[Vitamine D3 and 25-hydroxycholecalciferol therapy in renal osteopathy: influence on serum calcium, phosphate, calcium phosphate products and on intestinal calcium absorption].
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BACKGROUND: Mineral metabolism has emerged as an important predictor of morbidity and mortality in dialysis patients. Kidney Disease Outcomes Quality Initiative (K/DOQI) clinical practice guidelines for bone metabolism and disease in chronic kidney disease (CKD) recommend that, in Stage 5 CKD, the target levels for calcium (Ca) (corrected for serum albumin), phosphate (P), calcium x phosphate (CaxP) product and parathyroid hormone (PTH) levels should be maintained at 8.4-9.5 mg/dl, 3.5-5.5 mg/dl, <55 mg2/dl2 and 150-300 pg/ml, respectively. OBJECTIVES: To evaluate our ability to achieve K/DOQI guidelines for bone metabolism and disease targets in our patients and to compare them between patients on hemodialysis (HD) and peritoneal dialysis (PD) and also with those reported in the literature. METHODS: We reviewed bone metabolism laboratory parameters in 57 HD patients and 69 PD patients, who had been on dialysis for more than 9 months. RESULTS: The percentage of patients whose serum Ca, P, CaxP product and PTH were within K/DOQI recommended target ranges were 46%, 53%, 77% and 28% in HD patients and 52%, 65%, 77% and 23% in PD patients, respectively. There were no significant differences between HD and PD patients in the percentage of all parameters that were within K/DOQI recommended target ranges. The percentage of our HD patients who had Ca, P, and PTH levels within recommended target range was similar to those in previous reports. CONCLUSION: In our unit, the management of bone and mineral metabolism in HD and PD patients is still far short of meeting K/DOQI guidelines. These findings appear similar in HD and PD patients. Our findings resemble those reported in the literature.
The object of this study was to determine the effect of calcium carbonate or calcium phosphate supplements on dietary protein and fat utilization from low and high manganese diets. During the 63-day study, the 14 human adult subjects ate a constant laboratory controlled diet. In separate periods, subjects consumed the basal diet alone or with supplements of calcium carbonate, calcium carbonate plus manganese gluconate, calcium phosphate, or calcium phosphate plus manganese gluconate. Contrast analyses of data indicated that manganese gluconate supplementation of diets, when combined with either calcium phosphate or calcium carbonate, increased fecal losses of fat. When used as single supplements, both calcium phosphate and calcium carbonate depressed fecal fat loss in comparison with values when no supplements were used. Manganese gluconate supplements depressed fecal nitrogen losses calculated as a percentage of dry fecal weight.
The adjuvant activity of three adjuvants, aluminium phosphate (AIPO4), calcium phosphate (CaHPO4) and stearyl tyrosine for tetanus toxoid (TT) were compared to soluble TT in mice at a dose of 0.5 Lf (1/10th of the single human dose) and in guinea-pigs at a dose of 7.5 Lf (1.5 times the single human dose). Three TT preparations varying in purity were used: (1) ammonium sulphate precipitated formalin detoxified tetanus toxin (AS-TT); (2) AS-TT ultrafiltered to remove low molecular weight peptides (UF-TT): and (3) chromatographically purified tetanus toxin subsequently detoxified with formalin (CP-TT). After primary immunization of mice, AIPO4 absorbed TTs induced higher toxin-neutralizing and IgG (by ELISA) antibodies than CaHPO4, stearyl tyrosine adsorbed or soluble TT preparations, but this difference was no longer present after secondary immunization. TT preparations of varying purities showed similar antibody responses after primary and secondary immunizations when adsorbed on each adjuvant. CP-TT preparation showed the highest neutralizing antibody level amongst soluble preparations after the first dose. All the preparations induced mainly IgG1 antibodies. However, stearyl tyrosine adsorbed TT induced relatively higher IgG2a and IgG2b responses than AIPO4, CaHPO4 adsorbed or soluble TTs particularly after booster injection. No preparation induced detectable IgG3 or IgM antibodies. AIPO4 adsorbed preparations induced higher IgE antibodies than CaHPO4 and stearyl tyrosine adsorbed vaccines. Among the soluble preparations, CP-TT induced lower anti-TT IgE antibodies than standard AS-TT. All these preparations were also tested in the US potency test for adsorbed TT in guinea-pigs. While all the preparations passed this test, AIPO4 adsorbed TT preparations induced higher neutralizing and IgG antibodies than CaHPO4 and stearyl tyrosine adsorbed or soluble TT preparations. In these animal models, purified TT was a strong immunogen and traditional AIPO4 adjuvant gave the highest antibody responses.
This study examines the possibilities for removing heavy metal cations from water with calcium phosphate, calcium hydrogen phosphate, and calcium dihydrogen phosphate at 293 K. It was reported that immobilization of aqueous heavy metal cations, which is known to be one of the characteristic properties of calcium hydroxyapatite, proceeded favorably with these phosphates. Calcium phosphate, calcium hydrogen phosphate, and calcium dihydrogen phosphate could favorably remove Pb2+ from aqueous solution. Calcium hydrogen phosphate also removed aqueous Cu2+, Co2+, and Cd2+, whereas these cations were not immobilized by calcium phosphate and calcium dihydrogen phosphate. A contribution of the dissolution-precipitation mechanism to immobilization with these phosphates is suggested.
Calcium phosphate bioceramics, such as hydroxyapatite, have long been used as bone substitutes because of their proven biocompatibility and bone binding properties in vivo. Recently, a zirconia-hybridized pyrophosphate-stabilized amorphous calcium phosphate (Zr-ACP) has been synthesized, which is more soluble than hydroxyapatite and allows for controlled release of calcium and phosphate ions. These ions have been postulated to increase osteoblast differentiation and mineralization in vitro. The focus of this work is to elucidate the physicochemical properties of Zr-ACP and to measure cell response to Zr-ACP in vitro using a MC3T3-E1 mouse calvarial-derived osteoprogenitor cell line. Cells were cultured in osteogenic medium and mineral was added to culture at different stages in cell maturation. Culture in the presence of Zr-ACP showed significant increases in cell proliferation, alkaline phosphatase activity (ALP), and osteopontin (OPN) synthesis, whereas collagen synthesis was unaffected. In addition, calcium and phosphate ion concentrations and medium pH were found to transiently increase with the addition of Zr-ACP, and are hypothesized to be responsible for the osteogenic effect of Zr-ACP.
Calcium phosphates were electrochemically deposited on titanium plates at temperatures from 4 degrees C to 92 degrees C in a solution of NaCl, K2HPO4 and CaCl2 x 2H20. Scanning electron microscopic studies showed that granular deposits formed on the electrode at electrolyte temperatures of 4 degrees C, 22 degrees C, and 37 degrees C; needle-like deposits formed at 52 degrees C-92 degrees C. The width and length of the needles increased with the temperature of the electrolyte. Based on the results of characterization by electron diffractometry. Fourier transform infrared spectroscopy and X-ray diffractometry, the granular deposits were identified as carbonate-containing calcium phosphate with low crystallinity, and the needle-like deposits as carbonate-containing apatite crystals elongated along the c-axis. Crystallinity of the deposits increased with the temperature of the electrolyte, whereas the orientation indices of the apatite increased with temperature up to 82 degrees C and slightly decreased at 92 degrees C.
OBJECTIVE: To examine the effects of Tamm-Horsfall protein (THP) of normal and low sialic acid content on urinary crystallization, and establish whether there are changes conducive to the formation of kidney stones. MATERIALS AND METHODS: Purified samples of THP were recovered from the urine of non-stone forming individuals. A portion of each THP sample was treated with the enzyme neuraminidase to yield the low sialic acid form of the protein. The two forms of THP were added separately to ultrafiltered urine and crystallization was then induced in the urine by evaporation at 37 degrees C. Two types of experiment were then conducted with the crystals that formed; the rate at which the resulting calcium phosphate or calcium oxalate crystals sedimented in the evaporated urine was determined and the proportion of these crystals and protein which was retained when the urine was passed through a 75 microns sieve was measured. RESULTS: Calcium phosphate and calcium oxalate crystals remained in stable colloidal suspension in ultrafiltered urine when in the presence of normal THP; these suspensions passed freely through the 75 microns sieves. When crystals formed in the presence of low sialic acid THP, the sedimentation was rapid and the crystals were readily retained with protein on the sieves. CONCLUSIONS: These results indicate that whilst normal THP inhibits urinary crystal aggregation, the properties of the low sialic acid form are consistent with the promotion of crystal aggregation and hence stone formation.
Approximate estimates of the ion-acitivity products of calcium phosphate and calcium oxalate in distal tubular urine were derived from the 16-h urinary excretion of calcium, oxalate, citrate, magnesium and phosphate. Urine variables were obtained from 96 normal subjects and 277 calcium stone formers and the calculations were carried out with iterative approximation using the EQUIL2 program. With respect to other ions of importance for the ion-activity products, the urine was assumed to have a fixed composition with pH 6.45. Significantly higher ion-activity products of both calcium phosphate and calcium oxalate were recorded in stone formers. It was concluded that diurnal variations in urine composition and pH might result in peaks of calcium phosphate supersaturation in distal tubular urine whereby a crystallization can occur. In association with abnormalities in terms of promotion and inhibition of calcium salt crystallization, such a precipitation can be of importance for the subsequent formation of calcium renal stones.
In this study, rat bone marrow cells (RBM) were used to evaluate two biodegradable calcium phosphate bone cements and bioactive calcium phosphate ceramics. The substances investigated were: two novel calcium phosphate cements, Biocement F and Biocement H, tricalcium phosphate (TCP), surface-modified alpha-tricalcium phosphate [TCP (s)] and a rapid resorbable calcium phosphate ceramic consisting of CaKPO(4) (sample code R5). RBM cells were cultured on disc-shaped test substrates for 14 days. The culture medium was changed daily and also examined for calcium, phosphate, and potassium concentrations. Specimens were evaluated using light microscopy, and morphometry of the cell-covered substrate surface, scanning electron microscopy, and energy dispersive X-ray analysis and morphometry of the cell-covered substrate surface. Areas of mineralization were identified by tetracyline labeling. Except for R 5, rat bone-marrow cells attached and grew on all substrate surfaces. Of the different calcium phosphate materials tested, TCP and TCP (s) facilitated osteoblast growth and extracellular matrix elaboration to the highest degree, followed by Biocements H and F. The inhibition of cell growth encountered with R 5 seems to be related to its high phosphate and potassium ion release.
The ability of calcium phosphate (CaP) and calcium pyrophosphate (CaPPi) to mediate matrix metalloproteinase-2 and -9 (MMP-2 and MMP-9) binding to fibrin was evaluated. Substrate gel electrophoresis (gelatin zymography) revealed that CaP bound MMP-2 and MMP-9, forming a high molecular weight aggregate with lowered electrophoretic mobility. Formation of the CaP : MMP aggregate was necessary for fibrin binding. In contrast, CaPPi did not aggregate MMPs and did not promote uptake of MMPs into fibrin. Scatchard analysis (Ca/P ratio) revealed that CaPPi (1.96) was chemically similar to calcium pyrophosphate dihydrate (2.00) compared to amorphous CaP (1.50) or crystalline CaP, hydroxyapatite (1.66). MMP : CaP interaction appeared to be electrostatic in nature as high salt concentration (NaCl > 150 mm) reduced binding. In contrast, two non-ionic detergents (Brij-35 and Tween-20) did not prevent MMP : CaP binding. MMP : CaP interaction did not involve the C-terminal MMP region because the specific tissue inhibitor of metalloproteinases (TIMPs) also did not block MMP : CaP interaction and fibrin binding. Although MMP : CaP binding could be decreased with albumin, this effect appeared non-specific due to the high albumin concentration required. High albumin concentration could also partially dissociate preformed MMP : CaP complexes. Interestingly, type I and type IV collagen substantially increased MMP : fibrin-binding activity, whereas denatured collagen, gelatin, did not. Inflammatory joint fluid from five patients also demonstrated similar MMP fibrin-binding activity consistent with CaP mediation. The relevance of these findings to CaP and CaPPi in the pathogenesis of crystal arthropathies such as basic calcium phosphate (BCP) and calcium pyrophosphate dihydrate crystal disease (CPPD) is discussed.
Defects in an intracellular chloride channel CLC-5 cause Dent's disease, an inherited kidney stone disorder. Using a collecting duct model, mIMCD-3 cells, we show expression of dimeric mCLC-5. Transient transfection of antisense CLC-5 reduces CLC-5 protein expression. Binding of both calcium phosphate (hydroxyapatite) and calcium oxalate monohydrate (COM) crystals overlaid onto mIMCD-3 cultures was affected by altered CLC-5 expression. Calcium phosphate crystal agglomerations (>10 microm) were minimal in control (9%) and sense (13%) CLC-5-transfected cells, compared to 66% of antisense CLC-5-transfected cells (P<0.001). Small calcium phosphate crystals (<10 microm) were found associated with 45% of sense CLC-5-treated cells, of which the majority (11/14 cells) appeared to be internalised within the cell. Calcium oxalate agglomerations (>10 microm) were also largely absent for controls or sense mCLC-5 transfectants (11% and 9% of cells, respectively) with COM crystal agglomerates predominating in antisense CLC-5 transfectants (66%, P<0.0001). We conclude that collecting duct cells with reduced CLC-5 expression lead to a tendency to form calcium crystal agglomeration, which may help explain the nephrocalcinosis and nephrolithiasis seen in Dent's disease.
Using an assay which allows continuous monitoring of the mixing of aqueous contents during membrane fusion, we have investigated the kinetics of calcium-phosphate-induced fusion of erythrocyte ghosts. In the presence of 10 mM phosphate, the threshold concentration for Ca2+-induced fusion was 1.25 mM, while the optimal concentration was approx. 1.75 mM Ca2+. Further enhancement of the cation concentration (greater than or equal to 2 mM) inhibited fusion of the ghosts. Initiation of fusion required the addition of phosphate prior to the addition of Ca2+, indicating that the combined interaction of Ca2+ and phosphate in or at the plane of the bilayer was a prerequisite for the induction of fusion. Furthermore, fusion was greatly facilitated upon transformation of calcium phosphate in the bulk medium from an amorphous to a solid, crystalline phase. It is suggested that membrane aggregation, and hence fusion, is facilitated by the formation of crystalline calcium phosphate nucleating on the ghost membrane. La3+, Mg2+ and Mn2+ did not trigger the fusion process, although aggregation of the ghosts did occur. Under conditions where calcium phosphate precipitation was inhibited, lanthanum phosphate precipitates facilitated fusion after prior treatment of ghosts with phosphate and Ca2+. These results indicated that fusion-prone conditions were induced prior to calcium phosphate precipitation. It is proposed that prior to calcium phosphate precipitation membrane changes are induced by separate interaction of Ca2+ and phosphate with the ghost membrane. Such an interaction could then render the ghosts susceptible to fusion and as soon as conditions are provided allowing close contact between adjacent membranes, fusion will be observed.
The occlusion of dentinal tubules with calcium phosphate, by a calcium phosphate precipitation method (CPP method), was investigated in vitro for evaluation of the potential value of this method for the treatment of dentin hypersensitivity. The method consists of treating the dentinal surface with a CPP solution, i.e., an acidic solution that contains both calcium and phosphate, followed by neutralization with basic post-treatment solution. The CPP solutions used in this study ([Ca] = 0.2 - 1.0 mol/L, [PO4] = 0.2 - 4.0 mol/L) were prepared by dissolving Ca(OH)2 or CaHPO4 x 2H2O in H3PO4 or HCl, and 1 mol/L NaOH solution was used for the post-treatment solution. Sections of human dentin disks treated by the CPP method were observed by scanning electron microscopy, and the precipitate in the dentinal tubules was subjected to x-ray micro-analysis. After treatment by the CPP method, dentinal tubules were occluded to a distance of approximately 15 microns from the surface, and the precipitate showed a Ca/P molar ratio of 1.03 +/- 0.01 To obtain a larger amount of precipitated mineral for further analysis, we used the same procedure with glass tubes (inside diameter, 1 mm). Powder x-ray diffraction analysis and FT-IR measurement revealed that the precipitate was dicalcium phosphate dihydrate (CaHPO4 x 2H2O). The instant precipitation of calcium phosphate mineral in the dentinal tubules demonstrated the potential value of the CPP method for the occlusion of dentinal tubules, this occlusion may be useful for the treatment of dentin hypersensitivity.
The use of biodegradable bone substitutes is advantageous for alveolar ridge augmentation because it avoids second-site surgery for autograft harvesting. This study examines the effect of novel, rapidly resorbable calcium phosphates and a calcium phosphate bone cement on the expression of bone-related genes and proteins by human bone-derived cells (HBDCs) and compares this behavior to that of tricalciumphosphate (TCP). Test materials were alpha-TCP, two materials with a crystalline phase Ca(2)KNa(PO(4))(2) and with a small amorphous portion containing either magnesium potassium phosphate (material denominated GB14) or silica phosphate (material denominated GB9), and a calcium phosphate bone cement (material denominated Biocement D). HBDCs were grown on the substrata for 3, 7, 14, and 21 days, counted, and probed for various mRNAs and proteins (type I collagen, osteocalcin, osteopontin, osteonectin, alkaline phosphatase, and bone sialoprotein). All substrates supported continuous cellular growth for 21 days. In the presence of GB14 and Biocement D specimens cell proliferation was reduced and cell differentiation increased. At day 21, the greatest number of cells was found on GB9 expressing significantly higher levels of bone-related proteins than cells grown on all other surfaces. Because all novel materials facilitated the expression of the osteoblastic phenotype at least as much as TCP and the polystyrene control, these biomaterials can be regarded as excellent candidate bone substitute materials. GB9 induced the highest proliferation and cellular differentiation after 21 days of incubation, suggesting that this material may possess a higher potency for enhancing osteogenesis than TCP.
Uric acid (UA), a waste product of purine metabolism, may be involved in calcium phosphate crystallization and deposition. Rats, which develop nephrocalcinosis on high-fat or magnesium-deficient diets, and patients with idiopathic calcium urolithiasis have hyperproteinuria, especially of nonalbumin protein, and a shift toward elevated serum UA. In rats, an increase in UA precursors and renal UA suggests hypoxemia, which stimulates xanthine oxidase. In patients, a primary increase in renal xanthine oxidase would explain the low urine UA in the presence of an elevated serum concentration. For calcium phosphate deposition (rats) or incorporation into stones (humans) to occur, a crucial factor may be xanthine oxidase-mediated overproduction of free radical species and subsequent tissue damage. Another factor may be whether sufficient UA is synthesized to neutralize these free radicals. Allopurinol use, which inhibits xanthine oxidase and has long been favored for the treatment of idiopathic calcium urolithiasis, may not prevent stones, because it also diminishes the availability of UA. An investigation of the factors that control serum UA homeostasis may be rewarding in research into the etiology of idiopathic calcium urolithiasis.
Calcium was determined in calcium phosphate samples by dissolving the sample in hydrochloric acid, adding hydroxynaphthol blue indicator and triethanolamine, adjusting the pH to 12.3--12.5 with potassium hydroxide solution, and titrating with standard disodium ethylenediaminetetraacetate solution. Time can be saved and the formation of a precipitate (which dissolves readily during the titration) can be avoided by adding at least 85% of the amount of complexing agent required for titration before adjusting the pH.
Radiographs and synovial fluids from 66 knees representing 59 patients with symptomatic osteoarthritis were evaluated to determine the pattern of radiographic abnormalities associated with basic calcium phosphate (BCP), calcium pyrophosphate dihydrate (CPPD), or both crystals together. Crystals were found in 71% of fluids. In general, CPPD crystals correlated with patient age, while BCP crystals correlated with joint degeneration. Synovial fluid BCP and CPPD crystals were found together more often than either alone. Joint compartment narrowing and osteophytes in three compartments are often associated with BCP crystals.