Initial bone tissue reactions of hydroxyapatite/collagen-(3-glycidoxypropyl)trimethoxysilane injectable bone paste.

We have previously reported that a novel bioresorbable self-setting injectable bone paste composed of hydroxyapatite/collagen bone-like nanocomposite (HAp/Col) and (3-glycidoxypropyl)trimethoxysilane (GPTMS) was successfully prepared and was replaced with new bone within 3 months of implantation in defects created in porcine tibia. In this study, the HAp/Col-GPTMS paste was implanted into bone defects in rat tibiae to investigate the initial kinetics and bone tissue response. Even though more than 35% of GPTMS molecules should be eluted rapidly from directly injected pastes according to previously reported cell culture tests, in this study, energy-dispersive X-ray spectrometry did not detect Si (GPTMS) deposition in tissues surrounding the paste at 1 day postimplantation. Further, no abnormal inflammatory responses were observed in the surrounding tissues over the test period for both directly injected and prehardened pastes. Companying these observations with the results of the previous animal test (in which the paste was fully resorbed and was substituted with new bone), the eluted GPTMS resolved in no harm in vivo from the initial to final (completely resorbed) stages. Material resorption rates calculated from X-ray microcomputed tomography (µ-CT) images decreased with increasing in GPTMS concentration. Histological observations indicated that tartrate-resistant acid phosphatase (TRAP) active cells, (assumed to be osteoclasts), exist on the periphery of pastes. This result suggested that the paste was resorbed by osteoclasts in the same way as the HAp/Col. Since a good correlation was observed between TRAP active areas in histological sections and material resorption rate calculated from µ-CT, the TRAP activity coverage ratio offers the possibility to estimate the osteoclastic resorption ratio of materials, which are replaced with bone via bone remodeling process.

Functional evaluation of mineral trioxide aggregate cement with choline dihydrogen phosphate.

To improve the cytocompatibility of mineral trioxide aggregate (MTA) cement and its ability for reparative dentin formation, the effect of adding choline dihydrogen phosphate (CDHP), which is reported to be biocompatible, to MTA cement was investigated. The L929 cell proliferation showed that the addition of CDHP improved cell viability. The addition of CDHP shortened the setting time of MTA cement, with a significant decrease in consistency above 0.4 g/mL. Diametral tensile strength of the set cement was improved by the addition of 0.4 g/mL CDHP. Solubility was judged to be within the range of clinical application. The spontaneous precipitation of low crystalline hydroxyapatite was examined by immersing the set cement in phosphate buffer saline, and it was found that the ability of the cement with 0.4 g/mL of CDHP was significantly improved compared with that of the cement without CDHP.

Physical Properties and Antimicrobial Release Ability of Gentamicin-Loaded Apatite Cement/α-TCP Composites: An In Vitro Study.

Apatite cement (AC), which has excellent osteoconductive ability, and alpha-tricalcium phosphate (α-TCP), which can be used for bone replacement, are useful bone substitute materials. The objective of this study was to clarify the physical properties and antimicrobial release ability of antibiotic-loaded AC/α-TCP composites in vitro. Gentamicin-loaded, rapid setting AC/α-TCP composites were prepared in 2 mixing ratios (10:3 and 10:6). The cement paste of AC/α-TCP composites was prepared in a plastic mold and dried in a thermostatic chamber at 37 °C and 100% relative humidity for 24 h. A diametral tensile strength test, powder X-ray diffraction analysis, and gentamicin release test were performed. The diametral tensile strengths of the AC/α-TCP composites were significantly less than that of AC alone. Powder X-ray diffraction patterns exhibited the characteristic peaks of hydroxyapatite in the AC/α-TCP composites and gentamicin-loaded AC/α-TCP composites. The concentration of the released gentamicin was maintained above the minimum inhibitory concentration of Staphylococcus aureus until Day 30 in both the gentamicin-loaded AC/α-TCP composites (10:3 and 10:6). Our results suggest that a gentamicin-loaded AC/α-TCP composite has potential as a drug delivery system. Further study is essential to investigate the antimicrobial activity and safety of the gentamicin-loaded AC/α-TCP composites in animal models.

Fabrication of bioresorbable hydroxyapatite bone grafts through the setting reaction of calcium phosphate cement.

We prepared hydroxyapatite (HAp) bone grafts by the setting reaction of calcium phosphate cement, and investigated the effects of the porosity and crystallinity on the osteoconductivity and bioresorbability. We examined the effect of the water-mixing ratio, pressure, and post-heat treatment temperature during preparation on the crystallite size and porosity of the HAp blocks. The quantity of protein adsorption increased with increasing porosity and specific surface area (SSA) of the HAp blocks, whereas the initial cell attachment was similar despite the different porosities and crystallinities. In in vitro dissolution tests with a pH 5.5 buffer, which mimics an osteoclast-created Howship's lacuna, both the porosity and SSA of the HAp blocks affected the solubility; most likely due to the increased contact area with the buffer. Thus, HAp blocks prepared by the setting reaction of calcium phosphate cement could be applicable for bioresorbable HAp bone grafts because of the high porosity and SSA.

Deletion of epithelial cell-specific p130Cas impairs the maturation stage of amelogenesis.

Amelogenesis consists of secretory, transition, maturation, and post-maturation stages, and the morphological changes of ameloblasts at each stage are closely related to their function. p130 Crk-associated substrate (Cas) is a scaffold protein that modulates essential cellular processes, including cell adhesion, cytoskeletal changes, and polarization. The expression of p130Cas was observed from the secretory stage to the maturation stage in ameloblasts. Epithelial cell-specific p130Cas-deficient (p130CasΔepi-) mice exhibited enamel hypomineralization with chalk-like white mandibular incisors in young mice and attrition in aged mouse molars. A micro-computed tomography analysis and Vickers micro-hardness testing showed thinner enamel, lower enamel mineral density and hardness in p130CasΔepi- mice in comparison to p130Casflox/flox mice. Scanning electron microscopy, and an energy dispersive X-ray spectroscopy analysis indicated the disturbance of the enamel rod structure and lower Ca and P contents in p130CasΔepi- mice, respectively. The disorganized arrangement of ameloblasts, especially in the maturation stage, was observed in p130CasΔepi- mice. Furthermore, expression levels of enamel matrix proteins, such as amelogenin and ameloblastin in the secretory stage, and functional markers, such as alkaline phosphatase and iron accumulation, and Na+/Ca2++K+-exchanger in the maturation stage were reduced in p130CasΔepi- mice. These findings suggest that p130Cas plays important roles in amelogenesis (197 words).

In vitro investigation of the cell compatibility and antibacterial properties of titanium treated with calcium and ozone.

The purpose of this study was to evaluate the surface modification of calcium ions on roughened titanium as a surface treatment of dental implants for cell attachment, growth, and initial bacterial adhesion. When a surface-roughened, pure titanium disk was immersed in a calcium chloride solution (100 mM) containing 20 ppm ozone for 24 h at 25ºC, calcium was detected on the surface by X-ray photoelectron spectroscopy. The calcium-modified, roughened titanium disk had a significantly greater concentration of the initially adhered cells as well as cells cultured over 7 days compared with titanium disks without surface modification. Furthermore, the initial bacterial adhesion on the calcium-ozone treated titanium disk was statistically less than on a pure titanium disk or titanium disk treated without ozone. Dissolved ozone was useful for modifying the surface of roughened titanium with calcium ions and the surface modification may be applicable for dental implants.

Fabrication of porous carbonate apatite granules using microfiber and its histological evaluations in rabbit calvarial bone defects.

Carbonate apatite (CO3 Ap) granules are known to show good osteoconductivity and replaced to new bone. On the other hand, it is well known that a porous structure allows bone tissue to penetrate its pores, and the optimal pore size for bone ingrowth is dependent on the composition and structure of the scaffold material. Therefore, the aim of this study was to fabricate various porous CO3 Ap granules through a two-step dissolution-precipitation reaction using CaSO4 as a precursor and 30-, 50-, 120-, and 205-µm diameter microfibers as porogen and to find the optimal pore size of CO3 Ap. Porous CO3 Ap granules were successfully fabricated with pore size 8.2-18.7% smaller than the size of the original fiber porogen. Two weeks after the reconstruction of rabbit calvarial bone defects using porous CO3 Ap granules, the largest amount of mature bone was seen to be formed inside the pores of CO3 Ap (120) [porous CO3 Ap granules made using 120-µm microfiber] followed by CO3 Ap (50) and CO3 Ap (30). At 4 and 8 weeks, no statistically significant difference was observed based on the pore size, even though largest amount of mature bone was formed in case of CO3 Ap (120). It is concluded, therefore, that the optimal pore size of the CO3 Ap is that of CO3 Ap (120), which is 85 µm.

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.

Fabrication of carbonate apatite honeycomb and its tissue response.

Carbonate apatite (CO3 Ap) block can be used as a bone substitute because it can be remodeled to new natural bone in a manner conforming with the bone remodeling process. Among the many porous structures available, honeycomb (HC) structure is advantageous for rapid replacement of CO3 Ap to bone. In this study, the feasibility to fabricate a CO3 Ap HC was studied, along with its initial tissue response in rabbit femur bone defect. First, a mixture of Ca(OH)2 and a wax-based binder was extruded from a HC mold. Then the fabricated HC was heated for binder removal and carbonation at 450°C in a mixed O2 -CO2 atmosphere, forming a CaCO3 HC. When the CaCO3 HC was immersed in 1 mol/L Na3 PO4 solution at 80°C for 7 days, its composition changed from CaCO3 to CO3 Ap, maintaining the structure of the original CaCO3 HC. Compressive strengths of the CaCO3 and CO3 Ap HCs were 65.2 ± 7.4 MPa and 88.7 ± 4.7 MPa, respectively. When the rabbit femur bone defect was reconstructed with the CO3 Ap HC, new bone penetrated the CO3 Ap HC completely. Osteoclasts and osteoblasts were found on the surface of the newly formed bone and osteocytes were also found in the newly formed bone, indicating ongoing bone remodeling. Furthermore, blood vessels were formed inside the pores of CO3 Ap HC. Therefore, CO3 Ap HC has good potential as an ideal bone substitute. © 2019 Wiley Periodicals, Inc. J Biomed Mater Res Part A: 107A: 1014-1020, 2019.

Fabrication and evaluation of interconnected porous carbonate apatite from alpha tricalcium phosphate spheres.

Carbonate apatite (CO3 Ap) blocks have attracted considerable attention as an artificial bone substitute material because CO3 Ap is a component of and shares properties with bone, including high osteoconductivity and replacement by bone similar to autografts. In this study, we fabricated an interconnected porous CO3 Ap block using α-tricalcium phosphate (TCP) spheres and evaluated the tissue response to this material in a rabbit tibial bone defect model. Interconnected porous α-TCP, the precursor of interconnected porous CO3 Ap, could not be fabricated directly by sintering α-TCP spheres. It was therefore made via a setting reaction with α-TCP spheres, yielding interconnected porous calcium-deficient hydroxyapatite that was subjected to heat treatment. Immersing the interconnected porous α-TCP in Na-CO3 -PO4 solution produced CO3 Ap, which retained the interconnected porous structure after the dissolution-precipitation reaction. The diametral tensile strength and porosity of the porous CO3 Ap were 1.8 ± 0.4 MPa and 55% ± 3.2%, respectively. Both porous and dense (control) CO3 Ap showed excellent tissue response and good osteoconductivity. At 4 weeks after surgery, approximately 15% ± 4.9% of the tibial bone defect was filled with new bone when reconstruction was performed using porous CO3 Ap; this amount was five times greater than that obtained with dense CO3 Ap. At 12 weeks after surgery, for porous CO3 Ap, approximately 47% of the defect was filled with new bone as compared to 16% for dense CO3 Ap. Thus, the interconnected porous CO3 Ap block is a promising artificial bone substitute material for the treatment of bone defects caused by large fractures or bone tumor resection. © 2018 Wiley Periodicals, Inc. J Biomed Mater Res Part B: Appl Biomater, 107B: 269-277, 2019.

Enhanced Osseointegration Capability of Poly(ether ether ketone) via Combined Phosphate and Calcium Surface-Functionalization.

Biomedical applications of poly(ether ether ketone) (PEEK) are hindered by its inherent bioinertness and lack of osseointegration capability. In the present study, to enhance osteogenic activity and, hence, the osseointegration capability of PEEK, we proposed a strategy of combined phosphate and calcium surface-functionalization, in which ozone-gas treatment and wet chemistry were used for introduction of hydroxyl groups and modification of phosphate and/or calcium, respectively. Surface functionalization significantly elevated the surface hydrophilicity without changing the surface roughness or topography. The cell study demonstrated that immobilization of phosphate or calcium increased the osteogenesis of rat mesenchymal stem cells compared with bare PEEK, including cell proliferation, alkaline phosphatase activity, and bone-like nodule formation. Interestingly, further enhancement was observed for samples co-immobilized with phosphate and calcium. Furthermore, in the animal study, phosphate and calcium co-functionalized PEEK demonstrated significantly enhanced osseointegration, as revealed by a greater direct bone-to-implant contact ratio and bond strength between the bone and implant than unfunctionalized and phosphate-functionalized PEEK, which paves the way for the orthopedic and dental application of PEEK.

Surface plasma treatment and phosphorylation enhance the biological performance of poly(ether ether ketone).

Poly(ether ether ketone) (PEEK) has emerged as an alternative endosseous material to metal implants mainly because of its lack of allergic sensitivity and radiolucency, while maintaining similar mechanical properties with bone. However, a disadvantage of PEEK is its weak osseointegration ability compared with metal implants. To overcome this, we prepared a phosphate group-modified PEEK by plasma treatment and subsequent phosphorylation reaction. Plasma treatment and phosphate modification of PEEK changed its hydrophobic surface to a hydrophilic surface while maintaining the original surface topography and roughness. Phosphate modification increased the bioactivity of rat bone marrow stromal cells (BMSCs), including proliferation, alkaline phosphatase activity, and bone-like nodule formation; however, this effect was negligible in plasma-treated PEEK. In addition, phosphate modification attenuated the phenotypic polarization of lipopolysaccharide-primed RAW264.7 macrophages to an inflammatory phenotype, based on the finding that macrophages on phosphate-modified PEEK produced decreased levels of the inflammatory cytokine and increased levels of the anti-inflammatory cytokine. Finally, in an animal study, phosphate-modified PEEK exhibited a doubled pullout force from the femur bone cavity compared with bare PEEK. Thus, we conclude that phosphate modification can significantly improves the implant-bone bonding strength of PEEK by enhancing BMSCs activity and reducing excessive inflammation.

Synergistic effect of surface phosphorylation and micro-roughness on enhanced osseointegration ability of poly(ether ether ketone) in the rabbit tibia.

This study was aimed to investigate the osseointegration ability of poly(ether ether ketone) (PEEK) implants with modified surface roughness and/or surface chemistry. The roughened surface was prepared by a sandblast method, and the phosphate groups on the substrates were modified by a two-step chemical reaction. The in vitro osteogenic activity of rat mesenchymal stem cells (MSCs) on the developed substrates was assessed by measuring cell proliferation, alkaline phosphatase activity, osteocalcin expression, and bone-like nodule formation. Surface roughening alone did not improve MSC responses. However, phosphorylation of smooth substrates increased cell responses, which were further elevated in combination with surface roughening. Moreover, in a rabbit tibia implantation model, this combined surface modification significantly enhanced the bone-to-implant contact ratio and corresponding bone-to-implant bonding strength at 4 and 8 weeks post-implantation, whereas modification of surface roughness or surface chemistry alone did not. This study demonstrates that combination of surface roughness and chemical modification on PEEK significantly promotes cell responses and osseointegration ability in a synergistic manner both in vitro and in vivo. Therefore, this is a simple and promising technique for improving the poor osseointegration ability of PEEK-based orthopedic/dental implants.

Fabrication of self-setting ß-TCP granular cement using ß-TCP granules and sodium hydrogen sulfate solution.

Bridging beta-tricalcium phosphate (ß-TCP) granules with dicalcium phosphate dihydrate (DCPD) creates a porous, interconnected ß-TCP granular cement (GC) that is useful for reconstructing bone defects: the interconnected pores can accelerate new bone ingrowth and the set cement prevents the loss of granules from the bone defect area. However, the setting time of ß-TCP GC in an acidic calcium phosphate solution is too short (<1 min) for handling in clinical applications, such as in orthopedic surgery. To address this issue, we sought to optimize the setting time of ß-TCP GC using ß-TCP granules and NaHSO4 solution, as [Formula: see text] is a known inhibitor of DCPD formation. Both DCPD and calcium sulfate dihydrate (CSD) precipitated on the surface of ß-TCP granules and bridged ß-TCP granules to one another. Increasing NaHSO4 concentration (from 0.5 mol/L to 5 mol/L) led to an increase in the amount of precipitant from 2.6 ± 0.2% to 21.6 ± 1.3% for DCPD and 1.3 ± 0.3% to 10.1 ± 0.5% for CSD. The diametral tensile strength was also increased from 0.03 ± 0.01 MPa to 2.0 ± 0.1 MPa with increasing NaHSO4 concentration. When 2 mol/L NaHSO4 solution was used as the liquid phase, setting time became 5.3 ± 0.2 min, which is suitable for handling in clinical applications to repair bone defects. In conclusion, ß-TCP GC using NaHSO4 solution as the liquid phase has good potential value as bone augmentation cement.

Physical and Histological Comparison of Hydroxyapatite, Carbonate Apatite, and ß-Tricalcium Phosphate Bone Substitutes.

Three commercially available artificial bone substitutes with different compositions, hydroxyapatite (HAp; Neobone®), carbonate apatite (CO3Ap; Cytrans®), and ß-tricalcium phosphate (ß-TCP; Cerasorb®), were compared with respect to their physical properties and tissue response to bone, using hybrid dogs. Both Neobone® (HAp) and Cerasorb® (ß-TCP) were porous, whereas Cytrans® (CO3Ap) was dense. Crystallite size and specific surface area (SSA) of Neobone® (HAp), Cytrans® (CO3Ap), and Cerasorb® (ß-TCP) were 75.4 ± 0.9 nm, 30.8 ± 0.8 nm, and 78.5 ± 7.5 nm, and 0.06 m²/g, 18.2 m²/g, and 1.0 m²/g, respectively. These values are consistent with the fact that both Neobone® (HAp) and Cerasorb® (ß-TCP) are sintered ceramics, whereas Cytrans® (CO3Ap) is fabricated in aqueous solution. Dissolution in pH 5.3 solution mimicking Howship's lacunae was fastest in CO3Ap (Cytrans®), whereas dissolution in pH 7.3 physiological solution was fastest in ß-TCP (Cerasorb®). These results indicated that CO3Ap is stable under physiological conditions and is resorbed at Howship's lacunae. Histological evaluation using hybrid dog mandible bone defect model revealed that new bone was formed from existing bone to the center of the bone defect when reconstructed with CO3Ap (Cytrans®) at week 4. The amount of bone increased at week 12, and resorption of the CO3Ap (Cytrans®) was confirmed. ß-TCP (Cerasorb®) showed limited bone formation at week 4. However, a larger amount of bone was observed at week 12. Among these three bone substitutes, CO3Ap (Cytrans®) demonstrated the highest level of new bone formation. These results indicate the possibility that bone substitutes with compositions similar to that of bone may have properties similar to those of bone.

Effect of setting atmosphere on apatite cement resorption: An in vitro and in vivo study.

OBJECTIVES: The aim of this present study was to investigate the effect of setting atmosphere on replacement of apatite cement with new bone both in vitro and in vivo. MATERIAL AND METHODS: Apatite cement consisting of an equimolar mixture of tetracalcium phosphate and anhydrous dicalcium phosphate was mixed with distilled water and allowed to set at 37 °C and 100% relative humidity under 0%, 5%, and 100% CO2 atmospheres. X-Ray diffraction and Fourier Transform Infrared Spectroscopy were employed to confirm the carbonate apatite formation. Micro-CT and histological evaluation was made at 1 and 6 month(s) using twelve 10-week-old specific-pathogen-free male Wistar rats. RESULTS: B-type carbonate apatite was found when the apatite cement was set under 100% CO2 and 5% CO2. More carbonate apatite was formed in the case of 100% CO2 when compared with 5% CO2, and none was formed under 0% CO2. Interestingly, unreacted tetracalcium phosphate was significant when apatite cement was set under 0% CO2, indicating the formation of Ca-deficient hydroxyapatite. When a bone defect of rat tibia was reconstructed in these conditions of apatite cement and sintered hydroxyapatite, replacement of the apatite cement was confirmed 6 months after implantation, whereas no replacement was observed in the case of sintered hydroxyapatite. The amount of replacement of apatite cement with bone was greater, on the order of 100% CO2 and 5% CO2, followed by 0% CO2. CONCLUSION: The results obtained in the present study demonstrated that setting atmosphere clearly plays an important role in the replacement of set apatite cement with bone.

In vivo stability evaluation of Mg substituted low crystallinity ß-tricalcium phosphate granules fabricated through dissolution-precipitation reaction for bone regeneration.

Although ß-tricalcium phosphate (ß-TCP) is widely used in clinical applications as a bone substitute owing to its positive tissue response and its ability to be replaced by new bone through a bone-remodeling process, it has the limitation of rapid resorption in vivo, which might become a reason for tissue atrophy and high crystallinity, which decrease biocompatibility. A reduction in the crystallinity might increase the biocompatibility of the bone substitute. To overcome the drawbacks of ß-TCP, decrease in crystallinity and solubility, both are required. Therefore, in this study, the feasibility of fabricating Mg substituted low crystalline ß-TCP (Mg-LC-ß-TCP) granules formed in aqueous solution was evaluated in vivo focusing long-term adsorption and bone formation in bone defects formed in the rabbit femur using sintered ß-TCP granules as a control. With Mg-LC-ß-TCP, the resorption of the substitute was suppressed, and no tissue atrophy was observed even at 24 weeks post-implantation, whereas a few granules with surrounding tissue atrophy were observed at 12 weeks post-implantation. Tartrate-resistant acid phosphatase-staining indicated that the density of osteoclasts type cells with Mg-LC-ß-TCP was significantly lower than that with ß-TCP, and also the numbers of osteoblasts type cells with Mg-LC-ß-TCP were significantly higher than that with ß-TCP. It is suggested that Mg substitution to form low crystallinity ß-TCP is a valuable way to overcome the limitations of ß-TCP as a bone substitute.

Fabrication and evaluation of carbonate apatite-coated calcium carbonate bone substitutes for bone tissue engineering.

Carbonate apatite-coated calcium carbonate (CO3 Ap/CaCO3 ) was fabricated through a dissolution-precipitation reaction using CaCO3 granules as a precursor to accelerate bone replacement based on superior osteoconductivity of the CO3 Ap shell, along with Ca2+ release from the CaCO3 core and quicker resorption of the CaCO3 core. In the present study, CaCO3 , 10% CO3 Ap/CaCO3 , 30% CO3 Ap/CaCO3 , and CO3 Ap granules were fabricated and examined histologically to evaluate their potential as bone substitutes. Larger contents of CaCO3 in the granules resulted in higher Ca2+ release and promoted cell proliferation of murine preosteoblasts at 6 days compared with CO3 Ap. Interestingly, in a rabbit femur defect model, 10% CO3 Ap/CaCO3 induced significantly higher new bone formation and higher material resorption compared with CO3 Ap at 8 weeks. Nevertheless, CO3 Ap showed a superior osteoconductive potential compared with 10% CO3 Ap/CaCO3 at 8 weeks. All tested granules were most likely resorbed by cell mediation including multinucleated giant cell functions. Therefore, we conclude that CO3 Ap/CaCO3 has a positive potential for bone tissue engineering based on well-controlled calcium release, bone formation, and material resorption.

Compositional and histological comparison of carbonate apatite fabricated by dissolution-precipitation reaction and Bio-Oss®.

Carbonate apatite (CO3Ap) is an inorganic component of bone. This study aimed to compare the composition and tissue response to of CO3Ap (CO3Ap-DP) fabricated by the dissolution-precipitation reaction using calcite as a precursor and Bio-Oss®, which is widely used in orthopedic and dental fields as a synthetic bone substitute. X-ray diffraction and Fourier transform infrared results showed that CO3Ap-DP and Bio-Oss® were both B-type carbonate apatite with low crystallinity. The average sizes of CO3Ap-DP and Bio-Oss® granules were 450 ± 58 and 667 ± 168µ m, respectively, and their carbonate contents were 12.1 ± 0.6 and 5.6 ± 0.1 wt%, respectively. CO3Ap-DP had a larger amount of CO3 than Bio-Oss® but higher crystallinity than Bio-Oss®. When a bone defect made at the femur of rabbits was reconstructed with CO3Ap-DP and Bio-Oss®, CO3Ap-DP granules were partially replaced with bone, whereas Bio-Oss® remained at 8 weeks after implantation. CO3Ap-DP granules elicited a significantly larger amount of new bone formation at the cortical bone portion than Bio-Oss® at 4 weeks after the implantation. The results obtained in the present study demonstrated that CO3Ap-DP and Bio-Oss® showed different behavior even though they were both classified as CO3Ap. The CO3 content in CO3Ap played a more important role than the crystallinity of CO3Ap for replacement to bone and high osteoconductivity.

Feasibility evaluation of low-crystallinity ß-tricalcium phosphate blocks as a bone substitute fabricated by a dissolution-precipitation reaction from α-tricalcium phosphate blocks.

Although sintered ß-tricalcium phosphate blocks have been used clinically as artificial bone substitutes, the crystallinity of ß-tricalcium phosphate, which might dominate biocompatibility, is extremely high. The objective of this study is to evaluate the feasibility of fabricating low-crystallinity ß-tricalcium phosphate blocks, which are expected to exhibit good biocompatibility via a dissolution-precipitation reaction of α-tricalcium phosphate blocks as a precursor under hydrothermal conditions at 200°C for 24 h. Although ß-tricalcium phosphate is a metastable phase, the presence of Mg2+ in the reaction solution inhibits the formation of its corresponding stable phase and induces ß-tricalcium phosphate formation under acidic conditions. It was found that low-crystallinity ß-tricalcium phosphate blocks could be fabricated from α-tricalcium phosphate blocks immersed in 1.0 mol/L MgCl2 + 0.1 mol/L NaH2PO4 solution while maintaining the shape of the α-tricalcium phosphate blocks. The crystallite size of the fabricated ß-tricalcium phosphate blocks was 42 nm, which was substantially smaller than that of the sintered ß-tricalcium phosphate blocks. When the fabricated ß-tricalcium phosphate blocks were implanted into bone defects in rabbit femurs, they exhibited excellent tissue responses. In particular, the initial osteoconductivity (two and four weeks) was substantially greater than that of sintered ß-tricalcium phosphate blocks.