Abstract:Control of the corrosion that occurs in magnesium alloys in vivo is a significant challenge for their use as resorbable orthopaedic implants. In this work, we report on the provision of bioactive calcium phosphate (CaP) coatings on magnesium alloys that can delay substrate corrosion while offering an attendant physiochemical environment with properties known to promote an osteoinductive response in vivo. RF magnetron sputtering from hydroxyapatite (HA) powder targets has been employed to create CaP coatings on… Show more
“…XPS can analyse a depth of ~5-10 nm (under the conditions employed in this study) and SEM-EDX can analyse a depth of ~0.9 µm. Both techniques confirm the presence of both Ca and P on the surface but the application of dry pressed HA targets in this study can lead to re-sputtering of the HA coating, resulting in a lower-thanexpected Ca/P ratio and possible disparities between methods [25][26][27]. In addition, said characterisation techniques provided information on coating uniformity on the Ti substrate; no Ti 2p signals were obtained by XPS (Fig.…”
There is continued focus on the development of new biomaterials and associated biological testing methods needed to reduce the time taken for their entry to clinical use. The application of Raman spectroscopy to the study of individual cells that have been in contact with biomaterials offers enhanced in vitro information in a potentially non-destructive testing regime. The work presented here reports the Raman spectral analysis of discreet U-2 OS bone cells after exposure to hydroxyapatite (HA) coated titanium (Ti) substrates in both the as-deposited and thermally annealed states. These data show that cells that were in contact with the bioactive HA surface for 7 days had spectral markers similar to those cultured on the Ti substrate control for the same period. However, the spectral features for those cells that were in contact with the annealed HA surface had indicators of significant differentiation at day 21 while cells on the as-deposited surface did not show these Raman changes until day 28. The cells adhered to pristine Ti control surface showed no spectral changes at any of the timepoints studied. The validity of these spectroscopic results has been confirmed using data from standard in vitro cell viability, adhesion, and proliferation assays over the same 28-day culture period. In this case, cell maturation was evidenced by the formation of natural bone apatite, which precipitated intracellularly for cells exposed to both types of HA-coated Ti at 21 and 28 days, respectively. The properties of the intracellular apatite were markedly different from that of the synthetic HA used to coat the Ti substrate with an average particle size of 230 nm, a crystalline-like shape and Ca/P ratio of 1.63 ± 0.5 as determined by SEM-EDX analysis. By comparison, the synthetic HA particles used as a control had an average size of 372 nm and were more-rounded in shape with a Ca/P ratio of 0.8 by XPS analysis and 1.28 by SEM-EDX analysis. This study shows that Raman spectroscopy can be employed to monitor single U-2 OS cell response to biomaterials that promote cell maturation towards de novo bone thereby offering a label-free in vitro testing method that allows for non-destructive analyses.
“…XPS can analyse a depth of ~5-10 nm (under the conditions employed in this study) and SEM-EDX can analyse a depth of ~0.9 µm. Both techniques confirm the presence of both Ca and P on the surface but the application of dry pressed HA targets in this study can lead to re-sputtering of the HA coating, resulting in a lower-thanexpected Ca/P ratio and possible disparities between methods [25][26][27]. In addition, said characterisation techniques provided information on coating uniformity on the Ti substrate; no Ti 2p signals were obtained by XPS (Fig.…”
There is continued focus on the development of new biomaterials and associated biological testing methods needed to reduce the time taken for their entry to clinical use. The application of Raman spectroscopy to the study of individual cells that have been in contact with biomaterials offers enhanced in vitro information in a potentially non-destructive testing regime. The work presented here reports the Raman spectral analysis of discreet U-2 OS bone cells after exposure to hydroxyapatite (HA) coated titanium (Ti) substrates in both the as-deposited and thermally annealed states. These data show that cells that were in contact with the bioactive HA surface for 7 days had spectral markers similar to those cultured on the Ti substrate control for the same period. However, the spectral features for those cells that were in contact with the annealed HA surface had indicators of significant differentiation at day 21 while cells on the as-deposited surface did not show these Raman changes until day 28. The cells adhered to pristine Ti control surface showed no spectral changes at any of the timepoints studied. The validity of these spectroscopic results has been confirmed using data from standard in vitro cell viability, adhesion, and proliferation assays over the same 28-day culture period. In this case, cell maturation was evidenced by the formation of natural bone apatite, which precipitated intracellularly for cells exposed to both types of HA-coated Ti at 21 and 28 days, respectively. The properties of the intracellular apatite were markedly different from that of the synthetic HA used to coat the Ti substrate with an average particle size of 230 nm, a crystalline-like shape and Ca/P ratio of 1.63 ± 0.5 as determined by SEM-EDX analysis. By comparison, the synthetic HA particles used as a control had an average size of 372 nm and were more-rounded in shape with a Ca/P ratio of 0.8 by XPS analysis and 1.28 by SEM-EDX analysis. This study shows that Raman spectroscopy can be employed to monitor single U-2 OS cell response to biomaterials that promote cell maturation towards de novo bone thereby offering a label-free in vitro testing method that allows for non-destructive analyses.
“…An example of ToF-SIMS 3D depth profiling has been presented for RF magnetron-sputtered CaP coatings of different thicknesses on Mg alloy implants to confirm the successful deposition and 3D distribution of species in coatings with a thickness of 70 nm and 210 nm [ 61 ]. Moreover, depth profiling, which indicates the different intensities of phase-associated ions over time, has been shown to be useful in detecting different phase transitions, starting with the surface of plasma-deposited coatings embedded with silver nanoparticles, followed by the implant material.…”
Section: Interaction and Morphology Studiesmentioning
The development of bioactive coatings for orthopedic implants has been of great interest in recent years in order to achieve both early- and long-term osseointegration. Numerous bioactive materials have been investigated for this purpose, along with loading coatings with therapeutic agents (active compounds) that are released into the surrounding media in a controlled manner after surgery. This review initially focuses on the importance and usefulness of characterization techniques for bioactive coatings, allowing the detailed evaluation of coating properties and further improvements. Various advanced analytical techniques that have been used to characterize the structure, interactions, and morphology of the designed bioactive coatings are comprehensively described by means of time-of-flight secondary ion mass spectrometry (ToF-SIMS), X-ray photoelectron spectroscopy (XPS), Fourier transform infrared spectroscopy (FTIR), atomic force microscopy (AFM), scanning electron microscopy (SEM), transmission electron microscopy (TEM), 3D tomography, quartz crystal microbalance (QCM), coating adhesion, and contact angle (CA) measurements. Secondly, the design of controlled-release systems, the determination of drug release kinetics, and recent advances in drug release from bioactive coatings are addressed as the evaluation thereof is crucial for improving the synthesis parameters in designing optimal bioactive coatings.
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