Biocompatibility of electrophoretical deposition of nanostructured hydroxyapatite coating on roughen titanium surface: in vitro evaluation using mesenchymal stem cells

A nano hydroxyapatite (HAp) layer was coated on a roughen titanium surface by means of electrophoretic deposition with an acetic anhydride solvent system. The objectives of this current study are to investigate whether nano-HAp can improve mechanical strength at a lower sintering temperature and biocompatibility. Densification temperature was lowered from usual 1000 to 800 degrees C. The coating interfacial bonding strength, phase purity, microstructure, and biocompatibility were investigated. Degradation of HA phase was not detected in XRD. A porous TiO2 layer acts as a gradient coating layer with an intermediate thermal expansion coefficient between hydroxyapatite and titanium that reduces the thermal stress. From SEM image, the coating does not contain any crack. Mesenchymal stem cell (MSC) is the progenitor cell for various tissues in mature animals, which can improve integration of bone tissue into implant. In this in vitro study, rabbit MSCs culture indicated that the HAp/Ti nanocomposite biomaterial had good biocompatibility and bioactivity. Around materials and on its surface cell grew well with good morphology. Proliferation of the MSCs on the nano-HAp coating was higher than its micron counterpart in XTT assay. These properties show potential for the orthopaedic and dental applications.     

Effect of ring-opening polymerization condition on the characteristic and mechanical properties of hydroxyapatite/poly(ethylene glutarate) biomaterials

Preparation of hydroxyapatite/poly(ethylene glutarate) (HAp/PEG) composites was carried out by ring-opening polymerization (ROP) of cyclic oligo(ethylene glutarate) in porous HAp scaffolds using various reaction temperatures and times. The content of ROP-PEG interpenetrated into the porous HAp scaffold was about 13-18 wt % with the values of number average molecular weight (overline_M{n}) and weight average molecular weight (overline_M{W}) of 2120-3630 and 2760-5250 g/mol, respectively. The increase in polymerization time and temperature brought about increase in molecular weight of ROP-PEG, but decrease in its content. Compressive strength and compressive modulus of the HAp/PEG composites were about 5.8-20.1 and 105-208 MPa, respectively. These mechanical properties depend upon the effects of distribution, content, and molecular weight of ROP-PEG in the composites. In vitro bioactivity of the HAp/PEG composites was studied by soaking them in simulated body fluid (SBF) for 28 days. The formation of HAp nanocrystal on the composite surfaces through the consumption of calcium and phosphorus from the SBF solution was observed after soaking, indicating the bioactivity of these HAp/PEG composites.     

Fabrication and cellular biocompatibility of porous carbonated biphasic calcium phosphate ceramics with a nanostructure

Microwave heating was applied to fabricate interconnective porous structured bodies by foaming as-synthesized calcium-deficient hydroxyapatite (Ca-deficient HA) precipitate containing H(2)O(2). The porous bodies were sintered by a microwave process with activated carbon as the embedding material to prepare nano- and submicron-structured ceramics. By comparison, conventional sintering was used to produce microstructured ceramics. The precursor particles and bulk ceramics were characterized by transmission electron microscopy (TEM), dynamic light scattering, scanning electron microscopy (SEM), X-ray diffraction (XRD), Fourier-transformed infrared spectroscopy (FTIR) and mechanical testing. TEM micrographs and assessment of the size distribution showed that the needle-like precursor particles are on the nanoscale. SEM observation indicated that the ceramics formed by microwave sintering presented a structure of interconnective pores, with average grain sizes of approximately 86 and approximately 167nm. XRD patterns and FTIR spectra confirmed the presence of carbonated biphasic calcium phosphate (BCP), and the mechanical tests showed that the ceramics formed by microwave sintering had a compressive strength comparable to that obtained by conventional methods. Rat osteoblasts were cultured on the three kinds of BCP ceramics to evaluate their biocompatibility. Compared with the microscale group formed by conventional sintering, MTT assay and ALP assay showed that nanophase scaffolds promoted cell proliferation and differentiation respectively, and SEM observation showed that the nanoscale group clearly promoted cell adhesion. The results from this study suggest that porous carbonated biphasic calcium phosphate ceramics with a nanostructure promote osteoblast adhesion, proliferation and differentiation. In conclusion, porous carbonated BCP ceramics with a nanostructure are simple and quick to prepare using microwaves and compared with those produced by conventional sintering, may be better bone graft materials.     

Compositional and microstructural changes of engineered plasma-sprayed hydroxyapatite coatings on Ti6Al4V substrates during incubation in protein-free simulated body fluid

Hydroxyapatite (HAp) coatings engineered for maximum surface roughness (coating type I), porosity (coating type II), and tensile adhesion strength (coating type III) were deposited by atmospheric plasma spraying (APS) onto Ti6Al4V substrates and characterized for their microstructure, phase composition, and design properties. The composition of the as-sprayed coatings changed during treatment with protein-free simulated body fluid (Hank's Balanced Salt Solution, HBSS) for up to 12 weeks by preferential dissolution of thermal decomposition products, and amorphous calcium phosphate (ACP). From solutions supersaturated with respect to calcium and phosphorus ions, a thin, very porous layer precipitated onto the leached surfaces of coating type II samples after an incubation time of 8 weeks, consisting of spherical agglomerates of a poorly crystallized bone-like Ca-deficient defect hydroxyapatite that is thought to accelerate in vivo bone apposition rates and, hence, may induce favorable osseoconductive conditions.     

Novel porous hydroxyapatite prepared by combining H2O2 foaming with PU sponge and modified with PLGA and bioactive glass

Porous hydroxyapatite (HA) scaffolds have been intensively studied and developed for bone tissue engineering, but their mechanical properties remain to be improved. The aim of this study is to prepare HA-based composite scaffolds that have a unique macroporous structure and special struts of a polymer/ceramic interpenetrating composite and a bioactive coating. A novel combination of a polyurethane (PU) foam method and a hydrogen peroxide (H(2)O( 2)) foaming method is used to fabricate the macroporous HA scaffolds. Micropores are present in the resulting porous HA ceramics after the unusual sintering of a common calcium phosphate cement and are infiltrated with the poly(D,L-lactic-co-glycolic acid) (PLGA) polymer. The internal surfaces of the macropores are further coated with a PLGA-bioactive glass composite coating. The porous composite scaffolds are characterized in terms of microstructure, mechanical properties, and bioactivity. It is found that the HA scaffolds fabricated by the combined method show high porosities of 61-65% and proper macropore sizes of 200-600 microm. The PLGA infiltration improved the compressive strengths of the scaffolds from 1.5-1.8 to 4.0-5.8 MPa. Furthermore, the bioactive glass-PLGA coating rendered a good bioactivity to the composites, evidenced by the formation of an apatite layer on the sample surfaces immersed in the simulated body fluid (SBF) for 5 days. The porous HA-based composites obtained from this study have suitable porous structures, proper mechanical properties, and a high bioactivity, and thus finds potential application as scaffolds for bone tissue engineering.     

Phase composition and in vitro bioactivity of porous implants made of bioactive glass S53P4

This work studied the influence of sintering temperature on the phase composition, compression strength and in vitro properties of implants made of bioactive glass S53P4. The implants were sintered within the temperature range 600-1000°C. Over the whole temperature range studied, consolidation took place mainly via viscous flow sintering, even though there was partial surface crystallization. The mechanical strength of the implants was low but increased with the sintering temperature, from 0.7 MPa at 635°C to 10 MPa at 1000°C. Changes in the composition of simulated body fluid (SBF), the immersion solution, were evaluated by pH measurements and ion analysis using inductively coupled plasma optical emission spectrometry. The development of a calcium phosphate layer on the implant surfaces was verified using scanning electron microscopy-electron-dispersive X-ray analysis. When immersed in SBF, a calcium phosphate layer formed on all the samples, but the structure of this layer was affected by the surface crystalline phases. Hydroxyapatite formed more readily on amorphous and partially crystalline implants containing both primary Na(2)O·CaO·2SiO(2) and secondary Na(2)Ca(4)(PO(4))(2)SiO(4) crystals than on implants containing only primary crystals.     

Preparation and characterization of hydroxyapatite/poly(ethylene glutarate) biomaterials

Hydroxyapatite/poly(ethylene glutarate) (HAp/PEG) biomaterial composites were prepared by ring-opening polymerization (ROP) of cyclic oligo(ethylene glutarate) (C-PEG) in porous HAp scaffolds. The HAp/C-PEG precomposites were prepared by immersing the porous HAp scaffolds in the mixture solution of C-PEG and dibutyl tinoxide catalyst overnight and polymerizing at 200 degrees C for 24, 48, and 72 h under vacuum. The successful ROP of C-PEG in the porous HAp scaffolds was corroborated by the signals of hydroxyl end-group of PEG shown in the (1)H NMR spectrum of the ROP-products extracted from the composites. PEG in the composites was present as a thin layer coating on the HAp grains and was evenly distributed throughout the samples. The PEG content was about 13-16 wt % and decreased with increasing polymerization time. Its molecular weight (M(w), weight average) measured by gel permeation chromatography was in the range of 4300-6800 g/mol. Compressive strength of the HAp/PEG composites was significantly increased from 3 MPa of the porous HAp scaffold to 11-15 MPa, depending on the PEG content in the composites. In vitro bioactivity of the HAp/PEG composites was studied by soaking in simulated body fluid (SBF) at 36.5 degrees C for 7-28 days. After prolonged soaking, the HAp nanocrystals precipitated from the SBF solution and formed as a layer of globular aggregates, coated on the composite surfaces. This result suggested that the HAp/PEG composite was a bioactive material.     

In vivo response of porous hydroxyapatite and beta-tricalcium phosphate prepared by aqueous solution combustion method and comparison with bioglass scaffolds

Pure hydroxyapatite (HAp) and a biphasic calcium phosphate [containing 90% of beta-tri-calcium phosphate (beta-TCP) and 10% HAp] were tailored through an aqueous solution combustion synthesis. Porous struts were prepared using all the powders along with bioglass, a known bioactive material, and subsequently characterized. Sterilized struts were implanted to the lateral side of radius bone of 24 black Bengal goats of either sex, in which a blank hole was left unfilled in a group of six specimens to act as control. The bone formation response of the three implanting materials in vivo has been studied using scanning electron microscope and histological analysis in contrast with positive controls. Push-out tests were used to assess the mechanical strength at the bone-biomaterial interface. It was observed that interfacial response was strongly dependent on combinations of different physical and chemical parameters. The surface of beta-TCP exhibited similar characteristics of bone and was distinct from those of intervening apatite layer of bioglass. Lower bone ingrowth and reduced strength was observed with HAp compared to beta-TCP/bioglass-based implants. Bone formation response of the Ca-P material varied according to the composition of the implanting material, which could be tailored through this novel synthesis.     

Activity of plasma sprayed yttria stabilized zirconia reinforced hydroxyapatite/Ti-6Al-4V composite coatings in simulated body fluid

Hydroxyapatite (HA)/yttria stabilized zirconia/Ti-6Al-4V bio-composite coatings deposited onto Ti-6Al-4V substrate through a plasma spray technique were immersed in simulated body fluid (SBF) to investigate their behavior in vitro. Surface morphologies and structural changes in the coatings were analyzed by scanning electron microscopy, thin-film X-ray diffractometer, and X-ray photoelectron spectroscopy. The tensile bond strength of the coatings after immersion was also conducted through the ASTM C-633 standard for thermal sprayed coatings. Results showed that carbonate-containing hydroxyapatite (CHA) layer formed on the surface of composite coatings after 4 weeks immersion in SBF solution, indicating the composite coating possessed excellent bioactivity. The mechanical properties were found to decrease with immersion duration of maximum 56 days. However, minimal variation in mechanical properties was found subsequent to achieving supersaturation of the calcium ions, which was attained with the precipitation of the calcium phosphate layers. The mechanical properties of the composite coating were found to be significantly higher than those of pure HA coatings even after immersion in the SBF solution, indicating the enhanced mechanical properties of the composite coatings.     

Poly(D,L-lactic acid) coated 45S5 Bioglass-based scaffolds: processing and characterization

A comparative investigation has been carried out on the mechanical properties and bioactivity of Bioglass-based foams, before and after applying a poly(D,L-lactic acid) (PDLLA) coating layer on the foam struts. It was found that the bioactivity of foams upon immersion in simulated body fluid (SBF) was maintained in the PDLLA-coated foams; however, the transformation kinetics in SBF of the crystalline phase (Na(2)Ca(2)Si(3)O(9)) in the foam struts to an amorphous calcium phosphate phase was retarded by PDLLA coating. The compressive and three-point bending strengths of the Bioglass-based foams were slightly improved by the PDLLA-coating, and the work-of-fracture of the foams was considerably enhanced, as indicated by stress-strain curves. Immersion in SBF for 4 weeks led to a large decrease of the mechanical strength of as-sintered foams decreased (from 0.3 to 0.03 MPa), because of the transformation of the crystalline phase to an amorphous calcium phosphate. On the other hand, the mechanical strength was well-maintained in PDLLA-coated foams after immersion in SBF for 8 weeks. This behavior was attributed to the in-situ formation of a nanocomposite PDLLA/calcium phosphate film on the strut surfaces upon immersion in SBF.     

Preparation and characterization of hydroxyapatite/poly(ethylene adipate) hybrid composites

Hydroxyapatite/poly(ethylene adipate) (HAp/PEA) composites were prepared by in situ ring-opening polymerization of cyclic oligo(ethylene adipate) (C-OEA) within the porous HAp templates. HAp was firstly prepared by a co-precipitation method using calcium hydroxide and phosphoric acid and then shaped as a rectangular porous template. PEA precursor was synthesized by bulk polymerization of dimethyl adipate and ethylene glycol in the presence of tetraisopropyl orthotitanate. C-OEA was obtained by cyclo-depolymerization of the PEA precursor under high dilution condition using dibutyl tinoxide as a catalyst. The HAp/PEA composites were prepared by immersing the porous HAp templates in the mixture solution of C-OEA and dibutyl tinoxide catalyst overnight and ring-opening polymerizing at 180, 200 and 220 degrees C for 24 h. The ring-opening polymerized PEA formed as a thin film coating on the surface of porous HAp template. The HAp/PEA composites contained PEA in the range of 20-26 wt%. The weight-average molecular weights of ring-opening polymerized PEA were in the range of 3800-4450 g/mol. Compressive strength of the HAp/PEA composite was significantly increased from 25 MPa in the porous HAp template to 140 MPa in the composite.     

Conversion of sea urchin spines to Mg-substituted tricalcium phosphate for bone implants

The skeleton of sea urchin spines is composed of large single crystals of Mg-rich calcite, which have smooth, continuously curved surfaces and form a three-dimensional fenestrated mineral network. Spines of the echinoids Heterocentrotus trigonarius and Heterocentrotus mammillatus were converted by the hydrothermal reaction at 180 degrees C to bioresorbable Mg-substituted tricalcium phosphate (beta-TCMP). Due to the presence of Mg in the calcite lattice, conversion to beta-TCMP occurs preferentially to hydroxyapatite formation. The converted beta-TCMP still maintains the three-dimensional interconnected porous structures of the original spine. The main conversion mechanism is the ion-exchange reaction, although there is also a dissolution-reprecipitation process that forms some calcium phosphate precipitates on the surfaces of the spine network. The average fracture strength of urchin spines and converted spines (beta-TCMP) in the compression tests are 42 and 23MPa, respectively. In vivo studies using a rat model demonstrated new bone growth up to and around the beta-TCMP implants after implantation in rat femoral defects for 6 weeks. Some new bone was found to migrate through the spine structural pores, starting from the outside of the implant through the pores at the edge of the implants. These results indicate good bioactivity and osteoconductivity of the porous beta-TCMP implants.     

A novel akermanite bioceramic: preparation and characteristics

Akermanite (Ca(2)MgSi(2)O(7)) ceramics are prepared by sintering akermanite powder compacts at 1370 C for 6 h. The sintering behavior and mechanical properties of akermanite ceramics are investigated. The bioactivity of akermanite ceramics is evaluated by soaking in simulated body fluid (SBF), and hydroxyapatite (HAp) formation on the surface of akermanite ceramics after soaking is characterized by X-ray diffraction (XRD), energy dispersive spectrometry (EDS), and scanning electron microscopy (SEM). The results show that the bending strength of akermanite ceramics can reach 176 MPa, the fracture toughness is 1.83 MPa m(1/2), and akermanite ceramics can induce HAp formation on their surface when soaked in SBF. Our results indicate that akermanite ceramics are bioactive, and possess improved mechanical properties compared with those of HAp ceramics and may be used as bioactive implant materials.     

On the feasibility of phosphate glass and hydroxyapatite engineered coating on titanium

In this report, bioactive calcium phosphate (CaP) coatings were produced on titanium (Ti) by using phosphate-based glass (P-glass) and hydroxyapatite (HA), and their feasibility for hard tissue applications was addressed in vitro. P-glass and HA composite slurries were coated on Ti under mild heat treatment conditions to form a porous thick layer, and then the micropores were filled in with an HA sol-gel precursor to produce a dense layer. The resultant coating product was composed of HA and calcium phosphate glass ceramics, such as tricalcium phosphate (TCP) and calcium pyrophosphate (CPP). The coating layer had a thickness of approximately 30-40 microm and adhered to the Ti substrate tightly. The adhesion strength of the coating layer on Ti was as high as 30-33 MPa. The human osteoblastic cells cultured on the coatings produced by the combined method attached and proliferated favorably. Moreover, the cells on the coatings expressed significantly higher alkaline phosphatase activity than those on pure Ti, suggesting the stimulation of the osteoblastic activity on the coatings. On the basis of these observations, the engineered CaP coating layer is considered to be potentially applicable as a hard tissue-coating system on Ti-based implants.     

Calcium phosphate ceramic coatings on porous titanium: effect of structure and composition on electrophoretic deposition, vacuum sintering and in vitro dissolution

Bioactive calcium phosphate ceramics (CPC) guide bone formation along their surface. This property is conceptually attractive from the viewpoint of enhancing early bone tissue formation in porous metal coatings. The various studies conducted to exploit this idea, however, reveal a considerable variability of the effect. This suggests material- and processing-induced parametric influences. Thus this study focuses on the formulation of model porous metal-CPC materials for use in one-parametric analyses of material factors. Easily reproducible, porous metals with a uniform porous structure and CPC coating are made with orderly oriented wire mesh (OOWM) porous metal coatings and electrophoretically deposited CPC films. The deposition of the ceramic can be hampered by adsorbed water. Subsequent vacuum sintering leads to several phase transformations: hydroxyapatite is transformed to a mixture of oxyhydroxyapatite and tetracalcium phosphate; the underlying titanium promotes the beta- to alpha-tricalcium phosphate transformation; and Ca-deficient hydroxyapatite is transformed to a mixture containing oxyhydroxyapatite and alpha- and beta-tricalcium phosphate. These phase transformations provoke a considerable increase of in vitro dissolution in 0.05 M tris buffered physiological solution.     

Calcium phosphate-coated porous titanium implants for enhanced skeletal fixation

Porous titanium fiber implants for cementless skeletal fixation by bone ingrowth were treated with a calcium phosphate coating applied by a plasma flame-spray technique. In a paired experiment, treated and control implants were inserted in the humeri and olecranons of 36 adult dogs for periods of 1, 2, 4, and 6 weeks. After the animals were sacrificed, a biomechanical evaluation of the strength of skeletal fixation of the implants and a histologic evaluation of bone ingrowth was done. The mean shear strength of skeletal fixation at four weeks for the calcium phosphate-coated implants was 24% greater (P less than .01) than for paired controls. No difference in strength of fixation between treated and control implants was present at other time periods. The osteoconductive properties of the ceramic coating were demonstrated by bone forming in direct contact with the calcium phosphate coating on the metal fibers of the treated implants. No significant increase for the volume of bone ingrowth was established for treated implants compared to paired controls at any time period.     

Strengthening of bone-implant interface by the use of granule coatings on alumina ceramics

Three different granule coatings (a granular alumina ceramic coating, a granular hydroxyapatite coating, and a polished granular hydroxyapatite coating) applied to alumina ceramic substrate were evaluated for their strengthening effects of the bone-implant interface in rabbit tibiae. For a comparison, noncoated alumina ceramics, and dense hydroxyapatite were assessed in the same way. The granular alumina ceramic coating, creating a bioinert, porous surface, was most effective due to a strong mechanical bond between the bone and implant. The interface strength was even higher than that of the dense hydroxyapatite. The granular hydroxyapatite coating, creating a bioactive, porous surface, was less effective than the granular alumina ceramic coating because of the brittleness of the hydroxyapatite granules, although it formed a direct and mechanical bond with bone tissue. The polished granular hydroxyapatite coating, creating a bioactive, smooth surface, was least effective because of the brittleness of the hydroxyapatite granules, though it presented an improved interface strength compared with that of the noncoated alumina ceramics due to a direct bond between the bone and hydroxyapatite granules.     

Synthesis and characterization of macroporous chitosan/calcium phosphate composite scaffolds for tissue engineering

Chitosan scaffolds reinforced by beta-tricalcium phosphate (beta-TCP) and calcium phosphate invert glass were fabricated with a low-cost, bioclean freeze-drying technique via thermally induced phase separation. The microstructure, mechanical performance, biodegradation, and bioactivity of the scaffolds were studied. The composite scaffolds were macroporous, and the pore structures of the scaffolds with beta-TCP and the glass appeared very different. Both the compressive modulus and yield strength of the scaffolds were greatly improved, and reinforced microstructures were achieved. The bioactivity tests showed a continuous decrease in both Ca and P concentrations of a simulated body fluid (SBF) after the scaffolds with beta-TCP were immersed in the SBF for more than 20 h, which suggests that an apatite layer might be formed on the scaffolds. However, the same was not observed for the pure chitosan scaffolds or the scaffolds incorporated with the glass. This was further confirmed by micrographs from scanning electron microscopy. This study suggests that the desirable pore structure, biodegradation rate, and bioactivity of the composite scaffolds might be achieved through controlling the ratio of chitosan and calcium phosphates or beta-TCP and the glass.     

Hydroxylapatite coating of porous implants improves bone ingrowth and interface attachment strength.

The effect of a plasma-sprayed hydroxylapatite (HA) coating on the degree of bone ingrowth and interface shear attachment strength was investigated using a canine femoral transcortical implant model. Cylindrical implants were fabricated by sintering spherical Co-Cr-Mo particles 500-710 microns in diameter; the nominal implant dimensions were 5.95 +/- 0.05 mm diameter by 18 mm in length. One half of each implant was coated with hydroxylapatite, 25-30 microns in thickness, by a plasma-spray technique. Using strict aseptic technique, the implants were placed through both femoral cortices into defects approximately 0.05 mm undersized. After 2, 4, 6, 8, 12, 18, 26, and 52 weeks, the implants were harvested and subjected to mechanical pullout testing and undecalcified histologic evaluation. The application of the HA coating to porous implants enhanced both the amount of bone ingrowth and the interface attachment strength at all time periods. These differences were statistically significant for the percent of bone ingrowth at the 4-, 6-, 12-, 18-, 26-, and 52-week time periods, and interface shear strength values were significantly different at the 6-, 8-, 12-, 18-, and 26-week time periods. The rate of development of interface strength and bone ingrowth was also more rapid for the HA-coated implants. No evidence of any disruption, mechanical failure, or biologic resorption of the HA coating was observed. The results of the present study--demonstrating a beneficial effect of the HA coating at all time periods--are believed to be due to the use of paired comparisons, which allow assessment of subtle differences that might otherwise have been obscured by normal biological variability.     

Biocompatibility of dense hydroxyapatite prepared using an SPS process

In the present study, the preparation of dense hydroxyapatite (HAp) materials at relative low temperatures using a spark plasma sintering was carried out. The bioactivity of HAp samples prepared by a spark plasma sintering method was investigated by in vitro tests and compared with HAp obtained by a conventional hot-pressing method. No growth of bone-like HAp crystals on surface of HAp sintered by a conventional hot-pressing method at 1200 degrees C was observed after immersion in a simulated body fluid (SBF) for 2 days. However, many large bone-like HAp crystals were observed on the surface of HAp samples prepared by a spark plasma sintering at 1200 degrees C after 2 days in the SBF immersion test. Especially, the negatively charged surface of the HAp samples prepared by spark plasma sintering was covered with larger HAp crystals compared with the positively charged surface. The electric poling of HAp was measured using a thermally simulated current technique. This rapid growth of bone-like HAp crystals of the HAp samples made by spark plasma sintering was believed to be related with the OH(-) and/or Ca(2(+) ) ion deficiency at the grain boundaries of the HAp matrix grains as well as a small electric poling effect resulting during the spark plasma sintering process.