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PubMed1995

Structural evaluation of human and sheep bone and comparison with synthetic hydroxyapatite by FT-Raman spectroscopy.

The composition of whole human and sheep cortical bone tissue, and of a synthetic hydroxyapatite (P120), were compared using Fourier transform Raman (FT-Raman) spectroscopy. Deproteination procedures to remove the bulk of the collagen present in bone tissue allowed isolation of the mineral phase. A comparison of the spectra obtained from both whole and deproteinated bone with those of synthetic hydroxyapatite showed direct correlation only in the region of 952 cm-1 (symmetric P-O mode). In contrast, human and sheep bone were very closely matched in both, the organic and inorganic structures. The results demonstrate that deproteination of bone is not a necessary precursor to obtain spectral information.

Animals↗
PubMed1995

Effects of gamma-alumina and hydroxyapatite coatings on the growth and metabolism of human osteoblasts.

The behavior of human osteoblasts cultivated on hydroxyapatite or alumina-coated disks of Ti6AL4V was studied in vitro. Cell anchorage and spreading were observed by scanning electron microscopy. Cell growth was monitored by counting cells and measuring DNA at 5 h and 2,5, and 10 days after cell seeding. Cells grown for 10 days were labeled with 14C-proline and total protein and collagen synthesis were measured; the type of collagen was also determined. Both ceramics showed excellent biocompatibility. At 10 days of culture the cells showed a higher rate of proliferation on alumina than on hydroxyapatite. Neither ceramic altered the collagen biosynthesis or the osteoblast-like properties of the cells, as indicated by the high percentage of type I collagen.

Aluminum Oxide↗
PubMed2003

Dry mechanochemical synthesis of hydroxyapatites from dicalcium phosphate dihydrate and calcium oxide: a kinetic study.

Calcium phosphate ceramics have been used successfully as synthetic bone substitutes in orthopedics, dentistry, and maxillofacial surgery. One way of preparing these ceramics is the sintering of a calcium-deficient hydroxyapatite (CDHA), which can be obtained in different ways. Mechanochemistry is one possible means of synthesizing CDHA, with an expected molar calcium-to-phosphate (Ca/P) ratio +/- 0.005. The grinding can be carried out under dry or wet conditions. To optimize the experimental conditions of CDHA preparation by dry mechanosynthesis and for a better understanding of the DCPD/CaO mechanochemical reaction, we performed a kinetic study in which some of the experimental parameters were varied. Carried out with two different vertical rotating ball mills, this kinetic study showed that (1) experiments are reproducible and give as a final product a hydroxyapatite powder, formed of nano-sized crystals of around 20 nm, with a controlled Ca/P ratio; (2) the time for complete disappearance of DCPD and the time for complete reaction are in direct proportion to the mass of the ground powder; but (3) the time for complete disappearance of DCPD is independent of the Ca/P ratio while the time for complete reaction increases exponentially with the Ca/P ratio; and (4) the time for complete disappearance of DCPD corresponds to the time for complete reaction solely for Ca/P = 1.5. These observations suggest a reaction mechanism in two well differentiated stages: (First stage) CaO reacts with DCPD to give first an amorphous calcium phosphate (ACP) with a low Ca/P ratio that transforms into CDHA when its Ca/P ratio reaches 1.5. At the same time, CaO is hydrated into Ca(OH)(2) by the water produced by the reaction. (Second stage) If the Ca/P > 1.5 in the initial mixture, the excess Ca(OH)(2) is added to CDHA 1.5 by reacting with the HPO(4) group of CDHA until its Ca/P ratio reaches the expected value. The slower the reaction, the higher the Ca/P in the initial mixture.

Biocompatible Materials↗
PubMed2005

Effects of bone morphogenetic protein-2 and hyaluronic acid on the osseointegration of hydroxyapatite-coated implants: an experimental study in sheep.

Research efforts aim at enhancing early osseointegration of cementless implants to improve early fixation and, thus, reduce the risk of loosening. The aim of the present study was to investigate whether bone morphogenetic protein (BMP) 2 had a positive effect on the osseointegration of hydroxyapatite-coated implants. Hydroxyapatite (HA) implants (perforated hollow cylinders and solid rods) were coated with BMP-2 and hyaluronic acid (HY) as the carrier or with HY alone. Uncoated HA implants served as controls. The osseointegration of the implants was evaluated either by light microscopy or by pullout tests after 1, 2, and 4 weeks of unloaded implantation in the cancellous bone of 24 sheep. The BMP-2 coating significantly increased bone growth into the implant perforations compared with HA-coated implants at 2 and 4 weeks. Bone-implant contact and interface shear strength of BMP-2 implants were lower than HA implants at 2 weeks. At 4 weeks, there was no significant difference in bone-implant contact and shear strength between BMP-2 and HA-coated implants. The BMP-2 coating enhanced gap healing but had no positive or even an inhibitory effect (at 2 weeks) on bone-implant contact and interface shear strength. In the clinical situation, a perfect press-fit implantation cannot be achieved, and BMP-2 may be beneficial for enhancing bone growth into gaps around cementless implants.

Animals↗
PubMed2003

Adsorption of catechol and comparative solutes on hydroxyapatite.

Contemporary medical and dental adhesives often have difficulty sticking to wet surfaces or weaken with long-term exposure to water. Substantial research has been dedicated to finding a means of achieving adhesion in an aqueous environment. A study evaluates the adsorption of catechol relative to other chemical groups as means of gauging how effective they may be as adsorptive groups in adhesives. Contact angle and surface-tension measurements of solutions of catechols and other chemical groups were used to determine their works of adhesion. Adsorption isotherms were also constructed to ascertain Langmuir constants. Solutes containing catechol groups were compared to solutes containing other polar groups to see how well catechol adsorbs to hydroxyapatite, the mineral component of bones and teeth, relative to other chemical groups found in adhesives. The results of this study show that catechol and molecules containing catechol groups have higher rates and energies of adsorption to hydroxyapatite than do groups such as alcohols, amines, and carboxylic acids.

Adsorption↗
PubMed1987

Bone ingrowth into polymer coated porous synthetic coralline hydroxyapatite.

Porous hydroxyapatite, converted hydrothermally from the calcium carbon exoskeleton of the coral genus Goniopora (CHAG), has been shown to be effective as a scaffold for bone ingrowth (2,3,5-7,9). However, the large pores in the material resulted in low compressive strengths. In a previous study, we found that microcoating the internal surfaces of CHAG with dilactic-polyactic acid (DL-PLA) improved significantly its compressive properties. The objective of this study was to determine the effect of PLA microcoating on bone ingrowth into CHAG plugs. Plugs of thick- (3:1 chloroform to DL-PLA by weight), medium- (10:1), and thin- (30:1) coated as well as uncoated CHAG were implanted transcortically in the proximal third of the diaphysis of the rabbit tibia. Specimens were harvested at 3, 12, and 24 weeks for mechanical testing and contralaterally for histological and histomorphometric assessment. At 12 weeks, uncoated CHAG plugs developed an average ultimate interface shear stress of 26.7 MPa, compared with 17 MPa for 30:1 and 8 MPa for 10:1 and 3:1 coated specimens. At 24 weeks, there were no significant differences in shear stress among any of the specimens. Histomorphometric assessments showed that the ratio of area fractions of new bone to area fractions of new bone and void space increased from 68-70% for 3:1 and 10:1 coated specimens at 3 weeks, and to 85.5-89.5% at 24 weeks. In comparison, uncoated and 30:1 specimens had area fraction ratios of about 82% at 3 weeks and 93% at 24 weeks. Histologic sections demonstrated direct apposition of new bone to both the coating and the hydroxyapatite as well as degradation of the coating.

Animals↗
PubMed1992

Treatment of proteoglycan aggregates with physeal enzymes reduces their ability to inhibit hydroxyapatite proliferation in a gelatin gel.

In vitro, cartilage proteoglycans (PGs) are effective inhibitors of hydroxyapatite formation and growth. Their inhibitory ability decreases with decreasing PG size and charge density. It has been suggested that the enzyme-mediated alteration in the size and conformation of PGs in the growth plate may similarly facilitate the calcification process. In this study, a gelatin gel system was used to monitor hydroxyapatite formation and growth in the presence of proteoglycan aggregates, before and after enzyme treatment. To reproduce the physeal degradation cascade, an enzyme preparation was used that contained all of the growth plate enzymes. At a concentration of 500 micrograms/ml, the untreated proteoglycan aggregates reduced the amount of mineral formed by 30%. When the aggregates were treated with the heat-inactivated enzyme, the same extent of inhibition was found. In contrast, treating the aggregates with the crude growth plate enzyme preparation removed all the inhibitory ability, such that 500 micrograms/ml of proteoglycan preparation yielded 10% more mineral than the controls. Treatment of the aggregates with chondroitinase ABC and trypsin, similarly removed all the inhibitory ability. These data, suggest that enzymatic degradation of proteoglycans may contribute to the regulation of growth plate calcification.

Animals↗
PubMed1992

Slow release of anticancer drugs from porous calcium hydroxyapatite ceramic.

We have developed a new delivery system for sustained release of an anticancer drug (cis-platinum) by enclosure into blocks of porous calcium hydroxyapatite ceramic. The slow release of this drug from this system was confirmed in in vitro experiments. When this system was implanted into normal back muscle, or the tibia, sustained release of cis-platinum was observed during a 12-week period after implantation. The diffusion rate of cis-platinum into blood and other organs (liver, kidney, brain) was less than 10% of that at the implanted site. This delivery system placed into experimental tumors of mice also showed a uniform release of anticancer drug for more than 3 months. Inhibition of tumor growth was more marked after local implantation of this system than after intraperitoneal administration of cis-platinum. These results indicate that this new approach to a drug delivery system may well have an important role in cancer chemotherapy. In bone tumors it is attractive because the mechanical strength of calcium hydroxyapatite ceramic permits partial surgical excision and replacement of the bone defect at the same time.

Animals↗
PubMed1981

Fluoride remineralization of demineralized bovine tooth enamel and hydroxyapatite pellets.

Both bovine enamel and hydroxyapatite pellets were remineralized in a fluoride-containing remineralization solution after prior demineralization for various lengths of time. In both the enamel and pellet systems, the degree of remineralization attainable was directly related to the extent of prior demineralization, although the demineralized material was never 100% recovered in remineralization. In some cases, fluoride levels up to several thousand parts per million were found at depths as great as 50 micrometers from the surface. The stoichiometry of the remineralized material and electron microprobe examination were consistent with the formation of fluoridated hydroxyapatite rather than calcium fluoride.

Animals↗
PubMed1983

Solution activity product (KFAP) and simultaneous demineralization-remineralization in bovine tooth enamel and hydroxyapatite pellets.

The effects of changing the ion activity product of the remineralization solution at pH 4.5 (pKFAP 108-118) on the remineralization behavior of demineralized bovine tooth enamel and hydroxyapatite pellets have been studied. Solutions containing calcium-4.5, phosphate, and fluoride in acetate buffers were used. The 45Ca/F molar ratios indicated the formation of fluoridated hydroxyapatite in the enamel or the pellet when the pKFAP values for remineralizing solutions were less than 112. When the pKFAP values were greater than 112, the 45Ca/F ratios were found to be much less than 5. Also, when the pKFAP values were large (greater than 112), the remineralization patterns based on the fluoride distribution in the tooth (or pellet) were found to be different than when the pKFAP values were small (less than 112). The hypothesis that a pKFAP value of 112 is the demarcation between remineralization only and simultaneous dissolution-remineralization has been proposed based on these results.

Animals↗
PubMed1992

Self-setting hydroxyapatite cement: a novel skeletal drug-delivery system for antibiotics.

A novel approach using a self-setting hydroxyapatite (HAP) cement as a skeletal drug-delivery system has been proposed to solve the problem of delivering drugs to skeletal tissue at sufficiently high local concentrations for desirable therapeutic effects. HAP cements loaded with antibiotics can be formed in situ and can be used as bonding materials between bone and prostheses, as well as drug-release devices. The cement also possesses sufficient mechanical strength to be a potential bone grafting material. Using cephalexin and norfloxacin as model drugs, we observed continuous in vitro release profiles of these compounds from cement pellets loaded 0.9-4.8% by weight with one of the drugs. This drug-release pattern correlated well with the Higuchi model. This hydroxyapatite cement drug-delivery system can be applied in the treatment of osteomyelitis and infected compound fractures.

Anti-Bacterial Agents↗
PubMed1997

Complete removal and exchange of sodium dodecyl sulfate bound to soluble and membrane proteins and restoration of their activities, using ceramic hydroxyapatite chromatography.

Up to now, removal of sodium dodecyl sulfate (SDS) from proteins in terms of restoration of their activity was an unsolved problem. A general procedure using ceramic hydroxyapatite (HAP) chromatography was developed for the complete removal of SDS bound to soluble or membrane proteins. This procedure involves (i) the binding of the SDS-protein complexes onto the ceramic hydroxyapatite column, (ii) extensive washing of bound proteins with phosphate buffer containing a mild detergent to exchange SDS, (iii) elution of the retained protein by increasing the phosphate concentration. Using this approach, complete exchange of [35S]SDS into a nonionic detergent such as dodecyl maltoside was achieved with a 90-100% protein recovery. The efficiency of protein-bound SDS removal is very likely due to the combined effect of phosphate ions and the hydrophobic tail of nonionic detergent: acting together, they are able to displace SDS molecules from their protein-binding sites. The advantages of this HAP-mediated SDS removal method include high efficiency, rapidity, simplicity and general applicability to a wide variety of detergents and soluble or membrane proteins. Of utmost importance, SDS-treated P-glycoprotein, glutamate dehydrogenase, and lysozyme fully recovered their enzymatic activities after HAP chromatography, including lysozyme electroeluted from SDS-polyacrylamide gel electrophoresis. This demonstrates that reactivation of SDS-treated protein can be achieved, provided that SDS is completely removed under mild conditions.

ATP Binding Cassette Transporter, Subfamily B, Mem↗
PubMed1984

Solubility of calcium salts, enamel, and hydroxyapatite in aqueous solutions of simple carbohydrates.

The solubility of various Ca(II) salts, hydroxyapatite, and powdered human dental enamel in the presence of simple carbohydrates was studied by determining the complex strength between Ca(II) and the carbohydrates. In 1.0 M CaSO4, the following simplified sequence of complex strengths was obtained for the more common carbohydrates: Na-citrate greater than D-sorbitol greater than xylitol greater than D-mannitol greater than D-fructose greater than D-glucose greater than D-xylose. Whereas the more soluble Ca(II) compounds (like CaSO4) exerted measurable complexation with xylitol, no such complexation was found with hydroxyapatite and enamel powder. This also concerned other alditols. Calculation of the stability constants (K) showed sorbitol (K = 0.81 M) and xylitol (K = 0.67 M) to form stronger complexes in saturated CaSO4 than other alditols. The most suitable coordination site appeared to be a vicinal cis-cis-triol. Precipitation studies showed that 0.5 M xylitol and 0.5 M sorbitol significantly retarded the formation of calcium phosphate precipitates from a solution of Ca(II) and phosphate, compared with the effect caused by glucose, sorbose, or xylose. The effect caused by xylitol and sorbitol was explained in terms of partial displacement of water molecules in the primary hydration layer of Ca(II) ions, caused by competition between polyol and water molecules. In the presence of aldoses and ketoses, virtually instantaneous precipitation occurred. These results suggest that open-chain alditols may influence the chemical reactions of Ca(II) in plaque, saliva, and caries lesions. Alditols do not function as demineralizing agents of the teeth, however. Through the retarding effect on calcium phosphate precipitation, alditols may favorably govern remineralization of carious lesions.

Calcium↗
PubMed1979

Lattice parameters and cation distribution of solid solutions of calcium and strontium hydroxyapatite.

Solid solutions of strontium and calcium hydroxyapatite were synthesized by solid-state reaction. Lattice parameters of these compounds were determined using two types of Guinier cameras. They vary linearly with the molar percentage of strontium hydroxyapatite. The distribution of Ca and Sr ions over the fourfold and sixfold positions in the apatite structure was determined by comparing experimental and calculated values for the intensity ratios of suitable reflections. A slight, although significant, preference of Sr for the sixfold position was found. An ideal behavior is predicted for these solid solutions.

Calcium↗
PubMed1980

Ion migration energies and color center absorption in hydroxyapatite.

By modelling a hydroxyapatite crystal lattice as a finite cluster of atoms surrounded by an array of point ions, molecular orbital MS-Xalpha calculations point to differences in fluoride and hydroxyl migration energies. Despite the fluoride ion being more tightly bound to the lattice, it is in fact more mobile. Color center optical absorption is predicted to be in the infrared region of the spectrum for hydroxyapatite, whereas for fluorapatite the absorption peak of a vacancy is predicted to within 2% of the observed peak in the optical region. This result is discussed in terms of experimental studies of bonding between the calcified and organic phases of bone.

Absorption↗
PubMed1981

Lattice parameters and cation distribution of solid solutions of calcium and lead hydroxyapatite.

Solid solutions of calcium hydroxyapatite (CaOHA) and lead hydroxyapatite (PbOHA) of the formula Ca10-x Pbx (PO4)6 (OH)2 were prepared by coprecipitation followed by heating at 800 degrees C in a steam of CO2-free water vapor of 1 atm. The samples were apatitic in the range 0 less than x less than 6 and contained lead phosphates as a second phase at higher Pb/Ca ratios. Lattice parameters and cation distribution of the apatitic samples were determined by X-ray diffraction. The lattice parameters varied linearly with x in the range considered, whereas all Pb2+ were located in the six-fold position for cations. There was a miscibility gap in the apatite series of solid solutions in the range 1 less than x less than 4, whereas apatites in the range 6 less than x less than 10 were not stable under the conditions of preparation. It is concluded that apatites in the range 4 less than x less than 6 represent a minimum in the free energy of solid solutions between CaOHA and PbOHA.

Calcium↗
PubMed1982

Structural factors influencing the ability of compounds to inhibit hydroxyapatite formation.

For a compound to inhibit potently the transformation of amorphous calcium phosphate into hydroxyapatite, it is suggested that the minimum structural requirement is a phosphate group and, at some other position, either another phosphate group preferably or a carboxylic moiety. Primary amino groups abolish inhibitor potential. Inhibitor potency is modified by various secondary factors, including the number and proximity of active groups, their stereochemistry, steric factors, the lability of the molecule, and in special instances its lipophilicity. Parameters used to monitor the transformation indicate that inhibitors can be grouped into two classes, and it is suggested that this is because one class acts as a hydroxyapatite crystal growth inhibitor. The close proximity of two phosphate groups or of a phosphate and multiple carboxylic groups is proposed to determine in part whether or not a compound acts as a crystal growth inhibitor. Further, bulky side groups render a molecule inactive as a crystal growth poison, although it will still inhibit by other mechanisms.

Adenosine Triphosphate↗
PubMed1979

Studies on the incongruent solubility of hydroxyapatite.

The solubility of hydroxyapatite was examined in calcium phosphate mixtures produced by combining Ca(OH)2 and H3PO4 in varying proportions under a wide variety of conditions. Analyses of supernatants for calcium, phosphorus, and pH provided the data for solubility calculations. Alteration of the solid/liquid ratio by removing supernatant or by evacuating water from equilibrated mixtures had little effect on the solubility product. However, when precipitates separated from some mixtures were reequilibrated at various solid/solution ratios in either water, acetate buffer, or supernatants obtained from previous equilibrations, incongruent solubility was demonstrated and was attributed to the formation of other calcium phosphate phases which coated the apatite. The experiments indicated that hydroxyapatite has a thermodynamic solubility product between 10(-57) and 10(-60), but the exact value is difficult to determine because of the formation of surface coats.

Calcium Phosphates↗