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PubMed1997

Effect of electrochemically deposited apatite coating on bonding of bone to the HA-G-Ti composite and titanium.

The surfaces of hydroxyapatite-glass-titanium (HA-G-Ti) functionally gradient composite and titanium bars were treated with electrochemical apatite deposition, and a cathodic current was applied at 62 degrees C in a solution containing calcium and phosphate ions. Specimens with and without the electrochemical surface treatment were implanted in the femurs of Japanese white rabbits. The rabbits were sacrificed at 3, 6, and 9 weeks after implantation, and the bonding strengths of bone to these specimens were determined by a pull-out method. At 3 and 6 weeks after implantation the specimens with the electrochemical surface treatment showed larger values for the Weibull modulus and characteristic strengths than those of untreated specimens, whereas there was no remarkable difference in the results at 9 weeks. Especially the pull-out strengths of surface-treated specimens were significantly larger than the untreated ones at 3 weeks after implantation. Scanning electron microscopy and Fourier transform infrared absorption spectroscopy of the specimen surface after implantation demonstrated that formation of new bone was enhanced by the electrochemical surface treatment. It can be concluded that the electrochemical surface treatment undoubtedly contributes to the early stage fixation between bone and implant.

Animals↗
PubMed1998

Development of porous apatite ceramic for local delivery of chemotherapeutic agents.

An experimental study was conducted on a drug delivery system (DDS), using porous apatite ceramics (PAC): hydroxyapatite block (HAb) [Ca10(PO4)6(OH)2] having a porosity of 35-48% and pore size range of 50-300 microm, and beta-tricalcium phosphate block (TCP) [Ca3(PO4)2] having a porosity of 75-80% and pore size range of 100-400 microm, for sustained release of a chemotherapeutic agent. Methotrexate (MTX) was loaded in the pores of PAC blocks by centrifuging the blocks in MTX solution. Impregnation of MTX in PAC blocks (1 cm3) was confirmed by a magnetic resonance imaging (MRI) study using Gadolinium-DTPA enhancement. The MRI showed high signal intensity in the PAC, which was confirmed by dye loading into the pores. To estimate the MTX-releasing capability of the PAC, the blocks were stored in 3 mL of phosphate-buffered saline (PBS) at 37 degrees C and the PBS was replaced every 48 h. The amount of MTX released was assayed by high-performance liquid chromatography. This study showed that MTX-impregnated PAC (0.63-2.25 mg/block) released the drug in a steady manner and maintained its concentration (0.1-1.0 microg/mL) up to 12 days. This concentration is high enough to be effective against tumor cells. Chemotherapeutic agent-impregnated PAC, prepared by simple centrifugation, could be a valuable form of local chemotherapy when used as a strut graft to repair bone defects. This new DDS material could also be used as an adjuvant to extended curettage and provide a means to reduce the recurrence of tumors without risk of systemic toxicity.

Antimetabolites, Antineoplastic↗
PubMed1998

In vitro influence of apatite-granule-specific area on human growth hormone loading and release.

Although calcium phosphate biomaterials often are used as drug delivery systems (DDS) at bone sites, the conditions affecting the loading of the therapeutic agent (TA) have not been well documented. A human growth hormone (hGH) adsorption method was used in this study to investigate the influence of the formulated apatite (AP)-specific area on loading and release. AP powders were formulated with a 200-500 microm granulometry and various specific areas. Two milligrams of hGH in solution were deposited for 24 h at 37 degrees C on 100 mg of AP with different specific areas. The amount of hGH loaded was determined by immunoradiometric assay (IRMA) and eluted stain bioassay (ESTA) using Nb2 lymphoma rat cells. Although loading was not greatly influenced by a specific area between 3 and 25 m2/g, dependency was noted for higher specific areas. Human GH release was measured by IRMA and ESTA over a 33-day period, with half-time release between 25 and 79 h. Comparison of IRMA and ESTA measurements for the hGH amounts loaded showed that hGH biologic activity was conserved. Results indicate that it is feasible to control the quantity of TA loading on AP by modifying specific areas for in vivo applications.

Animals↗
PubMed1998

Apatite precipitation after incubation of biphasic calcium-phosphate ceramic in various solutions: influence of seed species and proteins.

The dissolution-precipitation process for calcium-phosphate ceramics in contact with biological fluid was studied by incubating blocks of biphasic calcium phosphate composed of hydroxyapatite (HA) and beta-tricalcium phosphate (beta-TCP) in different solutions: ionic simulated body fluid (SBF) without protein or SBF that contained various proteins and macromolecules separately (fibronectin, vitronectin, albumin, and poly-L-glutamic acid). Transmission electron microscopy studies revealed that apatite-precipitated microcrystals appeared around ceramic crystals as a result of secondary nucleation; microcrystals were in continuity with the lattice planes of the HA crystals but in a different direction from that of beta-TCP; the size of the precipitates was smaller when fibronectin, vitronectin, and poly-(L-glutamic acid) were present in SBF as compared to SBF without protein; and fibronectin and vitronectin initiated crystal nucleation in the void spaces between the ceramic crystals.

Apatites↗
PubMed1998

Electrophoretic coating of multilayered apatite composite on alumina ceramics.

By means of an electrophoretic deposition technique followed by sintering, alumina and zirconia ceramics were coated with apatitic composites composed of porous surface and intermediate layers of hydroxyapatite and an adhesive calcium phosphate layer. The electrophoretic deposition of these layers was attained by the use of a mixed solvent of acetylacetone and alcohol as well as the mixed powders of the calcium phosphates and alumina. The adhesive layer was formed by the codeposition of calcium phosphate glass powders (Ca/P = 1/2) with hydroxyapatite, while the open porosity of the surface layer was increased with the addition of alumina to the hydroxyapatite layers. The resultant phases of sintered composite layers were tricalcium phosphate and alumina with a small amount of hydroxyapatite.

Aluminum Oxide↗
PubMed1999

Al2O3 doped apatite-wollastonite containing glass ceramic provokes osteogenic differentiation of marrow stromal stem cells.

Fresh marrow cells were obtained from femora of Fischer rats and cultured in a medium containing 15% fetal calf serum (FCS) until confluence. After trypsinization, cells were subcultured at a cell density of 100 x 10(3)/35-mm well in the presence of FCS, beta-glycerophosphate, and ascorbic acid phosphate on four different culture substrata. The period of subculture was 2 weeks; the substrata used were the culture dish, apatite-wollastonite containing glass ceramic (AW), hydroxyapatite coated AW (HA/AW), and Al2O3 doped AW (Al/AW). The HA coating was attained by the incubation of AW in simulated physiological solution. The glass matrix of AW and HA/AW contained MgO, CaO, P2O5, and SiO2; Al/AW contained Al2O3 in addition to these components. The subculture on Al/AW substratum showed many alkaline phosphatase (ALP) positive nodules and the highest ALP activity. On a Northern blot analysis the housekeeping gene of beta-actin mRNA was evenly detected from the cells cultured on all substrata; however, bone-specific osteocalcin mRNA was only detected from the cells on Al/AW. These results indicate that Al/AW provokes the osteoblastic differentiation of marrow stromal stem cells.

Alkaline Phosphatase↗
PubMed1999

The structural characteristics and mechanical behaviors of nonstoichiometric apatite coatings sintered in air atmosphere.

Two types of nonstoichiometric apatite coatings on Ti6Al4V substrates were prepared in our laboratory by electrodeposition and hydrothermal synthesis. One was composed of highly pure needle-like calcium-deficient hydroxyapatite, and the other consisted of needle-like calcium-deficient hydroxyapatite and plate-like brushite. In this paper, the morphology, phase transition, and bonding strength of these coatings sintered at 450( degrees ) approximately 980 degrees C in an air atmosphere were studied. In addition, the residual stresses in pure calcium-deficient hydroxyapatite coatings and coatings sintered at 800 degrees C were measured by a hole-drilling method. The results show that after being sintered at a higher temperature, the coating/Ti6Al4V samples exhibited structural characteristics such as biphasic [hydroxyapatite + beta-Ca3(PO4)2] coatings and dense TiO2 film/Ti6Al4V in which the residual stresses in the coatings slightly increased. In addition, the results indicate that bonding strength was improved.

Air↗
PubMed2000

Dissolution of dense carbonate apatite subcutaneously implanted in Wistar rats.

Hydroxyapatite (HA) is used as a biomaterial in orthopedic applications because it is similar in composition to bone mineral; however, carbonate apatite (CHA) is closer in chemical composition to bone mineral because bone mineral contains significant amounts of carbonate, yet there have been few reports comparing biological responses to HA and CHA. It is generally agreed that bone forms a bond of some kind to HA, and there is conflicting evidence as to whether HA is resorbed in vivo or not. However, comparative reports generally agree that beta-tricalcium phosphate (beta-TCP) is removed faster than HA from an implant site by an undetermined combination of resorption and/or dissolution. The work reported here attempts to provide the first directly comparable subcutaneous dissolution data for dense sintered HA, beta-TCP, and CHA in rats. The weight losses of HA, beta-TCP, and 3.2 wt % CHA were approximately 60, 520, and 320 mg m(-2) day(-1), respectively. Histological sections did not show any evidence of giant cells, and all of the samples were encapsulated with fibrous tissue. beta-TCP and (to a lesser degree) CHA were found to be resistant to dissolution around the edges of the sample. An X-ray analysis did not indicate that any phase transformation had occurred in the dissolution resistant region.

Animals↗
PubMed2000

Evolution of porosity during in vitro hydroxycarbonate apatite growth in sol-gel glasses.

A bioactive glass of composition (mol %) SiO(2) 58, CaO 36, P(2)O(5) 6 was obtained using the sol-gel method. Changes in porosity during the growth of a hydroxycarbonate apatite (HCA) phase were studied at different stages of the process. The high Ca (II) content led to a higher degree of porosity and enhancement of Ca(2+) released from surface and bulk when the glass was soaked into simulated body fluid (SBF). Saturation of the media (SBF) together with the porosity of the material led to fast growth of HCA on the surface and into the pores.

Apatites↗
PubMed2000

Bioactive polymethyl methacrylate-based bone cement: comparison of glass beads, apatite- and wollastonite-containing glass-ceramic, and hydroxyapatite fillers on mechanical and biological properties.

A new bioactive bone cement (designated GBC) consisting of polymethyl methacrylate (PMMA) as an organic matrix and bioactive glass beads as an inorganic filler has been developed. The bioactive beads, consisting of MgO-CaO-SiO(2)-P(2)O(5)-CaF(2) glass, have been newly designed, and a novel PMMA powder was selected. The purpose of the present study was to compare this new bone cement GBC's mechanical properties in vitro and its osteoconductivity in vivo with cements consisting of the same matrix as GBC and either apatite- and wollastonite-containing glass-ceramic (AW-GC) powder (designated AWC) or sintered hydroxyapatite (HA) powder (HAC). Each filler added to the cements amounted to 70 wt %. The bending strength of GBC was significantly higher than that of AWC and HAC (p < 0.0001). Cements were packed into intramedullar canals of rat tibiae in order to evaluate osteoconductivity as determined by an affinity index. Rats were sacrificed at 2, 4, and 8 weeks after operation. An affinity index, which equaled the length of bone in direct contact with the cement expressed as a percentage of the total length of the cement surface, was calculated for each cement. At each time interval studied, GBC showed a significantly higher affinity index than AWC or HAC up to 8 weeks after implantation (p < 0.03). The value for GBC increased significantly with time up to 8 weeks (p < 0.006). The handling property of GBC was comparable with that of PMMA bone cement. Our study revealed that the higher osteoconductivity of GBC was due to the higher bioactivity of the bioactive glass beads at the cement surface and the lower solubility of the new PMMA powder to MMA monomer. In addition, it was found that the smaller spherical shape and glassy phase of the glass beads gave GBC strong enough mechanical properties to be useful under weight-bearing conditions. GBC shows promise as an alternative with improved properties to the conventionally used PMMA bone cement.

Animals↗
PubMed2001

Biologically and chemically optimized composites of carbonated apatite and polyglycolide as bone substitution materials.

We report on the development and characterization of a new composite material consisting of amorphous carbonated apatite, Ca(5)(PO(4), CO(3))(3)(OH), and microstructured poly(hydroxyacetic acid), polyglycolide (PGA). This material is able to keep the pH of a surrounding solution within the physiological range (7.2-7.6). This was achieved by chemical fine-tuning of the counterplay between the acidic degradation of the polyester and the basic dissolution of calcium phosphate. Microporous samples with pore sizes of <1 microm and compact samples were prepared. The biological behavior was assayed in vitro by long-term osteoblast culture. Morphological and biochemical analyses of cell differentiation revealed excellent biocompatibility, leading to cell attachment, collagen and osteocalcin expression, and mineral deposition. This material could be of use as a biodegradable bone substitution material and as a scaffold for tissue engineering.

Animals↗
PubMed2001

Effects of apatite and wollastonite containing glass-ceramic powder and two types of alumina powder in composites on osteoblastic differentiation of bone marrow cells.

Previously we developed a composite consisting of apatite and wollastonite containing glass-ceramic (AW-GC) powder and bisphenol-a-glycidyldimethacrylate (Bis-GMA)-based resin (designated AWC), and demonstrated that AWC showed direct contact with living bone. Another new composite consisting of mainly the delta-crystal phase of alumina bead powder and Bis-GMA-based resin (designated ABC) was developed. Although alumina ceramics are bioinert and a composite filled with the pure alpha-crystal phase of alumina powder (designated alphaALC) did not allow direct bone formation in vivo, ABC was shown to have excellent osteoconductivity. One purpose of this study was to investigate whether AW-GC powder in a composite promotes osteoblastic differentiation of rat bone marrow cells as AW-GC bulk did. Another purpose was to evaluate the effects of the delta-crystal phase of alumina powder in a composite on osteoblastic differentiation. In a cell culture with dexamethasone, alkaline phosphatase (AP) activity at both days 7 and 14, and the levels of osteocalcin mRNA and alpha1(I) collagen mRNA at day 14 and osteopontin mRNA at day 7, were highest on AWC, followed by ABC, and finally alphaALC. Scanning electron microscopy showed more abundant mineralized globules and a fibrous collagen matrix on AWC at day 14, followed by ABC. In a cell culture without dexamethasone, AP activity at both days 7 and 14, and the level of osteopontin mRNA at day 7, were higher on ABC than on any other composite, whereas osteocalcin mRNA could not be detected. These results indicate that AW-GC powder in a composite promotes osteoblastic differentiation of bone marrow cells intensively when supplemented with dexamethasone. The delta-crystal phase of alumina powder in a composite promotes greater osteoblastic differentiation than the alpha-crystal phase of alumina powder.

Alkaline Phosphatase↗
PubMed2001

Ultrastructure of the interface between bioactive composite and bone: comparison of apatite and wollastonite containing glass-ceramic filler with hydroxyapatite and beta-tricalcium phosphate fillers.

We have developed a bioactive bone cement that consists of apatite and wollastonite containing glass-ceramic (AW-GC) powder and bisphenol-a-glycidyl dimethacrylate (Bis-GMA)-based resin. In this study, we made three types of composite (designated AWC, HAC, and TCPC) consisting of AW-GC, hydroxyapatite (HA,) or beta-tricalcium phosphate (beta-TCP) powder as the inorganic filler and Bis-GMA-based resin as the organic matrix. The proportion by weight of the filler mixed into the cement was 70%. Rectangular plates (10 x 15 x 2 mm) of each composite were made and abraded with 2000 alumina powder. These composites were implanted into tibial metaphyses of rabbits. Specimens were prepared 10 and 25 weeks after implantation and examined using transmission electron microscopy (TEM). AWC was in direct contact with bone 10 weeks after implantation, and AW-GC particles were partially absorbed at the surface. HAC was in contact with partially mineralized extracellular matrix 10 weeks after implantation. In TCPC-implanted specimens, randomly oriented mineral was observed 10 weeks after implantation; however, collagenous extracellular matrix rarely was observed. In 25-week specimens, AW-GC particles were completely absorbed and replaced by new bone, and there was no intervening soft tissue. Both HAC and TCPC were in contact with bone at 25 weeks. These results indicate that AWC has higher bioactivity than either HAC or TCPC.

Animals↗
PubMed2002

Cells, rather than extracellular matrix, nucleate apatite in glutaraldehyde-treated vascular tissue.

Glutaraldehyde (GA) causes a large increase in [Ca(2+)](i) and [P(i)](i) and calcification of porcine aortic valve fibroblasts. Calcification in GA-treated vascular tissue is likely to begin intracellularly, but the potential role of extracellular matrix has not been taken into account in earlier studies. To compare the role of cells and matrix in calcification, intestinal pouches made of a lipid-extracted rat small intestine were prepared. Lipid-extracted porcine aortic valves, or cells cultured from those same valves, were placed in intestinal pouches, sealed, fixed with GA, and grafted in rat subcutis. Cells in the pouches calcified in 3 weeks whereas the valvular matrix did not calcify for 9 weeks. Cellular calcification spread to the wall of the intestinal pouches and grew heavier after 9 weeks. Similarly, smooth muscle cells calcified exclusively in GA-treated rat aorta grafted in rat subcutis for 3 weeks. Calcification of extracellular matrix was seen after 9 weeks. Cells initiate calcification and extracellular matrix serves as a substrate for the subsequent growth of apatite in GA-treated vascular tissue.

Animals↗
PubMed2002

Biodegradation of carbonate apatite/collagen composite membrane and its controlled release of carbonate apatite.

The aim of this study was to investigate the biodegradation of carbonate apatite (CO(3)Ap)/collagen composite membrane as a new guided tissue regeneration membrane in vivo and to estimate its controlled release of CO(3)Ap in vitro. To control the biodegradation of the guided tissue regeneration membrane and to promote hard tissue regeneration in the periodontal region, we added CO(3)Ap into the collagen membrane. To investigate the biodegradation of CO(3)Ap/collagen composite membranes, the prepared membranes (CO(3)Ap:0, 10 wt %) were cut into 5 x 5 x 0.1 mm and subcutaneously implanted into the backside of male rats under general anesthesia. The explanted membranes were investigated histologically. To estimate their controlled release of CO(3)Ap in vitro, the membranes (CO(3)Ap 0-10 wt %, 5 x 5 x 0.1 mm) were immersed into collagenase solution and compulsorily dissolved for 48 h. Histological results suggested that the membrane had a good biocompatibility and the biodegradable period was shortened with the presence of CO(3)Ap. In the solubility experiments of the membrane, eluted Ca concentrations gradually increased with total dependence on the dissolution of the collagen membrane. Our study demonstrated that the biodegradation time can be controlled by CO(3)Ap contents in the membrane and CO(3)Ap could be released from the membrane with the biodegradation period.

Absorbable Implants↗
PubMed2002

Effect of water vapor treatment on apatite formation on precalcified titanium and bond strength of coatings to substrates.

In previous investigations, a simple method, precalcification, was developed for bioactivating titanium. After a titanium sample was precalcified in a boiling saturated Ca(OH)(2) solution and then immersed in a calcium phosphate supersaturated solution, an apatite coating rapidly precipitated onto its surface. In the present study, heat-treatment in water vapor was carried out prior to precalcification. Heat-treatment in water vapor stimulated the chemical reaction between titanium, calcium, and phosphate. Coating properties were improved, and the bond strength of the coating to substrate was enhanced.

Apatites↗
PubMedDate not supplied

Bone bonding behavior of three kinds of apatite containing glass ceramics.

We have produced three kinds of apatite-containing glass ceramics of the same chemical composition: MgO (4.6), CaO (44.9), SiO2 (34.2), P2O5 (16.3), CaF2 (0.5) (in weight ratio). They contain different crystal combinations and have different mechanical properties. The first glass ceramic (A-GC) was prepared by heating a glass plate to 870 degrees C. It contains only oxy- and fluoroapatite (35 wt%). The second glass ceramic (A-W-GC), and the third (A-W-CP-GC), were prepared by heating glass powder compacts to 1050 degrees C and 1200 degrees C, respectively. A-W-GC contains oxyapatite and fluoroapatite (Ca10(PO4)6(O,F2] (35 wt%) and beta-wollastonite (40 wt%). A-W-CP-GC contains oxyapatite and fluoroapatite (20 wt%), beta-wollastonite (CaO X SiO2) (55 wt%), and beta-whitlockite (3CaO X P2O5) (15 wt%). The bending strengths of A-GC, A-W-GC, and A-W-CP-GC were 88MPa, 178MPa, and 213MPa, respectively, in air. Rectangular ceramic plates (15mm X 10mm X 2mm) were implanted into a rabbit tibia. Ten and 25 weeks after implantation, the segment of tibia with implant was excised for examination. The segment was held by a special jig and the traction breaking load (failure load) was measured by an autograph. A-GC showed a lower load than A-W-GC and A-W-CP-GC. The loads for A-W-GC and A-W-CP-GC were almost equal. The failure loads did not change significantly between 10 and 25 weeks for any of the materials. The interface was examined by Giemsa surface staining, contact micro-radiography, and SEM-EPMA. Giemsa surface staining and CMR revealed direct bonding between the materials and the bone for all the three materials. SEM-EPMA showed that Si and Mg content decreased, Ca content did not change, and P content increased at the reaction zone between all three glass ceramics and bone. This was observed at 10 weeks, as well as at 25 weeks, after implantation. The reaction zone was narrowest with A-GC, wider with A-W-GC, and widest with A-W-CP-GC.

Animals↗