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Biocompatibility and resorption of a brushite calcium phosphate cement.

A hydraulic calcium phosphate cement with beta-tricalcium phosphate (TCP) granules embedded in a matrix of dicalcium phosphate dihydrate (DCPD) was implanted in experimentally created defects in sheep. One type of defect consisted of a drill hole in the medial femoral condyle. The other, partial metaphyseal defect was located in the proximal aspect of the tibia plateau and was stabilized using a 3.5 mm T-plate. The bone samples of 2 animals each per group were harvested after 2, 4, 6 and 8 weeks. Samples were evaluated for cement resorption and signs of immediate reaction, such as inflammation, caused by the cement setting in situ. Differences regarding these aspects were assessed for both types of defects using macroscopical, radiological, histological and histomorphometrical evaluations. In both defects the brushite matrix was resorbed faster than the beta-TCP granules. The resorption front was followed directly by a front of new bone formation, in which residual beta-TCP granules were embedded. Cement resorption occurred through (i) extracellular liquid dissolution with cement disintegration and particle formation, and (ii) phagocytosis of the cement particles through macrophages. Signs of inflammation or immunologic response leading to delayed new bone formation were not noticed at any time. Cement degradation and new bone formation occurred slightly faster in the femur defects.

Absorbable Implants↗

In vivo cell interactions with calcium phosphate bioceramics.

Biointegration, resorption process, and solubility in physiological environments of calcium phosphate materials are scarcely described by ultrastructural studies. In vivo cells interactions with calcium phosphate materials are scarcely described by ultrastructural studies. In vivo cells interactions with calcium phosphate biomaterials are mediated by different proteins from physiological fluid, and in order to observe at the ultrastructural level the cell colonization, the resorption, process and the biointegration, we used in these experiments calcium phosphate materials precoated with fibronectin or not precoated. Two kinds of well determined materials were used for this study, Beta-tricalcium phosphate (B-TCP) and hydroxyapatite (HAP). The implants were soaked in human fibronectin diluted solution and were implanted in the connective tissue of rabbit abdomen. Our results showed that the fibroblasts and macrophagous++ cells interaction with the calcium phosphate crystal (B-TCP and HAP) was more important in the experiments with a fibronectin bilayer. In the presence of fibronectin at the grains surface of the material, cystic cavities' or fibrous encapsulation was suppressed and cells with fibers were in close contact with the material. The presence of fibronectin immediately after implantation seemed to increase the adhesion and the cell colonization. Fibronectin creates an organic interface between crystals and cells, and can promote interactions from cells and biomaterials.

Abdomen↗

Setting, hardening and resorption of calcium phosphate hydraulic cements.

Two examples of calcium phosphate hydraulic cements (CPHC) are presented. Type I cements consist in mixtures of beta-tricalcium phosphate (beta-TCP) and monocalcium phosphate monohydrate (MCPM), to which some plaster of Paris (CSH) is added as a setting retardant. Type II cements consist in mixtures of beta-TCP, dicalcium phosphate dihydrate (DCPD) and calcium carbonate (CC), to which some hydroxyapatite (HAP) is added as a setting accelerator. The setting time of these cements ranges from a few minutes up to a few hours, according to their composition and the amount of mixing water. Tensile strengths ranging from 1.2 up to 3.5 MPa have been recorded on cements which porosity ranged from 38 up to 55 vol%. In-vivo experiments on dogs have shown the perfect biocompatibility and resorbability of cement I.

Animals↗

Effect of temperature on electrochemical deposition of calcium phosphate coatings in a simulated body fluid.

Calcium phosphate coatings were deposited on titanium plate by an electrochemical method in simulated body fluid at 5-62 degrees C. X-ray diffractometry and FTIR studies demonstrated that the deposits at 5, 22 and 37 degrees C were amorphous and those at 52 and 62 degrees C contained Mg(OH)2, CaCO3 and carbonate apatite of low crystallinity. The calcium, magnesium and phosphorus contents of deposit increased in direct proportion to the square root of loading time of cathodic potential. Induction periods, which might be thought to be the time required to decrease the pH of the electrolyte around the cathode by the formation of H2 gas and to start deposition of calcium phosphate, were observed on all the regression lines. It is concluded that in the electrochemical synthesis of calcium phosphate in this temperature range the diffusion process is a rate-determining step.

Body Fluids↗

Calcium, phosphate, and PTH levels in the hemodialysis population: a multicenter study.

BACKGROUND: Hyperphosphatemia is one of the main factors in the pathogenesis of secondary hyperparathyroidism. In addition, in dialysis patients elevated levels of phosphate and calcium times phosphate (Ca x P) ion product are associated with extraskeletal calcifications, as well as an increased risk of death. Cardiovascular calcifications may possibly be related to the high cardiovascular mortality seen in dialysis patients. In the USA the prevalence of hyperphosphatemia among dialysis patients is very high, over 60% of patients having serum P levels > 5.5 mg/dl, but no data are available for the Italian population, which follows different diet and dialysis schedules. METHODS: We looked at a random sample of 638 patients enrolled in 29 outpatient dialysis units in Southern Italy (Campania and Sicily). Data were centrally analyzed after extraction from a computerized database. In each patient the average of two or more values obtained in the six-month observation period (from Jan. 99 to June 99) was calculated. Mean age was 61.2 years (95% CI 59.9-62.5) years, dialytic age 72.2 months (95% CI 67.4-77.0). RESULTS: Mean +/- SD (95% CI) levels of phosphate were 5.74 +/- 1.57 (5.62-5.86) mg/dl; total calcium 9.06 +/- 0.98 (8.98-9.14) mg/dl; Ca x P 51.4 +/- 14.4 (50.3-52.5) mg2/dl2; alkaline phosphatase was 167 +/- 119 (157-177) IU/L; intact parathyroid hormone (PTH) levels, available in a subset of 239 patients, were 318 +/- 413 (265-370) pg/ml. Among these patients 51.6% had serum P > 5.5 mg/dl, 36% > 6 mg/dl; Ca x P levels were > 55 mg2/dl2 in 35.5% of the population, 24% > 60 mg2/dl2; iPTH levels were < 100 pg/ml in 43%; 25.4% had hyperparathyroidism (defined as iPTH > 400 pg/ml), and only 19.5% of patients had PTH in the desired interval of 100 to 250 pg/ml. CONCLUSIONS: Compared to data reported from the USA, mean levels of phosphate, Ca x P, and PTH seem better controlled in this Italian hemodialysis population. However, in a significant number of patients these parameters were still outside the normal range. Both over-suppression of PTH levels and hyperparathyroidism are present, although surprisingly the former was more frequent. Treatment of hyperphosphatemia should be more aggressive in hemodialysis patients; PTH levels are difficult to maintain in the desired range of 100-250 pg/ml.

Calcium↗

Micro-architecture of calcium phosphate granules and fibrin glue composites for bone tissue engineering.

Calcium phosphate ceramics are currently used as bone graft substitutes in various types of clinical applications. Fibrin glue is also used in surgery due to its haemostatic, chemotactic and mitogenic properties. By combining these two biomaterials, new composite scaffolds were prepared. In this study, we attempt to analyse whether thrombin concentration in the fibrin glue could influence the properties of the composite. The association between fibrin glue and calcium phosphate ceramic granules was characterized at the ultra structural level. Micro and macroporous biphasic calcium phosphate ceramic granules with a diameter of 1-2mm composed of hydroxyapatite and beta-tricalcium phosphate (60/40) were associated to fibrin glue. The composites were observed by scanning and transmission electron microscopy and microcomputed tomography. Fibre thickness, porosity and homogeneity of the fibrin clot were modified by increased the thrombin concentration. Mixing fibrin glue with calcium phosphate granules (1:2) did not modify the microstructure of the fibrin clot in the composite. Nevertheless, thrombin interacted with the bioceramic by inducing the nucleation of crystalline precipitate at the ceramic/fibrin glue interface. Combining fibrin sealant and calcium phosphate ceramics could lead to new scaffolds for bone tissue engineering with the synergy of the properties of the two biomaterials.

Bone Substitutes↗

Hip screw augmentation with an in situ-setting calcium phosphate cement: an in vitro biomechanical analysis.

OBJECTIVES/HYPOTHESIS: This study was performed to determine whether a new, in situ-setting calcium phosphate cement would have sufficient mechanical integrity to reinforce compression screw fixation of unstable intertrochanteric fractures. We compared the cut-out resistance of screws augmented with calcium phosphate cement to the cut-out resistance of screws augmented with polymethylmethacrylate (PMMA). We used PMMA as the standard for comparison because it is currently used clinically. Our hypothesis was that initial fixation strength with PMMA and calcium phosphate cement augmentation would not be significantly different from one another. STUDY DESIGN: Cut-out testing of compression hip screws in paired human cadaveric proximal femurs was performed before and after augmentation with PMMA or calcium phosphate cement. Bilateral testing was performed to allow pairwise comparisons of the materials used for augmentation, and repeated testing was done to provide an internal control for the effects of bone quality. The initial fixation of screws augmented with calcium phosphate cement was compared with that of screws augmented with PMMA. METHODS: Ten paired human femurs (mean age, 75 +/- 9.2 years) were implanted with Richards AMBI compression hip screws. Basicervical osteotomies were then performed, yielding isolated proximal fragments for mechanical testing. Preaugmentation cut-out tests were performed under displacement control, with cut-out continuing to five millimeters at two millimeters per second. The screws were then removed, and the screw tracks were filled with 2.0 cubic centimeters of PMMA (one side) or calcium phosphate cement (contralateral side). After augmentation, the screws were reinserted and the cements were allowed to harden for twenty-four hours. Postaugmentation testing followed the protocols for preaugmentation testing, and the initial fixation strength of screws augmented with calcium phosphate cement was compared with the initial fixation strength of screws augmented with PMMA using a two-way repeated measures analysis of variance. RESULTS: The cut-out behavior of screws augmented with calcium phosphate cement was not significantly different from the cut-out behavior of screws augmented with PMMA. With calcium phosphate cement, yield strength increased by 15.8 percent (from 1,354 +/- 632 newtons to 1,568 +/- 320 newtons); with PMMA, the yield strength increased by 26.8 percent (from 1,477 +/- 526 newtons to 1,834 +/- 225 newtons). However, only the increase with PMMA augmentation was significant at p < 0.05). The energy to yield increased significantly (41 percent, p < 0.05) with both types of augmentation (from 2,399 +/- 1,186 newton-millimeters to 3,378 +/- 857 newton-millimeters for calcium phosphate cement, and from 2,635 +/- 1,113 newton-millimeters to 3,741 +/- 426 newton-millimeters for PMMA), whereas the stiffness increased only slightly with PMMA augmentation (6.2 percent, from 481 +/- 180 newtons per millimeter to 511 +/- 92 newtons per millimeter) and fell slightly with calcium phosphate cement augmentation (10 percent, from 457 +/- 201 newtons per millimeter to 411 +/- 663 newtons per millimeter). CONCLUSIONS: The in situ-setting calcium phosphate cement investigated in this study compared favorably with PMMA in a single-cycle cut-out test of augmented compression hip screws in senile trabecular bone. Our results suggest that these materials may have promise as substitutes for PMMA in the salvage of compression hip screw fixation in elderly osteopenic patients with complex intertrochanteric fractures and that further study of their use in this application is warranted.

Aged↗

Clinical and histologic observation of replacement of biphasic calcium phosphate by bone tissue in monkeys.

Biphasic calcium phosphate, consisting of beta-tricalcium phosphate and hydroxyapatite, was implanted in wide and deep periodontal osseous defects of monkeys in combination with the guided tissue regeneration technique. After 12 weeks, sites treated with a combination of biphasic calcium phosphate and guided tissue regeneration maintained the shape of the ridge, but both guided tissue regeneration and control sites (untreated) showed extreme resorption. A histopathologic investigation revealed that numerous macrophages contained small particles of ceramic within their vesicles and the active bone replacement occurred from the surrounding bone. Biphasic calcium phosphate has osteoconductive potential and this potential may be related to degradation by macrophage phagocytosis.

Alveolar Bone Loss↗

Basic calcium phosphate deposition in the joint: a potential therapeutic target in osteoarthritis.

PURPOSE OF REVIEW: Basic calcium phosphate crystals are responsible for a number of clinical syndromes. The study of basic calcium phosphate crystal deposition diseases has been hindered by a lack of readily available, accurate, diagnostic tests. Recent data have provided further understanding of the mechanisms by which basic calcium phosphate crystals induce inflammation and degeneration within the joint, as well as their potential role in other conditions such as cancer and atherosclerosis. RECENT FINDINGS: New information on the effects of basic calcium phosphate crystals on matrix metalloproteinases and mitogenesis further supports a role for basic calcium phosphate crystals in the pathogenesis of osteoarthritis. Phosphocitrate remains the most promising of the potential therapeutic agents, which could antagonize the effects of basic calcium phosphate crystals, although other therapies have also been examined. SUMMARY: Further work is needed to clarify the exact role basic calcium phosphate crystals play in the development of osteoarthritis.

Arteriosclerosis↗

Dissolution properties of calcium phosphate granules with different compositions in simulated body fluid.

Calcium phosphate granules were produced through the calcination of three hydroxyapatites (HAs) at 1150 degrees C: nearly-stoichiometric (NS-HA), calcium-deficient (CD-HA), and carbonated (Carb-HA). The characterization using scanning electron microscope (SEM), X-ray diffraction (XRD), and Fourier-transformed infrared spectroscopy (FT-IR) showed that those granules exhibit strong differences in chemical composition, surface texture, and dissolution behavior. Sample dissolution in a simulated body fluid (SBF) and precipitation of a calcium phosphate layer on the granule surface were followed up for 7 days by chemical analysis of calcium content in SBF. After 80-min reaction, precipitation of new calcium phosphate phases predominates over the dissolution of original granules. Comparison between SEM images and XRD/FT-IR analysis revealed that the composition of the calcium phosphate layer and its precipitation rate depend on the HA composition and its heat treatment. Calcined carbonated apatite induces the precipitation of an octacalcium phosphate (OPC) layer, whereas a carbonated apatite layer is formed from calcined-deficient HA. The calcined nearly-stoichiometric HA exhibits low efficiency in producing carbonated apatite layer.

Biocompatible Materials↗

TEM study of calcium phosphate precipitation on bioactive titanium surfaces.

Formation of calcium phosphate on alkali- and heat-treated titanium surfaces was investigated using transmission electron microscopy (TEM). The samples were prepared by immersing the alkali- and heat-treated titanium in a revised simulated body fluid (R-SBF) with the same HCO(3)(-) concentration level as in human blood plasma. The deposition and precipitation on treated titanium surfaces were extracted for TEM examination without thinning process. Electron diffraction of the precipitates revealed that octacalcium phosphate (OCP), instead of hydroxyapatite (HA), directly nucleates from amorphous calcium phosphate. The OCP crystals continuously grew on the titanium surfaces rather than transforming into apatite. Calcium titanate was also identified by electron diffraction. Its role in the formation bioactive calcium phosphate, however, is not clear from this experiment.

Biomimetic Materials↗

Morphological analysis of renal cell culture models of calcium phosphate stone formation.

Cell culture models of calcium phosphate renal stone formation were established using the MDCK cell line. Renal microliths were detected within pseudocysts in three-dimensional soft agar cultures, and were also observed in the basal region of cells lining the cell sheet, and immediately beneath domes or blisters in monolayers and collagen gel cultures. Light and scanning electron microscopy indicated that these microliths had a similar lamellated and spherical appearance to those in humans. These microliths were first detected microscopically after 21 days of culture, and were found to be composed of calcium phosphate by X-ray and micro-infrared spectroscopic analyses. These culture models may provide a powerful new tool to study the pathogenesis of renal stone diseases and/or calcium phosphate stone formation in humans and animals.

Animals↗

Constant composition dissolution of mixed phases. II. Selective dissolution of calcium phosphates.

Characterization of the dissolution kinetics of individual synthetic and biological calcium phosphates is of considerable importance since these phases often coexist in biological minerals. The constant composition method has been used to study the dissolution kinetics of a series of synthetic calcium phosphates, brushite (DCPD), beta-tricalcium phosphate (TCP), octacalcium phosphate (OCP), hydroxyapatite (HAP), and carbonated apatite (CAP) in the presence and absence of citric acid, as a function of pH and thermodynamic driving force. While citric acid markedly accelerates the dissolution of TCP, HAP dissolution is significantly inhibited. Moreover, this additive has almost no influence on the dissolution of DCPD, OCP, and CAP. Dual constant composition dissolution studies of mixed calcium phosphates in the presence of citric acid have also been made. Another factor, pH, also plays an important role in the dissolution of these calcium phosphates. In suspensions of calcium phosphate mixtures, specific phases can be selectively dissolved by changing experimental parameters such as pH and the presence of rate modifiers. This result has important applications for the dissolution control of dental hard tissues such as dentin, enamel, and calculus.

Calcium Phosphates↗

Elucidation of factors determining formation of calcium phosphate stones.

PURPOSE: In this study we compared the effect of the urinary concentration of calcium with that of oxalate on the urinary relative saturation ratio (RSR) of calcium oxalate and brushite in patients with pure or mixed calcium phosphate stones. MATERIALS AND METHODS: A retrospective data analysis was performed of urinary stone risk analysis in 133 patients with pure or mixed calcium phosphate stones (30% or greater). Correlation was sought between stone events and urinary stone risk factors, and between brushite and urinary constituent RSRs before and after adjustment for variables. RESULTS: The formation rates of total events and stone surgeries weakly but significantly correlated with RSR urinary brushite (total events r = 0.22, p = 0.01) but not with RSR calcium oxalate. Urinary RSR brushite significantly correlated positively with urinary calcium (r = 0.67, p <0.0001), phosphorus and pH. After adjustment RSR brushite inversely correlated with citrate (r = -0.3, p = 0.0006). However, it did not correlate with oxalate. CONCLUSIONS: In stone formers with calcium phosphate or mixed calcium oxalate-calcium phosphate stones the stone formation rate depends on the urinary saturation of brushite but not of calcium oxalate. RSR brushite is determined by the urinary concentration of calcium, phosphate, citrate and pH but not by oxalate. Hypercalciuria should be controlled in patients with idiopathic calcareous renal calculi who form stones of pure or mixed calcium phosphate.

Adult↗

A simplified estimate of the ion-activity product of calcium phosphate in urine.

A computerized program was used to calculate the activities of the calcium, phosphate, and hydrogen phosphate ions. The most important determinants for ion-activity products of calcium phosphate and brushite were calcium, phosphate, citrate, urine volume, and pH. These urine variables were used to derive indices corresponding to ion-activity products of calcium phosphate (AP[CaP] index) and calcium hydrogen phosphate (AP[Bru] index). Factors were given to adapt these indices to collection periods shorter than 24 h. Relationships between the AP(CaP) index and ion-activity products of hydroxyapatite and octacalcium phosphate were also formulated. With urine electrolytes within the normal range a very good correlation was obtained between these indices and corresponding ion-activity products. The coefficient of correlation was better than 0.99. The presented indices might be useful in the evaluation of patients with renal calcium stone formation.

Calcium Phosphates↗

Influence of annealing temperature on RF magnetron sputtered calcium phosphate coatings.

The effect of different annealing temperatures on the characteristics of thin calcium phosphate coatings fabricated by radiofrequency magnetron sputtering was studied. Annealing of the as-sputtered films was necessary to change the amorphous coating to a crystalline coating. The films were annealed for 2 and 4 h at 400, 600, 800, 1000 and 1200 degrees C under dry argon or argon and water vapour flow. After annealing, the structure and the chemical composition of these films were characterized with incident light microscopy, Rutherford backscattering spectrometry (RBS), X-ray diffraction (XRD), and Fourier transform infrared absorption spectrometry (FTIR). Incident light microscopy showed cracks in the coatings annealed at a higher temperature than 400 degrees C. RBS revealed that the as-sputtered coatings had a high Ca/P ratio which decreased with increasing annealing temperature. After annealing at a temperature of 600 degrees C or more the XRD showed crystalline hydroxyapatite (HA) coatings. However, the second phase, present in the coatings, changed from tetra-calcium phosphate to calcium oxide to beta-tri-calcium phosphate with increasing annealing temperature. FTIR measurements showed the existence of OH- and PO- bonds in all coatings, although the PO- bonds varied for different annealed coatings, from the PO- bonds due to HA to PO- bonds due to other calcium phosphates. From the results of this study we suggest that 600 degrees C is probably the best annealing temperature to obtain a better characterization and understanding of the coating.

Biocompatible Materials↗

Behaviour of an injectable calcium phosphate cement with added tetracycline.

A calcium phosphate cement containing an antibiotic can be used for filling bone defects and to ensure local antibiotherapy. A calcium phosphate cement (already marketed under the name of Cementek can become injectable thanks to the addition of silicone. For dental applications, the behaviour of this injectable cement with added tetracycline was investigated. The tetracycline hydrochloride does not allow maturation of the cement: the tetracycline has to be treated with a calcium sulphate solution. The treated tetracycline (TTC) allowed maturation of the cement towards hydroxyapatite. But the setting time was longer and the mechanical properties decreased. Study in a continuous flow cell showed that the tetracycline is released in a continuous manner: thus, after 6 days, 60% of the antibiotic was released into the surrounding medium.

Calcium Phosphates↗

A preliminary study on osteoinduction of two kinds of calcium phosphate ceramics.

With respect to the effect of material factors on calcium phosphate biomaterial-induced osteogenesis, the osteoinductive property of two kinds of porous hydroxyapatite ceramics, which were made by different producers, was investigated in dorsal muscles of dogs. One hydroxyapatite ceramic (S-HA), macroporous implants with rough pore walls containing abundant micropores, was made by Sichuan Union University (Chengdu, China); the other hydroxyapatite ceramic (J-HA), porous implants with smooth macropore walls composed of regularly aligned crystal grains, was provided by Mitsubishi Ceramic Int. (Japan). Different tissue response was detected histologically and microradiographically after the ceramic samples had been implanted in dorsal muscles of dogs for 3 and 6 months. Bone formation was found in S-HA at 3 months, which increased at 6 months. In contrast, no bone formation was detected in J-HA at both 3 and 6 months. These results indicate that with the special architecture, calcium phosphate ceramic can induce bone formation in soft tissue. As both materials were very similar in their chemical and crystallographic structures, but varied in their microstructures, the latter seem to be an important factor affecting the osteoinductive capacity of calcium phosphate ceramics. These data suggest that, by controlling the preparation of calcium phosphate ceramic, bone substitutes with intrinsic osteoinductive property can be developed from calcium phosphates.

Animals↗