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[In vitro study on the cytotoxicity of strontium substituted hydroxyapatite].

OBJECTIVE: To evaluate the cytotoxicity of strontium substituted hydroxyapatite. METHODS: Cell Relative Growth Rate(RGR) method, MTT assay and Flow Cytometry(FCM) method were used, and the strontium substituted hydroxyapatite contained different strontium concentration(0%,1%,5%,10%,100%). RESULTS: It was found that there's no apparent cytotoxicity of all the strontium substituted hydroxyapatite,but the cytotoxicity increased as the strontium concentration raised. As the FCM method appeared, there was no apparent difference between the pure hydroxyapatite and 1%,5% strontium substituted hydroxyapatite. CONCLUSION: Strontium substituted hydroxyapatite has good biocompatibility, and 10%,100% strontium substituted hydroxyapatite has weak cytotoxicity.

English Abstract↗

[Experimental study on the osseointegration of nanophase hydroxyapatite biograde-coated implants].

OBJECTIVE: To study the osseointegration of the nanophase hydroxyapatite biograde-coated implants and host bone. METHODS: Nanophase hydroxyapatite biograde-coated implants, hydroxyapatite biograde-coated implants and noncoated Ti-6Al-4V implants were inserted into both femoral of Beagle canines the tissue response, dynamic osteogensis and SEM were studied at 4, 8 and 12 weeks. RESULTS: The degradation of nanophase hydroxyapatite was slower than hydroxyapatite, dynamic osteogensis and the form of osteoblast were better than the control groups. CONCLUSION: The biological reaction of Nanophase hydroxyapatite biograde-coated implants is better than hydroxyapatite coated implant.

Animals↗

Application of hydroxyapatite-sol as drug carrier.

The application of hydroxyapatite-sol as a drug carrier is being developed. Hydroxyapatite-sol which is a suspension consisting of hydroxyapatite nano-crystals, was synthesized using an ultrasonic homogenizer. The size of the crystals was 40 x 15 x 10 mm3 on average and their specific surface area was 100 m2/g. An amount of a glycoside antibiotics adsorbed onto hydroxyapatite nano-crystals was measured. The drug adsorbed 0.2 mg per 1 mg of hydroxyapatite. The affect of the drug adsorbed onto the hydroxyapatite was investigated using cancer cells. The drug, adsorbed onto the hydroxyapatite nano-crystals, inhibited cancer cell growth.

Adsorption↗

Bone augmentation with hydroxyapatite: histological findings in 55 cases.

Bone augmentation was performed for 55 implant patients (21 male, 34 female) with hydroxyapatite granules. In 38 patients the hydroxyapatite was covered with lyophilized dura mater to avoid its loss during the healing phase. Sixty-seven punch biopsies were obtained between 14 days and 7 years. The undecalcified specimens were embedded in poly(methyl methacrylate), sectioned and stained to demonstrate bone formation. Increasing metaplastic formation of woven bone trabeculae was observed beginning with the earliest specimens. The hydroxyapatite was in intimate contact with the bone and later was partially included within its matrix. In some areas, the woven bone was resorbed by osteoclasts together with the hydroxyapatite, and mature lamellar bone was deposited by dense layers of osteoblasts. After 20 months, sufficient new bone formation occurred to permit implant insertion. In a whole mandible specimen obtained 5.6 years after implant placement, there was almost complete ossification of the hydroxyapatite. The inclusion of the titanium implants within the newly formed bone was evident, and the hydroxyapatite granules applied during second-stage surgery were firmly embedded within the newly formed trabecular bone. The bone augmentation technique described has been used successfully in over 600 cases and can be considered as safe. Bone formation in contact with hydroxyapatite was demonstrated histologically in a large number of human biopsies.

Aged↗

Hydroxyapatite cement. II. Obliteration and reconstruction of the cat frontal sinus.

Frontal sinus obliteration and reconstruction can be performed with autogenous grafts or synthetic implants, each of which has significant limitations. Hydroxyapatite cement, which can be shaped intraoperatively and sets to a microporous hydroxyapatite implant, was applied to this problem. Nine cats had the anterior table of their frontal sinus unilaterally removed and the sinus cavity stripped of its mucosa. Hydroxyapatite cement was used to obliterate the cavity and reconstruct the overlying anterior table defect. The unoperated side served as the control, and the animals were sacrificed up to 18 months postoperatively. There were no adverse reactions, infections, mucoceles, or implant extrusions. The normal anatomic contour of the forehead region overlying the hydroxyapatite cement implants was maintained in all animals. Histologic examination of undecalcified whole sinus sections revealed progressive replacement of the implants with woven bone without a loss of volume. Replacement of the hydroxyapatite cement by woven bone is postulated to occur through a combination of implant resorption coupled with osteoconduction. The use of hydroxyapatite cement proved successful for the reconstruction and obliteration of cat frontal sinuses, and may be appropriate for the same application in humans.

Animals↗

Facial skeletal augmentation using hydroxyapatite cement.

This study investigates the use of a new calcium phosphate cement, which sets to solid, microporous hydroxyapatite, for facial bone augmentation. In six dogs, the supraorbital ridges were augmented bilaterally with this hydroxyapatite cement. On one side, the hydroxyapatite cement was placed directly onto the bone within a subperiosteal pocket. On the opposite side, the cement was contained within a collagen membrane tubule and then inserted into a subperiosteal pocket. The use of collagen tubules facilitated easy, precise placement of the cement. All implants maintained their original augmented height throughout the duration of the study. They were well tolerated without extrusion or migration, and there was no significant sustained inflammatory response. Histologic studies, performed at 3, 6, and 9 months revealed that when the cement was placed directly onto bone, progressive replacement of the implant by bone (osseointegration of the hydroxyapatite with the underlying bone) without a loss of volume was observed. In contrast, when the cement-collagen tubule combination was inserted, primarily a fibrous union was noted. Despite such fibrous union, the hydroxyapatite-collagen implant solidly bonded to the underlying bone, and no implant resorption was observed. Hydroxyapatite cement can be used successfully for the experimental augmentation of the craniofacial skeleton and may be applicable for such uses in humans.

Animals↗

Glass-reinforced hydroxyapatite composites: secondary phase proportions and densification effects on biaxial bending strength.

CaO-P(2)O(5) glasses with additions of MgO and CaF(2) were used as a sintering aid of hydroxyapatite, and glass-reinforced hydroxyapatite composites obtained. Glasses promoted significant changes in the microstructure of the composites, namely with the formation of tricalcium phosphate secondary phases, beta and alpha-TCP. Quantitative phase analysis was performed by the Rietveld method using General Structure Analysis Software. Grain size measurements were carried out on SEM photomicrographs, using a planimetric procedure according to ASTM E 112-88. Flexural bending strength was determined from concentric ring-on-ring testing. Flexural bending strength (FBS) of glass-reinforced hydroxyapatite composites was found to be about twice or three times higher than that of unreinforced hydroxyapatite and tended to depend more on porosity and beta and alpha-TCP secondary phases, rather than on grain size. Traces of alpha-tricalcium phosphate significantly enhanced the strength of the composites. Using the rule of mixtures to estimate the zero porosity bending strength, the Duckworth-Knudsen model applied to the composites gave a porosity correction factor, b, with a value of 4.02. Weibull statistics were also used to analyze biaxial strength data and the level of reinforcement obtained by comparing failure probability for the composites and for the unreinforced hydroxyapatite. Lower activation energies for grain growth were observed for the composites compared to unreinforced hydroxyapatite, which should be attributed to the presence of a liquid glassy phase that promotes atomic diffusion during the sintering process.

Biocompatible Materials↗

In vivo hydroxyapatite formation induced by lipids.

Proteolipids and complexed acidic phospholipids that cause in vitro hydroxyapatite formation, similarly cause hydroxyapatite deposition in 10-mu pore Millipore chambers when implanted in rabbit muscle pouches. The amount of mineral deposited during a 3-week period, based on the calcium and phosphate contents of the chambers, was directly related to the dry weight of the lipid implanted in the chamber. Chambers containing total lipid extract from rabbit bone from which the complexed acidic phospholipids had been removed, acidic phospholipids from which the the proteolipids had been removed, and empty chambers did not accumulate any detectable mineral during the course of the study. Chambers implanted with synthetic hydroxyapatite served as controls for chemical analyses. The presence of hydroxyapatite in the chambers was established 3 weeks after implantation based on electron microscopic, compositional, and wide-angle X-ray diffraction analyses of the deposits. In the cell-free chambers, lipid-induced hydroxyapatite deposition, but not bone matrix formation occurred. This study demonstrates that proteolipids and complexed acidic phospholipids can cause hydroxyapatite mineral deposition in a physiologic environment. To date, these lipids are the only materials isolated from mineralizing tissues, other than reconstituted collagen, that have been shown capable of causing in vivo mineralization in the absence of cells.

Animals↗

Powder suspension method for critically re-examining the two-site model for hydroxyapatite dissolution kinetics.

A powder dissolution method has been developed, and experiments with the hydroxyapatite suspensions confirm earlier conclusions based on dissolution from hydroxyapatite disks. Although a quantitative assessment of the properties of site 1 was not possible from the data obtained in the present study, a rather accurate and independent evaluation of the properties and the behavior of site 2 of the two-site model for hydroxyapatite dissolution was possible, and the results clearly validate the original two-site model. The present work together with the earlier disk studies show that dissolution from site 2 is well described by a first-order expression, rate = kc2 (Cs2-C), where kc2 is a first-order rate constant, Cs2 is the apparent solubility for site 2 (defined by an ion activity product, KHAP, of the form a10Ca2+PO43-a2OH-, and the solution conditions), and C is the microenvironmental solution concentration of hydroxyapatite. For four different precipitated hydroxyapatite preparations, a single KHAP value of 1 X 10(-128) +/- 1 was found to be consistent with experiments using solutions covering wide ranges of partial saturation and calcium-phosphate ratios. The hydroxyapatite powder and pellet methods (including the data evaluation procedures) now offer a powerful combination for investigating the complex kinetics associated with dental enamel dissolution in particular and enamel chemistry in general.

Calcium↗

The inhibitory effect of cartilage proteoglycans on hydroxyapatite growth.

The calcification of connective tissues, including cartilage, is under the control of many interacting systems. Proteoglycans are thought to retard the deposition of hydroxyapatite crystals, and modification of the proteoglycans presumably facilitates mineralization in those tissues that are actively calcifying. The mechanism underlying these regulations remains speculative. This study investigates this question by comparing the inhibitory effectiveness of several macromolecules at neutral pH and approximately physiological ionic strengths. Inhibitors tested include bovine nasal proteoglycan monomer A1D1D1 and aggregate-containing A1 fractions, glycosaminoglycan chains (chondroitin 4-sulfate), and neutral dextran (as an uncharged analog). Hydroxyapatite growth was assessed either by measuring the time-dependent decreases in solution calcium and phosphate concentrations, or by determining utilization of hydroxyl ion in a pH-Stat. All species studied inhibit hydroxyapatite growth, and the extent of inhibition for each class is concentration-dependent. The proteoglycan aggregate-containing A1 fraction is more effective than the proteoglycan monomer at the same concentration, and the proteoglycan monomer is more effective than chondroitin 4-sulfate. Neutral dextran inhibits hydroxyapatite growth less effectively than proteoglycans. These results suggest that inhibition of hydroxyapatite growth by proteoglycans critically depends on both status (aggregate, monomer, etc.) and hydrodynamic size of this macromolecule, supporting the hypothesis that modification of proteoglycans in vivo functions to modulate the effectiveness of proteoglycans as a hydroxyapatite growth inhibitor.

Animals↗

Osseous metaplasia with functioning bone marrow in hydroxyapatite orbital implants.

BACKGROUND: Bone formation within the hydroxyapatite implant has been reported in explanted spheres in humans. Bone-specific differentiation was observed to occur earlier in the pores of spherical hydroxyapatite implants enhanced with osteogenin within the rabbit socket. We observed previously unreported bone marrow formation in a coralline hydroxyapatite implant placed into the rabbit orbit after evisceration. METHODS: One eye of each of 10 New Zealand white rabbits weighing between 2 and 3 kg was eviscerated and implanted with hydroxyapatite spheres. The explanted hydroxyapatite spheres 20 weeks after surgery were examined under the microscope. RESULTS: Histopathologic examination of the excised implants showed the presence of trabeculae of mature bone with fatty marrow and hematopoietic elements. Scattered throughout the fatty tissue were bone marrow elements consisting of precursors of the granulocytic and erythroid series and also megakaryocytes. CONCLUSION: The osseous metaplasia with functioning bone marrow was incidentally observed in the coralline hydroxyapatite implant without the addition of any osteogenesis-inducing agents.

Animals↗

Transmission electron microscopic observation of heterogeneous fluoridated hydroxyapatites.

Two different types of heterogeneous fluoridated hydroxyapatites previously synthesized at 80 degrees C and pH 7.4, which might be composed of hydroxyapatite covered with fluorapatite (H-FAp) and fluorapatite covered with hydroxyapatite (F-HAp), were observed by high-resolution transmission electron microscopy. The transverse-sectional shapes of both apatites were clearly hexagonal. This confirmed that both apatites were highly crystallized. However, their shapes differed slightly from each other in detail:H-FAp was more slender and F-HAp was typically hexagonal, probably reflecting the shapes of the original core apatites, hydroxyapatite and fluorapatite. The portions thought to be synthesized with fluoride-free solution showed clear electron beam damage. The expanded lattice image observed at the edge of the F-HAp crystal coincided approximately with the a-axis dimension of 0.9443 nm obtained from the X-ray diffraction data of synthetic hydroxyapatite. After incubation in 0.5 mol l-1 acetate buffer solution (37 degrees C, pH 4.0) for one month, the fluorapatite-covered H-FAp maintained its sharp corners, while the hydroxyapatite-covered F-HAp showed rounded corners.

Hydroxyapatites↗

Mechanical failure of hydroxyapatite- and polysulfone-coated titanium rods in a weight-bearing canine model.

Three types of material that have shown potential as coatings for orthopaedic implants were studied. Using a weight-bearing canine model, Ti-6A1-4V femoral intramedullary rods coated with (1) sintered titanium beads, (2) plasma-sprayed hydroxyapatite, and (3) silyl coupled polysulfone beads were evaluated for mechanical strength and bone ingrowth. The model was designed to secure optimal prosthetic stability by obtaining maximal bony ingrowth during an initial non-weight-bearing phase, then stressing the implant during a full-weight-bearing phase. None of the rods coated with titanium beads failed. All 17 polysulfone-coated rods failed, 13 of them at the interface between the polysulfone coating and the titanium core. Of 18 rods coated with hydroxyapatite, 15 suffered implant breakdown at the interface between the hydroxyapatite coating and the titanium core. This may be due to dissolution of the plasma-sprayed hydroxyapatite in vivo. Testing of retrieved specimens from both hydroxyapatite- and polysulfone-coated implants showed that the shear strength at the coating-rod interface had decreased to less than 40% of the shear strength at manufacture. Despite mechanical failure, histologic study showed extensive bone ingrowth or apposition onto both the polysulfone and hydroxyapatite coatings.

Animals↗

Apparent solubility distributions of hydroxyapatite and enamel apatite.

Samples of human dental enamel and hydroxyapatite were equilibrated at 5 mg/40 ml for 9 days at 37 degrees C with acetate buffers adjusted to a range of saturations with respect to hydroxyapatite. Sigmoidal apparent solubility distributions, in which the fraction dissolved was plotted against--log(ion activity product for hydroxyapatite) (pIHA), were constructed. About 10% of the hydroxyapatite and 14% of the enamel was very soluble, dissolving even at pIHA 55. The apparent solubility distributions for both solids were invariant with pH (4.5, 5.0, 5.5), within experimental error, showing that solubility was controlled by a phase with the stoichiometry of hydroxyapatite, probably in the form of a surface layer or complex on the crystals, in agreement with other studies on carbonate-apatites and bone mineral. The pIHA at 50% dissolution was employed as an average value. The pIHA (50%) values for pooled data (58.76 for enamel and 60.17 for hydroxyapatite) were lower than the respective pIHA previously measured by conventional equilibration techniques. However, the average pIHA measured for enamel was higher than that obtained by the same technique in another study, possibly because of differences in specimen preparation and equilibration time. The possible implications of the findings for understanding the process of dental caries are discussed.

Dental Enamel↗

Synthesis of Si, Mg substituted hydroxyapatites and their sintering behaviors.

Si, Mg-substituted hydroxyapatites, alone and co-substituted, have been prepared to obtain biomaterials having an improved biocompatibility. From FT-IR, XRD and ICP analyses, it was confirmed that single phases of hydroxyapatite substituted by Si alone or co-substituted by Si, Mg. The XRD data indicated the absence of extra phases related to silicon and magnesium oxide or other calcium phosphate species. Si-substituted hydroxyapatite of up to 2 wt% for Si and Si, Mg co-substituted hydroxyapatite of 1 wt% for the each ion keep their original structures intact for the sintering temperatures of up to 1200 degrees C. However, it is observed that ion substitutions by an amount higher than the above ratios for each hydroxyapatite lead to destabilization of original structures of the hydroxyapatite and to the production of tricalcium phosphate and calcium phosphate silicate phases when the samples were sintered at 1100 degrees C or higher.

Biocompatible Materials↗

Exposed hydroxyapatite orbital implants. Report of six cases.

Six patients with complications of primary or secondary hydroxyapatite implants were studied. Complications included socket infection and/or conjunctival dehiscence. Complications were detected during regular follow-up examinations, and various treatment approaches were used. The hydroxyapatite implant exposure occurred 4 to 6 weeks (mean, 4.5 weeks) after implantation. Three of the six implants were wrapped in preserved donor sclera before implantation. One of the implants showed wide exposure and chronic infection and was removed. In two cases, scleral patch grafts with a conjunctival pedicle graft were performed, resulting in successful coverage of the implant without further conjunctival dehiscence. In one of the patients, a Tenon's conjunctival flap was advanced to cover the defect, and was unsuccessful with the spicules of the hydroxyapatite eroding through the vascular flap after 1 month. Three of the patients demonstrate a persistent conjunctival epithelial defect. These three patients with chronically exposed hydroxyapatite have remained stable with follow-up intervals ranging from 8 to 12 months. Early exposure of hydroxyapatite orbital implants is a potential problem despite meticulous technique. Implant coverage is difficult, although chronic exposure seems to be tolerated often in the hydroxyapatite orbital implant without migration or extrusion.

Adolescent↗

Defect repair in rat mandible with hydroxyapatite cement compared to small intestine submucosa.

AIM: The aim of this study was to evaluate the bone formation in surgically created defects of rabbit mandibles by synthetic hydroxyapatite of calcium compared to small Intestine Submucosa. MATERIAL AND METHOD: 24 mice lineage Wisthar-Furth were used. A bony defect of 0,75 cm x 1,5 cm in mandibular ramus was accomplished in all animals. The hydroxyapatite implants were placed on the left hemimandiblein group I, small Intestine submucosa in group II, and the right served as control. The euthanasia was accomplished in the 40 degrees postoperative day, it was proceeded the macroscopic and histological analysis. RESULTS: medium length in millimeters of the hemimandibless in the hydroxyapatite group was of 3,75, in the small intestine submucosa 3,03 and the control group was of 2,63 (p: 0,022). Histomorphometry study revealed new bone grown in 76,64% of the total area in hydroxyapatite group (p: 0,022). In Small Intestinal submucosa group new bone grown in 63,64% do total (p: 0,0022). DISCUSSION: satisfactory bone integration was observed of the synthetic hydroxyapatite in that experimental model. Small intestinal submucosa cause osteoinduction CONCLUSION: using hydroxyapatite of calcium resulted in formation of significantly larger volume fractions of new bone when compared to small intestinal submucosa group.

Animals↗

Exposure rate of hydroxyapatite spheres in the anophthalmic socket: histopathologic correlation and comparison with silicone sphere implants.

We retrospectively reviewed enucleations and secondary anophthalmic socket sphere implantations for a 3 year period. We compared the incidence of exposure of hydroxyapatite implants to the incidence of exposure of silicone implants. We found that the incidence of hydroxyapatite exposure following enucleation was 3 of 27 (11.1%), and following secondary anophthalmic socket implantation was 3 of 32 (9.4%). The incidence of silicone sphere exposure following enucleation was 0 of 48 (0%), and following secondary implantation was 1 of 30 (3.3%). The difference in exposure rate between hydroxyapatite and silicone reached statistical significance in the enucleation group (p = 0.043) and in the combined enucleation and secondary implantation group (p = 0.033), but not in the secondary implantation group when considered separately. Osteoinduction and fibrovascular infiltration were found in all hydroxyapatite specimens examined histopathologically. In the exposed implants, liquefaction necrosis of the implant occurred. In the nonexposed implant, complete fibrovascular ingrowth was noted at 7 months. We believe that the hydroxyapatite anophthalmic sphere is associated with a higher incidence of exposure and postoperative inflammation when compared to silicone anophthalmic spheres. Patient selection and technique modification may reduce the incidence of hydroxyapatite implant exposure.

Adult↗