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Inhibition of hydroxyapatite crystal growth by lipoteichoic acid.

Lipoteichoic acids were purified from freshly isolated strains of Streptococcus sanguis and Lactobacillus casei, and from a laboratory strain of Strep. rattus BHT. Their effects on the seeded growth of hydroxyapatite in 1 mM CaCl2, 7.5 mM KH2PO4, 50 mM HEPES (pH 7.2) were determined. At lipoteichoic acid concentrations of 25 microM organic phosphate (Po) and higher, the growth of hydroxyapatite appeared to be inhibited, at 100 microM Po inhibition was complete for several hours. In contrast, low concentrations of lipoteichoic acids (5-10 microM Po) may have a stimulatory effect on the seeded growth of hydroxyapatite. It was concluded that lipoteichoic acid may be one of the bacterial products that inhibit enamel lesion repair in vivo.

Animals↗

Interactions of streptococcal glucosyltransferases with alpha-amylase and starch on the surface of saliva-coated hydroxyapatite.

The salivary pellicle consists of various proteins and glycoproteins which may interact with one another. Experiments were performed to elucidate the interactions of streptococcal glucosyltransferase (Gtf) enzymes with human salivary alpha-amylase in solution and on the surface of saliva-coated hydroxyapatite (SHA) beads. The Gtf enzymes -B, -C and -D, when immobilized on to SHA beads, reduced the activity of adsorbed amylase; GtfD showed the highest inhibition of salivary amylase activity. The presence of glucan produced by immobilized GtfD did not further reduce amylase activity. The amount of amylase adsorbed on to hydroxyapatite beads was reduced when salivary amylase was added simultaneously with any of the Gtf enzymes, suggesting that amylase and Gtfs may compete with each other for binding sites on hydroxyapatite. Starch hydrolysates produced by SHA-surface-bound salivary amylase were tested for their effect on glucan production from sucrose by Gtf enzymes in solution and on SHA beads; glucan production by SHA-immobilized GtfB was stimulated in the presence of starch hydrolysates. Glucan synthesized by SHA-immobilized GtfB in the presence of starch hydrolysates was less susceptible to hydrolysis by the fungal enzyme mutanase than was glucan made by SHA-immobilized GtfB in the absence of starch hydrolysates. Glucan production by GtfB associated with streptococci immobilized on to SHA was also enhanced in the presence of starch hydrolysates. The adhesion of oral micro-organisms to SHA coated with glucan made in the presence and absence of starch hydrolysates was investigated, and some bacteria displayed higher adhesion activities for the glucan made in the presence of the hydrolysates. Therefore, the interaction of amylase and Gtf enzymes on a SHA surface may modulate the formation of glucan and the adherence of oral micro-organisms.

Adsorption↗

Adsorption of saliva-coated and plain streptococcal cells to the surfaces of hydroxyapatite beads.

This study was conducted to observe the differences between the adsorption of Streptococcus sanguis by saliva-coated and non-saliva-coated cells (plain cells) to the surfaces of saliva-coated, bovine serum albumin-coated or buffer-washed hydroxyapatite beads. Saliva treatment of bacterial cells altered the manner of saturation due to changes of affinity and/or the maximal number of adsorption sites on the hydroxyapatite beads. This result indicates that saliva treatment of bacterial cells might be a necessary step in testing bacterial adsorption to saliva-coated hydroxyapatite beads or saliva-coated teeth.

Adsorption↗

Induction and morphology of hydroxyapatite, precipitated from metastable simulated body fluids on sol-gel prepared silica.

Hydroxyapatite crystallization is induced at 37 degrees C by sol-gel prepared silica from metastable calcium phosphate solutions. The morphology of the apatite forming on the silica surface depends on the nature of the solutions. For example, apatite grew in a flake-like form at pH 7.4. The morphology changed to plate-like when the pH was adjusted to pH 7.2. At this lower pH, the apatite plate even exhibited a hexagonal feature, reflecting the unique hexagonal structure of hydroxyapatite. An increase in either Mg or P ion concentration of the fluid can cause apatite to grow in a rod-like shape while addition of F ions to the fluid leads to a perfect needle pattern. The flake geometry of apatite was not altered by increasing Ca concentration from 2.5 to 3.8 mM in the solution. From this we conclude that sol-gel prepared silica is an efficient apatite inducer and the morphology of the hydroxyapatite deposit is determined by factors of the fluid such as pH, Ca/P molar ratio, Mg and F concentrations.

Body Fluids↗

Relationship between the colour change of hydroxyapatite and the trace element manganese.

Hydroxyapatite containing the trace element manganese turns blue after sintering at high temperature in an oxidizing atmosphere. This phenomenon is related to the oxidation of manganese ions in the special crystal structure of hydroxyapatite. Combination of the crystal structure with electron spin resonance spectra gives an explanation for this oxidation of Mn2+ ions at Ca(l) sites into Mn5+ ions and the formation of MnO4(3-) ions at PO4(3-) sites, corresponding to the colour change of hydroxyapatite.

Calcium↗

Bone marrow induced osteogenesis in hydroxyapatite and calcium carbonate implants.

In this experimental study, blocks of natural coral (calcium carbonate) and its structurally similar derivate in the form of hydroxyapatite (calcium phosphate) were implanted in rat latissimus dorsi muscle with autogenous bone marrow to compare their bone-forming capability. A block without marrow placed in the opposite latissimus muscle served as a control. The animals were killed at 3, 6 and 12 weeks and, in the hydroxyapatite group, also at 24 weeks. The sections were analysed histologically and histomorphometrically. Bone was found only in implants containing bone marrow. Bone formation was significantly (p < 0.05) higher in coral than in hydroxyapatite implants at 3 weeks (10.8% versus 4.8%) and at 12 weeks (13.7% versus 6.3%, bone/total original block area). At 12 weeks all the coral implants had lost their original structure, and the cross-sectional area of the block had diminished to 40% of the original area.

Animals↗

Uptake and biodistribution of 99mtechnetium methylene-[32P] diphosphonate during endosteal healing around titanium, stainless steel and hydroxyapatite implants in rat tibial bone.

Early evaluation of intraosseous implant success and failure is critical, but, until now, there have been no reliable systems of measurement. The present study assessed whether the use of 99mtechnetium methylene-[32P]diphosphonate (99mTcMD32P), a marker for both bone formation and mineralization, can indicate if an implant is bone-bonding or non-bonding. Moreover, this study examined how bone-bonding (titanium and hydroxyapatite) and non-bonding (stainless steel) implants affected the normal healing of bone after marrow ablation, as measured by uptake of 99mTc and 32P. Titanium, hydroxyapatite and stainless steel implants were placed in the right tibiae of Sabra strain rats following ablation of the marrow, and 99mTcMD32P was injected 18 h before harvest. AT 3, 6, 14, 21 and 42 d (and in some experiments, on days 28 and 35) post-injury, the treated and contralateral tibiae were removed and cleaned of soft tissue. The uptake of 99mTc and 32P was measured in the whole bone, as well as in its organic and inorganic phases. Effects of the implants were assessed by comparing the treated to the untreated tibia in each rat. The distribution of 99mTc and 32P varied with each implant. After the insertion of titanium, increased 99mTc uptake was seen in whole bone and in the inorganic and organic phases at days 6-14. 32P uptake in whole bone and in the inorganic phase increased only at day 6, and 32P uptake was decreased in the organic phase at that time. In tibiae implanted with hydroxyapatite, 99mTc and 32P uptake was seen in the whole bone at days 6 and 14. While 99mTc uptake was increased in both the organic and inorganic phases, 32P uptake into the organic phase was decreased at both day 6 and day 14. In tibiae implanted with stainless steel, effects were observed only on day 6. The increased 99mTc uptake in whole bone reflected increases in both the organic and mineral phases. Increased 32P uptake was observed in whole bone as well, due to an increase in the 32P uptake in the mineral phase only; incorporation of 32P in the organic phase was comparable to that found in the contralateral limb. The results of this study indicate that implants alter bone healing, as indicated by the uptake of 99mTc and 32P in the different bone compartments. Moreover, decreased 32P uptake by the organic phase in the presence of bone-bonding implants suggests that cleavage of 99mTcMD32P into its technetium and methylene diphosphonate moieties was inhibited, perhaps as a function of the onset of calcification in the newly synthesized osteoid. The effect of the implants on bone healing was observed on days 6-14, when active bone formation and mineralization were occurring, supporting the hypothesis that these materials events associated with initial calcification. Uptake of 99mTc varies as a function of time, and uptake of 32P varies with time and distribution in the mineral or organic phase of bone, suggesting that these parameters may be useful as indicators of bone-bonding.

Animals↗

Primary placement of a hydroxyapatite-coated sleeve in bioceramic orbital implants.

PURPOSE: To study a new surgical option of primary placement of a hydroxyapatite-coated sleeve into the Bioceramic implant during enucleation or evisceration. DESIGN: Retrospective, observational case series. METHODS: A standard enucleation or evisceration was performed, followed by the preplacement of a hydroxyapatite-coated sleeve into the Bioceramic implant. Care must be taken to ensure the sleeve has been positioned centrally when the implant is put inside the orbital socket. Complications such as sleeve exposure, Bioceramic implant exposure, and infection were closely observed. RESULTS: Twenty-seven patients were treated in above fashion with five enucleation and 22 evisceration procedures. Five of the sleeves have exposed spontaneously during 1 to 4 months after original surgery. They had no further complication, except for one sleeve around which there were visible Bioceramic spicules attributable to long-term corticosteroid usage. The remaining 22 sleeves that did not spontaneously expose pursued secondary exposure of the sleeve and peg insertion by the conjunctival cutdown procedure 3 months postoperatively. One sleeve was medially positioned far away from the implant center. Re-insertion of new sleeve and peg was scheduled 2 weeks later. One additional sleeve was obliquely positioned after conjunctival cutdown procedure. Fortunately, all 27 patients were successfully fitted with a peg-coupled prosthesis with good motility. CONCLUSIONS: Primary placement of a hydroxyapatite-coated sleeve into the Bioceramic implants has several advantages, including high patient acceptance, technical simplicity, and an office-based conjunctival cutdown pegging procedure. By avoiding the expense of postoperative imaging studies and additional prosthetic modification, a more rapid and efficient rehabilitation is possible.

Adult↗

Slow crack growth behaviour of hydroxyapatite ceramics.

Among materials for medical applications, hydroxyapatite is one of the best candidates in orthopedics, since it exhibits a composition similar to the mineral part of bone. Double torsion technique was here performed to investigate slow crack growth behaviour of dense hydroxyapatite materials. Crack rate, V, versus stress intensity factor, K(I), laws were obtained for different environments and processing conditions. Stress assisted corrosion by water molecules in oxide ceramics is generally responsible for slow crack growth. The different propagation stages obtained here could be analyzed in relation to this process. The presence of a threshold defining a safety range of use was also observed. Hydroxyapatite ceramics appear to be very sensitive to slow crack growth, crack propagation occurring even at very low K(I). This can be explained by the fact that they contain hydroxyl groups (HAP: Ca(10)(PO(4))(6)(OH)(2)), favouring water adsorption on the crack surface and thus a strong decrease of surface energy in the presence of water. This study demonstrates that processing conditions must be carefully controlled, specially sintering temperature, which plays a key role on V-K(I) laws. Sintering at 50 degrees C above or below the optimal temperature, for example, may shift the V-K(I) law towards very low stress intensity factors. The influence of ageing is finally discussed.

Biocompatible Materials↗

Early bone apposition in vivo on plasma-sprayed and electrochemically deposited hydroxyapatite coatings on titanium alloy.

Three different implants, bare Ti-6Al-4V alloy, Ti-6Al-4V alloy coated with plasma-sprayed hydroxyapatite (PSHA), and Ti-6Al-4V alloy coated with electrochemically deposited hydroxyapatite (EDHA), were implanted into canine trabecular bone for 6 h, 7, and 14 days, respectively. Environmental scanning electron microscopy study showed that PSHA coatings had higher bone apposition ratios than those exhibited by bare Ti-6Al-4V and EDHA coatings after 7 days; however, at 14 days after implantation, EDHA and PSHA coatings exhibited similar bone apposition ratios, much higher than that for bare Ti-6Al-4V. The ultrastructure of the bone/implant interface observed by transmission electron microscope showed that the earliest mineralization (6 h-7 days) was in the form of nano-ribbon cluster mineral deposits with a Ca/P atomic ratio lower than that of hydroxyapatite. Later-stage mineralization (7-14 days) resulted in bone-like tissue with the characteristic templating of self-assembled collagen fibrils by HA platelets. Though adhesion of EDHA coatings to Ti-6Al-4V substrate proved problematical and clearly needs to be addressed through appropriate manipulation of electrodepositon parameters, the finely textured microstructure of EDHA coatings appears to provide significant advantage for the integration of mineralized bone tissue into the coatings.

Alloys↗

Self-assembly of synthetic hydroxyapatite nanorods into an enamel prism-like structure.

The application of surfactants as reverse micelles or microemulsions for the synthesis and self-assembly of nanoscale structures is one of the most widely adopted methods in nanotechnology. These synthesized nanostructure assemblies sometimes have an ordered arrangement. The aim of this research was to take advantage of these latest developments in the area of nanotechnology to mimic the natural biomineralization process to create the hardest tissue in the human body, dental enamel. This is the outermost layer of the teeth and consists of enamel prisms, highly organized micro-architectural units of nanorod-like calcium hydroxyapatite (HA) crystals arranged roughly parallel to each other. In particular, we have synthesized and modified the hydroxyapatite nanorods surface with monolayers of surfactants to create specific surface characteristics which will allow the nanorods to self-assemble into an enamel prism-like structure at a water/air interface. The size of the synthetic hydroxyapatite nanorods can be controlled and we have synthesized nanorods similar in size to both human and rat enamel. The prepared nanorod assemblies were examined using transmission electron microscopy (TEM) and atomic force microscopy (AFM). The specific Langmuir-Blodgett films were shown to be comprised of enamel prism-like nanorod assemblies with a Ca/P ratio between 1.6 and 1.7.

Air↗

In vitro growth and differentiation of osteoblast-like cells on hydroxyapatite ceramic granule calcified from red algae.

PURPOSE: The purpose of this study was to analyze the interaction between osteoblast-like cells isolated from mandibular bone and hydroxyapatite ceramic bone substitute obtained from calcified red algae to assess the growth and differentiation of adherent cells on this biomaterial. MATERIALS AND METHODS: The macroporous ceramic material C GRAFT/Algipore (The Clinician's Preference LLC, Golden, CO) is composed of 100% hydroxyapatite and possesses specific mechanical and physiochemical properties. Osteoblast-like cells were seeded on 200 mg of biomaterial and cultured for 6 and 21 days under osteogenic differentiation conditions. Specific alkaline phosphatase activity, DNA, and protein content of the proliferating cells were analyzed. The morphology of the cells in contact with the biomaterial was examined by scanning electron microscopy. The osteoblastic phenotype of the cells was confirmed by analysis of the expression of bone-specific genes (osteocalcin, osteopontin and collagen type I) by semi-quantitative reverse transcriptase polymerase chain reaction. RESULTS: The DNA and protein content increased over the culture period. Scanning electron microscopy showed cells spreading on the surface of the biomaterials, covering the macropores, and colonizing the depth of the particles. The analysis of the expression patterns of bone-related genes confirmed the osteoblastic phenotype of the cultured cells. CONCLUSION: The results of this study showed that hydroxyapatite ceramic bone substitute obtained from calcified red algae support the proliferation and differentiation of human osteoblast-like cells on its surface in vitro and might be suitable for use as scaffolds in tissue engineering strategies in vivo.

Bone Substitutes↗

An investigation of the effects of hydroxyapatite coatings on the fixation strength of cortical screws.

Hydroxyapatite (HA) are commonly applied to orthopaedic implants for acceleration of osteointegration and so overcoming the loosening problems such as in cortical screws. Electrophoretic deposition (EPD) of hydroxyapatite was applied for coating of cortical screws in this work. The effects of hydroxyapatite-coated and uncoated cortical screws on insertion and extraction torque were investigated through in vivo experiments. Three groups of screws were undertaken: first group with no coating, second group coated with HA and the third group coated with HA+interlayer, a synthetic calcium silicate compound. Five sheep were operated, and 60 cortical (20 x 3) screws from those of groups were implanted in cortical femurs to observe the effect of HA and interlayer on screws. Results show that as an alternative to plasma spray coating method, the EPD process enables to produce a quick, easy, cheap and uniform adjustable coating layer. Also from biomechanical and SEM examinations, HA coating by EPD method of cortical screws resulted in extremely improved fixation with reduced risk of loosening problem.

Animals↗

Nanoscale characterization of the interface between bone and hydroxyapatite implants and the effect of silicon on bone apposition.

Silicon plays an important role in bone mineralization and formation and is therefore incorporated into a wide variety of medical implants and bone grafts used today. The significance of silicon (Si) can be understood through an analysis of the mechanisms of bone bonding to calcium containing biomaterials and through comparisons of hydroxyapatite (HA) and silicon-substituted hydroxyapatite (Si-HA). The addition of Si to HA causes a decrease in grain size that subsequently affects surface topography, dissolution-reprecipitation rates and the bone apposition process. Through the use of high-resolution transmission electron microscopy (HR-TEM) studies, the interactions between bone and silicon hydroxyapatite (Si-HA) at interfaces are reviewed and related to their impact on bone apposition and ultimately the performance of medical implants.

Animals↗

Histologic evaluation of a 12-year-old threaded hydroxyapatite-coated implant placed in conjunction with subantral augmentation procedure: a clinical report.

This clinical report presents a histologic evaluation of a threaded hydroxyapatite-coated root-form implant retrieved from the posterior maxillary area of a patient after being in function for 12 years. The implant had been placed in conjunction with subantral augmentation. Histologically, the portion of the implant that was examined under light microscopy appeared to be integrated with the surrounding bone; 74.3% of the surface of the implant had close bone apposition at the interface. The residual graft particles were present and in close proximity to the implant surface. These observations suggest that subantral augmentation procedure performed simultaneously with the placement of a hydroxyapatite-coated implant by using hydroxyapatite as a graft material may result in osseointegration between the implant and the surrounding bone that can be maintained under long-term function.

Aged↗

Osteogenesis in a human hydroxyapatite orbital implant 5.5 years after implantation.

PURPOSE: We report the histological findings of a hydroxyapatite orbital implant 5.5 years after implantation. METHODS: The hydroxyapatite orbital implant was excised from the left orbit of a 17-year-old female, and it was decalcified and processed for routine light microscopic evaluation. RESULTS: The implant showed complete fibrovascularization with nearly complete osteogenesis with hematopoiesis. CONCLUSION: Bone may biointegrate throughout the pores of hydroxyapatite orbital implants.

Adolescent↗

Bone formation in hydroxyapatite orbital implants.

PURPOSE: We examined the histopathologic features of two hydroxyapatite orbital implants that were removed from two patients. METHODS: Two surgically removed hydroxyapatite orbital implants were decalcified and processed for routine light microscopic examination. RESULTS: Both implants demonstrated fibrovascular ingrowth and foci of bone formation. CONCLUSION: Bone may form in hydroxyapatite orbital implants.

Adult↗

Wrapping hydroxyapatite orbital implants with posterior auricular muscle complex grafts.

PURPOSE: To report the use of posterior auricular muscle complex grafts as a wrapping material for hydroxyapatite orbital implants in enucleation surgery. METHOD: In a retrospective multicenter study, autogenous posterior auricular muscle complex grafts were used to cover hydroxyapatite orbital implants in 83 patients with a mean age of 38.6 years (range, 1 to 85 years), of whom 63 had primary unilateral enucleation and 20 had secondary orbital implants following enucleation. The mean follow-up period after posterior auricular muscle complex grafts was 36 months (range, 14 to 60 months). RESULTS: Of the 83 patients, two (2.4%) developed limited orbital implant exposure, which was treated with a second posterior auricular muscle complex "patch graft." No patient developed postoperative orbital infection or implant extrusion. All patients were fitted with an acceptable prosthesis and had satisfactory cosmetic and functional results. No patient developed vascular compromise or a wound defect associated with the posterior auricular donor site. CONCLUSION: Autogenous posterior auricular muscle complex grafts should be considered as an appropriate wrapping material for hydroxyapatite orbital implants for primary enucleation and for secondary orbital implants after enucleation.

Adolescent↗