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High early strength calcium phosphate bone cement: effects of dicalcium phosphate dihydrate and absorbable fibers.

Calcium phosphate cement (CPC) sets in situ to form resorbable hydroxyapatite with chemical and crystallographic similarity to the apatite in human bones, hence it is highly promising for clinical applications. The objective of the present study was to develop a CPC that is fast setting and has high strength in the early stages of implantation. Two approaches were combined to impart high early strength to the cement: the use of dicalcium phosphate dihydrate with a high solubility (which formed the cement CPC(D)) instead of anhydrous dicalcium phosphate (which formed the conventional cement CPC(A)), and the incorporation of absorbable fibers. A 2 x 8 design was tested with two materials (CPC(A) and CPC(D)) and eight levels of cement reaction time: 15 min, 30 min, 1 h, 1.5 h, 2 h, 4 h, 8 h, and 24 h. An absorbable suture fiber was incorporated into cements at 25% volume fraction. The Gilmore needle method measured a hardening time of 15.8 min for CPC(D), five-fold faster than 81.5 min for CPC(A), at a powder:liquid ratio of 3:1. Scanning electron microscopy revealed the formation of nanosized rod-like hydroxyapatite crystals and platelet crystals in the cements. At 30 min, the flexural strength (mean +/- standard deviation; n = 5) was 0 MPa for CPC(A) (the paste did not set), (4.2 +/- 0.3) MPa for CPC(D), and (10.7 +/- 2.4) MPa for CPC(D)-fiber specimens, significantly different from each other (Tukey's at 0.95). The work of fracture (toughness) was increased by two orders of magnitude for the CPC(D)-fiber cement. The high early strength matched the reported strength for cancellous bone and sintered porous hydroxyapatite implants. The composite strength S(c) was correlated to the matrix strength S(m): S(c) = 2.16S(m). In summary, substantial early strength was imparted to a moldable, self-hardening and resorbable hydroxyapatite via two synergistic approaches: dicalcium phosphate dihydrate, and absorbable fibers. The new fast-setting and strong cement may help prevent catastrophic fracture or disintegration in moderate stress-bearing bone repairs.

Bone Substitutes↗

Accurate coronary calcium phosphate mass measurements from electron beam computed tomograms.

Precipitates of calcium phosphate in coronary arteries always indicate the presence of atherosclerosis. The mass of these precipitates is related to the severity of atherosclerosis. To determine the accuracy of electron beam computed tomographic (CT) mass estimates, we imaged 21 human hearts inside an anthropomorphic chest phantom using an Imatron C-100 electron beam CT scanner (Imatron, San Francisco, CA). We then incinerated the 63 imaged coronary arteries and weighed the mineral ash. We calculated the mass estimates from the images using an algorithm derived from a model that assumes simple radiographic properties of the coronary arteries. We also calculated the currently used coronary calcium score for each artery. Although both the mass estimates and the scores correlated with the actual mass of the incinerated specimens (r = .97 and r = .93), the correlation with the mass estimates was better (P = .02; William's test). The regression equation relating the actual mass to the mass estimates was y = 1.37 x + 14, indicating that the CT mass estimates consistently underestimate actual coronary calcium phosphate mass. We conclude that relative mass estimates using electron beam CT scanning are accurate and that both these and the currently employed calcium scores reflect the actual mass of precipitated calcium phosphate in diseased coronary arteries.

Algorithms↗

Effects of a rhubarb (Rhei rhizoma) solution and its fractions on the formation of calcium phosphate precipitates.

Inhibitory effects of a rhubarb (Rhei rhizoma) solution and its fractions on the formation of calcium phosphate precipitates were studied. The rhubarb solution inhibited both the amorphous calcium phosphate (ACP) formation and the rate of hydroxyapatite (HAP) transformation, and extended the induction time. When the solution was fractionated using membrane filters, a filtrate with the molecular weight between 3 and 10 kDa (with 2/3 recovery of polyphenols) was found to be responsible for both the ACP formation and the extension of the induction time. Another filtrate with the molecular weight below 3 kDa (with 1/3 recovery of polyphenols) may be responsible for the inhibition of both the ACP formation and the rate of HAP transformation, and the extension of the induction time. When the extract of rhubarb was fractionated using a Sephadex LH-20 column chromatography, fraction IV greatly inhibited the formation of calcium phosphate precipitates, while fractions I, II and III slightly inhibited that reaction. Our finding suggests that fraction IV may contain useful substance(s) for the prevention of oral calcium phosphate precipitation (calculus formation). However, strong calcium chelating properties would limit the concentration that could be safely employed.

Analysis of Variance↗

Formation and transformation of calcium phosphates: relevance to vascular calcification.

Only one phase, carbonate apatite, is present in the mineral phase of normal calcification while different calcium phosphate phases are found in the mineral phases of pathological calcifications. In vascular calcification, the mineral phase has been described as hydroxyapatite; carbonate apatite, amorphous calcium phosphate. It has also been speculated that octacalcium phosphate may be a precursor for the apatite. This paper provides a brief review of: (1) the different calcium phosphates in normal and pathological calcifications; (2) the conditions for the formation and transformation of biologically relevant calcium phosphates in synthetic systems and a concise discussion on the relevance of results from synthetic systems to the calcium phosphate phase(s) involved in vascular calcification.

Apatites↗

Amorphous calcium phosphate.

ACP is a unique calcium phosphate in that it lacks long-range crystalline order. Yet the constancy in the composition of ACP over a wide range of solution conditions suggests a well-defined local structural unit. Although this order within disorder is the most distinguishing feature of ACP, the solution instability of ACP and its ready transformation into crystalline phases such as OCP and apatite may be of greater biological relevance. In particular, the initiating role ACP plays in matrix vesicle mineralization raises the importance of this phase from a mere laboratory curiosity to that of a key intermediate in skeletal calcification.

Animals↗

Biomimetic calcium phosphate composite coating of dental implants.

PURPOSE: The aim of the present study was to test the hypothesis that calcium phosphate coating of titanium screw-type implants enhances peri-implant bone formation in the jaw. MATERIALS AND METHODS: Ten adult female foxhounds received experimental titanium screw-type implants in the mandible 3 months after removal of all premolar teeth. Four types of implants were evaluated in each animal: implants with machined titanium surface (the control group), implants coated with collagen I (the collagen-only group), implants with a composite coating of calcium phosphate and mineralized collagen I (the composite group), and implants with calcium phosphate (hydroxyapatite [HA]) coating (the HA-only group). Peri-implant bone regeneration was assessed histomorphometrically after 1 and 3 months in 5 dogs each by measuring bone-implant contact (BIC) and the volume density of the newly formed peri-implant bone (BVD). RESULTS: After 1 month, BIC was significantly enhanced only in the group of implants with composite coating of calcium phosphate and mineralized collagen (P = .038). Volume density of the newly formed peri-implant bone was significantly higher in all coated implants after 1 month. No significant difference from baseline was found in BIC for the collagen-only and HA-only groups, but BVD was significantly higher in implants with composite coating (P = .041). After 3 months, BIC and BVD were significantly higher in all coated implants than in the controls with machined surfaces. CONCLUSION: It was concluded that composite coating of dental screw-type implant surfaces using calcium phosphate and collagen can enhance BIC and peri-implant bone formation.

Animals↗

Endocytosis is required for the mitogenic effect of basic calcium phosphate crystals in fibroblasts.

Basic calcium phosphate (BCP) crystals stimulate mitosis of cultured fibroblasts and synoviocytes in vitro. Although intimate crystal-cell contact is required for mitogenesis, and lysosomotropic agents such as chloroquine and ammonium chloride block the mitogenic effect of crystals, the requirement of endocytosis has not been demonstrated. Synthetic BCP crystals, uniformly trace-labeled with 45Ca, were added to cultured, quiescent, confluent human foreskin fibroblasts in the presence or absence of ammonium chloride. After 4 or 24 hours, cultures were pulsed with 3H thymidine. The cells were then released with EDTA and trypsin, and fractionated on preformed Percoll density gradients. Fractions were analyzed for incorporation of 45Ca and [3H]thymidine and cell number. The cells containing 45Ca crystals also were heavily labeled with thymidine. Ammonium chloride decreased the amount of crystals endocytosed, and inhibited mitogenesis. These data suggest that mitogenesis induced by BCP crystals is preceded by endocytosis and dissolution in the acidic environment of phagolysosomes.

Calcium Phosphates↗

[Latest progress in studies of self-setting calcium phosphate cement].

Self-setting calcium phosphate cement(CPC) is a non-ceramic form of hydrxyapatitic artificial bone material (HAP) which was first exclusively produced in America several years ago. CPC is free from the drawbacksto ceramic HAP, including sintering and difficulties in shaping. CPC has the characteristics of simple-producing and easy applyication. In 1991, the clinical application of CPC to repairing the calvarial bone defects was approved, and the results reportedly so far were good or excellent. This paper presents the latest progress in the studies of CPC, concerning the probing into the setting process, the producing of fast-setting and non-decayed types of CPC, the studies of organic compound CPC, the in vitro results of CPC as a drug delivery system, etc. As the research goes on deeply and broadly, CPC is hopefully becoming a standard material in repairing bone defects at the non- or low-bearing site in the future.

Artificial Organs↗

Laser-induced crystallization of calcium phosphate coatings on polyethylene (PE).

Calcium phosphate (CaP) coatings are used for obtaining a desired biological response. Usually, CaP coatings on metallic substrates are crystallized by annealing at temperatures of at least 400-600 degrees C. For polymeric substrates, this annealing is not possible due to the low melting temperatures. In this work, we present a more suitable method for obtaining crystalline coatings on polymeric substrates, namely laser crystallization. We were successful in obtaining hydroxyapatite coatings on polyethylene. Because of the UV transmission characteristics of the CaP coatings, the use of a low wavelength (157 nm) F(2) laser was necessary for this. As a result of the laser treatment, the CaP coating broke up into islands. The cracks between the islands became larger and the surface became porous with increasing laser energy. The mechanism behind the formation of this morphology did not become clear. However, the fact that crystalline CaP coatings can be obtained on polymeric substrates in an easy way, possibly allows for the development of new products.

Calcium Phosphates↗

The reinforcement of cancellous bone screws with calcium phosphate cement.

The ability of calcium phosphate cement (CPC) to reinforce cancellous screws placed in previously stripped holes was studied in vitro. The distal end of canine femurs were harvested. A total of 15 screws were placed in six femurs. The pullout strength (failure force), failure displacement, stiffness, and energy absorbed were determined for the screws in the intact cancellous bone. Next, these stripped screw holes were packed with CPC. The pullout test was repeated, and the results were compared using a paired, Student's t test. We found that the CPC was able to reinforce the previously stripped holes and significantly increase the pullout strength (1,159 +/- 278 N versus 678 +/- 297 N) and the stiffness (1,990 +/- 569 N/mm versus 1,519 +/- 609 N/mm) of the constructs, as well as the energy absorbed by the constructs until failure (467 +/- 180 N.mm versus 278 +/- 140 N.mm). There was no difference in the failure displacement (0.94 +/- 0.23 versus 0.85 +/- 0.51 mm). This study documents the ability of CPC to acutely reinforce cancellous bone screws in a region with no or poor-quality cancellous bone.

Animals↗

Remineralization of enamel subsurface lesions by sugar-free chewing gum containing casein phosphopeptide-amorphous calcium phosphate.

Casein phosphopeptide-amorphous calcium phosphate nanocomplexes (CPP-ACP) exhibit anticariogenic potential in laboratory, animal, and human in situ experiments. The aim of this study was to determine the ability of CPP-ACP in sugar-free chewing gum to remineralize enamel subsurface lesions in a human in situ model. Thirty subjects in randomized, cross-over, double-blind studies wore removable palatal appliances with six human-enamel half-slabs inset containing sub-surface demineralized lesions. The appliances were inserted immediately before gum-chewing for 20 min and then retained for another 20 min. This was performed four times per day for 14 days. At the completion of each treatment, the enamel half-slabs were paired with their respective demineralized control half-slabs, embedded, sectioned, and subjected to microradiography and densitometric image analysis, for measurement of the level of remineralization. The addition of CPP-ACP to either sorbitol- or xylitol-based gum resulted in a dose-related increase in enamel remineralization, with 0.19, 10.0, 18.8, and 56.4 mg of CPP-ACP producing an increase in enamel remineralization of 9, 63, 102, and 152%, respectively, relative to the control gum, independent of gum weight or type.

Adult↗

Mechanical properties of porous, electrosprayed calcium phosphate coatings.

Mechanical properties of calcium phosphate coatings (CaP), deposited using the electrostatic spray deposition (ESD) technique, have been characterized using a range of analytical techniques, including tensile testing (ASTM C633), fatigue testing (ASTM E855), and scratch testing using blunt and sharp scratch styli. Moreover, a simple explantation procedure was successfully introduced using ESD-coated, threaded dental implants to characterize the mechanical performance of CaP coatings qualitatively under conditions that mimic clinical situations as close as possible. Generally, all analysis techniques revealed that ESD coatings need to be crystallized in order to ensure interfacial adhesion to the substrate and sufficient mechanical strength of the superficial reticular structure. Crystalline carbonated hydroxyapatite coatings (CHA, heat-treated at 700 degrees C) were resistant to fatigue as well as to plastic ploughing deformation by means of various scratch styli, and the fragile surface structure of ESD coatings was maintained to a large extent after unscrewing CHA-coated dental implants from femoral condyles of goat cadavers. From these experiments, it was concluded that interfacial adhesion of crystalline CHA ESD coatings to the titanium substrate was sufficient, but that mechanical strength of the superficial architecture of ESD coatings need to be optimized for applications where high shear and compressive stresses are imposed onto the rather fragile coating surface of reticular ESD morphologies.

Animals↗

Relationship between osteogenic characteristics of bone marrow cells and calcium phosphate surface relief and solubility.

The capacity of mouse bone marrow cells to adhere to calcium phosphate surfaces and form tissue plates depending on the surface relief and solubility was studied in ectopic bone formation test. Calcium phosphate coating of titanium disks, made by the anodic spark (microarch) oxidation in 10% orthophosphoric acid with hydroxyl apatite particles, differed by the structure (thickness of coating, size of pores, and roughness) and solubility (level of in vitro oxidation of 1-week extracts of implants). Chemical (phasic and element) composition of the studied calcium phosphate coatings was virtually the same. The findings indicate that histogenesis is regulated by physicochemical characteristics of the implant surface. It seems that the osteogenic potential of calcium phosphate surfaces is largely determined by their relief, but not by pH of degradation products.

Animals↗

Calcium phosphate deposition in human dental plaque microcosm biofilms induced by a ureolytic pH-rise procedure.

The objectives were to develop and characterize a procedure based on a ureolytic pH rise to deposit calcium phosphate into microcosm dental plaque biofilms and to test the importance of the plaque pH range. Plaque biofilms were cultured in a multiplaque culture system ('artificial mouth') with a continuous supply of a simulated oral fluid (basal medium mucin; BMM) with 146 mmol/l (5% w/v) sucrose periodically applied over 6 min every 8h. After initial plaque growth, the biofilms were periodically exposed for up to 16 days to 6-min applications of calcium phosphate monofluorophosphate urea (CPMU) solution containing 20 mmol/l CaCl(2), 12 mmol/l NaH(2)PO(4), 5 mmol/l monofluorophosphate and 500 mmol/l urea (pH 5.0). Three application regimes were examined, one included a sucrose-induced acidic pH fluctuation. Plaque hydrolysis of the urea in CPMU caused the pH to rise to between 8.2 and 8.8, depositing fluoridated and carbonated calcium phosphates, and possibly some calcium carbonate, into the plaque. Calcium, phosphate and fluoride deposition was rapid for about 4 days and then slowed. After 10 days' treatment under standard conditions (BMM containing 1 mmol/l urea and 1 mmol/l arginine), plaque calcium and phosphate concentrations had increased up to 50-fold and 10-fold to approximately 2-4 and 1-2 mmol/g plaque protein, respectively. The calcium, phosphate and fluoride content increased steadily. Calcium phosphate deposition was proportional to the plaque resting pH, increasing over four-fold when the BMM urea concentration was increased from 0 to 20 mmol/l, which raised the resting pH from 6.4 to 7.2 and yielded a mean plaque calcium concentration of 14.3 mmol/g protein, one subsample reaching 20.8 mmol/g protein. Supplementation of BMM with 20% human serum inhibited deposition. These results support the hypothesis that an alkaline pH in plaque is critical in promoting plaque mineralization and that mineral deposition is modulated by serum. These factors are likely to be important in regulating calculus formation.

Biofilms↗

The use of an injectable, biodegradable calcium phosphate bone substitute for the prophylactic augmentation of osteoporotic vertebrae and the management of vertebral compression fractures.

STUDY DESIGN: A biomechanical study comparing two materials for augmentation of osteoporotic vertebral bodies and vertebral bodies after compression fracture. OBJECTIVES: To compare an injected, biodegradable calcium phosphate bone substitute with injected polymethylmethacrylate bone cement for strengthening osteoporotic vertebral bodies and improving the integrity of vertebral compression fractures. SUMMARY OF BACKGROUND DATA: Injection of polymethylmethacrylate bone cement into fractured vertebral bodies has been used clinically. However, there is concern about thermal damage to the neural elements during polymerization of the polymethylmethacrylate bone cement as well as its negative effects on bone remodeling. Biodegradable calcium phosphate bone substitutes have been studied for enhancement of fixation in fractured vertebrae. METHODS: Forty fresh osteoporotic thoracolumbar vertebrae were used for two separate parts of this study: 1) injection into osteoporotic vertebrae: intact control (n = 8), calcium phosphate (n = 8), and polymethylmethacrylate bone cement (n = 8) groups. Each specimen then was loaded in anterior compression until failure; 2) injection into postfractured vertebrae: calcium phosphate (n = 8) and polymethylmethacrylate bone cement (n = 8) groups. Before and after injection, the specimens were radiographed in the lateral projection to determine changes in vertebral body height and then loaded to failure in anterior bending. RESULTS: For intact osteoporotic vertebrae, the average fracture strength was 527 +/- 43 N (stiffness, 84 +/- 11 N/mm), 1063 +/- 127 N (stiffness, 157 +/- 21 N/mm) for the group injected with calcium phosphate, and 1036 +/- 100 N (stiffness, 156 +/- 8 N/mm) for the group injected with polymethylmethacrylate bone cement. The fracture strength and stiffness in the calcium phosphate bone substitute group and those in the polymethylmethacrylate bone cement group were similar and significantly stronger than those in intact control group (P < 0.05). For the compression fracture study, anterior vertebral height was increased 58.5 +/- 4.6% in the group injected with calcium phosphate and 58.0 +/- 6.5% in the group injected with polymethylmethacrylate bone cement as compared with preinjection fracture heights. No significant difference between the two groups was found in anterior vertebral height, fracture strength, or stiffness. CONCLUSION: This study demonstrated that the injection of a biodegradable calcium phosphate bone substitute to strengthen osteoporotic vertebral bodies or improve vertebral compression fractures might provide an alternative to the use of polymethylmethacrylate bone cement.

Absorbable Implants↗

Effects of calcium phosphate bioceramics on skeletal muscle cells.

With advances in ceramics technology, calcium phosphate bioceramics have been applied as bone substitutes. The effects of implants on bony tissue have been investigated. The effects upon adjacent skeletal muscles have not been determined. The focus of this work is to elucidate the biological effects of various calcium phosphate bioceramics on skeletal muscles. Four different kinds of powder of calcium phosphate biomaterials including beta-tricalcium phosphate (beta-TCP), hydroxyapatite (HA), beta-dicalcium pyrophosphate (beta-DCP) and sintered beta-dicalcium pyrophosphate (SDCP), were tested by myoblast cell cultures. The results were analyzed by cell count, cell morphology and concentration of transforming growth factor beta 1 (TGF-beta 1) in culture medium. The cell population and TGF-beta 1 concentration of the control sample increased persistently as the time of culture increased. The changes in cell population and TGF-beta 1 concentration in culture medium of the beta-TCP and HA were quite low in the first 3 days of culture, then increased gradually toward the seventh day. The changes in cell population and TGF-beta 1 concentration in culture medium of the silica, beta-DCP, and SDCP were quite similar. They were lower during the first day of culture but increased and reached that of the control medium after 7 days' culture. Most cells on B-TCP and HA diminished in size with radially spread, long pseudopods. We conclude that HA and beta-TCP are thought to have an inhibitory effect on growth of the myoblasts. The HA and beta-TCP may interfere with the repair and regeneration of injured skeletal muscle after orthopedic surgery.

Animals↗

Precipitation of nucleotides by calcium phosphate.

Earlier it has been shown that nucleic acids of high molecular weight can be be introduced into cells by coprecipitation with calcium phosphate. We have studied the requirements for calcium phosphate coprecipitation of shorter nucleotides. The degree of coprecipitation of dodecanucleotides lacking terminal phosphate varied between 25 and 72%. Tetramers with a 5'-monophosphate were coprecipitated to 29-87% by calcium phosphate. A high content of guanosine residues and an increased number of terminal phosphate groups increased the degree of coprecipitation of nucleotides. The trinucleotide pppA2'p5'A2'p5'A was effectively precipitated by calcium phosphate but the monophosphate and the core structure were not.

Calcium Phosphates↗

Comparison of urinary tract infection in calcium oxalate and calcium phosphate stone formers.

To compare the frequency of urine infection in calcium oxalate and calcium phosphate stone formers, we reviewed charts from patients whose last renal stone submitted for analysis was predominantly composed of calcium phosphate in 118 and of calcium oxalate in 223. Positive cultures were commoner, but not significantly, in the phosphate than the oxalate stone formers, both in men (17 vs. 7.6%) and women (22 vs. 15%). Bacteria frequently producing urease were found in only 4% of the phosphate group. Urine leucocytes were slightly more frequent in the oxalate group for men and significantly so for women. The results do not support the concept that calcium phosphate stones are mainly due to infection with urease-producing or other bacteria.

Calcium Oxalate↗