Growth of calcium phosphate on surface-modified cotton

A study of the growth of amorphous calcium phosphate on surface-modified cotton fibres by a combination of scanning electron microscopy/electron diffraction X-ray analysis, micro-FTIR and X-ray photoelectron spectroscopy is reported. Cotton fibres phosphorylated by the urea/phosphorous acid method and then soaked in saturated Ca(OH)₂ for approximately one week were found to stimulate the growth of a calcium phosphate coating on their surfaces after soaking in 1.5×SBF for as little as 1 day. Ca(OH)₂ soaking of the fibres is found to produce highly crystalline clusters lodged in the fibres which were confirmed by micro-FTIR to be calcium phosphite monohydrate (CaHPO₃·H₂O). In contrast, phosphorylated fibres not subjected to the Ca(OH)₂ treatment did not exhibit calcium phosphate growth upon immersion in 1.5×SBF solution. Soaking of the Ca(OH)₂-treated fibres with time in the 1.5×SBF solution produced progressively thicker layers of calcium phosphate on the fibres as confirmed by scanning electron microscopy and X-ray photoelectron spectroscopy. In general, calcium phosphate coatings formed over 1 1–5 day period soaking in 1.5×SBF solution appeared to consist of agglomerations of a large number of small spherical particles, while coatings formed after 17 days of soaking were distinctly chunky, thick and non-uniform in appearance. Micro-FTIR indicated that CaHPO₃·H₂O clusters were still present in cotton samples even after 4 days of soaking, while after 17 days, only the infrared spectrum typical of calcium phosphate was observed. EDX-measured Ca:P ratios of the coatings, although variable, suggested amorphous calcium phosphate. The mechanism of formation of the coating is believed to involve dissolution of the CaHPO₃.H₂O clusters upon introduction of the Ca(OH)₂-treated phosphorylated cotton into the 1.5×SBF solution which elevates the Ca²⁺ ion concentration in the vicinity of the fibres so stimulating calcium phosphate formation. It is postulated that phosphite groups chemically bound to the cotton fibre surface or a calcium phosphite coating on the fibres act as nucleation sites for calcium phosphate growth in 1.5×SBF solution.     

Further studies of calcium phosphate growth on phosphorylated cotton fibres

Further studies using scanning electron microscopy/energy dispersive X-ray analysis (SEM/EDX), micro-Fourier transform infrared spectroscopy (FTIR), X-ray photoelectron spectroscopy (XPS) and solid state magic angle spinning nuclear magnetic resonance (MAS NMR) techniques of calcium phosphate growth on Ca(OH)₂-treated urea/H₃PO₃- and urea/H₃PO₄-modified cotton fibres are reported. In the case of the Ca(OH)₂-treated urea/H₃PO₃-modified fibres which have been reported in an earlier paper, further experiments subjecting the urea/H₃PO₃-modified cotton to alternative soaking treatment procedures to Ca(OH)₂ as well as different calcium phosphate growth media such as the alkaline phosphatase-catalysed hydrolysis of disodium p-nitrophenylphosphate to free phosphate have reaffirmed the importance of the Ca(OH)₂ treatment step for the stimulus and growth of calcium phosphate growth on the fibres. Studies on cotton phosphorylated by a slightly different method using urea/H₃PO₄ instead of urea/H₃PO₃ show that a phosphorylated cotton with similar properties to the urea/H₃PO₃-modified fibres can be produced. Soaking of these fibres in saturated Ca(OH)₂ solution leads to cotton coated with thin layers of calcium phosphate formed by partial hydrolysis of the PO₄ functionalities in the phosphorylated cotton which are believed to act as nucleation layers for further calcium phosphate deposition when the fibres are subsequently soaked in 1.5×SBF solution. SEM/EDX studies of the calcium phosphate coatings formed on the Ca(OH)₂-treated urea-H₃PO₄ fibres as a function of soaking time in 1.5 × SBF show that coatings deposit and become noticeably thick after approximately 9 days. XPS studies indicated the presence of carbonate species in the calcium phosphate coating deposited. In common with the calcium phosphate coated Ca(OH)₂-treated urea/H₃PO₃-modified fibres studied earlier, the average EDX-measured Ca: P ratios of the coatings formed on the Ca(OH)₂-treated urea/H₃PO₄ fibres are 1.60 and give very similar micro-FTIR spectra with evidence of carbonate which suggests that amorphous calcium deficient apatite has deposited.     

In-vitro apatite formation on phosphorylated bamboo

Natural self-reinforced composite, bamboo, was surface modified by phosphorylation with urea–H3PO4 and NaOH–H3PO4 methods; then precalcification was performed by immersing samples in saturated Ca(OH)2 solution. After that, calcium phosphate can be formed on the surface of bamboo samples in calcification media: simulated body fluid (1.5 SBF) and accelerated calcification solution (ACS). Experimental results reveal that pre-calcification is an inevitable step for the formation of calcium phosphate. The calcium phosphate formed in 1.5 SBF was identified by thin-film X-ray diffraction as apatite which was not well crystallized. Compared with the urea–H3PO4 method, the NaOH–H3PO4 method has the advantages of quicker and continuous apatite formation and stronger adhesive between apatite and bamboo.     

Calcium phosphate fibres synthesized from a simulated body fluid

The biomimetic coating method was used for fabricating calcium phosphate fibres for biomedical applications such as bone defect fillers. Natural cotton substrate was pre-treated with phosphorylation and a Ca(OH)₂ saturated solution. The pre-treated samples were then soaked in simulated body fluid (SBF) of two different concentrations, 1.5 times and 5.0 times the ion concentration of blood plasma. The cotton was then burnt out via sintering of the ceramic coating at 950^∘C, 1050^∘C, 1150^∘C, and 1250^∘C. The results demonstrated that osteoblastic cells were able to cover the entire surface cotton fibres, and the cell coverage appeared to be independent of surface roughness and Ca/P ratio of fibres.     

In–vitro calcium phosphate growth over functionalized cotton fibers

Biomimetic growth of calcium phosphate compound on cotton sheets treated with tetraethoxy silane and soaked in simulated body fluid solution was studied using scanning electron microscopy (SEM), energy dispersive X-ray analysis (EDAX), micro-Fourier transform infrared spectroscopy (FTIR) and X-ray diffractometry (XRD). Micro-FTIR and EDAX results show that silicon was coupled to the cotton fiber when cotton was treated with tetra-ethoxy silane (TEOS) at 125°C for 1 h. Calcium phosphate nucleation started to occur on the surface of TEOS-treated cotton fibers upon immersion in 1.5×SBF (simulated body fluid solution) within 3 days and after 20 days, all the fiber surfaces were found covered with a thick and porous coating of calcium phosphate. The Ca and P determined by inductively coupled plasma spectroscopy (ICP) analysis revealed that the Ca/P ratio as well as the amount of calcium phosphate coating depends on the soaking time in SBF solution. © 1999 Kluwer Academic Publishers     

超声电沉积碳/碳复合材料磷灰石涂层的工艺优化

通过声电沉积在碳/碳复合材料表面制备钙磷生物活性涂层,采用SEM(带EDAX)或FE-SEM,XRD,FTIR研究电解质浓度和初始pH值对钙磷生物活性涂层的形貌、结构和组成的影响,并采用拉伸测试评价涂层与基体的结合力.结果表明:随着电解质浓度的降低,其组成由羟基磷灰石与磷酸三钙组成的混合涂层转变为含碳酸根的羟基磷灰石涂层,n(Ca)/n(P)和碳酸根的含量逐渐增加,片状晶体颗粒减小.随着初始电解液pH值的升高,涂层致密,均为片状含碳酸根的羟基磷灰石,n(Ca)/n(P)呈增加的趋势,片状晶体的厚度在30～40nm之间.涂层拉伸实验表明:小电解质浓度时,其涂层与基体的结合强度(5.62MPa)大于高电解质浓度时涂层与基体的结合强度(3.85MPa).     

致密磷酸钙陶瓷在动态SBF中类骨磷灰石层形成研究

磷酸钙陶瓷植入体内后其表面类骨磷灰石层的形成是诱导成骨的先决条件．本实验在模拟体液(ｓｉｍｕｌａｔｅｄ ｂｏｄｙ ｆｌｕｉｄ,ＳＢＦ)以人体骨骼肌组织内体液的正常生理流率(２ｍＬ/１００ｍＬ·ｍｉｎ)和偏离正常生理流率流动的动态条件下,研究在动态ＳＢＦ中影响致密磷酸钙陶瓷表面类骨磷灰石层形成的因素．结果表明:在生理流率条件下,材料的粗糙表面有利于类骨磷灰石的形成,加大ＳＢＦ中Ｃａ^２＋、ＨＰＯ_４^２－离子浓度,类骨磷灰石层的形成速度加快．比起通常使用的静态浸泡试验,ＳＢＦ以生理流率流动的动态试验能够更好地模拟类骨磷灰石生长的体内环境．动态ＳＢＦ对了解类骨磷灰石形成,进而了解磷酸钙陶瓷在体内诱导成骨机理是十分有用的．     

通过仿生法在硅橡胶表面制备磷灰石薄膜的研究

用CaCl2的乙醇溶液和K2HPO4溶液对硅橡胶进行预处理,将处理过的硅橡胶分别浸渍于模拟体液和钙磷饱和溶液中来制备磷灰石薄膜.利用薄膜X射线衍射、红外吸收光谱和扫描电子显微镜对形成的薄膜进行了表征.结果表明,分别在模拟体液中7d和在钙磷饱和溶液中5d后,硅橡胶表面形成了一层磷灰石薄膜;在模拟体液中的薄膜表面呈网状并分布有许多球状晶粒,在钙磷饱和溶液中的薄膜为结晶良好的片状晶体.     

NaOH treatment of vacuum-plasma-sprayed titanium on carbon fibre-reinforced poly(etheretherketone)

Carbon fibre-reinforced polyetheretherketone (CF-PEEK) substrates were coated with titanium by vacuum-plasma-spraying and chemically treated in 10 M sodium hydroxide (NaOH) solution. After NaOH treatment, the specimens were immersed in simulated body fluid (SBF) containing ions in concentrations similar to those of human blood plasma. Scanning electron microscopy, energy-dispersive X-ray analysis and diffuse reflectance Fourier transformed–infrared spectroscopy were used to analyse the NaOH-treated VPS-Ti surface and the calcium phosphate layer formed during immersion in SBF. It was observed that a carbonate-containing calcium phosphate layer was formed on the NaOH-treated VPS-Ti surface during immersion in SBF, whereas no calcium phosphate precipitation occurred on the untreated surfaces. It is therefore concluded that vacuum-plasma-spraying with titanium and subsequent chemical modification in 10 M NaOH solution at 60°C for 2 h is a suitable method for the preparation of bioactive coatings for bone ongrowth on CF-PEEK.     

Synthesis and Characterization of Bioceramic Calcium Phosphates by Rapid Combustion Synthesis

Calcium hydroxyapatite(Ca10(PO4)6(OH)2) has been synthesized in short duration by rapid solution combustion by employing different fuels.Calcium nitrate was taken as source of calcium and diammonium hydrogen phosphate served as the source of phosphate ions.Citric acid,tartaric acid,sucrose,glycine and urea were used as the fuels and nitrate ions and nitric acid were used as oxidizers.The influence of fuels on the morphology of the phase formed was studied.Results of the studies by powder X-ray diffraction a...     

Bioactivity of calcium phosphate coatings prepared by electrodeposition in a modified simulated body fluid

The purpose of this study was to investigate bioactivity of calcium phosphate coatings prepared by electrodeposition in a modified simulated body fluid (SBF). Calcium phosphates were electrodeposited on commercially pure titanium substrates in the modified SBF at 60 °C for 1 h maintaining the cathodic potentials of − 1.5 V, − 2 V, and − 2.5 V (vs. SCE). Subsequently, the calcium phosphate coatings were transformed into apatites during immersion in the SBF at 36.5 °C for 5 days. The apatites consisted of needle-shaped crystallites distributed irregularly with different grain sizes. As the coatings were electrodeposited at higher cathodic potential, the crystallite of the apatites got denser and the grain sizes of the apatites became bigger during subsequent immersion in the SBF. However, as the coatings were electrodeposited at higher cathodic potential, the coatings were transformed into apatites with lower crystallinity and the Ca/P atomic ratio of the apatites got higher than 1.67, that of stoichiometric hydroxyapatite, after subsequent immersion in the SBF. In addition, CO₃²⁻ ions contained in the modified SBF were incorporated in the calcium phosphate coating during electrodeposition and had an influence on transforming the calcium phosphate into bonelike apatite during subsequent immersion in the SBF showing that CO₃²⁻ incorporated in the apatites disturbed crystallization of the apatites. These results revealed that the coating electrodeposited at − 2.0 V (vs. SCE) in the modified SBF containing CO₃²⁻ ions was the most bioactive showing transformation into carbonate apatite similar to bone apatite.     

Deposition of Bioactive Layer on NiTi Alloy by Chemical Treatment

A simple chemical method was developed for inducing bioactivity on NiTi alloys(50 at.pct by Ni/Ti).A layer of calcium phosphate was deposited on the surface to improve biocompatibility of thealloy.NiTi alloys were first etched in HNO3 aqueous solution,and then treated with boiling diluted NaOH solution.A rough surface was created and a thin TiO2 layer was formed on the surface.Pre-calcification was then introduced by immersing the treated NiTi alloys in supersaturated Na2HPO4 solution and supersaturated Ca(OH)2 solution in turn before calcification in simulated body fluid (SBF).A dense and uniform bonelike calcium phosphate(Ca-P) bioactive layer was formed on the surfaces of the specimen,which would improve their biocompatibility.Morphology and element analysis on NiTi surfaces during the treatments were investigated in detail by means of environment scanning electron microscopy(ESEM),energy dispersion X-ray spectroscopy(EDXS),and X-ray diffraction (XPD).     

Surface modification of Ti–Nb–Zr–Sn alloy by thermal and hydrothermal treatments

Surface modifications by thermal and hydrothermal treatments in solution with calcium ions were investigated with the aim of improving bioactivity and wear resistance of a Ti–Nb–Zr–Sn alloy. The results showed that the first step of thermal treatment at 600 °C significantly increases the surface hardness and energy by forming oxides of Ti and Nb. The second step of hydrothermal treatment in a boiled supersaturated Ca(OH)₂ solution induces a bioactive layer containing CaTiO₃, CaCO₃, Ca(OH)₂ and TiO₂. Using this treatment, a complete Ca–P layer can be formed within 3 days of soaking in simulated body fluid (SBF). The origin of such fast apatite formation was analyzed by comparison with single step thermal or hydrothermal treatment and with thermal plus hydrothermal treatment without calcium ions. The results suggest that the increase of surface energy by thermal treatment and the incorporation of calcium ions by the hydrothermal treatment in calcium ion solution play important roles in the formation of bioactive apatite.     

The mechanism of biomineralization of bone-like apatite on synthetic hydroxyapatite: an in vitro assessment.

The mechanism of biomineralization of bone-like apatite on synthetic hydroxyapatite (HA) has been investigated in vitro, in which the HA surface was surveyed as a function of soaking time in simulated body fluid (SBF). In terms of surface structure by transmission electron microscopy with energy-dispersive X-ray spectrometry, the HA whose Ca/P atomic ratio was 1.67 revealed three different characteristic soaking periods in SBF, i.e. the first soaking period, in which the HA surface increased the Ca/P ratio up to 1.83 to form an amorphous phase of Ca-rich calcium phosphate; the second soaking period, in which the HA surface decreased the Ca/P ratio up to 1.47 to form an amorphous phase of Ca-poor calcium phosphate; and the third soaking period, in which the HA surface gradually increased the Ca/P ratio up to 1.65 to eventually produce the bone-like nano-cerystallites of apatite, which grew forming complex crystal assemblies with a further increase in immersion time. Analysis using electrophoresis spectroscopy indicated that, immediately after immersion in SBF, the HA revealed a highly negative surface potential, which increased to reach a maximum positive value in the first soaking period. The surface potential then decreased to again reach a negative value in the second soaking period and thereafter converge to a constant negative value in the third soaking period. This implies that the HA induces biomineralization of apatite by smartly varying its surface potential to trigger an electrostatic interaction, first with positive calcium ions and second with negative phosphate ions in the SBF.     

Influence of experimental parameters on spatial phase distribution in as-sprayed and incubated hydroxyapatite coatings

In the present study, the behavior and properties of plasma-sprayed hydroxyapatite coatings [Ca(10)(PO(4))(6)(OH)(2), HAp] were investigated in relation to the spraying process. The experiments were focused on the influence of type of feedstock and spray power on the phase composition and distribution within the coatings. Depth profiles of the coatings were investigated before and after incubation in revised simulated body fluid (SBF) by X-ray diffraction and infrared spectroscopy. Besides HAp, the coatings contain oxyapatite (OAp) and carbonate apatite (CAp). Additionally, tricalcium phosphate (TCP), tetracalcium phosphate (TTCP), CaO, and an amorphous phase were detected in the coatings. The HAp content directly depends on the used spray powder and spray power, where the influence of spray powder is much higher than the influence of the spray power. The grain size range of the spray powder strongly influences the HAp content in the coating and the formation of CaO. The in vitro behavior of the coatings in simulated body fluid mainly depends on the contents of CaO and amorphous calcium phosphate, respectively. The formation of portlandite due to the reaction of the coating with the SBF is strongly influenced by the porosity of the coatings and can be used as an indicator for the depth of interaction between fluid and coating.     

Biomimetic synthesis of hydroxyapatite/bacterial cellulose nanocomposites for biomedical applications

Hydroxyapatite (HAp) and bacterial cellulose (BC) are both excellent materials for use in biomaterial areas. The former has outstanding osteoconductivity and bioactivity and the latter is a high-strength nano-fibrous and extensively used biomaterial. In this work, the HAp/BC nanocomposites with a 3-dimensional (3-D) network were synthesized via a biological route by soaking both phosphorylated and unphosphorylated BCs in 1.5 simulated body fluid (SBF). Scanning electron microscopy (SEM), X-ray diffraction (XRD), Fourier transformed infrared spectroscopy (FTIR), and transmission electron microscopy (TEM) were employed to characterize the HAp/BC nanocomposites. SEM observations demonstrated that HAp crystals were uniformly formed on the phosphorylated BC fibers after soaking in 1.5 SBF whereas little HAp was observed on individual unphosphorylated BC fibers. Our experimental results suggested that the unphosphorylated BC did not induce HAp growth and that phosphorylation effectively triggered HAp formation on BC. Mechanisms were proposed for the explanation of the experimental observations. XRD and FTIR results revealed that the HAp crystals formed on the phosphorylated BC fibers were carbonate-containing with nano-sized crystallites and crystallinities less than 1%. These structural features were close to those of biological apatites.     

Laser‐Raman and Nuclear Magnetic Resonance (NMR) studies on plasma‐sprayed hydroxylapatite coatings: Influence of bioinert bond coats on phase composition and resorption kinetics in simulated body fluid

The structure and phase composition of HAp coatings deposited onto Ti6Al4V coupons (50x20x2mm) by atmospheric plasma spraying (APS) were studied by laser‐Raman spectroscopy, ³¹P‐ and ¹H‐MAS‐NMR and 2D‐³¹P/¹H HETCOR‐CP‐NMR spectroscopy, and XRD with Rietveld refinement. The samples investigated comprised APS HAp coatings with and without a TiO₂ bond coat as well as coatings incubated for different lengths of time (up to 12 weeks) in simulated body fluid (SBF) under physiological conditions. In APS coatings the presence of a bond coat increased the proportion of well‐ordered crystalline HAp at the expense of distorted apatite‐like structures such as oxyHAp and oxyapatite, and thermal decomposition products such as tricalcium phosphate (TCP) and tetracalcium phosphate (TTCP), and also decreased the amount of amorphous calcium phosphate (ACP). Incubation in SBF further advanced the proportion of crystalline HAp since the disordered structures, the thermal decomposition products, and ACP exhibit substantially higher solubility.     

Growth of calcium phosphate on ion-exchange resins pre-saturated with calcium or hydrogenphosphate ions: an SEM/EDX and XPS study

Calcium phosphate formed on the surfaces of ion-exchange resins pre-saturated with either Ca²⁺ or HPO₄ ^2- ions has been studied using a combination of scanning electron microscopy (SEM)/energy dispersive X-ray (EDX) analysis and X-ray photoelectron spectroscopy (XPS). Calcium phosphate was formed at a temperature of 36.5°C via two methods. On Ca²⁺ or HPO₄ ^2--saturated resins, 1.5xSBF (simulated body fluid) solution was used while on Ca²⁺-saturated resins only, a novel biomimetic growth medium using the alkaline phosphatase-catalysed hydrolysis reaction of disodium p-nitrophenylphosphate as a source of inorganic phosphate was employed. SEM micrographs showed that the use of 1.5xSBF growth medium solution led to extensive coverage of the resins with calcium phosphate. In contrast, calcium phosphate coatings formed via the alkaline phosphatase-catalysed reaction were of a more variable quality whose morphology could be influenced by adding albumin and collagen to the growth medium. Average Ca:P ratios determined by EDX for coatings formed from the 1.5xSBF growth medium were in the range 1.62–1.74 suggesting that hydroxyapatite had formed. In contrast, Ca:P ratios for the calcium phosphate compounds formed on resins from the alkaline phosphatase reaction were lower at 1.50 suggesting that calcium-deficient hydroxyapatite had formed which was confirmed by inductively coupled plasma (ICP) analysis and X-ray diffraction of isolated amorphous and crystallized powder samples, respectively. Evidence from X-ray photoelectron studies supports a mechanism of formation of the coatings which involves diffusion of the ion out of the interior of the resin to create a high local concentration at the surface thus stimulating precipitation of the coating material on the resin beads.     

Biomimetic mineralisation of apatites on Ca2+ activated cellulose templates

We investigated the activation of regenerated cellulose 2D model thin films and 3D fabric templates with calcium dihydroxide. The Langmuir–Blodgett (LB) film technique was applied for manufacturing of the model thin films using a trimethylsilyl derivative of cellulose (TMS-cellulose). Regenerated cellulose films were obtained by treating the TMS-cellulose LB-films with hydrochloric acid vapours. For 3D templates, regenerated cellulose fabrics (Lyocell®) were used. The regenerated cellulose templates were activated with a Ca(OH)₂-suspension and subsequently exposed to 1.5 × SBF to induce the in situ formation of biomimetic calcium phosphate phases. FTIR and Raman spectroscopy showed that the Ca(OH)₂ and calcite present from reaction with HCO₃⁻ on the template surface were dissolved in the initial stage of exposure to the 1.5 × SBF. After 1 day, the formation of apatitic phases in 1.5 × SBF was observed. According to detailed calculations, high supersaturation levels S in close vicinity to the template surface (S > 16) resulting from the Ca²⁺ diffusion induced the formation of biomimetic calcium phosphate. The biomimetic calcium phosphates were identified by FTIR and Raman spectroscopy as highly carbonated apatites (HCA) lacking hydroxyl ions. 3D fabric templates of regenerated cellulose covered with a biomimetic coating of apatite might be of particular interest for novel scaffold architectures in bone repair and tissue engineering.     

Biomimetic calcium phosphate coatings on nitric-acid-treated titanium surfaces

This study describes biomimetic calcium phosphate (Ca-P) coatings formation under simulated physiological conditions on Ti surfaces that go through nitric acid treatment (NT). In the present study, nitric acid treatment was used to treat Ti specimens so that Ti specimens could have the ability to induce Ca-P formation. After careful selection of the NT parameters, Ca-P coatings success fully formed on the nitric-acid-treated Ti surfaces in a supersaturated calcium phosphate solution (SCPS) and in the simulated body fluid (SBF). Before NT, the Ti specimen should go through mixed acid etching to increase its surface roughness because rough surfaces lead to good adherence between coatings and substrates. Amorphous Ca-P coatings were formed on the Ti surfaces by immersing the NT Ti specimens in SBF, while octacalcium phosphate (OCP) coatings were formed in the SCPS after 3 days of immersion. The study firstly proved that nitric acid treatment is not only just for surface passivation but also is another bioactive treatment as an alternative to the alkaline treatment and two-step method. The experimental results also confirmed that the conventional nitric acid treatment of a titanium surface does not increase the titanium oxide on the Ti surfaces. However, extending the nitric acid treatment time and enhancing the nitric acid treatment temperature help to increase Ti surface ability of Ca-P induction in simulated physiological environments. Ti specimens that had 600 min of NT at 60 °C had the best Ca-P induction ability under biomimetic conditions.