Oxidized NiTi surfaces enhance differentiation of osteoblast-like cells (original) (raw)
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Journal of Materials Science: Materials in Medicine, 2012
The influence of 6 % sodium hypochlorite (NaClO) treatment on adhesion and proliferation of MC3T3 pre-osteoblast cells seeded on electropolished (EP) and magnetoelectropolished (MEP) nitinol surfaces were investigated. The chemistry, topography, roughness, surface energy, wettability of EP and MEP nitinol surfaces before and after NaClO treatment were studied with X-ray photoelectron spectroscopy (XPS), profilometry, and contact angle meter. In vitro interaction of osteoblast cell and NaClO treated EP and MEP nitinol surfaces were assessed after 3 days of incubation by scanning electron microscopy. The XPS analysis shows that NaClO treatment increases oxygen content especially in subsurface oxide layer of EP and MEP nitinol. The changes of both basic components of nitinol, namely nickel and titanium in oxide layer, were negligible. The NaClO treatment did not influence physico-morphological surface properties of EP and MEP nitinol to a big extent. The osteoblast cells show remarkable adherence and proliferation improvement on NaClO treated EP and MEP nitinol surfaces. After 3 days of incubation they show almost total confluence on both NaClO treated surfaces. The present study shows that NaClO treatment of EP and MEP nitinol surfaces alters oxide layer by enriching it in oxygen and by this improves bone cell-nitinol interaction.
Journal of Biomedical Materials Research Part A, 2013
In this study we evaluated the effect of new plasma-nitrided Ti surfaces on the progression of osteoblast cultures, including cell adhesion, proliferation and differentiation. Ti surfaces were treated using two plasma nitriding protocols, hollow cathode for 3 h (HC 3h) and 1 h (HC 1h) and planar (Planar) for 1 h. Untreated Ti surfaces were used as Control. Cells derived from human alveolar and rat calvarial bones were cultured on Ti surfaces for periods of up to 14 days and the following parameters were evaluated: cell morphology, adhesion, spreading and proliferation, alkaline phosphatase (ALP) activity, extracellular matrix mineralization, and gene expression of key osteoblast markers. Plasma nitriding treatments resulted in Ti surfaces with distinct physicochemical characteristics. The cell adhesion and ALP activity were higher on plasma-nitrided Ti surfaces compared with untreated one, while cell proliferation and extracellular matrix mineralization were not affected by the treatments. In addition, the plasma-nitrided Ti surfaces increased the ALP, reduced the osteocalcin and did not affect the Runx2 gene expression. We have shown that HC 3h and Planar Ti surfaces slightly favored the osteoblast differentiation process and then these surfaces should be considered for further investigation using preclinical models.
PLoS ONE, 2014
An investigation was made of the adhesion, growth and differentiation of osteoblast-like MG-63 and Saos-2 cells on titanium (Ti) and niobium (Nb) supports and on TiNb alloy with surfaces oxidized at 165uC under hydrothermal conditions and at 600uC in a stream of air. The oxidation mode and the chemical composition of the samples tuned the morphology, topography and distribution of the charge on their surfaces, which enabled us to evaluate the importance of these material characteristics in the interaction of the cells with the sample surface. Numbers of adhered MG-63 and Saos-2 cells correlated with the number of positively-charged (related with the Nb 2 O 5 phase) and negatively-charged sites (related with the TiO 2 phase) on the alloy surface. Proliferation of these cells is correlated with the presence of positively-charged (i.e. basic) sites of the Nb 2 O 5 alloy phase, while cell differentiation is correlated with negatively-charged (acidic) sites of the TiO 2 alloy phase. The number of charged sites and adhered cells was substantially higher on the alloy sample oxidized at 600uC than on the hydrothermally treated sample at 165uC. The expression values of osteoblast differentiation markers (collagen type I and osteocalcin) were higher for cells grown on the Ti samples than for those grown on the TiNb samples. This was more particularly apparent in the samples treated at 165uC. No considerable immune activation of murine macrophage-like RAW 264.7 cells on the tested samples was found. The secretion of TNF-a by these cells into the cell culture media was much lower than for either cells grown in the presence of bacterial lipopolysaccharide, or untreated control samples. Thus, oxidized Ti and TiNb are both promising materials for bone implantation; TiNb for applications where bone cell proliferation is desirable, and Ti for induction of osteogenic cell differentiation.
In vitro studies on the influence of surface modification of NiâTi alloy on human bone cells
Journal of Biomedical Materials Research Part A, 2009
The in vitro cell behavior on Nitinol TM after different surface treatments was investigated. As references samples, commercially pure titanium (cpTi) and bioactive titanium were used. The surface treatments influenced the topography, surface energy, crystallographic structure, ion release, chemistry, and ability to form apatite layer from simulated body fluids. Regardless of the surface treatment, the bioactivity study showed that the kinetics of apatite film formation was similar for all tested samples. No clear indication of the surface characteristics influence on the ability for calcium-phosphate precipitation was evident. Cell activity studies showed that ground nickel titanium, spark oxidized and thermally oxidized (at 4008C and below) had higher cellular activity and caused increased alkaline phosphatase (ALP) and osteocalcin (OC) expression which was comparable to control tissue culture plastic and titanium reference samples. Regardless of surface modifications, preimmersion of the samples in media for 72 h resulted in cell proliferation at the same level for all samples. Therefore, it can be concluded that preconditioning of samples alters surface properties and modulates the cell response regardless of the initial surface treatment and its properties. Moreover, a detrimental effect on cell response was observed after 7 and 14 days in culture for alkali treated samples. This was attributed to a high surface nickel concentration and a high nickel ion release rate from these surfaces.
The phase state of NiTi implant material affects osteoclastic attachment
Journal of Biomedical Materials Research Part A, 2005
In the present work, the responses of mature osteoclasts cultured on austenite and martensite phases of NiTi shape memory implant material were studied. We used the sensitivity of osteoclasts to the underlying substrate and actin ring formation as an indicator of the adequacy of the implant surface. The results showed osteoclasts with actin ring on both NiTi phases. However, significantly more osteoclasts were present on the austenitic NiTi than on the martensitic NiTi. We also analyzed the surface free energy of the samples but found no significant difference between austenite and martensite phases. The results revealed that osteoclasts tolerated well the austenite phase of NiTi. The chemically identical martensitic NiTi was not as well tolerated by osteoclasts (e.g., indicated by diminished actin ring formation). This leads to the conclusion that certain physical properties specific to the martensitic NiTi have an adverse effect to the surviving of osteoclasts on this NiTi phase. These results confirm that mature, authentic osteoclasts can act as cell probes in experiments concerning aspects of biocompatibility of bone implant materials.
Effect of a niobium-containing titanium alloy on osteoblast behavior in culture
Clinical Oral Implants Research, 2009
The chemistry of titanium is a key factor in determining implant-tissue interactions. Reports that a vanadium-based titanium alloy (Ti-6Al-4V) exhibits some cytotoxicity led to a search for an alternative. The aim of the present study was to investigate the behavior of human osteoblast-like cells (Saos-2) cultured on Ti-6Al-4V or Ti-6Al-7Nb disks with a rough or a machined surface. In all four groups, the cells proliferated rapidly between days 1 and 3, and then plateaued. On day 1 of culture, the highest proliferation rate was of cells cultured on disks containing Nb with a machined surface. On day 7, there was no significant difference in cell density on all the tested surfaces. Alkaline phosphatase (ALP) activity was lower on the machined surfaces, regardless of the material used, suggesting that cells on rough surfaces exhibit a more mature phenotype. On day 3, cells cultured on rough disks made of Ti-6Al-7Nb showed the highest ALP activity; the lowest activity was observed on the machined Ti-6Al-4V surface. The highest level of osteocalcin (day 7) was found in the cells cultured on rough Ti-6Al-7Nb disks. Also, higher levels of transforming growth factor (TGFbeta) were noted for cells cultured on the rough Ti-6Al-7Nb disks, suggesting that the Nb-containing alloy supports more rapid maturation of the osteoblast. The results of the present study suggest that according to our cell culture preclinical model, Ti-6Al-7Nb may replace the Ti-6Al-4V alloy as an implant material.
Bone Cell-materials Interactions and Ni Ion Release of Anodized Equiatomic NiTi Alloy
Laser processed NiTi alloy was anodized for different durations in H 2 SO 4 electrolyte with varying pH to create biocompatible surfaces with low Ni ion release as well as bioactive surfaces to enhance biocompatibility and bone cell-materials interactions. The anodized surfaces were assessed for their in vitro cell-materials interactions using human fetal osteoblast (hFOB) cells for 3, 7 and 11 days, and Ni ion release up to 8 weeks in simulated body fluids. The results were correlated with surface morphologies of anodized surfaces characterized using field-emission scanning electron microscopy (FESEM). The results show that the anodization creates a surface with nano/micro roughness depending on anodization conditions. The hydrophilicity of NiTi surface was found to improve after anodization due to lower contact angles in cell media, which dropped from 32° to < 5°. The improved wettability of anodized surfaces is further corroborated by their high surface energy comparable to that of cp Ti. Relatively high surface energy, especially polar component, and nano/micro surface features of anodized surfaces significantly increased the number of living cells and their adherence and growth on these surfaces. Finally, a significant drop in Ni ion release from 268 ± 11 to 136 ± 15 ppb was observed for NiTi surfaces after anodization. This work indicates that anodization of NiTi alloy has a positive influence on the surface energy and surface morphology, which in turn improve bone cell-materials interactions and reduce Ni ion release in vitro.
Biomaterials, 2004
Nickel-titanium shape-memory alloys (NiTi-SMA) were coated with calcium phosphate by dipping in oversaturated calcium phosphate solution (CaP-coating). Polymorphonuclear neutrophil granulocytes (PMN) belong to the first cells which will adhere to implant materials. We analyzed the apoptosis of isolated human PMN after cell culture on non-coated and CaP-coated NiTi-SMA by light and scanning electron microscopy (cell morphology) and by flow cytometry (DNA-fragmentation). In contrast to PMN adherent to non-coated NiTi-SMA, the apoptosis of PMN adherent CaP-coated samples was inhibited. Cell culture media obtained from cultured leukocytes with CaP-coatings (conditioned media, CM) were able to transfer the apoptosis inhibiting activities to freshly isolated PMN. There was a significant ðpo0:01Þ increase in GM-CSF, IL-8, IL-6, and TNF-a within CM obtained from coated versus non-coated NiTi-SMA as was determined by ELISA. r
Biological Trace Element Research
The present review is based on a survey of 21 studies on the cytocompatibility of medical biomaterials containing nickel, as assessed by cell culture of human and animal osteoblasts or osteoblast-like cells. Among the biomaterials evaluated were stainless steel, NiTi alloys, pure Ni, Ti, and other pure metals. The materials were either commercially available, prepared by the authors, or implanted by various techniques to generate a protective layer of oxides, nitrides, acetylides. The observation that the layers significantly reduced the initial release of metal ions and increased cytocompatibility was confirmed in cell culture experiments. Physical and chemical characterization of the materials was performed. This included, e.g., surface characterization (roughness, wettability, corrosion behavior, quantity of released ions, microhardness, and characterization of passivation layer). Cytocompatibility tests of the materials were conducted in the cultures of human or animal osteoblasts and osteoblast-like cells. The following assays were carried out: cell proliferation and viability test, adhesion test, morphology (by fluorescent microscopy or SEM). Also phenotypic and genotypic markers were investigated. In the majority of works, it was found that the most cytocompatible materials were stainless steel and NiTi alloy. Pure Ni was rendered and less cytocompatible. All the papers confirmed that the consequence of the formation of protective layers was in significant increase of cytocompatibility of the materials. This indicates the possible further modifications of the manufacturing process (formation of the passivation layer).