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Magnetron co-sputtered silicon-containing hydroxyapatite thin films--an in vitro study.

The use of silicon-substituted hydroxyapatite (Si-HA) as a biomaterial has been reported recently. In vivo testing has shown that Si-HA promotes early bonding of the bone/implant interface. In order to extend its usage to major load-bearing applications such as artificial hip replacement implants, it has been proposed that the material could be used in the form of a coating on implant surfaces. This paper reports a preliminary study of the biocompatibility of magnetron co-sputtered silicon-containing hydroxyapatite (Si-HA) coatings on a metallic substrate. Magnetron co-sputtered Si-HA films of thickness 600 nm with a Si content of approximately 0.8 wt% were produced on titanium substrates. X-ray diffraction analysis showed that the as-deposited Si-HA films were either amorphous or made up of very small crystals. The crystallinity of Si-HA films was increased after post-deposition heat treatment at 700 degrees C for 3 h, and the principal peaks were attributable to HA. The formation of nano-scale silicon-calcium phosphate precipitates was noted on the heat-treated films. In vitro cell culture has demonstrated that human osteoblast-like cells attached and grew well on all films, with the highest cell growth and signs of mineralisation observed on the heat-treated Si-HA films. In addition, many focal contacts were produced on the films and the cells had well-defined actin cytoskeletal organisation. This work shows that as-deposited and heat-treated Si-HA films have excellent bioactivity and are good candidates when rapid bone apposition is required. Furthermore, heat-treated Si-HA films have improved biostability compared to as-deposited films under physiological conditions.

Actins↗

Ectopic bone formation associated with mesenchymal stem cells in a resorbable calcium deficient hydroxyapatite carrier.

Bone substitute materials can induce bone formation in combination with mesenchymal stem cells (MSC). The aim of the current study was to examine ectopic in vivo bone formation with and without MSC on a new resorbable ceramic, called calcium deficient hydroxyapatite (CDHA). Ceramic blocks characterized by a large surface (48 m2/g) were compared with beta-tricalcium phosphate (beta-TCP), hydroxyapatite (HA) ceramics (both ca. 0.5 m2/g surface) and demineralized bone matrix (DBM). Before implantation in the back of SCID mice carriers were freshly loaded with 2x10(5) expanded human MSC or loaded with cells and kept under osteogenic conditions for two weeks in vitro. Culture conditions were kept free of xenogenic supplements. Deposits of osteoid at the margins of ceramic pores occurred independent of osteogenic pre-induction, contained human cells, and appeared in 416 MSC/CDHA composites compared to 216 MSC/beta-TCP composites. ALP activity was significantly higher in samples with MSC versus empty controls (p<0.001). Furthermore, ALP was significantly (p<0.05) higher for all ceramics when compared to the DBM matrix. Compared to previous studies, overall bone formation appeared to be reduced possibly due to the strict human protocol. Ectopic bone formation in the novel biomaterial CDHA varied considerably with the cell pool and was at least equal to beta-TCP blocks.

Aged↗

Natural materials for treatment of industrial effluents: comparative study of the retention of Cd, Zn and Co by calcite and hydroxyapatite. Part I: batch experiments.

This work explores the heavy metal retention capacity of materials developed from minerals that are abundant in nature, with low cost and minimum environmental impact. To accomplish this objective we have: (a) characterized commercial samples of calcite (CA) and hydroxyapatite (HAP)--including their surface properties (BET area, electrophoretic mobility, SEM, and X-ray energy dispersive spectroscopy); and, (b) qualified and quantified the interaction of Cd, Zn and Co with calcite (CaCO3) and hydroxyapatite [Ca5(PO4)3OH] through batch experiments, in a range of metal concentrations (4 Zn>Co and Cd>Zn approximately Co, respectively. Retention increased with pCa and pH and could be modeled by: (a) a non-ideal ion exchange mechanism (Me/Ca) for the adsorption of Cd, Zn and Co onto CA; and, (b) a mechanism of non-ideal ion exchange and specific adsorption (Me/Ca and identical with PO4O-Me) in the case of HAP. The pH dependence is indirect in CA and is related to its solubility changes (pCa increases with pH, and so does sorption of Cd, Zn and Co). Both materials, HAP and CA, can be used for heavy metal retention. The former has better performance for water treatment due to its greater efficiency for the retention of Cd, Zn and Co (over two orders of magnitude per gram of material) and its lower solubility in a wide range of pH (6<pH<9).

Adsorption↗

Use of Mimix hydroxyapatite bone cement for difficult ossicular reconstruction.

OBJECTIVE: To describe the advantages of using Mimix hydroxyapatite (HA) bone cement in reconstructing a variety of ossicular chain abnormalities. STUDY DESIGN AND SETTING: Case series at a tertiary medical center. RESULTS: Twenty-five cases of HA reconstruction are included in this series (ages 23-74; mean, 47 years). The examples presented include (1) HA as the sole reconstructive material for incus erosion, (2) HA for securing a total or partial ossicular replacement prosthesis, (3) incus augmentation after crimping for revision stapedotomy with incus erosion, (4) HA in primary stapedotomy to fix the crimped prosthesis to an intact incus, and (5) other unique situations. Preoperative and postoperative audiograms were evaluated for 4-tone pure tone average (PTA), speech reception thresholds, word recognition scores, and air-bone gaps (AB gaps). Mean follow-up was 11 months (range 2 to 22 months). The mean PTA improved from 57 dB to 37 dB, whereas the mean AB gaps decreased from 33 dB to 16 dB. There were no cases of infection or extrusion. CONCLUSIONS: Hydroxyapatite bone cement is an excellent adjunct or alternative to ossiculoplasty with preformed prostheses. Easily malleable, rapidly setting, and rapidly hardening, Mimix is particularly well suited for middle ear work. SIGNIFICANCE: Definitive fixation with bone cements during difficult ossicular chain reconstruction may ensure a more enduring successful outcome.

Adult↗

Interaction of DNA with hydroxyapatite. Studies on the effect of the phosphate concentration of the column equilibration and washing buffer.

The ability of hydroxyapatite to bind DNA effectively in phosphate solutions used for column equilibration, sample loading and column washing has been examined. It was demonstrated that substantial amounts of DNA (up to 40%) were eluted in the washing buffer when the phosphate concentration in the lysing solution or urea-phosphate used for column equilibration, sample loading and column washing was 0.24 M. A reduction in the phosphate concentration from 0.24 to 0.15 M in urea-phosphate solution led to almost 100% binding, whereas a similar reduction in the lysing solution did not. A modified method for loading and eluting DNA from hydroxyapatite columns is presented.

Absorption↗

Separation of high-molecular mass RNAs by high-performance liquid chromatography on hydroxyapatite.

High-molecular-mass RNAs [transfer-(t-), 5S-, 18S- and 28S-RNA] in 25 mM sodium acetate buffer (pH 6.0) were separated by high-performance liquid chromatography (HPLC) on hydroxyapatite using a linear gradient (120 min-duration) from 0.03 to 0.147 M of phosphate buffer (pH 7.0) containing 0.3 M potassium chloride and 1 mM sodium azide with a slope of 2 mM/ml at a flow-rate of 0.5 ml/min. When the RNAs were dissolved in 4 M guanidine isothiocyanate-25 mM sodium acetate buffer (pH 6.0)-0.1 M beta-mercaptoethanol (4 M GIT), t-, 5S- and 18S- or 28S-RNAs but not 18S- and 28S-RNAs were separated. RNAs extracted from rat superior cervical ganglia with 4 M GIT could be separated. Thus, HPLC on hydroxyapatite is a rapid and accurate means of quantifying and/or preparing high-molecular-mass RNAs such as t- and ribosomal RNAs.

Animals↗

High-performance liquid chromatography of transfer ribonucleic acids on spherical hydroxyapatite beads.

High-performance liquid chromatography (HPLC) on newly developed spherical beads of hydroxyapatite was applied to the analysis of purified E. coli tRNAs (Val, Met, Tyr and Phe). tRNAs were eluted from the column separately with appreciable differences in retention time by a 45-min gradient of phosphate buffer (pH 6.8) of concentration from 64 to 123 mM; both the retention times and peak areas of respective tRNAs were highly reproducible. Total tRNA (tRNA(Total)) preparations obtained from E. coli and B. subtilis were also analysed on the column. It is possible even to elute tRNA(Total) which may, in general, contain 60 or more tRNA species with a relatively shallow gradient such as 75-132 mM. The recovery of tRNA from the column was as high as 90%. Owing to the complicated composition, the elution profile of tRNA(Total) had a wide spread irregular shape but, with a 1-h gradient more than ten peaks were easily detected. When the amino acid-accepting activity of tRNA in the eluate of tRNA(Total) was determined, for ten specific tRNAs, each activity peak was eluted sharply from the column. In addition, several tRNA activities were eluted in different fractions. This indicates that isoacceptors were separated by the column. The results show that HPLC on hydroxyapatite beads is useful for the purification and characterization of tRNA.

Bacillus subtilis↗

Improved pontic/tissue relationships using porous coralline hydroxyapatite block.

Porous coralline hydroxyapatite blocks were implanted in the anterior region of several patients to improve the pontic/ridge relationship for a fixed prosthesis, thereby improving esthetic appearance and oral hygiene performance. A beveled incision on the palate was used to displace the flap coronofacially without leaving an area of denuded bone or graft. Flap margins were sutured to the adjacent palatal tissues. At the 2 1/2- and 3-year postgrafting follow-up, the implant area was healthy and ridge contours have been maintained. The permanent fixed partial denture is functional, and radiographic evaluation indicates the continued presence of the porous coralline hydroxyapatite implant. Three-year results have been promising enough to warrant continuing the procedure in patients requiring localized ridge augmentation.

Aged↗

Solubility of calcium oxalate monohydrate and hydroxyapatite in EDTA solutions.

The effect of the addition of a buffer to an ethylenediaminetetracetic acid (EDTA) litholytic solution on the solubility of calcium oxalate monohydrate and hydroxyapatite is investigated. The experiments show that the addition of triethanolamine as a buffer to EDTA solutions at pH 8 and 8.5 enhances the solubility of these salts, confirming theoretical predictions. The solubility of calcium oxalate monohydrate at 25C in solutions with varying ratios of buffer to EDTA concentrations and an osmolality of 0.9 was determined. The solutions resulting in the highest solubility were finally tested for their litholytic ability at 37C. At pH 8 solubilities of 22.5 g X l-1 for calcium oxalate monohydrate and 20.2 g X l-1 for hydroxyapatite were obtained. The corresponding solubilities at pH 8.5 were respectively 24.5 g X l-1 and 19.2 g X l-1.

Calcium Oxalate↗

Inhibitors of crystal growth of hydroxyapatite: a constant composition approach.

Pyrophosphate, citrate and magnesium, inhibitors of hydroxyapatite crystal growth, were studied using a seeded crystal growth system of constant composition at pH 5.80, 6.60 and 7.40. With this technique, crystal growth was studied at constant supersaturation at different pH values without the induction of other calcium phosphate phases. One inhibitor unit (that concentration of material that results in a reduction of 50 per cent in the growth rate from control) was calculated using the Langmuir adsorption isotherm. Pyrophosphate and citrate increased inhibitor activity with decreasing pH, whereas magnesium increased inhibitor activity with increasing pH. These data suggest that, at the urinary concentrations of these inhibitors, pyrophosphate is the most potent inhibitor, citrate less, and magnesium least. Pooled urine collections were studied using the same system and were found to have decreased inhibitor activity as pH decreased. This suggests that other modulators of hydroxyapatite, either promoters or inhibitors, are active in this system at the pH values studied.

Citrates↗

In vitro evaluation of an hydroxyapatite root canal system filling material.

An objective of endodontics is the three-dimensional filling of the root canal system, preventing leakage. The sealing ability of two hydroxyapatite formulations was compared with that of a gutta-percha/sealer combination placed with a vertical compaction technique. Zones of lysis tests of the hydroxyapatites showed no bacteriocidal effect of the formulations. Bacterial challenges of the sealing abilities of the formulations and gutta-percha/sealer combination showed that the formulations were not superior to the gutta-percha/sealer combination in their apical seal.

Bacillus subtilis↗

Biological evaluation of collagen gels containing calcium hydroxide and hydroxyapatite.

Dentinal chips and different biocompatible materials have been used in the past as apical plugs to enhance healing and simplify obturation. The purpose of this study was to evaluate bone reactions to implants of hydroxyapatite, collagen, and calcium hydroxide, alone or in different combinations. The materials were implanted bilaterally in the mandibles of 36 guinea pigs and the reactions were compared histologically over a period of 16 wk. No major inflammatory reactions were observed in any of the implant combinations. Hydroxyapatite was not resorbed over the examination periods, but calcium hydroxide and collagen implants were partially or totally resorbed and replaced by bony tissue. The results indicate that cylinders which may be useful as apical plugs can be performed and will heal with minimal biological reactions.

Animals↗

Electrodeposition of hydroxyapatite coatings in basic conditions.

Hydroxyapatite films have been grown in this work by an electrodeposition method involving both physical and chemical processes and presenting several differences with respect to other reported works. Description of the coating formation is based on the evolution of current through the sample placed as positive electrode in the basic electrolyte. The characterisation of hydroxyapatite films is of special importance since the bioactive properties related to HAP have been directly identified with its specific composition (Ca/P ratio) and crystalline structure. This characterisation has been traditionally fulfilled by the use of XRD, FTIR and SEM. Results of a further characterisation of the coatings by TEM and SFM, additional to the analysis by XRD, FTIR and SEM, are presented. Interpretation and comparison of our results with those obtained by other electrodeposition methods lead to arguments in favour of a deposition produced directly from ionic species.

Electrochemistry↗

Structural insights of glass-reinforced hydroxyapatite composites by Rietveld refinement.

Phase transformations and interstitial and/or substitution of trace elements during the liquid-phase sintering process of P2O5-CaO-MgO glass-reinforced hydroxyapatite (GR-HA) composites were examined by X-ray diffraction and Rietveld analyses. Using the Rietveld method for structure refinement, changes in the lattice parameters of the two main phases of the composites, hydroxyapatite (HA) and beta-tricalcium phosphate (beta-TCP), as well as changes in several bond lengths and in the occupancy of the hydroxyl oxygen site in the HA phase structure were assessed. The glasses gave rise to formation of between approximately 45 and 50% of beta-TCP, with evidence for the Mg2+ enhancing the formation of beta-TCP. Between 1,300 and 1,350 degrees C, the beta-TCP inverts to alpha-TCP, without further decomposition of the residual HA. The glasses showed evidence for stabilisation of the hydroxyl group located in the hydroxyl channels. This is supported by measurements of the hydroxyl channel radius (Rc), the Ca2-OH bond length and the hydroxyl oxygen occupancy (Oocc). Results showed that the Mg2+ containing glasses induced the beta-TCP phase formation in the structure of GR-HA composites and retarded the beta-TCP into alpha-TCP transformation at higher temperatures. The chemical composition of the P2O5 glasses also induces modifications in the lattice parameters of the crystallographic phases present in the microstructure of the composites. This suggests some substitution of Mg2 + -for-Ca2+ in the beta-TCP structure during the liquid-phase sintering process.

Biocompatible Materials↗

Fabrication of hydroxyapatite bodies by uniaxial pressing from a precipitated powder.

Hydroxyapatite (OHAp) has been obtained by using a precipitation method. Thermal stability of the precipitated powder has been checked to define an appropriate working range to process ceramic bodies without the appearance of odd phases. Pellets of hydroxyapatite have been manufactured by means of uniaxial pressing and convenient thermal treatments have been investigated to achieve high density OHAp bodies. Finally, factors that could affect the ultimate mechanical properties of the sintered bodies prepared by uniaxial pressing have been discussed.

Hydroxyapatites↗

Measurements of the solubilities and dissolution rates of several hydroxyapatites.

Calcium phosphate based materials, such as apatites, are increasingly being developed and used in implants for orthopedic and dental applications. Previous investigation of various calcium phosphate ceramics has demonstrated great variability in the solubility characteristics in solution between materials with similar stoichiometric composition. Therefore, in this study, the solubility and rate of dissolution of three apatite sources, BoneSource, Norian cranial repair system (CRS), and a sintered hydroxyapatite (Calcitite) are evaluated in a thermodynamically closed system. The measured solubility under physiological conditions (tris buffer solution, pH 7.4, 37 degrees C) of BoneSource, Norian CRS and Calcitite is 7.5, 7.4 and 1.4 ppm, respectively. Initial dissolution rates at 10 min of BoneSource, CRS, and Calcitite were 0.0465, 0.1589, and essentially 0 mg/min respectively. Solubility product constants at 37 degrees C were calculated to be 1.49 x 10(-35) for CRS, 1.19 x 10(-35) for BoneSource, and 2.92 x 10(-42) for Calcitite. The increased solubility of the BoneSource and Norian CRS materials over that of Calcitite is related to their poor crystallinity compared to sintered hydroxyapatite.

Biocompatible Materials↗

Aging effect on the phase evolution of water-based sol-gel hydroxyapatite.

In a number of recent reports on the synthesis of sol-gel hydroxyapatite, aging of the precursor solution has been found to be critical in developing an apatitic phase. Critical aging time is required to complete reaction between Ca and P molecular precursors to form a desired intermediate complex that permits a further transformation to apatite phase under appropriate thermal treatment. In this investigation, we employed a water-based sol-gel process recently developed to fabricate hydroxyapatite at relatively low temperatures. The aging effect on apatite formation was systematically studied in terms of aging time and temperature. Experimental results show that the aging time is considerably reduced as aging temperature rises. Long-term thermal aging was unfavorable for apatite formation. The optimal aging parameters for apatite formation were experimentally determined, which was further consolidated into a phase evolution map. Aging kinetic was investigated by monitoring the variation of solution pH, following the determination of an apparent activation energy, which has a value as high as 10.35 kcal/mol, for the chemical reaction occurring upon aging. Optimal solution chemistry was elucidated based on the corresponding phase evolution map.

Biocompatible Materials↗

Preparation, characterization and in vitro release of gentamicin from coralline hydroxyapatite-gelatin composite microspheres.

Composite microspheres have been prepared from bioactive ceramics such as coralline hydroxyapatite [CHA, Ca10(PO4)6(OH)2] granules, a biodegradable polymer, gelatin and an antibiotic, gentamicin. In our earlier work, we have shown a gentamicin release from CHA granules--chitosan composite microspheres. In the present investigation, an attempt was made to prepare the composite microspheres containing coralline hydroxyapatite and gelatin (CHA-G), which were prepared by the dispersion polymerization technique and the gentamicin was incorporated by the absorption method. The crystal structure of the composite microspheres was analyzed using X-ray powder diffractometer. The Fourier transformed infrared spectrum clearly indicated the presence of amide and hydroxyl groups in the composite microspheres. Scanning electron micrographs and optical micrographs show that the composite microspheres are spherical in shape and porous in nature. The particle size of composite microspheres was analyzed and the average size was found to be 16 microm. The thermal behavior of composite microspheres was studied using thermogravimetric analysis and differential scanning calorimetric analysis. The cumulative in vitro release profile of gentamicin from composite microspheres showed near zero order patterns.

Biocompatible Materials↗