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Aging test and dynamic fatigue test of apatite-wollastonite-containing glass ceramics and dense hydroxyapatite.

The purpose of this study is to examine the changes in mechanical strength of two bioactive ceramics in living tissue. An aging test and dynamic fatigue test were performed using apatite-wollastonite-containing glass ceramics (A X W-GC) and dense hydroxyapatite (HA). Specimens (5 mm X 5 mm X 25 mm, abraded with No. 2000 Al2O3 powder) were implanted into subcutaneous tissue of rats for varying periods of time. The bending strength of aged samples was measured by the three-point loading method. The bending strength of A X W-GC was greater than that of HA (P less than 0.001). There was no reduction in bending strength for both A X W-GC and HA in living tissue. The n value of both A X W-GC and HA did not decrease significantly after implantation as assessed by the results of the dynamic fatigue test according to analysis of covariance. SEM-EPMA showed that Si and Mg contents decreased, Ca content did not change, while P content increased in the surface of A X W-GC. The area where x-ray intensity changed increased moderately after implantation. There were no changes in Ca and P at the interface between HA and soft tissue. In macroscopic and microscopic observations, specimens were found to be encapsulated with a thin layer of connective tissue. Foreign body giant cells, osteoblasts, or osteoclasts were not observed in the soft tissue. There was no bonding between ceramics and soft tissue.

Animals↗

Apatite-wollastonite containing glass ceramic-fibrin mixture as a bone defect filler.

The purpose of this study was to evaluate the usefulness of a mixture of apatite-wollastonite containing glass ceramic (A-W.GC, 42-60 mesh in granule size) with fibrin as a bone defect filler. A bone defect was drilled in the proximal metaphysis of the rat tibia and was filled with (1) fibrin glue, (2) A-W.GC granules, or (3) A-W.GC-fibrin mixture. Nothing was placed in the defect of the controls. The animals were serially sacrificed until 8 weeks after the operation, and the defect site was histologically examined and histomorphometrically analyzed for quantitative evaluation of newly formed bone and blood vessels. The use of fibrin glue as the binder markedly increased the ease of handling the A-W.GC granules. In the controls, little trabecular regeneration was observed in the defect site. Early vascularization (confirmed by microangiography) increased and the repairing process was accelerated in the defects filled with fibrin. In these defects filled with A-W.GC granules, good bone formation was observed around the granules. Bone formation was accelerated in the defects filled with A-W.GC fibrin mixture. Thus, the mixture showed good osteoconductive potential as well as acceleration of the repair process. Therefore, A-W.GC-fibrin mixture is considered to be a useful bone defect filler.

Animals↗

Apatite--wollastonite containing glass ceramic granule-fibrin mixture as a bone graft filler: use with low granular density.

The purpose of this study was to investigate bone formation after implantation of a mixture of apatite--wollastonite containing glass ceramic granules (A-W.GC, 220-355 microns) and fibrin glue with a low A-W.GC granular density. The fibrinogen was prepared from rat blood plasma. A mixture of A-W.GC granules and fibrin glue 1:4 in volume ratio (1:4 mixture) was implanted in the proximal metaphysis of the rat tibia, and bone formation was evaluated quantitatively by histomorphometry in undecalcified specimens. Microangiography was performed in animals sacrificed 5 days after the operation. The results were compared with those of a previous study in which a 1:1 mixture of A-W.GC granules and fibrin glue was used. With the 1:4 mixture, the A-W.GC density was 39.7% because of concentration of fibrin at packing. Good early vascularization comparable to that after implantation of the 1:1 mixture was observed. Bone formation proceeded at the same rate as after implantation of 1:1 mixture, but the level of the plateau was higher than that for the 1:1 mixture. The osteoconductive potential of the 1:4 A-W.GC-fibrin mixture was similar to that of the 1:1 mixture, and was higher than that of A-W.GC without fibrin.

Animals↗

Mechanical properties of bone after implantation of apatite-wollastonite containing glass ceramic-fibrin mixture.

Mechanical properties of bone implanted with a mixture of apatite-wollastonite containing glass ceramic (A-W.GC) granules and fibrin were examined by compression testing. A 1:1 mixture of A-W.GC granules and fibrin (Group 1), 1:4 mixture of A-W.GC granules and fibrin (Group 2), and 1:1 mixture of hydroxyapatite (HA) granules and fibrin (Group 3) were implanted in the distal femoral metaphyses of rabbits. Histomorphometric analysis suggested that A-W.GC has a greater osteoconductive potential than HA. Trabeculalike structures were observed 24 weeks after the operation in all groups but were most notable in Group 2. Twenty-four weeks after the operation, in Groups 1 and 3, the compressive strength and compressive stiffness were higher than that of normal cancellous bone, and the fracture toughness was comparable with that of normal cancellous bone. In Group 2, all three values were similar to those of normal cancellous bone. Implantation of A-W.GC granules at a low density induces the formation of bone tissue which is similar to normal bone in both mechanical properties and morphology.

Animals↗

Bone-bonding behavior under load-bearing conditions of an alumina ceramic implant incorporating beads coated with glass-ceramic containing apatite and wollastonite.

Alumina ceramic with a porous surface coated with glass-ceramic containing apatite and wollastonite (AW-GC) was implanted in a state of press-fit under load-bearing conditions in the femoral condylus of the mongrel dog and compared with a non-glass-ceramic-coated alumina ceramic. A trapezoid alumina ceramic implant (7 x 10 x 5 mm) with a lateral recess (0.9 mm deep) coated with alumina ceramic beads (mean diameter, 750 microns) in a single layer was prepared. The alumina ceramic beads were bonded to the alumina ceramic substratum using an identical alumina binder. The thickness of coating was 10-50 microns (mean, 30 microns). The surface of the beads and the substratum of the alumina implant were coated with AW-GC. A pull-out test and histologic examination were performed at 4, 8, and 24 weeks after implantation. The interfacial shear load was significantly increased from 8 to 24 weeks in both groups. The shear load of the glass-ceramic-coated implant was significantly greater than that of the noncoated implant at every stage. The interface shear load of the noncoated implant was 12.13 +/- 2.76 kg at 4 weeks, 13.92 +/- 4.18 kg at 8 weeks, and 24.17 +/- 5.17 kg at 24 weeks after implantation. The interface shear load of the glass-ceramic-coated implant was 17.96 +/- 2.81 kg at 4 weeks, 24.92 +/- 9.87 kg at 8 weeks, and 34.83 +/- 4.12 kg at 24 weeks after implantation. Histologic examination showed more ingrown bone tissue in the glass-ceramic-coated implants. It is suggested that AW-GC stimulated the bone ingrowth.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗

Thromboresistance of alkali- and heat-treated titanium metal formed with apatite.

Fibrin deposition and platelet adhesion onto alkali- and heat-treated titanium metal (AH-Ti), alkali- and water-treated titanium metal (Wa-Ti), and alkali- and heat-treated titanium metal formed with apatite (Ap-Ti) in simulated body fluid (SBF) were evaluated by exposure to anticoagulated blood or washed platelet suspension (WPS) under static conditions and subsequent observation with scanning electron microscopy (SEM). The results were compared with those for commercially pure titanium metal (cp-Ti). Thrombus formation on AH-Ti and Wa-Ti, which were exposed to heparinized whole blood for 1 h, was significantly less than that on cp-Ti, on which pronounced depositions of fibrin-erythrocytes and lymphocytes were observed. No thrombus was observed on Ap-Ti, possibly because of a high adsorption of heparin. Morphological change of platelets attached to surfaces via adsorbed plasma proteins was found to a significant extent on AH-Ti and Wa-Ti exposed to WPS. However, there was almost no difference between cp-Ti and Ap-Ti in round morphology of adherent platelets. These findings suggested that Ap-Ti exhibits stronger antithrombogenic characteristics than cp-Ti and other materials examined in heparinized blood.

Adsorption↗

A hybrid coating of biomimetic apatite and osteocalcin.

A novel hybrid coating of biomimetic apatite(BAp) and osteocalcin (OC) was prepared by incubating BAp-coated Ti6A14V coupons in an osteocalcin-containing medium. A significant amount (up to 1.0 wt %) of OC was adsorbed by the BAp coating within 3 h of incubation as demonstrated by high-performance liquid chromatography. Characterizations of the hybrid coating with environmental scanning microscopy and X-ray diffraction indicated that protein adsorption does not alter the microstructure of the coating. The presence of OC in the hybrid coating was visualized with fluorescence microscopy using an immuno-labeling procedure. The affinity of OC to the BAp coating was examined using a 20-h elution test in phosphate-buffered saline and only a minimal amount (<10%) of the loaded OC was eluted out. When the coating was fully dissolved in hydrochloric acid solution after elution, about 78% of the loaded OC could be recovered. Enzyme-linked immunosorbent assay and peptide sodium dodecylsulfate-polyacrylamide gel electrophoresis confirmed the integrity and activity of OC molecules throughout the tests. The preliminary cell culture tests showed a significant effect of OC on the attachment and proliferation of osteoblasts. The quick loading profile and high affinity of OC to the BAp coating make it an ideal candidate for the hybrid coating preparation in clinical environment.

Animals↗

Effects of various sterilization methods on the setting and mechanical properties of apatite cement.

Sterilization capability is a necessary requirement for any material that is to be used in a medical application. Therefore, it is necessary for apatite cement (AC) to be sterilized. Because there is little information on the sterilization methods of AC, the aims of this investigation were to evaluate the effects of various sterilization methods, including steam, dry heat, ethylene oxide (EtO) gas, and gamma irradiation sterilizations, on the setting and mechanical properties of AC. In the case of steam sterilization, because AC powder aggregated before setting-time measurements, the setting time could not be measured. When the powder was sterilized by dry heat or EtO gas, the setting time was prolonged significantly and the wet diametral tensile strength (DTS) value decreased significantly. Therefore, sterilizations with steam, dry heat, or EtO gas were suggested to be inappropriate methods for AC. Accordingly, the following experiments focused on gamma sterilization. The setting time of AC was retarded with an increase in gamma irradiation dose. The wet DTS value decreased with the increase in gamma irradiation dose. There was no compositional change due to the gamma irradiation. The following tests were carried out in order to examine the effect of the gamma irradiation on the setting reaction of AC in detail. Tetracalcium phosphate [TTCP: Ca(4)(PO(4))(2)O] and dicalcium phosphate anhydrous (DCPA: CaHPO(4)) were separately irradiated, and the cements were produced with the use of irradiated powder and nonirradiated powder. Although the wet DTS value of AC produced from irradiated TTCP and nonirradiated DCPA decreased with increasing gamma irradiation dose, there was no significant difference. In contrast, the wet DTS value of AC produced from irradiated DCPA and nonirradiated TTCP significantly decreased with the increase in gamma irradiation dose. In conclusion, although the detailed mechanism of the delayed setting time and decreased DTS value was not clarified by the present study, it was found that gamma irradiation affected DCPA more than TTCP.

Adhesives↗

Osteointegration of biomimetic apatite coating applied onto dense and porous metal implants in femurs of goats.

Biomimetic calcium phosphate (Ca-P) coatings were applied onto dense titanium alloy (Ti6Al4V) and porous tantalum (Ta) cylinders by immersion into simulated body fluid at 37 degrees C and then at 50 degrees C for 24 h. As a result, a homogeneous bone-like carbonated apatitic (BCA) coating, 30 microm thick was deposited on the entire surface of the dense and porous implants. Noncoated and BCA-coated implants were press-fit implanted in the femoral diaphysis of 14 adult female goats. Bone contact was measured after implantation for 6, 12, and 24 weeks, and investigated by histology and backscattered electron microscopy (BSEM). After 6 weeks, bone contact of the BCA-coated Ti6Al4V implants was about 50%. After 12 and 24 weeks, bone contact was lower in comparison with the 6-week implantations at, respectively 24 and 39%. Regarding the BCA-coated porous Ta implants, bone contacts were 17, 30, and 18% after 6, 12, and 24 weeks, respectively. However, bone contact was always found significantly higher for BCA-coated dense Ti6Al4V and porous Ta cylinders than the corresponding noncoated implants. The results of this study show that the BCA coating enhances the bone integration as compared to the noncoated implants.

Alloys↗

Evidence of octacalcium phosphate and Type-B carbonated apatites deposited on the surface of explanted acrylic hydrogel intraocular lens.

Fourier-transform infrared (FTIR) microspectroscopy combining with attenuated total reflection (ATR) microsampling technique and micro-Raman spectrophotometer were used to detect the deposited materials on the surface of acrylic hydrogel intraocular lens (IOL) with or without ocular implantation. Surface morphology and the interface of this IOL were further examined by a confocal laser scanning microscope. The brand-new IOL exhibited a very smooth, transparent and featureless surface, but the explanted IOL had an irregular cerebriform-like opaque appearance. Both FTIR/ATR and Raman microspectroscopic analyses showed the deposits on the surface of acrylic hydrogel IOL after ocular implantation to consist of octacalcium phosphate (OCP) and Type B carbonated apatites, leading to the opalescence of acrylic hydrogel IOL. Both vibrational microspectroscopic examinations also confirmed the mineralization still in progress on the surface of acrylic hydrogel IOL after ocular implantation for 2 years.

Aged↗

Osteogenic protein-1 enhances osseointegration of titanium implants coated with peri-apatite in rabbit femoral defect.

This study evaluated the effect of osteogenic protein-1 (OP-1) carried by Peri-Apatite (PA) on bone healing in the gap surrounding implants in a rabbit model. Cylindrical titanium implants (3 x 9 mm) were uniformly coated with PA precipitated from a calcium and phosphate solution. OP-1 solution containing 60 microg OP-1 was directly loaded on the implants immediately before implantation for the experimental group, whereas buffer solution was loaded on the implants for the control. The implant was placed in the distal femur and surrounded by a 1-mm gap. The implants were retrieved and examined 6 weeks after implantation. Mechanical testing (push-out) data showed that OP-1 enhanced implant fixation by 80%. Histomorphometric measurements indicated that bone ingrowth in the initial gap expressed as a percentage of the whole gap was significantly higher in the specimens treated with OP-1 than the control group (25.4% vs. 8.9%, p < 0.05). The percentage of the surface of implants, which was covered by bone, was significantly higher in the OP-1-treated group compared to the control group (65% vs. 25%, p < 0.05). This study suggests that OP-1 can be loaded on orthopedic implants through PA to enhance the osseointegration of orthopedic implant.

Alloys↗

Calcium-deficient apatite synthesized by ammonia hydrolysis of dicalcium phosphate dihydrate: influence of temperature, time, and pressure.

In this work, calcium-deficient apatites (CDA) were synthesized by ammonia hydrolysis reaction of dicalcium phosphate dihydrate (DCPD; CaHPO4 x 2 H2O) to obtain biphasic calcium phosphates (BCP) without any extraionic substitution. The influence of three parameters was studied: temperature of the reaction (70 and 100 degrees C), time of the reaction (4 and 18 h), and the pressure (open and closed system). Experiments were made according to a factorial design method (FDM) allowing optimization of the number of samples as well as statistical analysis of results. Moreover, the influence of temperature (until 200 degrees C) was investigated. The crystal size of CDA was determined according to the Scherrer's formula and from Rietveld refinements taking the CDA anisotropy into account. The last method seems to be a reliable method to determine crystallite sizes of CDA, since crystallite sizes of CDA along <00l> and directions were accessible. The results describe the hydroxyapatite % (HA%) in BCP by a first-order polynomial equation in the experimental area studied and the HA content was found to increase by raising time and temperature of the reaction. Moreover, the type of reaction system (open/closed vessel) appeared to have little influence on HA%.

Ammonia↗

Ipriflavone does not alter bone apatite crystal structure in adult male rats.

We have previously found that a short-term treatment with high doses of ipriflavone increased bone density and improved the biomechanical properties of adult male rat bones, without altering their mineral composition. To determine whether this effect can be associated with alterations of bone crystal structure, we have performed X-ray diffraction analysis of bones obtained from rats treated with ipriflavone at doses that were effective in inducing favorable changes on bone density and biomechanics. Eighteen-week-old male Sprague Dawley rats were treated by oral route with either ipriflavone (200 or 400 mg/kg/day), or its vehicle for 12 weeks. The treatment was well tolerated and body weight increased to the same extent in all animals. As a measure of bone crystallinity, we examined the (310) and (002) reflections of the X-ray diffraction patterns, corresponding to the directions perpendicular and parallel to the c-axis of the crystals, respectively. No major differences were observed between ipriflavone-treated and control animals for the broadening parameter beta(1/2) for (310) and (002) peaks, as well as for lattice parameters. Therefore, a 12-week treatment with ipriflavone at high doses does not induce significant modifications of bone "crystallinity." Thus, the positive effect of ipriflavone on bone mineral density appears to be associated with an increased apatite crystal formation rather than an increase of crystal size. These results provide further evidence for the safety and usefulness of ipriflavone in the treatment of osteoporotic syndromes.

Animals↗

Experimental osteoinduction in rats: collagen-apatite versus osteogenin-containing gelatine.

An experimental study in rats was done to investigate the bone-regenerating properties of collagen apatite (Collapat) and to compare it with osteoinduction dependent on osteogenin-containing gelatine (OCG). The test substances were implanted orthotopically (calvarial defect--7 mm in diameter) and heterotopically (paravertebral muscles, abdominal muscles). The results were evaluated histologically and enzymatically (alkaline phosphatase). Collapat caused neither osteoinduction in the heterotopic site nor healing of the bone defects. Foreign body reaction without new bone formation was encountered. OCG implantation leads to new bone formation in the muscles within 3 weeks, associated with a significant increase in alkaline phosphatase activity, and to extensive new bone formation in the calvarial defect within 4 weeks. The defects did not heal if left empty. The value of clinical application of Collapat appears to be doubtful. Osteoinduction with OCG requires further experimental investigation.

Animals↗

Histological study of a case of recurrent olecranon bursitis with mixed calcium pyrophosphate dihydrate and apatite crystal deposits.

This is the report of a light and transmission electron microscopic study of an olecranon bursitis and of the adjacent distal tricipital tendon in an 83 year-old man. The data are compared with those of a similar study in the same patient performed 2 years ago. Calcium pyrophosphate dihydrate crystals were observed in the bursal fluid, in the inner part of the bursal wall (extracellular localization and intracellular phagocytosis) as well as in the peripheral part of the tendon. In addition, small apatite deposits were observed in the bursa and tendon by electron microscopy. The origin of these bursal deposits is discussed; it is suggested that they may be related to an exchange from the tendon to the remodelled bursal wall.

Aged↗

Inhibition of apatite formation by phosphorylated metabolites and macromolecules.

Apatite formation from synthetic extracellular fluids is rate-limited both at the initial amorphous precursor deposition step and at the amorphous-crystalline transformation reaction. Nucleotide diphosphates and triphosphates and low molecular weight metabolites containing two attached ester phosphate groups all inhibited amorphous-crystalline conversion at concentrations of 10(-5) to 10(-6)M. Both native and synthetic polynucleotides as well as the phosphoproteins from rat dentin or egg yolk also inhibited crystal formation from amorphous calcium phosphate. In all cases, substantial amounts of inhibitor molecules were incorporated into the stabilized amorphous precipitates. Treatment of isolated, inhibitor-stabilized amorphous precipitates with hydrolytic enzymes such as alkaline phosphatase or papain reversed the inhibitory effect and permitted crystallization to proceed normally.

Alkaline Phosphatase↗

Radiation induced free radicals as molecular probes in synthetic apatites.

Free radicals generated in synthetic apatitic calcium phosphates by X-ray radiation were investigated by electron spin resonance (ESR) spectroscopy. Among the species stable enough at -188 degrees C to be identified were hydrogen atoms, phosphate radicals, and oxygen anion radicals. The ESR spectra were markedly dependent on the specific surface of the mineral. Oxygen radicals dominated the spectra of low specific surface samples while phosphate radicals were the predominant species at higher specific surfaces. Our studies suggest that the oxygen radicals are more stable in the bulk of the crystal while the hydrogen atoms and the phosphate radicals are stabilized at or near the crystal surface. It was concluded that the surface species are potentially capable of serving as probes of biologically relevant mineral-organic interfaces.

Apatites↗