health care and biotechnologies. The ability of metal alloys to be used in practice is significantly affected by interaction with a cell line in a given environment. Biological tests in vitro show the applicability of metal alloys on the implant surface. This work demonstrated the ability of the cell line MG63 to colonize the sample surface of different compositions of the material.
- Keywords
- in vitro,
- MeSH
- Biocompatible Materials MeSH
- Biomedical and Dental Materials MeSH
- Biotechnology * MeSH
- Cell Line * physiology MeSH
- Implants, Experimental MeSH
- Extracellular Matrix physiology MeSH
- Culture Techniques MeSH
- Metals, Light * MeSH
- Microscopy MeSH
- Publication type
- Research Support, Non-U.S. Gov't MeSH
In this study, we investigate the preparation of surface pattern of functional groups on poly(lactide) (PLA) surfaces through the controlled deposition of core-shell self-assemblies based on functionalized PLA-b-PEO amphiphilic block copolymers from selective solvents. Through grafting RGDS peptide onto the functionalized copolymer surface, the presented approach enables to prepare PLA surfaces with random and clustered spatial distribution of adhesive motifs. The proposed topography of the adhesion motif was proved by atomic force microscopy techniques using biotin-tagged RGDS peptide grafted on the surface and streptavidin-modified gold nanospheres which bind the tagged RGDS peptides as a contrast agent. The cell culture study under static and dynamic conditions with MG63 osteosarcoma cell line showed that the clustered distribution of RGDS peptides provided more efficient initial cell attachment and spreading, and resistance to cell detachment under dynamic culture compared to randomly distributed RGDS motif when with the same average RGDS peptide concentration.
- MeSH
- Biomimetics MeSH
- Cell Adhesion drug effects MeSH
- Metal Nanoparticles MeSH
- Lactates chemistry MeSH
- Humans MeSH
- Microscopy, Atomic Force MeSH
- Cell Line, Tumor MeSH
- Nanostructures chemistry MeSH
- Oligopeptides MeSH
- Polyethylene Glycols chemistry MeSH
- Surface Properties MeSH
- Streptavidin chemistry MeSH
- Protein Binding MeSH
- Gold MeSH
- Check Tag
- Humans MeSH
- Publication type
- Journal Article MeSH
- Research Support, Non-U.S. Gov't MeSH
NF-κB is activated in a variety of human cancers. However, its role in osteosarcoma (OS) remains unknown. Here, we have elucidated the implication of NF-κB in the oncogenic phenotype of OS tumor cells. We reported that activation of NF-κB was a common event in the human OS. Inhibition of NF-κB using inhibitor Bay 11-7085 repressed proliferation, survival, migration, and invasion but increased apoptosis in 143B and MG63 OS cells, indicating that NF-κB is critically implicated in the oncogenesis of OS. Notably, Bay 11-7085 not only inactivated NF-κB but also reduced the phosphorylation of AKT via its induction of PTEN, suggesting the existence of a novel NF-κB/PTEN/PI3K/AKT axis. In vivo, Bay 11-7085 suppressed tumor growth in the bone by targeting NF-κB and AKT. Interestingly, combined treatment with Bay 11-7085 and the PI3K inhibitor, LY294002, triggered an augmented antitumor effect. Our results demonstrate that NF-κB potentiates the growth and aggressiveness of OS. Pharmacological inhibition of NF-κB represents a promising therapy for the treatment of OS.
- Keywords
- Bay 11-7085 Sigma-Aldrich,
- MeSH
- Molecular Targeted Therapy MeSH
- Dimethyl Sulfoxide therapeutic use MeSH
- Disease Models, Animal MeSH
- Mice, Inbred NOD MeSH
- Tumor Cells, Cultured drug effects MeSH
- NF-kappa B drug effects MeSH
- Nitriles pharmacology therapeutic use MeSH
- Osteosarcoma * drug therapy genetics pathology MeSH
- Polymerase Chain Reaction methods MeSH
- Proto-Oncogene Proteins c-akt antagonists & inhibitors genetics metabolism MeSH
- RNA, Neoplasm isolation & purification drug effects MeSH
- Sulfones pharmacology therapeutic use MeSH
- In Vitro Techniques MeSH
- Animals MeSH
- Check Tag
- Animals MeSH
Fabrication of porous and biologically inspired biomaterials that mimic the formation of microstructural structures of nacre in the form of calcite (CaCO3) and evaluation of the biocompatibility of such organic-inorganic composite scaffold for bone tissue engineering, are focus of this paper. Nacre's self-assembly characteristics are concerned about the development of calcite filled biomineralized scaffold following the nature based biomineralization process and biomimetic applications. The PVP-CMC hydrogel film, comprised of PVP:0.2, CMC:0.8, PEG:1.0, Agar:2.0, Glycerene:1.0 and water:95.0 w/v%; acts as catalyst and template for the nucleation and growth of the inorganic CaCO3 within the scaffold. The PVP-CMC hydrogel (in the dry state) was immersed in ionic solutions (g/100 ml) of Na2CO3 and CaCl2·H2O in different concentrations sets i.e. Set-1: 10.50/14.70; Set-2: 5.25/7.35; Set-3: 4.20/5.88; Set-4: 2.10/2.94; Set-5: 1.05/1.47, Set-6: 0.55/0.55 for 90 min. As a result, "PVP-CMC-CaCO3" hydrogel scaffold was fabricated having bio-inspired structural and functional properties. Cell proliferation and cell viability were examined until 7 days in the presence of "PVP-CMC-CaCO3" scaffolds using permanent cell lines MG63 (human osteosarcoma), L929 (murine fibroblasts) as well as cultures from mouse bone explants (CC-MBE), confirmed that the said hydrogel scaffolds are biocompatible. But, from mechanical strength as well as biocompatibility point of view, scaffolds prepared in Set-1 to 3 ionic solutions were superior. In conclusion, these three calcite filled hydrogel scaffolds are recommended and can be used for osseointegration.
- MeSH
- Biocompatible Materials chemistry MeSH
- Cell Line MeSH
- X-Ray Diffraction MeSH
- Hydrogels chemistry MeSH
- Microscopy, Electron, Scanning MeSH
- Mice MeSH
- Osseointegration physiology MeSH
- Spectroscopy, Fourier Transform Infrared MeSH
- Tissue Engineering methods MeSH
- Tissue Scaffolds chemistry MeSH
- Cell Survival physiology MeSH
- Animals MeSH
- Check Tag
- Mice MeSH
- Animals MeSH
- Publication type
- Journal Article MeSH
