Apatite formation on Poly (vinyl alcohol)-coated Poly (?-caprolactone) films by incubation in simulated body fluids

In this study, biodegradable poly (?-caprolactone) (PCL) films were coated with poly (vinyl alcohol) (PVA) and then incubated in a simulated body fluid 1.5SBF to prepare an apatite (HA)/PCL composite. It was found that the bone-like apatite formability of PCL was enhanced by PVA coating. The changes of surface properties induced by PVA coating were effective for apatite formation. The apatite formability increased with increasing coating amount. After 24 h incubation, apatite was formed on PVA-coated PCL film but hardly any apatite was found on uncoated PCL plate. The surface chemistry of the specimens was examined using XPS, FT-IR-ATR. The apatite formed was characterized by using SEM, TF-XRD, FT-IR, EDS. The apatite formed was similar in morphology and composition to that of natural bone. This indicated that simple PVA coating on PCL substrate could serve as a novel way to accelerated apatite formation via biomimetic method.     

Compositional dependence of in-vitro bioactivity in sodium calcium borate glasses

Borate glasses with composition xCaO (100−x) B₂O₃ (20≤x≤50), where x is in mole percent) and 50CaO·45B₂O₃·5Na₂O have been prepared using conventional melt quench technique. Samples were submerged in simulating body fluid solution (SBF) at 37 °C for various periods of time. After storage the samples were analyzed in order to investigate if a surface layer of hydroxyl carbonate apatite layer (HCA layer/Ca-P layer) had formed. The analysis technique used included Fourier-Transform Infrared Spectroscopy (FTIR). The rate of HCA layer formation on the surface of exposed glass samples is determined by FTIR, percentage weight loss measurements of glass samples in SBF and variation of pH of SBF as a function of time. Increase in calcium content in the glass matrix has shown to decrease the rate of HCA formation on glass surfaces. The borate glass with x=20 has shown HCA layer formation on glass surface within two days of dipping. The bone like apatite formation of glass surface demonstrates the potential of glass for integration with bone.     

Low-temperature biomimetic formation of apatite/TiO2 composite coatings on Ti and NiTi shape memory alloy and their characterization

Through a low temperature process, a bilayer composite coating was formed on Ti and NiTi shape memory alloy (SMA). The composite coating consisted of a layer of titania, which was formed using a H₂O₂-oxidation and hot water aging technique, and a layer of apatite, which was formed through an accelerated biomimetic process by immersing as-oxidized metals in a high-strength simulated body fluid (5SBF). Various techniques including X-ray diffraction, scanning electron microscopy and energy dispersive X-ray spectroscopy were used to characterize the surfaces of samples at different stages of coating formation and the coatings formed. Bioactive apatite/TiO₂ coatings could be formed on NiTi SMA and firmly bonded to the metal substrate. But there were differences for the formation of the composite coating on Ti and NiTi SMA substrates. The composite coatings formed will render both metals bioactive and hence bone-bonding.     

Effect of employing ultrasonic waves during pulse electrochemical deposition on the characteristics and biocompatibility of calcium phosphate coatings

In the present work, we investigated the effect of employing ultrasonic waves during pulse electrochemical deposition on surface topography, chemical composition and biocompatibility of calcium phosphate (Ca-P) coatings. The SEM and 3D AFM images showed that the anodized titanium surface was covered with the uniform and refined size of plate-like Ca-P crystals, when the ultrasonic treatment of the electrolyte with power of 60 W was carried out during deposition. In contrast, for the Ca-P; 0 W coating applied under only the magnetic stirring of the electrolyte, the microstructure was non-uniform and some Ca-P crystals with the larger size were randomly observed in different regions, causing a rougher surface. The FTIR results also revealed that employing the ultrasound increases the deposition of a coating involved in only the most stable Ca-P phase of carbonated hydroxyapatite (CHA). However, in the absence of ultrasound, besides the prominent phase of CHA, some less stable Ca-P phases like octa calcium phosphate (OCP) and brushite were also formed in the Ca-P; 0 W coating. The Ca-P; 60 W coating showed the higher ability for apatite biomineralization after a 7-day immersion in the simulated body fluid (SBF). This coating also provided a better surface for the cellular activity, as compared to the Ca-P; 0 W coating.     

In vitro bioactivity and biocompatibility of calcium phosphate cements using Hydroxy-propyl-methyl-Cellulose (HPMC)

In this study, the bioactivity and biocompatibility of new calcium phosphate bone cements (CPC) using Hydroxy-propyl-methyl-Cellulose (HPMC) was evaluated to understand the effect of HPMC on bone-bonding apatite formation and biocompatibility. In vitro bioactivity was investigated by incubating the CPC samples containing different ratios of HPMC (0%, 2% and 4% HPMC) in simulated body fluid (SBF) for 2, 7, 14 and 28 days. The formation of bone like apatite was confirmed on CPC surfaces by SEM and XRD analysis. Higher HPMC content of CPC showed faster apatite deposition in SBF. A high Ca ion dissolution profile was also reported with an increase of pH in all samples in SBF. The apatite formation ability of these CPC samples was found to be dependent on both surface chemistry and immersion time in SBF. The In vitro cytotoxicity test showed that the CPC samples with 4% HPMC were fairly cytocompatible for fibroblast L-929 cells. SEM images showed that MG-63 cells were successfully attached to the CPC samples and well proliferated.     

Bioactivity of SiO2-CaO-P2O5-Na2O glasses containing zinc-iron oxide

Glasses with composition x(ZnO,Fe₂O₃)(65 − x)SiO₂20(CaO,P₂O₅)15Na₂O (6 ≤ x ≤ 21 mol%) were prepared by melt-quenching technique. Bioactivity of the glasses was investigated in vitro by examining apatite formation on the surface of glasses treated in acellular simulated body fluid (SBF) with ion concentrations nearly equal to those in human blood plasma. Formation of bioactive apatite layer on the samples treated in SBF was confirmed by using Fourier transform infrared reflection (FTIR) spectroscopy, grazing incidence X-ray diffraction (GI-XRD) and scanning electron microscope (SEM) equipped with energy dispersive X-ray spectrometer. Development of an apatite structure on the surface of the SBF treated glass samples as functions of composition and time could be established using the GI-XRD data. FTIR spectra of the glasses treated in SBF show features at characteristic vibration frequencies of apatite after 1-day of immersion in SBF. SEM observations revealed that the spherical particles formed on the glass surface were made of calcium and phosphorus with the Ca/P molar ratio being close to 1.67, corresponding to the value in crystalline apatite. Increase in bioactivity with increasing zinc-iron oxide content was observed. The results have been used to understand the evolution of the apatite surface layer as a function of glass composition and immersion time in SBF.     

In vitro evaluation of bioactivity of CaO-SiO2-P2O5-Na2O-Fe2O3 glasses

Glasses with compositions 41CaO(52 − x)SiO₂4P₂O₅·xFe₂O₃3Na₂O (2 ≤ x ≤ 10 mol.%) were prepared by melt quenching method. Bioactivity of the different glass compositions was studied in vitro by treating them with simulated body fluid (SBF). The glasses treated for various time periods in SBF were evaluated by examining apatite formation on their surface using grazing incidence X-ray diffraction, Fourier transform infrared reflection spectroscopy, scanning electron microscopy and energy dispersive spectroscopy techniques. Increase in bioactivity with increasing iron oxide content was observed. The results have been used to understand the evolution of the apatite surface layer as a function of immersion time in SBF and glass composition.     

Bioactive calcium phosphate coating formed on micro-arc oxidized magnesium by chemical deposition

In order to improve the bioactivity of the micro-arc oxidized magnesium, a calcium phosphate coating was formed on the surface of micro-arc oxidized magnesium using a chemical method. The microstructures of the substrate and the calcium phosphate coating before and after the simulated body fluids (SBF) incubation were characterized by X-ray diffraction, Fourier-transformed infrared spectroscopy and scanning electron microscopy. The results showed that the calcified coating was composed of calcium deficient hydroxyapatite (HA) and dicalcium phosphate dihydrate (DCPD). After SBF incubation, some new apatite formation on the calcified coating surface from SBF could be found. The corrosion behaviours of the samples in SBF were also investigated by potentiodynamic polarization curves and immersion tests. The results showed that calcium phosphate coating increased the corrosion potential, and decreased the hydrogen gas release.     

Structure and apatite induction of a microarc-oxidized coating on a biomedical titanium alloy

An oxide coating with nanostructure was prepared by micro-arc oxidation (MAO) on a biomedical Ti-24Nb-4Zr-7.9Sn alloy. Chemical composition of the coating mainly includes O, Ti, Nb, Ca, P, Na, Zr and Sn, where the ratio of Ca/P is about 1.6. Ti, Nb, Zr and Sn participate in the oxidation to form TO₂, Nb₂O₅, ZrO₂ and SnO₂ nanocrystals, while Ca, P and Na are present in the form of amorphous phases. After alkali treatment, the surface of the MAO coating becomes rough, and Na concentration increases remarkably while P disappears basically. The alkali treated coating shows better apatite forming ability than the untreated one, as evidenced by apatite formation after SBF immersion for 7 days. The improvement of apatite forming ability of the modified coating is attributed to the formation of a sodium titanate layer and numbers of submicron-scale network flakes. The enhancement of the surface wettability of the alkali treated coating also plays an important role in promoting the apatite forming ability.     

Osteoblastic cell response on magnesium-incorporated apatite coatings

Magnesium is one of the most important bivalent ions associated with biological apatite. A series of magnesium-substituted calcium apatite coatings (Ca_10−xMg_x)(PO₄)₆(OH)₂, where x = 0, 0.50, 1.00, 1.50 and 2.00, are synthesized onto Ti6Al4V substrate by sol-gel dip-coating method to determine how magnesium influences the synthesis and the resulting structural and biological properties. X-ray diffraction (XRD) analysis shows that the incorporation of magnesium helps formation of Mg-containing β-TCP (β-TCMP) phase. X-ray photoelectron spectroscopy (XPS) is used to study the chemical composition and the results show that the apatite structure can only host magnesium less than ∼2.4 wt.% beyond which magnesium aggregates on the surfaces. The incorporation of magnesium slows down the dissolution of Ca²⁺ from the coating. The in vitro behavior of the coatings is evaluated with human osteosarcoma MG63 cells for cell morphology and proliferation. Similar cell morphologies are observed on all coatings. The cell proliferation results show that the incorporation of magnesium up to x = 2 has no adverse effect on cell growth.     

Effects of Mg and Zn on the surface of doped melt-derived glass for biomaterials applications

Bioactive glasses in the system SiO₂-CaO-Na₂O-P₂O₅ were synthesized pure and doped with magnesium or zinc by melt-derived method. The bioactivity was studied during in vitro assays: the ability of hydroxycarbonate apatite (HCA) layer to form on the glass surface was examined after contact with simulated body fluid (SBF). The X-ray diffraction (XRD), Fourier transform infrared spectroscopy (FTIR) and scanning electron microscopy (SEM) studies were performed before and after immersion in vitro assays. The SBF solutions were also analyzed using inductively coupled plasma-optical emission spectroscopy (ICP-OES).Introduction of magnesium and zinc as trace element induces several modifications on the observed phenomena at the glass surface and in SBF solution after immersion of the samples. The chemical durability of the glasses, the formation of the silica-rich layer and the crystallization of the HCA layer were affected, but not present the same modifications as the introduced doping element.     

Investigation on bio-mineralization of melt and sol-gel derived bioactive glasses

The bio-mineralization properties of the melt-derived bioactive glass 45S5 and the sol-gel derived bioactive glasses 58S and 77S were investigated and compared using in vitro test combined with BET, XRD, FTIR and SEM techniques. It was found that the surfaces of the three bioactive glasses could be mineralized by immersion in a simulated body fluid (SBF) at 37 °C for several hours. The bio-mineralized products on the surfaces of the bioactive glasses were apatite microcrystals with a low crystallinity, but the composition and morphologies of the apatite microcrystals on three glasses were different.     

Effects of mesoporous structure and UV irradiation on in vitro bioactivity of titania coatings

Mesoporous titania coatings (MTCs) with a pore size of 4.75 nm were prepared on Ti6Al4V substrates by a sol-gel process, and then irradiated with UV light at room temperature for 2 h. The effects of mesoporous structure and UV irradiation on the in vitro bioactivity were investigated. Simulated body fluid (SBF) tests reveal that the MTCs exhibit a high apatite-forming ability, which may be attributed to the following reasons: (i) the BET surface area of the MTCs is ∼190 m²/g, resulting in a greater density of Ti-OH groups than that without mesoporous structure; (ii) theoretical analysis reveals that the mesoporous structure can improve the driving force and nucleation rate of apatite precipitation in SBF. As compared with the MTCs, the UV-irradiated coatings do not exhibit any change in phase components and surface morphologies. However, the apatite-forming ability is higher on the UV irradiation coatings than on the MTCs because of the increase of Ti-OH groups and the improvement of wettability after UV irradiation. In addition, the investigation of the MG63 cell proliferation on the both substrates was performed. The results indicate that the MTCs before and after UV irradiation exhibit a good biocompatibility and are fit for the MG63 cell proliferation.     

Si，Ag，F离子共修饰HA纳米生物薄膜的制备与表征

采用单电流阶跃电化学沉积技术,在商业纯钛(CP-Ti)表面构建硅、银、氟离子共修饰羟基磷灰石(Si-Ag-F-HA)纳米复合薄膜。Ag+的持续释放可以提供有效的抗菌性,Si4+作为生物活性元素可以有效地抵消Ag+的潜在细胞毒性。采用电感耦合等离子体质谱法(ICP-MS)测定涂层中硅和银元素的释放规律。采用傅里叶变换红外光谱(FTIR)、扫描电子显微镜(SEM)、能量弥散X射线谱(EDS)、X-射线衍射(XRD)等技术对得到的材料进行了表征。结果表明：Si,Ag和F三种元素均匀地掺杂到了HA的晶体结构中。Si-Ag-F-HA为纳米级的针状晶体结构,薄膜整体致密且均匀。Si-Ag-F-HA纳米生物薄膜可以在一周内很好地诱导类骨磷灰石的形成,具有优异的生物活性。塔菲尔曲线测试结果证实涂层的耐SBF腐蚀性较好。ICP-MS测试结果表明Si-Ag-F-HA纳米生物薄膜可以提供持续的Si和Ag离子释放。FTIR 和ICP-MS等光谱技术为开发新型抗菌硬组织修复材料提供了高效快速的检测手段。     

X-ray photoelectron spectroscopy study of the growth kinetics of biomimetically grown hydroxyapatite thin-film coatings

Hydroxyapatite (HA) thin-film coatings grown biomimetically using simulated body fluid (SBF) are desirable for a range of applications such as improved fixation of fine- and complex-shaped orthopedic and dental implants, tissue engineering scaffolds and localized and sustained drug delivery. There is a dearth of knowledge on two key aspects of SBF-grown HA coatings: (i) the growth kinetics over short deposition periods, hours rather than weeks; and (ii) possible difference between the coatings deposited with and without periodic SBF replenishment. A study centred on these aspects is reported. X-ray photoelectron spectroscopy (XPS) has been used to study the growth kinetics of SBF-grown HA coatings for deposition periods ranging from 0.5 h to 21 days. The coatings were deposited with and without periodic replenishment of SBF. The XPS studies revealed that: (i) a continuous, stable HA coating fully covered the titanium substrate after a growth period of 13 h without SBF replenishment; (ii) thicker HA coatings about 1 μm in thickness resulted after a growth period of 21 days, both with and without SBF replenishment; and (iii) the Ca/P ratio at the surface of the HA coating was significantly lower than that in its bulk. No significant difference between HA grown with and without periodic replenishment of SBF was found. The coatings were determined to be carbonated, a characteristic desirable for improved implant fixation. The atomic force and scanning electron microscopies results suggested that heterogeneous nucleation and growth are the primary deposition mode for these coatings. Primary osteoblast cell studies demonstrated the biocompatibility of these coatings, i.e., osteoblast colony coverage of approximately 80%, similar to the control substrate (tissue culture polystyrene).     

Adsorption and desorption of Ca and PO4 species from SBFs on RF-sputtered calcium phosphate thin films

RF magnetron sputtering of calcium phosphate (CaP) coatings is a promising technique to apply thin bioactive films on bulk implant materials. In this paper the properties of the interface between RF sputtered coatings and simulated body fluids (SBFs) are related to the ability to form CaP crystals on the coating surface. Two types of coatings were compared: coatings with a low Ca over P ratio (∼0.8; CaP_low), which remain inert when immersed in SBF₂ (i.e. SBF with twice the Ca and PO₄ concentrations), and coatings with a high Ca over P ratio (1.6; CaP_high), which show the formation of CaP crystals on their surface within 2 h. Low energy ion scattering (LEIS) and radioactive labeling of the SBFs combined with liquid scintillation counting (LSC) allowed us to study very accurately the composition of the adsorbates of both coating groups after 10 min of immersion in SBF₂. For the adsorbate on CaP_high and CaP_low coatings coverages were found consistent with ionic adsorption and Ca/P ratios of 1.24 ± 0.02 and 2.17 ± 0.10, respectively. Adsorption was found to be reversible over the studied immersion period. After an induction period of 40 min a CaP precipitate started to form on the CaP_high coatings with a Ca/P ratio of 1.30 ± 0.02. Further, no significant desorption of coating species was observed during this induction period.     

Hydroxyapatite/titania composite bioactivity coating processed by sol-gel method

Hydroxyapatite/titania composite material was coated onto a titanium (pure Ti) substrate by sol-gel method. The hydroxyapatite (HA) and titania (TiO₂) sol were made from precursor and mixed together. The insertion of TiO₂ enhanced the chemical affinity and the physical consistency between HA and Ti substrate. The HA/TiO₂ composite coating adhered tightly to the Ti substrate. Owing to the insertion of TiO₂, the crystallinity of HA has been delayed. The specimens with HA/TiO₂ composite coatings were soaked into SBF, and displayed good bone-like apatite forming ability. The bioactivity of the composite HA coatings were tested in vitro by cell culture.     

Improving the bioactivity of NiTi shape memory alloy by heat and alkali treatment

TiO₂ films were formed on an NiTi alloy surface by heat treatment in air at 600 °C. Heat treated NiTi shape memory alloys were subsequently alkali treated with 1 M, 3 M and 5 M NaOH solutions respectively, to improve their bioactivity. Then treated NiTi samples were soaked in 1.5SBF to evaluate their in vitro performance. The results showed that the 3 M NaOH treatment is the most appropriate method. A large amount of apatite formed within 1 day's soaking in 1.5SBF, after 7 day's soaking TiO₂/HA composite layer formed on the NiTi surface. SEM, XRD, FT-IR and TEM results showed that the morphology and microstructure are similar to the human bone apatite.     

Enhanced in vitro biocompatibility of ultrafine-grained titanium with hierarchical porous surface

Bulk ultrafine-grained Ti (UFG Ti) was successfully fabricated by equal-channel angular pressing (ECAP) technique in the present study, and to further improve its surface biocompatibility, surface modification techniques including sandblasting, acid etching and alkali treatment were employed to produce a hierarchical porous surface. The effect of the above surface treatments on the surface roughness, wettability, electrochemical corrosion behavior, apatite forming ability and cellular behavior of UFG Ti were systematically investigated with the coarse-grained Ti as control. Results show that UFG-Ti with surface modification had no pitting corrosion and presented low corrosion rate in simulated body fluids (SBF). The hierarchical porous surface yielded by surface modification enhanced the ability of UFG Ti to form a complete apatite layer when soaked in SBF and promoted osteoblast-like cells attachment and proliferation in vitro, which promises to have a significant impact on increasing bone-bonding ability and reducing healing time when implanted due to faster tissue integration.     

Effect of heat treatment on bioactivity of anodic titania films

Anodic oxidation could be employed to produce crystalline titania films on Ti6Al4 V surfaces for inducing apatite formation in simulated body fluid (SBF). In this work, the effect of further heat treatment on the bioactivity of anodic titania films was researched. The surface constitution, morphology, crystal structure and apatite-forming ability of titania films were characterized by X-ray photoelectron spectroscopy (XPS), scanning electron microscopy (SEM) and X-ray diffraction (XRD). The results indicated the apatite formation on the Ti6Al4 V surfaces could be attributed to abundance of Ti-OH groups formed via anodic oxidation, but subsequent heat treatment would decrease the amount of surface hydroxyl (OH) groups and result in the loss of the apatite-forming ability.