Characterization and thermal decomposition of synthetic carbonate apatite powders prepared using different alkali metal salts.

Two types of synthetic carbonate apatite [potassium-containing carbonate apatite (CAK) and sodium-containing carbonate apatite (CANa)] were prepared and characterized by thermogravimetric analysis, X-ray diffraction analysis (XRD) and Fourier transform infrared spectroscopy. The chemical formulas of carbonate apatite were determined to be Ca9.36K0.12(PO4)5.12(CO3)0.88(OH)1.73 and Ca8.72Na1.33(PO4)4.96(CO3)1.04(OH)1.80, respectively. Thermogravimetric analysis showed that the final weight loss at 1,200°C reached about 11.2% for CAK and 13.9% for CANa. Carbonate loss gradually occurred above 150°C and continued to 1,200°C. The crystallinity of the apatite phase was found to be much improved between 800 and 850°C for CAK and 750 and 800°C for CANa, where rapid carbonate loss occurred. A small amount of CaO formed above 900°C. For CANa, NaCaPO4 also formed above 700°C in both apatites. Although the lattice parameters of the carbonate apatites varied with temperature, the final a and c lattice parameters attained constant values of 0.9421 and 0.6881 nm.

Characterization and in vitro evaluation of biphasic α-tricalcium phosphate/ß-tricalcium phosphate cement.

Biphasic calcium phosphate consisting of hydroxyapatite (HA) and ß-tricalcium phosphate(ß-TCP) is an excellent bone substitute with controllable bioresorbability. Fabrication of biphasic calcium phosphate with self-setting ability is expected to enhance its potential application as bone substitute. In this study, mixtures of α-TCP and ß-TCP with various compositions were prepared through α-ß phase transition of α-TCP powder at 1000°C for various periods. These powders were mixed with 0.25M Na2HPO4 at a P/L ratio of 2, and then hardened at 37°C at 100% RH for up to 24h. Material properties of biphasic HA/ß-TCP cement with different α-TCP/ß-TCP composition were characterized. These cements were also evaluated with respect to cell response in vitro using MC3T3-E1 cell lines. In conclusion, mechanical and biological properties of HA/ß-TCP cement could be controlled by changing the heat treatment time of α-TCP powder at 1000°C. In vitro results indicated that cell proliferation and ALP activity increased with increase ß-TCP content.

Effects of the method of apatite seed crystals addition on setting reaction of α-tricalcium phosphate based apatite cement.

Appropriate setting time is an important parameter that determines the effectiveness of apatite cement (AC) for clinical application, given the issues of crystalline inflammatory response phenomena if AC fails to set. To this end, the present study analyzes the effects of the method of apatite seed crystals addition on the setting reaction of α-tricalcium phosphate (α-TCP) based AC. Two ACs, both consisting of α-TCP and calcium deficient hydroxyapatite (cdHAp), were analyzed in this study. In one AC, cdHAp was added externally to α-TCP and this AC was abbreviated as AC(EA). In the other AC, α-TCP was partially hydrolyzed to form cdHAp on the surface of α-TCP. This AC was referred to as AC(PH). Results indicate a decrease in the setting time of both ACs with the addition of cdHAp. Among them, for the given amount of added cdHAp, AC(PH) showed relatively shorter setting time than AC(EA). Besides, the mechanical strength of the set AC(PH) was also higher than that of set AC(EA). These properties of AC(PH) were attributed to the predominant crystal growth of cdHAp in the vicinity of the α-TCP particle surface. Accordingly, it can be concluded that the partial hydrolysis of α-TCP may be a better approach to add low crystalline cdHAp onto α-TCP based AC.

Fabrication of bone cement that fully transforms to carbonate apatite.

The objective of this study was to fabricate a type of bone cement that could fully transform to carbonate apatite (CO3Ap) in physiological conditions. A combination of calcium carbonate (CaCO3) and dicalcium phosphate anhydrous was chosen as the powder phase and mixed with one of three kinds of sodium phosphate solutions: NaH2PO4, Na2HPO4, or Na3PO4. The cement that fully transformed to CO3Ap was fabricated using vaterite, instead of calcite, as a CaCO3 source. Their stability in aqueous solutions was different, regardless of the type of sodium phosphate solution. Rate of transformation to CO3Ap in descending order was Na3PO4>Na2HPO4>NaH2PO4. Transformation rate could be affected by the pH of solution. Results of this study showed that it was advantageous to use vaterite to fabricate CO3Ap-forming cement.

Fabrication of carbonate apatite blocks from set gypsum based on dissolution-precipitation reaction in phosphate-carbonate mixed solution.

Carbonate apatite (CO3Ap), fabricated by dissolution-precipitation reaction based on an appropriate precursor, is expected to be replaced by bone according to bone remodeling cycle. One of the precursor candidates is gypsum because it shows self-setting ability, which then enables it to be shaped and molded. The aim of this study, therefore, was to fabricate CO3Ap blocks from set gypsum. Set gypsum was immersed in a mixed solution of 0.4 mol/L disodium hydrogen phosphate (Na2HPO4) and 0.4 mol/L sodium hydrogen carbonate (NaHCO3) at 80-200°C for 6-48 h. Powder X-ray diffraction patterns and Fourier transform infrared spectra showed that CO3Ap block was fabricated by dissolution-precipitation reaction in Na2HPO4-NaHCO3 solution using set gypsum in 48 h when the temperature was 100°C or higher. Conversion rate to CO3Ap increased with treatment temperature. CO3Ap block containing a larger amount of carbonate was obtained when treated at lower temperature.

Effect of precursor's solubility on the mechanical property of hydroxyapatite formed by dissolution-precipitation reaction of tricalcium phosphate.

The effect of the solubility of the precursors, alpha tricalcium phosphate (α-TCP) and beta tricalcium phosphate (ß-TCP) on the mechanical strength of hydroxyapatite (HAp) bone substitute was investigated. Uniaxially pressed block starting from these precursors were treated hydrothermally with 1 mol/L of ammonia solution at 200°C for various durations. XRD analysis revealed that α-TCP block took 3 h whereas ß-TCP block took 240 h for complete transformation to HAp. The porosity of HAp obtained from ß-TCP block was found to be lower than that of HAp from α-TCP block. Diametral tensile strength of HAp from ß-TCP block showed a significantly higher value than that of HAp from α-TCP block. It is therefore concluded that solubility of precursor affects the mechanical strength of the HAp block.

Fabrication of solid and hollow carbonate apatite microspheres as bone substitutes using calcite microspheres as a precursor.

Spherical carbonate apatite (CO3Ap) microspheres approximately 1 mm in diameter were fabricated by granulation of calcium hydroxide around a core followed by carbonation and phosphatization through dissolution-precipitation reaction. CO3Ap microspheres with high uniformity could not be achieved without using a core. Solid CO3Ap microspheres were obtained using a calcite core whereas hollow CO3Ap microspheres were obtained using a NaCl core. The obtained microsphere was identified as B-type CO3Ap by Fourier transform infrared analysis and the carbonate content was approximately 7-8 wt% regardless of the type of core used for sample preparation. The mechanical strength of both the solid and hollow CO3Ap microspheres was sufficient for practical use as a bone substitute.

Fabrication of microporous calcite block from calcium hydroxide compact under carbon dioxide atmosphere at high temperature.

Effects of carbonation temperature and compacting pressure on basic properties of calcite block were studied using Ca(OH)2 compact made with 0.2-2.0 MPa and their carbonation at 200-800ºC for 1 h. Microporous calcite was obtained only when carbonated at 600ºC using Ca(OH)2 compact made with 0.2 MPa even though thermogravimetry analysis showed that calcite powder was stable up to 920ºC under CO2 atmosphere. CaO formed by carbonation at 700ºC and 800ºC is thought to be caused by the limited CO2 diffusion interior to the Ca(OH)2 compact. Also, unreacted Ca(OH)2 was found for Ca(OH)2 compact prepared with 0.5 MPa or higher pressure even when carbonated at 600ºC. As a result of high temperature carbonation, crystallite size of the calcite, 58.0 nm, was significantly larger when compared to that of calcite prepared at room temperature, 35.5 nm. Porosity and diametral tensile strength of the microporous calcite were 39.5% and 6.4 MPa.

Preparation of Sr-containing carbonate apatite as a bone substitute and its properties.

Sr-containing carbonate apatite (SrCAp) specimens of varied Sr contents, ranging from 0 to 13.3 mol%, were prepared through a phosphate treatment of set gypsum-and-carbonate mixture at 100°C for 7 days. Effects of Sr content in SrCAp on microstructure, osteoblast-like cell (MC3T3-E1) attachment and proliferation, and alkaline phosphatase (ALP) activity were evaluated. Sr(2+) ion substituted Ca(2+) ion in the apatite lattice. Carbonate content was about 9-13.6 wt%, increasing in content level as Sr content increased. Sr addition benefited cell attachment but had no significant influence on cell proliferation, although the latter was inhibited at the highest Sr content. ALP activity reached a peak in specimen containing 3.4 mol% of Sr. The present study revealed that SrCAp is a promising candidate for use as a bone substitute material with good resorbabilty and osteoconductivity.

Hydrothermal calcium modification of 316L stainless steel and its apatite forming ability in simulated body fluid.

To understand the feasibility of calcium (Ca) modification of type 316L stainless steel (316L SS) surface using hydrothermal treatment, 316L SS plates were treated hydrothermally in calcium chloride (CaCl(2)) solution. X-ray photoelectron spectroscopic analysis revealed that the surface of 316L SS plate was modified with Ca after hydrothermal treatment at 200°C. And the immobilized Ca increased with CaCl(2) concentration. However no Ca-modification was occurred for 316L SS plates treated at 100°C. When Ca-modified 316L SS plate was immersed in simulated body fluid (SBF) with ion concentrations nearly equal to those of human blood plasma, low crystalline apatite was precipitated on its surface whereas no precipitate was observed on non Ca-modified 316L SS. The results obtained in the present study indicated that hydrothermal treatment at 200°C in CaCl(2) solution is useful for Ca-modification of 316L SS, and Ca-modification plays important role for apatite precipitation in SBF.

Fabrication of low-crystalline carbonate apatite foam bone replacement based on phase transformation of calcite foam.

Carbonate apatite (CO(3)Ap) foam may be an ideal bone substitute as it is sidelined to cancellous bone with respect to its chemical composition and structure. However, CO(3)Ap foam fabricated using α-tricalcium phosphate foam showed limited mechanical strength. In the present study, feasibility of the fabrication of calcite which could be a precursor of CO(3)Ap was studied. Calcite foam was successfully fabricated by the so-called "ceramic foam" method using calcium hydroxide coated polyurethane foam under CO(2)+O(2) atmosphere. Then the calcite foam was immersed in Na(2)HPO(4) aqueous solution for phase transformation based on dissolution-precipitation reaction. When CaO-free calcite foam was immersed in Na(2)HPO(4) solution, low-crystalline CO(3)Ap foam with 93-96% porosity and fully interconnected porous structure was fabricated. The compressive strength of the foam was 25.6 ± 6 kPa. In light of these results, we concluded that the properties of the precursor foam were key factors for the fabrication of CO(3)Ap foams.

Tissue-response to calcium-bonded titanium surface.

Our previous study demonstrated that calcium-bonded titanium surface (Ca-Ti) can be obtained by hydrothermal reaction between titanium (Ti) and CaCl(2) and that bone-apatite like formation was observed after immersion in simulated body fluid. The purpose of the study was to determine the in vivo response to Ca-Ti surface using a rodent tibia model. Cylinders of commercially pure Ti were divided into three groups: (1) untreated group; (2) NaOH+hTi group: soaked in 5 mol/L NaOH solution at 60 degrees C then heated at 400 degrees C for 1 h; and (3) Ca-Ti group: hydrothermally treated in the presence of 10 mmol/L CaCl(2) at 200 degrees C for 24 h. The cylinders implanted in surgically created defects in tibias of 8-week old male Wistar rats were retrieved after 1, 2, and 4 weeks. Histomorphometric evaluations were made on stained decalcified thin sections. Results showed that at 1, 2, and 4 week after implantation, respectively, bone contact was 55.2 +/- 16.4%, 88.1 +/- 9.9%, and 96.1 +/- 4.8% for Ca-Ti implants, 5.7 +/- 5.3%, 19.9 +/- 1.2%, 57.4 +/- 4.8% for untreated; and 27.2 +/- 0.7%, 70.9 +/- 7.7%, and 96.0 +/- 5.1% for NaOH+hTi implants. These results suggest that hydrothermal treatment with CaCl(2) provides a bioactive Ca-Ti bonded surface that allows bone formation greater than that obtained with NaOH+heat treated Ti surfaces.

Fabrication of carbonate apatite block based on internal dissolution-precipitation reaction of dicalcium phosphate and calcium carbonate.

In this study, we investigated a novel method for fabrication of carbonate apatite block without ionic movement between precursor and solution by using precursor that includes all constituent ions of carbonate apatite. A powder mixture prepared from dicalcium phosphate anhydrous and calcite at appropriate Ca/P ratios (1.5, 1.67, and 1.8) was used as starting material. For preparation of specimens, the slurry made from the powder mixture and distilled water was packed in a split stainless steel mold and heat - treated, ranging from 60 degrees C to 100 degrees C up to 48 hours at 100% humidity. It appeared that carbonate apatite could be obtained above 70 degrees C and monophasic carbonate apatite could be obtained from the powder mixture at Ca/P ratio of 1.67. Carbonate content of the specimen was about 5-7%. Diametral tensile strength of the carbonate apatite blocks slightly decreased with increasing treatment temperature. The decrease in diametral tensile strength is thought to be related to the crystal size of the carbonate apatite formed.

Effects of sintering temperature on physical and compositional properties of alpha-tricalcium phosphate foam.

Effects of sintering temperature on the physical and compositional properties of alpha-TCP foam fabricated using the polyurethane foam method were examined. When a polyurethane foam coated with alpha-TCP slurry was sintered at 1,400-1,550 degrees C, alpha-TCP foam having basically the same fully interconnected porous structure was produced although shrinkage occurred with increasing sintering temperature. On porosity of the alpha-TCP foam, a higher foam porosity of 95% was obtained when sintered at 1,400 degrees C as compared to the 90% porosity obtained at a higher sintering temperature of 1,550 degrees C. Further, at 1,500 degrees C or higher temperature, frame became dense with disappearance of micropores. On compressive strength, it increased from approximately 50 to 250 kPa when sintering temperature was increased from 1,400 to 1,550 degrees C. Nonetheless, no compositional changes were observed even when the alpha-TCP foam was cooled in the furnace without quenching process. In light of the results obtained, it was concluded that alpha-TCP foam fabricated using the polyurethane method was useful as a bone substitute and/or scaffolding material for tissue engineering. Besides, alpha-TCP foam could be useful as a precursor for the fabrication of other calcium phosphate foams.

Effects of added mannitol on the setting reaction and mechanical strength of apatite cement.

Apatite cement containing porogen can be a useful material for the fabrication of biporous (macro- and microporous) apatite, which has gained much attention as a bone substitute material because of its large surface area and that it improves cell penetration. In the present study, the effects of added mannitol on the setting reaction and mechanical strength of apatite cement were evaluated. Apatite cements containing 0-40 wt% of mannitol were prepared and allowed to set in 0.9% saline kept at 37 degrees C for 1-7 days. Although the diametral tensile strength (DTS) value increased with time, it decreased with the amount of added mannitol. SEM observation and XRD analysis revealed that mannitol had no inhibitory effect on the transformation reaction of apatite cement to apatite. It was thus concluded that mannitol was a good candidate for the fabrication of biporous apatite because it is biocompatible, exhibits satisfactory dissolution behavior, and that it caused no inhibitory effects on the compositional transformation to apatitic material.

Effect of temperature on crystallinity of carbonate apatite foam prepared from alpha-tricalcium phosphate by hydrothermal treatment.

The effect of temperature on crystallinity of carbonate apatite (CAp) foam prepared from alpha-tricalcium phosphate (alpha-TCP) foam by hydrothermal treatment was investigated in the present study. The alpha-TCP foams were prepared through a conventional sintering method using polyurethane foam as template. Then, the resultant alpha-TCP foams were hydrothermally treated with Na2CO3 aqueous solution at 100 degrees C, 150 degrees C and 200 degrees C for 72 h. After hydrothermal treatment, the cancellous bone-like macroporous structure of the alpha-TCP foams was maintained. However, microscopic morphology of the foams' frame significantly changed after the 72 h treatment period. The smooth surface of alpha-TCP foam disappeared and the whole surface was covered with plate-like deposits. The plate-like deposits treated at 150 degrees C and 200 degrees C had smooth surface while those treated at 100 degrees C were constructed from spherical particles of approximately 200 nm in diameter. The results of X-ray diffraction and Fourier transform infrared analysis showed that alpha-TCP was completely converted to CAp and the crystallinity of CAp prepared at 100 degrees C was significantly lower than those prepared at 150 degrees C and 200 degrees C. Hydrothermal treatment of alpha-TCP foam at 100 degrees C allowed the formation of low-crystalline CAp foam but complete conversion needs a longer treatment period.

Fabrication of freeform bone-filling calcium phosphate ceramics by gypsum 3D printing method.

Transformation of gypsum model fabricated by three-dimensional printing (3DP) into hydroxyapatite (HA) by treating in ammonium phosphate solution is possible. However, 3DP powder supplied by the manufacturer contains unknown additives which may be questionable for biomaterials. Accordingly, pure plaster of Paris (POP) powder was used for fabrication in the present study. For accurate fabrication, reduction of supplied binder ink to 80% of standard amount for 3DP powder supplied by the manufacturer was found to be the optimal condition for POP fabrication. Transformation from POP to HA was done by immersing into 1 mol/L ammonium phosphate solution. However, preheating of fabricated POP specimen at 200 degrees C for 30 min to change from calcium sulfate dihydrate into calcium sulfate hemihydrate could accelerate the transformation into HA effectively. To increase compressive strength, HA transformed specimen was sintering at 1150 degrees C for 3 h. The compressive strength increased four times comparing with as transformed HA specimen. However, crystal structure was transformed to beta-TCP due to the chemical reaction between the transformed HA and remained phosphate from ammonium phosphate solution at the sintering temperature. A sophisticated application of the present 3DP method to fabricate the freeform bioceramic for osseous defect was attempted, and jaw bone defect filling biomaterial of beta-TCP and scaffold with macroporous structures could be fabricated. Present 3DP method has possibility to fabricate freeform bioceramic for osseous defect or scaffold.

Fabrication of low-crystallinity hydroxyapatite foam based on the setting reaction of alpha-tricalcium phosphate foam.

Low-crystallinity hydroxyapatite (HAP) foam is an ideal material for bone substitutes and scaffolds for bone tissue regeneration, because its interconnected pores provide the space for cell growth and tissue penetration, and its composition induces excellent tissue response and good osteoconductivity. In this study, the feasibility of low-crystallinity HAP foam fabrication was evaluated based on the phase transformation reaction or the so-called dissolution-reprecipitation reaction of alpha-tricalcium phosphate (alpha-TCP) foam granules. When alpha-TCP foam granules were placed in water at 37 degrees C for 1 day, no reaction was observed. However, alpha-TCP foam granules set to form low-crystallinity HAP by treating it hydrothermally at 200 degrees C. The network of fully interconnected pores was retained, and porosity was as high as 82%. Pore size ranged from 50 to 300 mum with an average pore size of 160 mum. Compressive strength was 207 kPa. Although no setting reaction was observed at 37 degrees C, the setting reaction caused by the hydrothermal treatment of alpha-TCP foam granules allowed the fabrication of any shape of low-crystallinity HAP. Therefore, this method may be useful for the fabrication of bone substitutes and scaffolds in bone tissue regeneration.

Effects of liquid phase on basic properties of alpha-tricalcium phosphate-based apatite cement.

Effects of liquid phase on the basic properties of alpha-tricalcuim phosphate (alpha-TCP)-based cement, BIOPEX, were investigated by employing three liquid phases: distilled water, neutral sodium hydrogen phosphate solution, and succinic acid disodium salt solution containing sodium salt of chondroitin sulfate. When mixed with neutral sodium hydrogen phosphate or succinic acid disodium salt solution, the initial setting times of the cement were 19.4 +/- 0.55 and 11.8 +/- 0.45 minutes respectively. These setting times were much shorter than that of distilled water, 88.4 +/- 0.55 minutes. Formation of needle-like crystals typical of apatite was much faster when neutral sodium hydrogen phosphate solution was used, as compared to distilled water or succinic acid disodium salt solution. Moreover, at 24 hours after mixing, the largest amount of apatite was formed when neutral sodium hydrogen phosphate solution was used, whereas use of succinic acid resulted in the least. On the final mechanical strength of the cement, that yielded with neutral sodium hydrogen phosphate solution was the highest. In contrast, lower mechanical strength was observed--especially at the initial stage--when succinic acid sodium salt was used. It was thus concluded that alpha-TCP-based cement allowed accelerated transformation to apatite, and that higher mechanical strength since the initial stage was achieved when neutral sodium hydrogen phosphate solution was used as the liquid phase.

Fabrication of B-type carbonate apatite blocks by the phosphorization of free-molding gypsum-calcite composite.

B-type carbonate apatite (CO3Ap) block may be an ideal artificial bone substitute because it is closer in chemical composition to bone mineral. In the present study, the feasibility to fabricate CO3Ap blocks was investigated using compositional transformation, which was based on the dissolution-precipitation reaction of a gypsum-calcite composite with free-molding behavior. For the compositional change, or phosphorization, gypsum-calcite composites of varying CaCO3 contents were immersed in 1 mol/L (NH4)3PO4 aqueous solution at 100 degrees C for 24 hours. No macroscopic changes were found after the treatment, whereas microscopic change was observed at SEM level. X-ray diffraction, Fourier transform infrared spectroscopy and CHN analysis indicated that the composites were B-type CO3Ap containing approximately 6-7 wt% of CO3, a value similar to that of biological bone apatite. Diametral tensile strength of the CO3Ap block was approximately 1-3 MPa. Based on the results obtained, it was therefore concluded that gypsum-calcite was a good candidate for the fabrication of CO3Ap blocks, coupled with the advantage that the composite can be molded to any shape by virtue of the setting property of gypsum.