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PubMed2006

Antibody-conjugated soybean oil-filled calcium phosphate nanoshells for targetted delivery of hydrophobic molecules.

Hollow calcium phosphate nanoparticles capable of encapsulating poorly water-soluble molecules were produced by self-assembly. Previously reported were solid calcium phosphate nanoparticles and water-filled calcium phosphate nanocapsules suited for encapsulating mostly hydrophilic, but not hydrophobic compounds. Here, calcium phosphate was deposited around 100 nm diameter, 1,2-dioleoyl-sn-glycero-3-phosphate stabilized soybean oil nanoemulsions using either calcium chloride or NaOH titrations to achieve shell thickness between 20-70 nm. The surface was functionalized with carboxylic acid via the addition of carboxyethylphosphonic acid to attach Molecular Probes AB-594C antibody using sulpho-n-hydroxysuccinimide and 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide hydrochloride with an efficiency of approximately 70%, while retaining near complete antibody function. Hydrophobic pyrene was encapsulated with an efficiency of 95%, at concentrations much higher than its water solubility limit, and exhibited spectral features characteristic of a hydrophobic environment. These materials can be used in the targeted delivery of many useful, yet poorly water-soluble pharmaceutical and nutraceutical compounds.

Antibodies↗
PubMed1997

Osteoclastic resorption of calcium phosphate ceramics with different hydroxyapatite/beta-tricalcium phosphate ratios.

To study the influence of calcium phosphate ceramic solubility on osteoclastic resorption, neonatal rabbit bone cells were cultured for 2 days on hydroxyapatite (HA), beta-tricalcium phosphate (beta-TCP) and two types of biphasic calcium phosphate (BCP) with HA/beta-TCP ratios of 25/75 and 75/25. Solubility was regulated by varying the ratio of less-soluble HA and more-soluble beta-TCP. After removal of stromal cells by pronase E treatment, ceramic surfaces were observed by scanning electron microscopy. Osteoclasts resorbed BCP most extensively, with an HA/beta-TCP ratio of 25/75, producing typical lobulated, zig-zag track-like resorption lacunae. On pure beta-TCP, which had the highest solubility in acid, osteoclasts formed smaller discontinuous island-like lacunae. The resorption pattern may have been modified by the large number of calcium ions released into the acidic microenvironment at the osteoclast-ceramic interface. No resorption lacunae were found on the other specimens. The extent of osteoclastic resorption of calcium phosphate ceramics might, to a certain degree, be proportional to solubility, although this was not the case when solubility was very high. It would appear that ceramic solubility influences osteoclast resorption activity.

Animals↗
PubMed1999

Canine calcium phosphate uroliths. Etiopathogenesis, diagnosis, and management.

Uroliths composed predominantly of calcium phosphates have been infrequently identified in dogs. Factors incriminated in the etiopathogenesis of calcium phosphate urolithiasis include an alkaline urine pH, hypercalciuria, decreased urine concentrations of crystallization inhibitors, and increased urine concentrations of crystallization promoters. Disorders associated with calcium phosphate urolith formation in dogs include primary hyperparathyroidism, hyperadrenocorticism, and idiopathic hypercalciuria. Medical therapy of patients with recurring calcium phosphate uroliths should be directed at removing or minimizing factors contributing to urine supersaturation with calcium phosphate.

Animals↗
PubMedDate not supplied

Local tissue irritating effects and adjuvant activities of calcium phosphate and aluminium hydroxide with different physical properties.

Effects of calcium phosphate and aluminium hydroxide adjuvants with different physical properties were examined in guinea pigs for local histopathological reactions, electron-microscopical changes of macrophages and adjuvanticity on total IgG antibody response to subcutaneously administered ovalbumin (OVA) and tetanus toxoid (TT). Calcium phosphate gel (Ca-gel) induced active inflammatory reactions consisting of neutrophils (pseudoeosinophils) and foamy macrophages associated with many multinuclear giant cells for at least 4 weeks. Aluminium hydroxide gel (Al-gel) also elicited granulomatous inflammatory reactions consisting mainly of macrophages with foamy cytoplasm, small lymphocytes and giant cells at the injection sites for up to 8 weeks or longer. Severity of local tissue irritation due to calcium phosphate gel (Ca-gel) was similar to that due to Al-gel except for the duration of the inflammatory reactions. Calcium phosphate suspension (Ca-sus)-induced local reactions completely ceased by the 4th week, while aluminium hydroxide suspension (Al-sus)-induced reactions were seen up to the 8th week. Electron-microscopical observations showed that both Al-gel and Al-sus caused damage of macrophages. The adjuvant activity of Al-gel for OVA or TT was significantly stronger than that of any other adjuvant material, whereas those of Ca-gel and Ca-sus were not seen at a dose of 3 mg calcium phosphate per millilitre. Al-sus-TT at a dose of 3 mg aluminium hydroxide per millilitre induced very low levels of antibody. These results suggest that calcium phosphate adjuvant may not be an useful alternative to Al adjuvant.

Adjuvants, Immunologic↗
PubMed1993

Biodegradation and bioresorption of calcium phosphate ceramics.

The use of several calcium phosphate (Ca-P) materials for bone repair, augmentation, substitution and as coatings on metal implants has gained clinical acceptance in many dental and medical applications. These Ca-P materials may be of synthetic or natural origin, available in different physical forms (dense or macroporous, particles or blocks) and are used in bulk as coatings for metallic and non-metallic substrates or as components in composites, cements and bioactive glasses. Biodegradation or bioresorption of calcium phosphate materials implies cell-mediated degradation in vitro or in vivo. Cellular activity during biodegradation or bioresorption occurs in acid media; thus the factors affecting the solubility or the extent of dissolution (which in turn depends on the physico-chemical properties) of the Ca-P materials are important. Enrichment of the microenvironment due to the release of calcium and phosphate ions from the dissolving Ca-P materials affects the proliferation and activities of the cells. The increase in the concentrations of the calcium and phosphate ions promotes the formation of carbonate apatite which are similar to the bone apatite. The purpose of this invited paper is to discuss the processes of biodegradation or bioresorption of Ca-P materials in terms of the physico-chemical properties of these materials and the phenomena involved including the formation of carbonate apatite on the surfaces and in the vicinity of these materials. This phenomenon appears to be related to the bioactivity of the material and the ability of such materials to directly attach to bone and to form a uniquely strong material-bone interface.

Biodegradation, Environmental↗
PubMedDate not supplied

Calcium phosphate cement: review of mechanical and biological properties.

PURPOSE: Calcium phosphate cement is a bioactive and biodegradable grafting material in the form of powder and liquid, which when mixed, sets as primarily hydroxyapatite, sometimes mixed with unreacted particles and other phases. This material has been extensively investigated due to its excellent biological properties, potential resorbability, molding capabilities, and easy manipulation. Because the material can potentially be replaced with bone after a period of time, it could retain the short-term biological advantages of hydroxyapatite without the long-term disadvantages. Although little is known about this material in the dental community, in vivo and in vitro studies show calcium phosphate cement as a promising material for grafting applications. In the following article, the authors review the biological and mechanical properties of calcium phosphate cement, as well as its potential use in clinical applications. MATERIALS AND METHODS: A Medline search was performed (timeline: 1980 to 2003) using the following keywords: calcium phosphate cement, hydroxyapatite cement, HA cement, and hydroxyapatite. The search was limited to the English language. The patent literature as well as a limited number of master's theses and books were reviewed after using the electronic database search service from a dental school library. RESULTS: Calcium phosphate cement appears to have excellent biological properties. At only 2 weeks, spicules of living bone with normal bone marrow and osteocytes can be seen. Excellent moldability is a desired clinical characteristic; however, further research is necessary in order to improve the mechanical properties of the cement. The resorption/replacement by bone capability of the cement remains controversial. Further research is needed to clarify this issue. Due to poor mechanical properties, clinical applications are currently limited to craniofacial applications. Further research is necessary to take advantage of the excellent biological properties of this cement under clinical applications. CONCLUSION: Further research is necessary to understand and improve the behavior of this type of cement under clinical situations.

Absorbable Implants↗
PubMed1987

Study of calcium phosphate crystalluria.

When the frequency of heavy calcium phosphate crystalluria was compared among postprandial urine specimens collected from 152 healthy persons (group 1) and 108 patients with calcium urolithiasis (group 2), there was no significant difference between these groups. However, the frequency of crystalluria was significantly greater in 49 patients in group 2 who required or had required surgical intervention for calcium stone disease than in group 1 (p less than 0.05). Calcium phosphate precipitates collected from 7 urine specimens by repeated centrifugation were studied by infrared, X-ray, thermal and electron probe analyses. The principal component of the precipitates was poorly crystallized apatite associated with some organic material. The precipitates from the 7 samples appeared as aggregates of fine spherules less than 1 micron. in diameter, and the calcium-to-phosphate molar ratio of a spherule ranged from 1.02 to 1.19. We conclude that not only calcium oxalate crystalluria but calcium phosphate crystalluria may have an important role in the formation of urinary stones.

Adult↗
PubMed2001

First histological observations on the incorporation of a novel calcium phosphate bone substitute material in human cancellous bone.

Calcium phosphates are frequently used as bone substitute materials because of their similarity to the mineral phase of bone, absence of antigenicity, and excellent osteoconductivity. However, in most currently available mineral substitutes, resorption occurs slowly if at all. In contrast, calcium phosphate cements have shown rapid resorption and remodeling in animal studies. In two prospective studies, a novel amorphous calcium phosphate cement (Biobon) was implanted in human patients for the first time. After 2-12 months, ten biopsies were obtained from nine individuals during secondary surgical interventions, for example, for implant removal. In all specimens, partial replacement of the material by new bone was observed, while residues of the cement were still visible. Undecalcified sections revealed extensive bone formation in immediate contact to the cement without fibrous interface. Polynucleated cells and superficial lacunae were indicative of resorptive activity, but inflammatory tissue response was absent. The new bone displayed regular trabecular and osteonal patterns. The histologic findings are in accordance with the excellent biocompatibility observed in the clinical follow-up. Though still incomplete, the resorbability of this cement appears superior to sintered calcium phosphates in these biopsy specimens. Presumably this is due to its amorphous crystalline structure. Biobon merits further studies as a promising substance for bone defect reconstruction in non-stress-bearing areas.

Aged↗
PubMed1998

The influence of hyperinsulinaemia on calcium-phosphate metabolism in renal failure.

BACKGROUND: Patients with renal failure are characterized by impaired insulin-mediated glucose uptake. Insulin plays a major role in the maintenance of phosphate homeostasis but it remains to be determined whether in uraemia insulin-dependent renal and extrarenal phosphate disposal is also affected. METHODS: The effect of hyperinsulinaemia on serum concentrations of phosphate, ionized calcium and intact PTH as well as renal excretion of calcium and phosphate was studied under euglycaemic conditions (glucose clamp technique) in patients with advanced renal failure and in healthy subjects. Fifteen patients with renal failure (mean serum creatinine 917 micromol/l) and 12 control subjects were included. All subjects underwent a 3-h euglycaemic clamp with constant infusion of insulin (50 mU/m2/min) following a priming bolus. The urine was collected for 3 h before and throughout the clamp. RESULTS: The tissue insulin sensitivity (M/I) was lower in patients with renal failure than in control subjects (5.3+/-2.4 vs 6.7+/-1.8mg/kg/min per mU/ml, P= 0.001) but the phosphate lowering action of insulin was larger in patients with renal failure than in control subjects. Urinary calcium excretion increased (P < 0.05) and phosphate excretion did not change during the clamp in both groups. Despite a decrease of serum ionized calcium in the group of patients with renal failure and no change in the control group, plasma PTH fell significantly in both groups but this effect was still significant after 180 min only in the renal failure group. A significant correlation was observed between changes in serum phosphate and PTH induced by hyperinsulinaemia (r = 0.48, P < 0.01 ) CONCLUSIONS: Phosphate-lowering effect of insulin is well preserved in severe renal failure despite the resistance to insulin-stimulated glucose uptake. The decrease of serum PTH observed during hyperinsulinaemia appears to be independent of serum ionized calcium.

Adult↗
PubMed1991

The periapical tissue reactions to a calcium phosphate cement in the teeth of monkeys.

A calcium phosphate cement, Grossman sealer, and Sargenti N2 were compared under conditions where root canals of monkey incisors were deliberately overfilled and the apical tissue responses were evaluated histologically. The periapical tissues exposed to Sargenti N2 revealed severe irritation at all times through the 6-month experimental period. The reactions to Grossman sealer were milder but persisted throughout the observation period. The calcium-phosphate-cement treated animals showed mild tissue irritation after 1 month, but thereafter the adverse tissue reactions were minimal. New bone formation adjacent to the cement was also observed. These results point to the possibility that calcium phosphate cement might be used in simplified endodontic procedures. The compatibility of calcium phosphate cement with the periapical tissue suggests that the cement may have other applications in dentistry and medicine.

Animals↗
PubMed1997

Restoration of pedicle screw fixation with an in situ setting calcium phosphate cement.

STUDY DESIGN: Pedicle screws were pulled out of human cadaveric vertebrae before and after augmentation with polymethylmethacrylate or in situ-setting calcium phosphate cement. The fixation strength of screws augmented with calcium phosphate cement was compared with that of screws augmented with polymethylmethacrylate. OBJECTIVES: To determine whether a new in situ-setting calcium phosphate cement might be suitable for augmenting the fixation of pedicle screws. The principle objective was to compare the pull-out resistance of screws augmented with calcium phosphate cement with the pull-out behavior of screws augmented with polymethylmethacrylate. Polymethylmethacrylate augmentation was chosen as the standard because of its current clinical use. Five types of screws were tested to determine whether screw design had an effect on the efficacy of augmentation. SUMMARY OF BACKGROUND DATA: Although many factors affect the pull-out resistance of pedicle screws, a key determinant of their performance is the strength of their attachment to the spine. In elderly, osteopenic patients, the screw-bone interface is especially at risk for stripping during insertion or pull-out after surgery. In these patients, polymethylmethacrylate has been used to augment pedicle screw fixation, although its use is not without risk. In situ-setting calcium phosphate cements may provide an alternative to polymethylmethacrylate in this application. Like polymethylmethacrylate, calcium phosphate cements can be injected into the prepared screw hole. They have the added advantage of being resorbed and replaced during healing and normal bone remodeling. METHODS: Thirty human lower lumbar vertebrae (L3-L5) were implanted bilaterally with one of five types of pedicle screws (n = 6 for each screw type). The screws were pulled out 3.0 mm at 0.25 mm/sec with a servohydraulic materials testing machine. The 3.0-mm pull-out distance, which was slightly longer than one thread pitch, was designed to strip the screw-bone interface but to leave the pedicle otherwise intact. After the initial testing, the screws in each vertebrae were removed, and the screw tracks were filled with 2.0 cc of polymethylmethacrylate (one side) or calcium phosphate cement (contralateral side). After augmentation, the screws were reinserted, and the cements were allowed to harden for 24 hours. Postaugmentation testing followed the protocols for preaugmentation testing, and the pull-out resistance of screws augmented with calcium phosphate cement was compared with the pull-out resistance of screws augmented with polymethylmethacrylate. RESULTS: Mechanically, calcium phosphate cement compared favorably with polymethylmethacrylate for augmenting pedicle screws. Both restored the strength of the screw-bone interface: across all screw types, the average increase in pull-out strength was 147% with polymethylmethacrylate augmentation and 102% with calcium phosphate cement. There were no significant differences because of screw type with either type of augmentation. CONCLUSIONS: The in situ-setting calcium phosphate cement investigated in this study compared favorably with polymethylmethacrylate in a single-cycle, pull-out test of augmented pedicle screws in senile trabecular bone. With further evaluation, this cement may offer an alternative to polymethylmethacrylate for the enhancement of pedicle screw fixation clinically.

Bone Cements↗
PubMed2002

Bone repair in radii and tibias of rabbits with phosphorylated chitosan reinforced calcium phosphate cements.

Biocompatibility of two calcium phosphate cements (CPCs), reinforced with phosphorylated chitosan (P-chitosan), was investigated in rabbits in present study. The two CPCs are monocalcium phosphate monohydrate (MPCM) with calcium oxide (CaO) in 1 M phosphate buffer (i.e. MCPM/CaO/1 M phosphate buffer cement, CPC-I) and dicalcium phosphate dihydrate (DCPD) with calcium hydroxide [Ca(OH)2] in 1 M Na2HPO4 solution (i.e. DCPD/Ca(OH)2/1 M Na2HPO4 cement, CPC-II). Different amount of P-chitosan was added to the liquid phase before the power phase was mixed with the liquid phase. The MCPM/CaO/1 M phosphate buffer/P-chitosan cements (P-CPC-I) with neutral pH were filled into the holey defects of rabbit tibias. While the DCPD/Ca(OH)2/1 M Na2HPO4/P-chitosan cements (P-CPC-II) shaped as prehardened cylinders were implanted into rabbit radial defects. After operation, the two serial groups and CPC-II controls were observed for 1, 4, 12 and 22 weeks, respectively. Histological and histomorphological studies proved that P-chitosan containing cements are biocompatible, bioabsorbable and osteoinductive. The biodegradation rate has a negative relationship with the P-chitosan content. Progressive substitution took place at the interface of implants and host bones. No adverse effects were found in tissues around the bone defects. Thus, they could be used as bone substitutes in clinic.

Animals↗
PubMed1998

Histological, chemical, and crystallographic analysis of four calcium phosphate cements in different rabbit osseous sites.

Four calcium phosphate cement formulations were implanted in the rabbit distal femoral metaphysis and middiaphysis. Chemical, crystallographic, and histological analyses were made at 2, 4, and 8 weeks after implantation. When implanted into the metaphysis, part of the brushite cement was converted into carbonated apatite by 2 weeks. Some of the brushite cement was removed by mononuclear macrophages prior to its conversion into apatite. Osteoclastlike cell mediated remodeling was predominant at 8 weeks after brushite had converted to apatite. The same histological results were seen for brushite plus calcite aggregate cement, except with calcite aggregates still present at 8 weeks. However, when implanted in the diaphysis, brushite and brushite plus calcite aggregate did not convert to another calcium phosphate phase by 4 weeks. Carbonated apatite cement implanted in the metaphysis did not transform to another calcium phosphate phase. There was no evidence of adverse foreign body reaction. Osteoclastlike cell mediated remodeling was predominant at 8 weeks. The apatite plus calcite aggregate cement implanted in the metaphysis that was not remodeled remained as poorly crystalline apatite. Calcite aggregates were still present at 8 weeks. There was no evidence of foreign body reaction. Osteoclastlike cell remodeling was predominant at 8 weeks. Response to brushite cements prior to conversion to apatite was macrophage dominated, and response to apatite cements was osteoclast dominated. Mineralogy, chemical composition, and osseous implantation site of these calcium phosphates significantly affected their in vivo host response.

Animals↗
PubMed1999

Incorporation of bovine serum albumin in calcium phosphate coating on titanium.

Calcium phosphate (Ca-P) and bovine serum albumin (BSA) were coprecipitated as a coating on commercially pure titanium (cpTi) with a high protein loading (15 wt %) by employing a recently developed wet-chemistry technique. It was observed that the incorporation of BSA significantly modified the morphology, composition, and crystallinity of the Ca-P coating. The Ca-P coating without BSA is a mixture of hydroxyapatite (HA) and octacalcium phosphate (OCP) with sharp-edged thin OCP crystal plates on the top layer, whereas only an HA phase was detected in the Ca-P/BSA coating. The crystal plates in the latter had a more rounded appearance. The Ca-P/BSA coatings were immersed respectively in neutral (pH 7.4) and acidic (starting pH 4.0) phosphate-buffered saline (PBS) at 37 degrees C over a 14-day period. No protein release was detected in the neutral PBS during the immersion; however, a continuous release of BSA was measured in the acidic PBS, subsequently leading to the formation of a very dense and well-adherent composite coating of BSA and Ca-P on cpTi. The present study provides the possibility to achieve a long-term effective release of biologically active proteins from a Ca-P-coated metallic implant.

Calcium Phosphates↗
PubMed2004

Cellular investigations on electrochemically deposited calcium phosphate composites.

Electrochemically deposited calcium phosphate (CaP) coatings are fast resorbable and existent only during the first period of osseointegration. In the present study, composite coatings with varying solubility (hydroxyapatite (HA), brushite with less HA and monetite (M) with less HA) were prepared and the influence of the degradation and the reprecipitation of CaP on osteoblastic cells were investigated. On the brushite composite coating a new precipitated, finely structured CaP phase was observed during immersion in cell culture medium with or without osteoblastic cells. The surface morphology of monetite and HA coatings were entirely unmodified under the same conditions. So it could be assumed that electrochemically deposited brushite with less HA acts as a precursor for new precipitated CaP. On this surface osteoblastic cells revealed a well-spread morphology with pronounced actin cytoskeleton and demonstrated good proliferation behaviour. Thus we suggest that brushite seems to be especially suitable for coating of implants as a matrix for nucleation and growth of new bone.

Actin Cytoskeleton↗
PubMed1996

Differences of bone bonding ability and degradation behaviour in vivo between amorphous calcium phosphate and highly crystalline hydroxyapatite coating.

Three types of calcium phosphate coating were formed on polyethersulphone (PES) rectangular plates using a biomimetic method: a 20 microns thick amorphous calcium phosphate (ACP 20) coating, a 50 microns thick amorphous calcium phosphate (ACP 50) coating, and a 50 microns thick highly crystalline hydroxyapatite (hHA 50) coating. Uncoated PES plates were used as a control group. These materials were implanted in the tibiae of rabbits and subcutaneously in rats, and the samples were harvested 8 and 16 weeks thereafter, and were examined histologically. The tensile failure loads at the bone-implant interfaces were determined using the detaching test. Each ACP coating was more degradable than the hHA 50 coating. However, newly formed bone came into direct contact with underlying materials as the coating degraded. No coating degraded in subcutaneous tissue. Soft tissue intervening was seen in uncoated samples. Failure load of ACP 20-, ACP 50- and hHA 50-coated samples were all relatively higher than that of the uncoated samples at each period. Significant increase of failure load was seen in hHA 50-coated samples by 16 weeks, however, no increase was seen in either the uncoated or ACP-coated samples. If coating longevity is desired, then the hHA coating is preferable. However, if only the osteoconducive property of calcium phosphate coating is desired for initial fixation of porous materials, the ACP coating may be advantageous.

Animals↗
PubMed1999

Hydrothermal-electrochemical deposition of calcium phosphates on various metals.

Calcium phosphates were formed on five kinds of substrate metals using a hydrothermal-electrochemical method in an autoclave with two electrodes. The electrolyte dissolving NaCl, K2HPO4, CaCl2.2H2O, tris (hydroxymethyl) aminomethane, and hydrochloric acid was maintained at 100 degrees C, 150 degrees C and 200 degrees C. The counter electrode, the anode, was platinum plate, 20 x 20 x 0.5 mm, and the working electrodes as cathode were pure titanium, pure zinc, pure nickel, pure iron, and stainless steel plates. A constant direct current at 12.5 mA/cm2 was loaded for 1 hr. Hydroxyapatite [Ca10(PO4)6(OH)2] only deposited at three temperatures on pure titanium plate. On pure zinc plate, both parascholzite [CaZn2(PO4)2.2H2O] and hydroxyapatite were formed at 150 degrees C and both parascholzite and ZnO were formed at 200 degrees C. Both hydroxyapatite and beta-TCP [beta-Ca3(PO4)2] were formed on pure nickel, pure iron, and stainless steel plates at 200 degrees C. It seems that the incorporation of the metal ions released from the electrode decreased the Ca/P ratio of the deposit due to the formation of other compounds except hydroxyapatite.

Calcium Phosphates↗
PubMed2002

Effect of porosity reduction by compaction on compressive strength and microstructure of calcium phosphate cement.

Hydroxyapatite (HA) calcium phosphate cements (CPCs) are attractive materials for orthopedic applications because they can be molded into shape during implantation. However their low strength and brittle nature limits their potential applications to principally non-load-bearing applications. Little if any use has been made of the HA cement systems as manufacturing routes for preset HA bone grafts, which although not moldable pastes, are resorbable, unlike HA sintered ceramic. It is known that the strength of cements can be increased beyond that attainable from slurry systems by compaction, and this study investigates whether compaction significantly alters the specific surface area and pore-size distribution of CPC prepared according to the method of Brown and Chow. Compaction pressures of between 18 and 106 MPa were used to decrease the porosity from 50 to 31%, which resulted in an increase in the wet compressive strength from 4 to 37 MPa. The Weibull modulus was found to increase as porosity decreased; in addition the amount of porosity larger than the reactant particle size increased as porosity decreased. It is proposed that this was caused by a combination of voids created by the aqueous solvent used in fabrication and shrinkage that occurs on reaction. The specific surface area was unchanged by compaction.

Biocompatible Materials↗