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PubMed1998

Evaluation of cytotoxicity of calcium phosphate cement consisting of alpha-tricalcium phosphate and dicalcium phosphate dihydrate.

A newly developed calcium phosphate cement, MM, was evaluated for tissue irritability by means of cell cultures. In this study, MM showed extensive mild cytotoxicity compared with other cements before setting. Inhibition of adhesion of L-929 cells was not observed after contact with MM for 24 hours. The influence of MM on colony formation was approximately the same as that of another calcium phosphate cement and less than that of a glass ionomer cement. Toxicity of MM after setting was compared with four cements; another calcium phosphate cement, glass ionomer cement, silicate cement and zinc oxide eugenol cement, but MM showed the least influence on cell morphology. Judging from these results, MM appears to be less cytotoxic than the cements in current use.

Animals↗

PubMed2003

Reconstruction of the immature craniofacial skeleton with a carbonated calcium phosphate bone cement: interaction with bioresorbable mesh.

Calcium phosphate cements have been recently introduced for use in craniofacial reconstruction. In the clinical setting, however, pulsations of the underlying brain and dura may interfere with the crystallization of these cements, thereby rendering their use in cranioplasty problematic. To circumvent such problems, many clinicians have interposed synthetic resorbable plates or mesh between the dura and the cement. At the present time, however, little is known about the influence of such materials or their breakdown products on the fate of calcium phosphate cements. The specific aim of this project was to evaluate the biocompatibility, osteoconductivity, and remodeling capacity of a calcium phosphate cement after implantation into experimental calvarial defects when combined with a resorbable mesh underlay. Four 10-mm diameter full-thickness calvarial defects (two frontal, two parietal) were created in each of six 3-week-old Yorkshire pigs. The defects were treated as follows: 1) empty control, 2) macroporous polylactic acid (70/30 L/DL polylactic acid [PLA]) mesh, 3) Norian CRS calcium phosphate cement, and 4) Norian CRS over PLA mesh underlay. Animals were divided into two groups. Half of the animals were killed 30 days after surgery, and half were killed 180 days after surgery, and the graft recipient sites were examined histologically. At 30 days, minimal bone ingrowth was observed in untreated calvarial defects or in those that were treated with PLA plates alone. Defects treated with the cement alone demonstrated a modest amount of new woven bone deposition, primarily at the periphery of the implants. Defects treated with calcium phosphate cement over PLA mesh underlays were characterized by remodeling and woven bone deposition at 30 days, with complete or near-complete osseous bridging of the ectocranial implant surfaces. Progressive bone ingrowth was noted in all defects at 180 days, with near-complete replacement of all Norian CRS implants by host bone. The PLA mesh remained incompletely resorbed at 180 days. No inflammatory response to the implants was observed at either time point. Calcium phosphate cement may be safely used for craniofacial reconstruction in the presence of PLA implants without compromise to its biocompatibility, osteoconductivity, or remodeling capacity.

Absorbable Implants↗

PubMed1990

New method of treatment for dentin hypersensitivity by precipitation of calcium phosphate in situ.

The hydrodynamic theory is generally accepted as an explanation of hypersensitive dentin and suggests that occlusion of exposed dentinal tubules should result in a reduction of hypersensitivity. We tried to block tubules by in situ precipitation of insoluble calcium phosphate. Calcium phosphate crystals precipitated in situ on patent dentin surfaces and formed in the tubules immediately upon serial application of sodium phosphate and calcium chloride solutions, thus occluding the tubules. The size of crystals, their degree of coverage, and the thickness of the precipitate depended on the method of application and concentration of the solutions. The application of 5% disodium phosphate solution followed by rubbing with 10% calcium chloride solution resulted in immediate relief from dental hypersensitivity in 84% of patients treated.

Adult↗

PubMed1995

Calcium phosphate supplementation results in lower rat fecal bile acid concentrations and a more quiescent colonic cell proliferation pattern than does calcium lactate.

Although there is general agreement that dietary calcium is protective against colon carcinogenesis, considerable controversy exists on the relative efficacy of the counterion in calcium supplements. We therefore conducted a comparative study in rats of four forms of calcium supplementation (calcium phosphate, casein, lactate, and a 50:50 phosphate-carbonate combination). The relative effects of these supplements on measurements of colon physiology, in vivo pH, fecal fat, individual bile acids, and in vivo cell proliferation were measured in the same animals. In contrast to results when amounts of calcium are varied, there was no effect of form of supplement on total fecal output or output of fecal fat. Calcium phosphate resulted in the most acidified cecal contents. Calcium phosphate and calcium casein resulted in lower fecal concentrations of lithocholate and lower amounts of total fecal bile acids than supplementation with the calcium lactate or combination diets. In addition, rats fed calcium phosphate had lower concentrations of fecal beta-muricholate than rats provided with the calcium combination supplement. In the proximal colon, calcium phosphate resulted in a significantly lower number of cells per crypt column and a lower labeling index than the calcium lactate diet. The position of the highest labeled cell was lower with calcium phosphate supplementation than with supplementation from the calcium combination or the calcium lactate diet. There was a highly significant correlation between the pH of cecal contents and labeling index in the proximal colon (r = 0.98, p = 0.003). The results suggest that calcium phosphate may inhibit colon tumor incidence more effectively than calcium lactate, because the calcium phosphate group had a lower colonic proliferative status than the calcium lactate group. Changes in the proliferative status of colonocytes are known to precede and accompany neoplasia.

Animals↗

PubMed2000

Osseointegration of composite calcium phosphate bioceramics.

The resistance of macroporous calcium phosphate ceramics to compressive strength generally is low and depends on, among other factors, porosity percentage and pore size. A compromise always is adopted between high porosity, required for a good integration, and mechanical strength, which increases with material density. We improved the strength of macroporous calcium phosphate ceramics of interconnected porosity by filling the pores with a highly soluble, self-setting calcium phosphate cement made of TCP and DCPD. Cylinders of the resulting material were implanted in sheep condyles and subjected to histological analysis after 20, 60, and 120 days. Microradiographs were made of the histological sections. The control material consisted of ceramic that had not been loaded with cement. Progressive ingrowth of bone into the ceramic pores occurred as the cement was degraded during the first implantation period. Marked degradation of the cement was apparent after 2 months, with fragmentation of the cement in most of the pores and the presence of bone tissue between the fragments. All the cement had been replaced by bone after 4 months. Some fragments of cement still were embedded in the newly formed bone. There was no significant difference between the integration of loaded and nonloaded ceramics. Filling the macroporous ceramic pores with a calcium phosphate cement significantly improved the mechanical strength of these ceramics without modifying their integration in the healing bone.

Animals↗

PubMed1996

Effects of hexafluorosilicate on the precipitate composition and dentine tubule occlusion by calcium phosphate.

OBJECTIVES: To improve the calcium phosphate precipitation (CPP) method for the occlusion of dentine tubules with calcium phosphate, the addition of calcium hexafluorosilicate (CaSiF6) to CPP solution was evaluated in vitro with respect to its occluding capacity and the composition of the precipitate. METHODS: The occlusion of dentine tubules was evaluated by SEM observations and by measurements of dentine permeability. The composition of the precipitate was determined by measuring the calcium to phosphorus (Ca/P) ratio of the precipitate in dentine tubules by energy dispersive X-ray spectroscopy (EDS). RESULTS: The addition of CaSiF6 to the CPP solution resulted in an increase of the Ca/P ratio in the precipitate not only on the dentine surface but also inside the dentine tubules; indicating that the precipitate became more apatitic in nature. The addition of CaSiF6 had no effect on occluding capacity in terms of measurements of dentine permeability or SEM observations. Dentine permeability decreased to approximately 4% of pretreatment values and the dentine tubules were occluded for approximately 10-15 microns from the dentine surface. CONCLUSION: It is concluded that the addition of CaSiF6 to CPP solution was desirable, since it provided a more apatitic precipitate in the dentine tubules, not only on the dentine surface but also inside the tubules, and there were no drawbacks with respect to its occluding capacity.

Apatites↗

PubMed1995

Rapid association of unconjugated bilirubin with amorphous calcium phosphate.

The association of unconjugated bilirubin (UCB) with amorphous calcium phosphate was studied in vitro. To this end UCB, solubilized in different micellar bile salt solutions, was incubated with freshly prepared calcium phosphate precipitate. It was demonstrated that amorphous calcium phosphate (ACP) rapidly binds and precipitates UCB in a dose-dependent way. The results indicate that binding of UCB to ACP is specific: binding to barium phosphate was negligible and addition of low amounts of Mg2+ before formation of the calcium phosphate precipitate (Ca:Mg = 5:1) inhibited binding by 80%. Free Ca2+ stimulated binding, whereas free phosphate ions inhibited binding of UCB in taurocholate solutions and to a lesser extent in glycocholate solutions. The apparent affinity of UCB for amorphous calcium phosphate was different in the various bile salt solutions. Binding of UCB decreased at pH > 8.5 in taurocholate solutions, but not in glycocholate solutions where binding of UCB was constant from pH 7.5-10.5. We propose a model in which UCB directly binds to amorphous calcium phosphate in the presence of bile salts that weakly interact with ACP, like taurocholate. In the presence of bile salts that strongly interact with ACP, such as glycochenodeoxycholate, binding of UCB may also occur via the bile salt. In conditions of unconjugated hyperbilirubinemia, such as the Crigler-Najjar syndrome, neonatal jaundice, and in the Gunn rat, considerable amounts of UCB diffuse across the intestinal mucosa. Binding of UCB to calcium phosphate in the intestine may stimulate its excretion and thereby constitute a relevant mechanism of excretion.

Bile Acids and Salts↗

PubMed1993

Calcium phosphates in pharmaceutical tableting. 2. Comparison of tableting properties.

Ten calcium phosphates suitable for direct compression (dibasic calcium phosphate dihydrate, dibasic calcium phosphate anhydrous and hydroxylapatite) were investigated with respect to their compressional behaviour. Except for Di-Cafos A all products gave tablets with sufficient to good mechanical strength. Nevertheless, there were differences between the products. All tablets prepared from the different products showed a high friability. This seems to be a problem of the calcium phosphates in general. On the other hand, the influence of magnesium stearate on the mechanical strength of the tablets was negligible for all products investigated. Moreover, a considerable effect of the particle size on the tensile strength of the tablets was found. The ejection forces and residual pressures were high in general, but critical only in the case of hydroxylapatites. Heckel plots were used to differentiate between plastic deformation and brittle fracture of the particles. In the case of calcium phosphates the slope of the Heckel plots indicated the hardness of the particles rather than their deformation behaviour.

Calcium Phosphates↗

PubMed2005

Physicochemical characterization of casein phosphopeptide-amorphous calcium phosphate nanocomplexes.

Milk caseins stabilize calcium and phosphate ions and make them available to the neonate. Tryptic digestion of the caseins yields phosphopeptides from their polar N-terminal regions that contain clusters of phosphorylated seryl residues. These phosphoseryl clusters have been hypothesized to be responsible for the interaction between the caseins and calcium phosphate that lead to the formation of casein micelles. The casein phosphopeptides stabilize calcium and phosphate ions through the formation of complexes. The calcium phosphate in these complexes is biologically available for intestinal absorption and remineralization of subsurface lesions in tooth enamel. We have studied the structure of the complexes formed by the casein phosphopeptides with calcium phosphate using a range of physicochemical techniques including x-ray powder diffraction, scanning electron microscopy, transmission electron microscopy, and equilibrium binding analyses. The amorphous nature of the calcium phosphate phase was confirmed by two independent methods: x-ray powder diffraction and selected area diffraction. In solution, the ion activity product of a basic amorphous calcium phosphate phase was the only ion product that was a function of bound phosphate independent of pH, consistent with basic amorphous calcium phosphate being the phase stabilized by the casein phosphopeptides. Detailed investigations of calcium and calcium phosphate binding using a library of synthetic homologues and analogues of the casein phosphopeptides have revealed that although the fully phosphorylated seryl-cluster motif is pivotal for the interaction with calcium and phosphate, other factors are also important. In particular, calcium binding and calcium phosphate stabilization by the peptides was influenced by peptide net charge, length, and sequence.

Animals↗

PubMed2003

[Cellular culture of osteoblasts and fibroblasts on porous calcium-phosphate bone substitutes].

PURPOSE OF THE STUDY: Calcium phosphate ceramics are synthetic bone substitutes able to fill in bone destruction as a support of the bone growth. This work consisted in an in vitro assessment of osteoblasts and fibroblasts cultures on macroporous calcium-phosphate bone substitutes to analyze the interaction between cells and bone substitute. MATERIALS AND METHODS: The macroporous ceramic was composed of 70% hydroxyapatite and 30% tri-calcium phosphate with known mechanical and physico-chemical properties. Three compounds were processed with different size of macropore and with or without microporosity on their surface. Cells were seeded on discs measuring 10 mm in diameter and 2 mm in thickness. Cellular viability was evaluated by the MTT test for every stage of observation. An histological study to observe the invasion in the depth of discs was performed. Scanning electron microscopy was used to analyze the cellular comportment in contact with the surface of substitutes. RESULTS: An exponential cellular growth was effective on each substitute with the two cellular types. Cells spread on the surface of the compounds covering macropores and colonized the depth of the discs. A size of macropore of 300 microm or more seemed to support this invasion. 15 microm sized interconnections appeared to be effective to allow cell migration between macropores. The cell proliferation was similar on substitutes with or without microporosity. CONCLUSION: Biomaterials currently used as bone substitute are more or less osteoconductive but they have no osteoinductive property. A hybrid association of calcium-phosphate ceramic with osteogenic cells should promote the development of a calcium phosphate compound with osteoinductive capacity.

Biocompatible Materials↗

PubMed1995

In vitro formation of "urinary stones": generation of spherulites of calcium phosphate in gel and overgrowth with calcium oxalate using a new flow model of crystallization.

Calcium phosphate (CaP) has been detected in the majority of urinary stones containing predominantly calcium oxalate (CaOx). Therefore, crystal phases of CaP might play an important role with respect to the formation of urinary calcium stones in general. Very often, CaP found in stones or tissue of human kidney occurs in the shape of small spherulites. In this paper, we report on a new flow model of crystallization (FMCG), which has been used to generate spherulites of CaP in a gel matrix of 1% agar-agar at 37 degrees C from a supersaturated, metastable solution continuously flowing over the gel surface. Scanning electron microscopy (SEM), X-ray diffraction and microscopic Fourier transformed infrared spectroscopy (FTIR) revealed that the particles formed (diameter: up to 200 microns) consisted of a poorly crystalline core of carbonatoapatite which was partly surrounded by a well-crystallized shell of octacalcium phosphate (OCP) showing radially oriented sheet-like structures. Subsequently, CaOx was grown on these spherulites from a flow of a correspondingly supersaturated solution conducted over the gel matrix. It could be shown by SEM that growth of calcium oxalate monohydrate (COM) was characteristically induced by the OCP shell. Radial sheet-like forms of OCP were directly continued by COM showing a certain radial orientation. The model of crystallization in gel matrices applied here should be well-suited to simulate the process of urinary stone formation under in vitro conditions.

Calcium Oxalate↗

PubMed1997

Growth of calcium phosphate on phosphorylated chitin fibres.

Calcium phosphate growth on chitin phosphorylated fibres was studied using scanning electron microscopy and energy dispersive X-ray analysis (SEM, EDX), micro-Fourier transform infrared spectroscopy (FTIR), and solid state magic angle spinning nuclear magnetic resonance (MAS NMR) techniques. The C6 chemical shift positions of 13C MAS NMR in the chitin fibres phosphorylated using urea and H3PO4 are obvious indicating that phosphorylation takes place not in the C1 but in the C6 region. Micro-FTIR and 31P MAS NMR suggested that ammonium hydrogen phosphate formed during the phosphorylation procedure. Chitin fibres phosphorylated using urea and H3PO4 and then soaked in saturated Ca(OH)2 solution at ambient temperature, which lead to the formation of thin coatings formed by partial hydrolysis of the PO4 functionalities, were found to stimulate the growth of a calcium phosphate coating on their surfaces after soaking in 1.5xSBF solution for as little as one day. The thin layer after Ca(OH)2 treatment functioned as a nucleation layer for further calcium phosphate deposition after soaking in 1.5xSBF solution. EDX-measured Ca : P ratios of the coatings of Ca(OH)2-treated phosphorylated chitin in 1.5xSBF solution suggested that calcium-deficient apatite was formed.

Journal Article↗

PubMed2006

Effects of additives on the rheological properties and injectability of a calcium phosphate bone substitute material.

An injectable calcium phosphate bone substitute material has been prepared by mixing amorphous calcium phosphate (ACP) and dicalcium phosphate dihydrate (DCPD) for use in noninvasive surgery, and the influence of additives, such as disodium hydrogen phosphate, polyethylene glycol (PEG), glycerin, and citric acid, on the rheological properties and injectability of the ACP + DCPD cement system have been studied in this work. Novel approach of thixotropy measurement has been used to characterize the stability of the pastes. The results show that the injectability and the setting time can be augmented by the addition of disodium phosphate solution to the paste but reduced by the addition of PEG 200, glycerin, or citric acid to the paste. This study suggests that the injectability and the setting time of the ACP + DCPD bone substitute material can be balanced, and the injectable calcium phosphate bone substitute material with satisfied fluidity and injectability for clinical operation can be prepared by optimizing the additives and their concentrations, according to different clinical requirements.

Biocompatible Materials↗

PubMed2005

Left ventricular function and calcium phosphate plasma levels in uraemic patients.

UNLABELLED: BACKGROUND. Recent investigations have focused on the pathogenetic role of disturbances of calcium phosphate metabolism in causing cardiovascular morbidity and mortality in haemodialysis patients. The aim of the present study was to assess left ventricular function and its relationship to phosphate and calcium plasma levels in stable uraemic patients on haemodialysis treatment. METHODS: Twenty uraemic patients (mean age 51+/-13 years) on maintenance haemodialysis and free from overt cardiac dysfunction, and 20 healthy volunteers underwent standard echocardiography, tissue Doppler-derived early (E(m)) and late (A(m)) diastolic velocities, tissue characterization with cyclic variations of integrated backscatter (CV-IBS), and serum biochemistry. RESULTS: With respect to tissue Doppler imaging (TDI), uraemic patients showed a lower E(m) peak, a higher A(m) peak, and a reduced E(m)/A(m) ratio of both interventricular septum and lateral wall (0.01>P<0.001) than controls. CV-IBS of both septum and posterior wall was significantly smaller in uraemic patients than in the control subjects (P<0.001). Moreover, the E(m)/A(m) ratio of septum and lateral wall were negatively related to serum phosphorus and to calcium phosphate product (P<0.001 for all). Accordingly, an inverse relationship was also found between CV-IBS of septum and lateral wall and calcium phosphate product and phosphorus (P<0.05 for all). CONCLUSIONS: These results showed early cardiac impairment of diastolic myocardial function evaluated by TDI and IBS analysis, and a close relationship between these changes and the calcium-phosphate plasma levels. These findings are well in keeping with the important role of hyperphosphataemia as a risk factor for cardiovascular damage, and justify the effort for optimal control of calcium phosphate metabolism in uraemic patients.

Adult↗

PubMed1993

Calcium phosphates in pharmaceutical tableting. 1. Physico-pharmaceutical properties.

Ten commercially available calcium phosphates used for direct tableting were evaluated. The particle size distributions, powder properties, sorption isotherms and pH values of aqueous slurries were compared. All samples showed good or at least sufficient flowability. Scanning electron micrographs illustrated the different kinds of manufacturing and gave hints on their expectable behaviour under compaction pressure. The sorption isotherms of identical chemical substances, which had been manufactured by different methods, differed strongly. This can be related to their specific surface areas. Most of the hydroxylapatites have large surface areas and can absorb up to more than 15% water at 93% relative humidity. Dibasic calcium phosphate dihydrate was non-hygroscopic and absorbed less than 1% water. With the exception of monobasic calcium phosphate monohydrate all calcium phosphates behaved quite neutral in water. Monobasic calcium phosphate monohydrate can be regarded as a solid acid. Although the calcium phosphates are usually stable substances, the role of crystal water in the case of dibasic calcium phosphate dihydrate and monobasic calcium phosphate monohydrate is problematic due to possible interactions with active ingredients.

Calcium Phosphates↗

PubMed2004

Proliferation, differentiation, and calcification of preosteoblast-like MC3T3-E1 cells cultured onto noncrystalline calcium phosphate glass.

The purpose of this study was to investigate calcium phosphate glass as a potential biomaterial for hard tissue repair. We prepared calcium phosphate glass using the system CaO-CaF(2)-P(2)O(5)-MgO-ZnO and cultured MC3T3-E1 cells onto the glass in alpha-MEM with beta-glycerophosphatase and ascorbic acid. Proliferation of the cells was determined to evaluate the biocompatibility of the prepared calcium phosphate glass. The alkaline phosphatase activity was measured to examine the osteoblast differentiation. Mineralization was evaluated by staining the calcium precipitates with Alizarin red. Culture onto the calcium phosphate glass exhibited no significant influence on cell proliferation compared to the polystyrene chosen as a control in this experiment (p > 0.05). The alkaline phosphatase activity in the experimental group, however, was enhanced by the calcium phosphate glass significantly at 10-18 days after incubation than that of the control group (p < 0.05). The promotion of bone-like tissue formation by the calcium phosphate glass was observed after 7 days and thereafter. The results of the present study indicate that the prepared calcium phosphate glass affects osteogenesis by increasing calcification of the extracellular matrix.

Alkaline Phosphatase↗

PubMedDate not supplied

Sodium d-fructose-1,6-diphosphate vs. sodium monohydrogen phosphate in total parenteral nutrition: a comparative in vitro assessment of calcium phosphate compatibility.

BACKGROUND: The supply of high amounts of calcium (Ca) and phosphorus (P) during total parenteral nutrition (TPN) is matter of concern because of the risk associated with calcium phosphate precipitation. The in vitro Ca-P compatibility in ready-for-use TPN solutions after the addition of different concentrations of inorganic phosphate or d-fructose-1,6-diphosphate (FDP) and calcium chloride was evaluated. METHODS: Four series of experiments for each Ca + P couple were carried out by varying amino acid concentrations (2% or 4%), temperature (25 degrees C or 37 degrees C), and pH. The extent of precipitation was estimated by visual inspection and particle count. The areas of maximal compatibility (ie, areas showing the complete absence of precipitates) were drawn from the precipitation curves. RESULTS: The precipitation extent was considerably higher in conditions mimicking body environment for both Ca + P couples. The compatibility area at 37 degrees C and 2% amino acid for CaCl2 + Na2HPO4 admixtures was included within 2.50 mmol/L CaCl2 and 2.22 mmol/L Na2HPO4, whereas that for CaCl2 + FDP was within 33.3 mmol/L CaCl2 and 10.0 mmol/L FDP (20 mEq/L of P). Unlike inorganic calcium phosphate, FDP dicalcium salt precipitation was kinetically delayed and was only minimally enhanced by decreasing amino acid concentration. CONCLUSIONS: Our data indicated that the use of FDP as the P source in parenteral nutrition solutions was effective in avoiding the life-threatening calcium phosphate precipitation. Thus, the addition of FDP to TPN admixtures represents a safe choice, allowing the simultaneous administration of high amounts of Ca and P in restricted fluid volumes, even at low amino acid concentrations.

Calcium↗

PubMed2005

[Calcium phosphate cements in medicine and dentistry--a review of literature].

Calcium phosphates represent the largest group of biominerals in vertebrate animals. They also have many uses in industry, agriculture, medicine and everyday life. The calcium phosphates containing the ionic species HPO4(2-) and PO4(3-) are biologically relevant. In medicine, calcium phosphates have been used for bone regeneration for several decades. The requirement of a mouldable, self-setting material has been fulfilled since the mid-1980s because of the development of calcium phosphate cements. Basically, they consist of a powder (e. g. di-, tri- or tetra-calcium phosphates) that is mixed with a liquid. Their properties depend on kind, amount, and location of each atom within the crystal structure. In dentistry calcium phosphate cements play a secondary role at the moment, although they often have an excellent biocompatibility. This review gives a general idea on development and chemistry of calcium phosphate cements and presents different cement types tested in vitro and in vivo.

Animals↗