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PubMed2003

In vitro calcium phosphate growth over surface modified PMMA film.

In vitro nucleation of calcium phosphate phase was studied over functionalized polymethyl methacrylate (PMMA) films using Fourier transform infrared spectroscopy, electron spectroscopy, scanning electron microscopy and energy dispersive X-ray analysis. PMMA films were prepared by dissolving commercial grade pellets in chloroform and cast into thin sheets. The films were immersed in a methanol solution of sodium hydroxide before treating with 1.5% solution of adenosine triphosphate (ATP) at a pH of 5.2 for 24 h. ATP treated films were then soaked in saturated lime solution for 4 days to initiate formation of calcium phosphate precursor phase over their surface. The above films immersed in simulated body fluid solution (1.5 x SBF) for more than 5 days led to the nucleation of apatitic calcium phosphate phase all over the film surface. The ATP coupled film not subjected to lime treatment did not show calcium phosphate nucleation behaviour upon immersion in SBF solution. The Ca/P ratio of the calcium phosphate phase increase with increase in soaking time in SBF solution.

Calcium Phosphates↗
PubMed1995

Anticariogenicity of calcium phosphate complexes of tryptic casein phosphopeptides in the rat.

Casein phosphopeptides (CPP) stabilize calcium phosphate through the formation of casein-phosphopeptide amorphous calcium-phosphate complexes (CPP-CP). The ability of CPP-CP to reduce caries activity was investigated by use of specific-pathogen-free rats inoculated with Streptococcus sobrinus. The animals consumed a defined cariogenic diet free of dairy products. Solutions (100 microL) of the CPP-CP (0.1, 0.2, 0.5, 1.0% w/v) were applied to the animals' molar teeth twice daily. Other groups of animals received solutions containing 500 ppm F, the non-phosphorylated peptides of a casein tryptic digest (0.5% w/v), or the calcium-phosphate complex of a synthetic octapeptide, Ac-Glu-Ser(P)-Ile-Ser(P)-Ser(P)-Ser(P)-Glu-Glu-NHMe, corresponding to the common sequence in the CPP. The CPP-CP significantly reduced caries activity in a dose-response fashion, with 1.0% CPP-CP producing 55% and 46% reductions in smooth surface and fissure caries activity, respectively, being similar to that of 500 ppm F. The anticariogenic effects of CPP-CP and fluoride were additive, since animals receiving 0.5% CPP-CP plus 500 ppm F had significantly lower caries activity than those animals receiving either CPP-CP or fluoride alone. The tryptic digest of casein with the phosphopeptides selectively removed showed no anticariogenic activity. The synthetic octapeptide-calcium phosphate complex significantly reduced caries activity, confirming that this calcium-phosphate-stabilizing portion of the casein phospho-peptides is associated with anticariogenicity. The CPP-CP did not significantly affect the level of S. sobrinus in fissure plaque.(ABSTRACT TRUNCATED AT 250 WORDS)

Amino Acid Sequence↗
PubMed1994

Osseous substance formation induced in porous calcium phosphate ceramics in soft tissues.

Porous calcium phosphate ceramics prepared according to the methods of laboratories in Leiden were implanted subcutaneously in rats for 3, 6 and 12 w. No bone formation could be observed. Porous calcium phosphate ceramics prepared according to the methods of laboratories in Sichuan were implanted subcutaneously/intramuscularly in dogs for 1, 3, 5 and 7 months. In particular, ceramics containing alpha-tricalcium phosphate evoked bone formation in the pores of the ceramics. The amount of bone was similar at the different time periods.

Animals↗
PubMed1998

[Osseointegration of 2 different types of calcium phosphate materials: ceramics and ionic cements].

UNLABELLED: Different kinds of calcium phosphate biomaterials can be used as bone substitutes. Ceramics are constituted by HA or TCP grains linked by grain boundaries. Their porosity depends on the powder characteristics and the sintering temperature. It can be very low with a pore size inferior to one micron. The setting of calcium phosphate hydraulic cements results from the precipitation of a calcium phosphate phase different from the one in suspension in the paste. The strength of the cement is given by the entanglement of the growing mineral crystals. Calcium phosphate hydraulic cements and ceramics have very different physico-chemical characteristics. We have studied the histological integration of both kinds of material. The first material was constituted by macroporous ceramics composed of 75% HA and 25% beta-TCP, the cement was made of beta-TCP grains dispersed in a DCPD matrix. The sequence of events which leads to the ceramic integration is always the same: a/ ingrowth of a loose connective tissue; b/ osteoblast differentiation from fibroblast-like cells of the connective tissue in close proximity to the implant surface; c/ osteoid synthesis at the ceramic surface toward the pore center; d/ remodeling of the immature bone and the ceramic itself. The cement is differently integrated. The osteoblasts differentiate at some distance from the implant and there is a trabeculae ingrowth toward the material. CONCLUSIONS: The early stages of both materials osteointegration are different. The integration is centrifugal for ceramics and centripetal for the cement.

Animals↗
PubMed2004

Cell growth and function on calcium phosphate reinforced chitosan scaffolds.

Macroporous chitosan scaffolds reinforced by calcium phosphate powders such as hydroxyapatite (HA) or calcium phosphate invert glass were fabricated using a thermally induced phase separation technique. Human osteoblast-like MG63 cells were cultured on the composite scaffolds for up to 11 days, and the cell growth and function were analyzed. The cell growth is much faster on the chitosan/HA scaffolds incorporated with the glass (CHG) than on the chitosan/HA scaffold without the glass (CH). The total protein content of cells were quantified and increased over time on both composites (CH, CHG) but was significantly higher on CHG after 7 days of culture. The cells on CHG also expressed significantly higher amount of alkaline phosphatase at days 7 and 11 and osteocalcin at day 7 than those on CH. The results suggested that the addition of glass in chitosan/hydroxyapatite composite scaffolds might enhance the proliferation and osteoblastic phenotype expression of MG63 cells. However, the chitosan-matrix scaffolds did not show higher phenotype expression of MG63 cells, in comparison with the TCPS plate, probably due to the degradation of chitosan and release of acidic byproducts. Larger amount of soluble calcium phosphate invert glasses should be added into the scaffolds to prevent chitosan from fast degradation that may affect the differentiation of osteoblast cells.

Alkaline Phosphatase↗
PubMed2003

In vitro behavior of HVOF sprayed calcium phosphate splats and coatings.

Hydroxyapatite (HA) coatings and splats deposited by high velocity oxy-fuel (HVOF) spray technique was investigated in vitro. HA coatings prepared from two different HA powder size range (30+/-5 and 50 +/-5 microm) were immersed in a simulated body fluid with various incubation periods of maximum 6 weeks. The dissolution/precipitation behavior was studied and the degradation of HA coatings caused by in vitro ageing was demonstrated by measuring the changes in flexural modulus through a 3-point bend test. It was found that the dissolution and precipitation behavior of the coatings was significantly dependent upon the incipient coating phase composition and the precipitation of bone-like hydroxyapatite on the coating's surface was found to be directly related to the dissolution process. Higher dissolution rates of tricalcium phosphate, tetracalcium phosphate and amorphous calcium phosphate relative to HA, resulted in accelerated precipitation. Furthermore, analysis of coatings' surface morphology demonstrated that advanced precipitation invariably occurred at regions where dissolution took place. Results showed that the changes in flexural modulus of investigated HA coatings accompanying different incubation duration was not systematic but was found to be dependent upon changes of coating structure and other factors brought about by in vitro ageing. In vitro investigation of individual HA splats collected from different HA particle sizes revealed, after 3 days ageing, that the rate ratio of precipitation to dissolution was directly determined by the local phase composition, and this phenomenon could be effectively used to explain the behavior of thermally sprayed HA coatings in vitro. It implied that the precipitation was strongly dependent on the first molecule attachment. To achieve rapid precipitation in vitro, partial molten state of HA particles during HVOF coating deposition was recommended.

Anions↗
PubMed2003

Microstructure analysis of calcium phosphate formed in tendon.

The surface of soft tendon tissue has been modified using calcium phosphate in order for the tendon to directly connect with hard bone and reconstruct an injured ligament. Calcium phosphate was coated onto the tendon in a soaking process using alternating a CaCl(2) (200 mM) and a Na(2)HPO(4) (120 mM) solution. According to SEM/EDX observations, calcium phosphate was formed, not only on the tendon surface, but also inside the tendon tissue. When the tendon was treated with seven soaking cycles, calcium phosphate was detected between 0-500 microm from the tendon surface. According to TEM observations, the crystal morphology of calcium phosphate depends on the distance from the surface. Hydroxyapatite crystals were observed near the surface, while octa-calcium phosphate crystals could be observed further from the surface, thus at initial soaking. The crystals were formed on collagen fibrils in spaces between the collagen fibrils with the c-axes of the crystals aligned parallel with the collagen fibrils. This finding suggests Ca(2+) ions to interact with the tendon surface, most probably with the carboxyl functional groups of collagen, and subsequently forming nucleation centers for the crystals.

Journal Article↗
PubMed2003

Noninvasive bone replacement with a new injectable calcium phosphate biomaterial.

The use of injectable calcium phosphate (CaP) biomaterials in noninvasive surgery should provide efficient bone colonization and implantation. Two different kinds of injectable biomaterials are presently under development: ionic hydraulic bone cements that harden in vivo after injection, and an association of biphasic calcium phosphate (BCP) ceramic granules and a water-soluble polymer vehicle (a technique particularly investigated by our group), providing an injectable CaP bone substitute (IBS). In our study, we compared these two approaches, using physicochemical characterizations and in vivo evaluations in light microscopy, scanning electron microscopy, and three-dimensional microtomography with synchrotron technology. Three weeks after implantation in rabbit bone, both biomaterials showed perfect biocompatibility and bioactivity, but new bone formation and degradation of the biomaterial were significantly greater for BCP granules than for ionic cement. Newly formed bone developed, binding the BCP granules together, whereas new bone grew only on the surface of the cement, which remained dense, with no obvious degradation 3 weeks after implantation. This study confirms that BCP granules carried by a cellulosic polymer conserve bioactivity and are conducive to earlier and more extensive bone substitution than a carbonated-hydroxyapatite bone cement. The presence of intergranular spaces in the BCP preparation, as shown on microtomography imaging, seems particularly favorable, allowing body fluids to reach each BCP granule immediately after implantation. Thus, the IBS functions as a completely interconnected ceramic with total open macroporosity. This new bone replacement approach should facilitate microinvasive bone surgery and local delivery of bone therapy agents.

Animals↗
PubMed2000

Tissue responses of calcium phosphate cement: a study in dogs.

The in vivo properties of a new kind of calcium phosphate cement were investigated in this study. Calcium phosphate cement was implanted as paste into femoral bone and dorsal muscle of dogs for 3 and 6 months, and as prehardened form into thigh muscles of dogs for 1, 2 and 6 months. Histology was performed on thin un-decalcified sections. No foreign body reaction, no inflammation and no necrosis were found both in bony site and in muscles. There was no connective tissue layer between the cement and bone when cement paste was implanted in the bone. A creeping substitution of cement by bone, in which osteoclast-like cells resorbed the cement as if the cement is a part of bone and new bone was formed directly on the resorption line of calcium phosphate cement, was found. Bone formation was found histomorphologically in pores and deep rugged surface of cement samples (both paste and prehardened form) implanted in muscles of dogs. The induced bone was also identified with backscattered scanning electron microscopy (BSE) and by energy-dispersive X-ray micro-analysis (EDX). The results suggest that the calcium phosphate cement used in this study is biocompatible, resorbable in a manner of creeping substitution, osteoconductive and osteoinductive. It seems that an ideal bone substitute can be developed by using this type of calcium phosphate cement.

Absorbable Implants↗
PubMedDate not supplied

Fully injectable calcium phosphate cement--a promise to dentistry.

Calcium phosphate cements (CPC) are self setting and biocompatible bone substitute materials with potential applications in dentistry. However, its clinical use has been challenged by poor rheological properties. A novel formulation of CPC has been developed, which gives a fully injectable and cohesive paste. This work investigates the suitability of the new "fully injectable calcium phosphate cement" (FI-CPC) for dental applications. The cementing properties, material characteristics, and the rheological properties were tested using a battery of material characteristics methods. The biocompatibility was also evaluated as per ISO 7405. The setting time (20 min) and compressive strength (>11 Mpa) of FI-CPC satisfy the clinical requirements. It underwent setting without any exothermic reaction, keeping good dimensional stability. The cement paste could be extruded through a 18-gauge needle, easily and fully. It showed excellent cohesion when immersed in water. FI-CPC was seen to set into a micro-porous mass of hydroxyapatite, the mineral part of human dentin. It showed good attachment to dentin walls, when filled in tooth perforations. FI-CPC was found non-toxic, non-allergic, non-pyrogenic, and soft-tissue compatible. The study shows that FI-CPC provides a self setting bio-compatible paste with excellent rheological properties for surgical applications. The set cement provides good and stable sealing. The osteoconductive property is an added advantage. FI-CPC proves to be an ideal material for endodontic sealing/filling and periodontic repair.

Animals↗
PubMed2002

Adsorption of proteins and calcium phosphate materials bioactivity.

The behaviour of calcium phosphate (CaP) based biomaterials in biological environments determines how they can be used in vivo. The prime requirement for CaP materials to be bioactive and bond to living bone is the formation of a bone-like apatite layer on their surface. This phenomenon can be reproduced in vitro using simulated body fluid (SBF): a protein-free solution with ion concentrations similar to those of human blood plasma. Although proteins are unanimously considered as important actors in CaP material bioactivity and biomineralization processes, a certain confusion exists about their role as promotors or inhibitors of mineral crystal formation or both. The adsorption of proteins on the mineral surface can alter the nucleation rate and lead to antagonistic effects depending on the concentration of the adsorbed proteins. The ambiguity of this general effect is illustrated by the action of albumin on calcium phosphate crystallization on type I collagen and several other examples found in the literature.

Adsorption↗
PubMed1989

[An experimental study on the osteoconductive properties of porous calcium phosphate glass ceramics].

Porous blocks of calcium phosphate glass ceramics (CaO 52 mol %, P2O5 47 mol %, Al2O3 1 mol %, mean pore size 200 microns, mean porosity 85%) were implanted in the distal femoral metaphysis of rats and were observed for up to one year. The materials were evaluated histologically and morphometrically using contact microradiography and Masson-Goldner staining. Comparative experiments were conducted employing similar methods as described above, with calcium phosphate glass ceramics, sintered hydroxyapatite and alumina ceramics, each having 70% porosity. The compressive strength of each implanted material was measured at a certain intervals up to 6 months after implantation. Bone formation was seen from the periphery to the center of the block. Mass of the regenerated bone and infiltrating depth of the new bone increased with the passage of time. New bone formation with the glass ceramics was observed to be almost equivalent to that obtained with hydroxyapatite, and was significantly higher than that with the alumina ceramics.

Animals↗
PubMed1990

[Effects of addition of calcium phosphates on the properties of gypsum].

Calcium hydrogen phosphate (CHP), calcium hydrogen phosphate dihydrate (CHPD) and calcium sulfate dihydrate (gypsum) crystals were added to calcium sulfate hemihydrate as gypsum model materials for improvement of the mechanical properties. The compressive and the bending strength of the hardened mixture were improved by the addition of CHPD, while the addition of CHP and gypsum crystals had no effect. The setting time was markedly reduced by the addition of CHPD, but remarkably increased in the case of CHP crystals. On the other hand, the setting expansion was decreased by the addition of CHP and CHPD crystals. Based on the observation of the surface profile of the calcium sulfate hemihydrate set in contact with glass, and scanning electron microscopy (SEM) of the fractured surface of the hardened mixture, the surface roughness was markedly decreased by the addition of CHPD. When the crystals were added to the metastable solution of calcium sulfate supersaturated with respect to gypsum, the presence of gypsum crystals deposited on the CHPD surface was confirmed by atomic absorption analysis, fourier transform infrared spectroscopy. SEM and electron X-ray microanalysis. Therefore, epitaxial growth of gypsum occurred on the CHPD surface and the addition of CHPD improved the mechanical properties of the gypsum materials.

Calcium Phosphates↗
PubMed1980

Compositional dependence of the formation of calcium phosphate films on bioglass.

Bioglass, which has a composition of sodium carbonate, calcium carbonate, phosphorous pentoxide and silica, has been shown to bond to living bone. This ability is dependent on controlled surface reactions. Investigators with 45S5 bioglass have demonstrated that the formation of a SiO2-rich layer and a calcium phosphate film on its surface in an aqueous environment is associated with the film bonding the bioglass to bone. The objects of this research were: 1. To study SiO2 dependence on the formation of a silica-rich layer and calcium phosphate films on a bioglass surface in a simulated physiological solution, and 2. To establish a correlation between in vitro surface reactions and in vivo bonding ability. It was discovered that three types of reactions occur in a simulated physiological solution depending on bioglass composition: 1. A calcium phosphate film and SiO2-rich layer form simultaneously and the reaction rate is fast for bioglasses which have a lower content of SiO2 (approximately 46 mol% SiO2). 2. A SiO2-rich layer forms first and a calcium phosphate film develops later between the aqueous environment and the SiO2-rich layer for bioglasses whose SiO2 content is between 46--55 mol %. 3. A calcium phosphate film does not form for glasses whose SiO2 content is more than 60 mol %.

Adhesiveness↗
PubMed1996

The nucleation and growth of calcium phosphate crystals at protein and phosphatidylserine liposome surfaces.

The kinetics of calcium phosphate crystal growth at the surfaces of proteins and phospholipids has been investigated using free drift and constant composition methods in supersaturated calcium phosphate solutions (relative supersaturations: with respect to hydroxyapatite, HAP, sigma HAP = 15.0, and with respect to octacalcium phosphate, OCP, sigma OCP = 1.9). Fibrinogen and collagen molecules adsorbed at hydrophobic surfaces as well as uncross-linked collagen fibrils induce ion binding and subsequent nucleation of calcium phosphate. The formation of OCP on phosphatidylserine vesicles introduced to highly supersaturated calcium phosphate solutions probably involves the interaction of the calcium ions with the ionized carboxylic groups of the phospholipid.

Calcium Phosphates↗
PubMed2000

Effect of external stresses on calcium phosphate glass investigated by IR spectroscopy.

The effect of external stress on calcium phosphate glasses was investigated because they experience high stress in practical uses such as mastication. In order to understand the structural change of calcium phosphate glasses caused by an applied stress, the IR reflection peak shift of the phosphate structural band of the calcium phosphate glass fiber due to a bending stress was measured. The peak shift represents the change of the P-O-P bond angle. The IR reflection peak of the phosphate structural bands near approximately 1300 and approximately 910 cm(-1) shifted to a higher wavenumber under tensile stress and to a lower wavenumber under compressive stress when calcium phosphate fiber with a uniform structure was employed. This indicates that the P-O-P bond angle increases under tensile stress and decreases under compressive stress. The extents of both peak shifts were larger with a lower [corrected] Ca/P ratio than with a higher [corrected] Ca/P ratio. Therefore, phosphate glass with a higher Ca/P ratio is expected to withstand loads under external stress.

Biocompatible Materials↗
PubMed1976

Presence or absence of inhibitors of crystal growth in bile. 1. Effect of bile on the formation of calcium phosphate, a constituent of gallstones.

When calcium and phosphate ions were mixed so that their final concentration was 4 mmol/1 and the pH was kept at 7-0, an amorphous precipitate immediately formed and this changed into crystalline material with an apatite-like structure after a period of time. The formation of either or both types of precipitate could be slowed down or prevented by adding to the crystallising medium trace amounts of pyrophosphate or citrate which are known inhibitors of the formation of calcium phosphate, or large quantities of sodium chloride which increased the ionic strength of the solution and hence the solubility of calcium phosphate, Both common duct and gallbladder bile from patients with gallstones composed of cholesterol and/or calcium carbonate had a very pronounced inhibitory action on the formation of these precipitates. Only very small amounts of bile were necessary to produce these effects, which therefore were not due to an increase in ionic strength. Ultrafiltration of bile showed that material with a molecular weight greater than 10 000 was mainly responsible for this activity. Because the inhibitor was present in both common duct and gallbladder bile, the liver is the likely source of origin. The possible identity of this material is examined. The powerful inhibitory effect of bile on the crystallisation of calcium phosphate is probably a contributory factor to the rare occurrence of the calcium phosphates, apatite and whitlockite, in gallstones.

Bile↗