In vivo setting behaviour of fast-setting calcium phosphate cement

The in vivo setting behaviour of fast-setting calcium phosphate cement (FSCPC) between femoral muscles of the rat was investigated to evaluate the possible value of FSCPC for medical and dental application. Conventional CPC (c-CPC) and FSCPC were implanted between femoral muscles, and various aspects of the setting behaviour such as setting time, mechanical strength and conversion ratio of cement into hydroxyapatite (HAP: Ca10(PO4)6(OH)2) were measured by the Vicat needle method, diametral tensile strength (DTS) measurement, and quantitative powder X-ray diffraction (XRD) analysis, respectively. The setting time of FSCPC in vivo was 5-7 min, in contrast to 48 min for c-CPC. As a result of its fast setting, set specimens of FSCPC showed higher mechanical strength from the initial stage than c-CPC. Higher DTS values were observed in FSCPC than c-CPC implanted after 24 h. Powder XRD analysis revealed faster conversion of FSCPC than c-CPC into HAP, which was responsible both for the faster setting and higher mechanical strength from the initial stage. We concluded, therefore, that FSCPC may be used for a wide range of clinical applications, i.e. fields where fast setting is required such as orthopaedic, plastic and reconstructive, and oral and maxillofacial surgery.     

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

Elimination of the data processing bottleneck in high-throughput sequencing will require both improved accuracy of data processing software and reliable measures of that accuracy. We have developed and implemented in our base-calling program phred the ability to estimate a probability of error for each base-call, as a function of certain parameters computed from the trace data. These error probabilities are shown here to be valid (correspond to actual error rates) and to have high power to discriminate correct base-calls from incorrect ones, for read data collected under several different chemistries and electrophoretic conditions. They play a critical role in our assembly program phrap and our finishing program consed.     

Setting reaction and hardening of an apatitic calcium phosphate cement

The combination of self-setting and biocompatibility makes calcium phosphate cements potentially useful materials for a variety of dental applications. The objective of this study was to investigate the setting and hardening mechanisms of a cement-type reaction leading to the formation of calcium-deficient hydroxyapatite at low temperature. Reactants used were alpha-tricalcium phosphate containing 17 wt% beta-tricalcium phosphate, and 2 wt% of precipitated hydroxyapatite as solid phase and an aqueous solution 2.5 wt% of disodium hydrogen phosphate as liquid phase. The transformation of the mixture was stopped at selected times by a freeze-drying techniques, so that the cement properties at various stages could be studied by means of x-ray diffraction, infrared spectroscopy, and scanning electron microscopy. Also, the compressive strength of the cement was measured as a function of time. The results showed that: (1) the cement setting was the result of the alpha-tricalcium phosphate hydrolysis, giving as a product calcium-deficient hydroxyapatite, while beta-tricalcium phosphate did not participate in the reaction; (2) the extent of conversion of alpha-TCP was nearly 80% after 24 hr; (3) both the extent of conversion and the compressive strength increased initially linearly with time, subsequently reaching a saturation level, with a strong correlation observed between them, indicating that the microstructural changes taking place as the setting reaction proceeded were responsible for the mechanical behavior of the cement; and (4) the microstructure of the set cement consisted of clusters of big plates with radial or parallel orientations in a matrix of small plate-like crystals.     

Comparison of calcium phosphate cement mixture and pure calcium hydroxide as direct pulp-capping agents

This review reports the different genetic factors that have been identified either as risk factor for Alzheimer's disease (AD) or directly causing the disease. First are reviewed epidemiological data and biological mechanisms about the apoplipoprotein E gene allele epsilon 4 that is a major risk factor for Alzheimer's disease. The second part describes the mutations responsible for early-onset autosomal dominant AD found in three different genes. The gene located on chromosome 21 encodes the amyloid precusor protein (APP). The presenilin 1 and presenilin 2 genes, located on chromosome 14 and 1 respectively, encode not yet known membrane proteins.     

Effect of calcium carbonate on clinical compliance of apatitic calcium phosphate bone cement

Incidentally discovered adrenal masses are common since the advent and application of sensitive noninvasive imaging methods. The significance of these so-called "incidentalomas" and the question of further evaluation or treatment remains elusive. This report describes a retrospective study of 86 patients with incidentaloma. Adrenalectomy was performed on 26 patients during initial admission. Histologically, two cortisol-producing adenomas, an adenoma with subclinical cortisol production, and two pheochromocytomas (all of the preceding detected during the preoperative hormonal evaluation), three cystic lesions, one myelolipoma, and one hematoma were found. One primary and two metastatic adrenal carcinomas were also found in this series. Sixty patients with a nonfunctioning incidentaloma smaller than 6 cm were observed in an average of 43 months with serial CT scans performed at 3, 9, and 18 months after the initial diagnosis. Enlargement of the mass was detected in two patients; both proved to be nonfunctioning adenomas. Based on these observations, it is concluded that the initial laboratory evaluation is mandatory in cases of incidentalomas, including parameters of adrenocortical and medullar function. Hormonally active incidentalomas and those suspected for malignancy should be treated surgically. Masses greater than 6 cm should also be removed. Smaller incidentalomas without endocrine activity or signs of malignancy should be followed by CT scan at 3, 9, and 18 months after the initial diagnosis.     

Incorporation of adenovirus in calcium phosphate precipitates enhances gene transfer to airway epithelia in vitro and in vivo

The anti-SRYD monoclonal antibody (mAbSRYD) raised against the IASRYDQL synthetic octapeptide, the 250-257 sequence of the Leishmania major surface glycoprotein gp63 recognizes both SRYD-containing peptides and the whole cognate major surface protein on intact parasites. Two SRYD-containing peptides, which antigenically and functionally mimic the RGDS sequence of fibronectin and efficiently inhibit parasite attachment to the macrophage receptors, were studied by two-dimensional transferred nuclear Overhauser effect experiments in the presence of mAbSRYD. The antibody-bound IASRYDQL octapeptide solution conformation was determined on the basis of 55 interproton-distance restraints, derived from NMR measurements. Eighteen structures which were first generated using an approach combining distance geometry and molecular dynamics, converge by energy minimization toward a folded structure with an average rmsd from the experimental data of less than 0.05 nm for the overall backbone and 0.025 nm for the SRYD motif. A distorted gamma-turn was found, stabilized by the backbone-backbone D255-NH to R253-CO hydrogen bond, while the R253 and D255 side chains are pointing in opposite directions. This latter antibody-bound structure is compared with that of the free octapeptide in dimethylsulfoxide solution, and with the crystal structure of the RYD fragment in OPG2 Fab, an antireceptor antibody that mimics the RGD cell adhesion site. On this basis, a mechanism for IASRYDQL-receptor interaction is discussed.     

Effect of calcium carbonate on the compliance of an apatitic calcium phosphate bone cement

Clinical requirements for calcium phosphate bone cements were formulated in terms of the initial setting time, the final setting time, the cohesion time and the ultimate compressive strength. Three cement formulations were tested. The previously developed Biocement H was made of a powder containing alpha-tertiary calcium phosphate and precipitated hydroxyapatite. Biocement B2 powder was made by adding some CaCO3 to Biocement H, whereas Biocement B1 was made by adding some CaCO3 but with simultaneous adjustment of the amount of precipitated hydroxyapatite.The liquid/ powder ratio of the cement paste and the accelerator concentrations (percentage Na2HPO4) in cement liquid were varied. For Biocement H there was no combination of L/P ratio and percentage Na2HPO4 for which all clinical requirements were satisfied. However, there was an area of full compliance for Biocements B1 and B2, of which that for B1 was the largest. Therefore, Biocement B1 may be applied in clinical situations as those in orthopaedics, plastic and reconstructive surgery and oral and maxillofacial surgery, even when early contact with blood is inevitable.     

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.     

The effect of pulpal fluid flow on tensile bond strength of a glass-ionomer cement: an in vivo and in vitro comparison

Previous studies of the bonding capabilities of glass-ionomer cements have concentrated on the use of in vitro testing conditions. Since early moisture contamination appears to have adverse effects on the physical properties of glass-ionomer cements, and with the probability of pulpally derived dentinal fluid being present under in vivo conditions, the objective of this study was to compare in vivo tensile bond strength with in vitro tensile bond strength of a glass-ionomer cement to dentin utilizing the same teeth under similar test conditions. A glass-ionomer lining cement was placed on freshly exposed labial dentin of the maxillary incisor on 10 Rhesus monkeys. Immediately following placement, an orthodontic button was placed over the cement and left undisturbed for 1 hour. The teeth were then extracted and stored in 100% relative humidity for 23 hours. An Instron testing machine was used to register in kilograms the force required to cause tensile bond failure of the cement. Identical methodology was then used on the same teeth for in vitro testing. The concluding results indicate that a statistically significant difference (P < or = 0.05) exists between in vivo and in vitro tensile bond strengths of the glass-ionomer lining cement and that the bond failure was cohesive in character for all cases both in vivo and in vitro. These findings suggest that clinically, tensile bond strengths of glass-ionomer cements to cut dentin can be expected to be weaker in vital teeth than in devital teeth.     

Initial histological evaluation of anti-washout type fast-setting calcium phosphate cement following subcutaneous implantation

Anti-washout-type fast-setting calcium phosphate cement using chitosan (aw-FSCPC(chi)), conventional CPC (c-CPC), CPC mixed with citric acid (CPC(citric)) and CPC mixed with polyacrylic acid (CPC(acrylic)) were implanted subcutaneously in rats immediately after mixing to shed some light on the understanding of the appearance of excellent tissue response to CPC. CPC(citric) and CPC(acrylic) set quickly, similar to aw-FSCPC(chi), but the former two stopped their transformation to apatitic minerals. The c-CPC, which required a long setting time, was found to be crumbled, but the other CPCs maintained the shape at implantation. The aw-FSCPC(chi) and CPC(citric) showed no inflammatory response whereas c-CPC and CPC(acrylic) showed an inflammatory response one week after implantation. A component of the aw-FSCPC(chi) and c-CPC was an apatitic mineral whereas CPC(citric) and CPC(acrylic) showed no transformation to apatite. We concluded that the non-crumbling property plays a more dominant role in the appearance of excellent tissue response to CPC than the transformation to apatite. Also, a non-crumbling property is not a sufficient condition, but a necessary condition for the appearance of the excellent tissue response to CPC.     

In vitro evaluation of a new injectable calcium phosphate material

BACKGROUND: Verbal autopsies have been widely used to determine the levels and causes of maternal death but few studies have assessed the reliability of various methods. METHODS: We compared the levels and causes of maternal mortality in three data sources from Matlab, Bangladesh: (1) maternal deaths identified through a unique demographic surveillance system (DSS); (2) maternal deaths identified as a result of a previous detailed investigation into the levels and causes of maternal mortality; and (3) maternal deaths identified in the current special study. All studies used lay reporting, but differed in terms of the nature of the study, the sex of the interviewer, the format of the questionnaire and the procedure to derive the diagnosis. RESULTS: There were substantial disagreements between the routine reporting and the special studies. The DSS identified 67.2% of all deaths occurring during pregnancy or within 42 days postpartum (82.3% of direct obstetric deaths, 70.0% of deaths due to induced abortions and 42.4% of indirect obstetric deaths). Extending the definition of maternal deaths to 90 days postpartum increased the numbers of maternal deaths between 1987 and 1993 from 174 to 196. The two special studies also disagreed in the ascertainment of the causes of maternal deaths and yielded different cause of death distributions; the proportion of direct obstetric deaths (excluding abortion) was 50.4% in the current system compared to 44.5% previously (P = 0.001). CONCLUSIONS: This study confirms the known difficulties in the ascertainment of the levels and causes of maternal mortality. The large disparities in the levels and causes of maternal mortality using three different methods of lay reporting in a population with an almost complete vital registration system add to the growing concern about the inaccuracies in the measurement of maternal mortality.     

The in vitro and in vivo indomethacin release from self-setting bioactive glass bone cement

The in vivo and in vitro drug release profiles from a self-setting bioactive CaO-SiO2-P2O5 glass bone cement containing indomethacin as a model drug were investigated. The cement containing 2% and 5% indomethacin (IMC) powder hardened within 5 min after mixing with ammonium phosphate buffer. After setting, in vitro drug release from drug-loaded cement pellets in a simulated body fluid (SBF) at pH 7.25 and 37 degrees C continued for two weeks. The hardened cement gradually formed low-crystallinity hydroxyapatite during the drug release test in SBF. An IMC-loaded cement device (2% and 5% drug) was implanted in the subcutaneous tissue on the back of rats. The in vivo IMC release from the cement increased and attained maximum levels (Cmax of 2% and 5% drug-loaded cements was 0.27 and 3.37 micrograms/ml, respectively) at Tmax, 3 and 0.5 d, respectively, upon subcutaneous (s.c.) administration in rats. This suggested that the s.c. administration of the cement provided IMC release for a much longer period than s.c. administration of the solution, and the plasma IMC concentration was dependent on the drug concentration in the cement. The plasma IMC concentration and the area under the curve from 2% and 5% IMC-loaded cements in rats were dependent on the concentration of IMC in the cements. The in vivo IMC concentration in plasma obtained by the deconvolution method was much lower than that delivered in SBF in vitro. Scanning electron microscopy and photomicrographs of cross sections showed that the bioactive bone cement had excellent biocompatibility with the surrounding soft tissues.     

Mechanical and histological evaluation of amorphous calcium phosphate and poorly crystallized hydroxyapatite coatings on titanium implants

The effect of amorphous calcium phosphate (Ca/P) and poorly crystallized (60% crystalline) hydroxyapatite (HA) coatings on bone fixation to "smooth" and "rough" (Ti-6A1-4V powder sprayed) titanium-6Al-4V (Ti) implants was investigated. Implants were evaluated histologically, mechanically, and by scanning electron microscopy (SEM) after 4 and 12 weeks of implantation in a rabbit transcortical femoral model. Histological evaluation of amorphous vs. poorly crystallized HA coatings showed significant differences in bone apposition (for rough-coated implants only) and coating resorption (for smooth- and rough-coated implants) that were increased within cortical compared to cancellous bone. The poorly crystallized HA coatings showed most degradation and least bone apposition. Mechanical evaluation, however, showed no significant differences in push-out shear strengths between the two types of coatings evaluated. Differences between 4 and 12 weeks were significant for coating resorption and push-out shear strength but not for bone apposition. Significant enhancement in interfacial shear strengths for bioceramic coated as compared to uncoated implants were seen for smooth-surfaced implants (3.5-5 times greater) but not for rough-surfaced implants at 4 and 12 weeks. Rough implants showed greater mean interfacial strengths than uncoated smooth implants at 4 and 12 weeks (seven times greater) and to coated smooth implants at 12 weeks only (two times greater). Mechanical failure of the bone/coating/implant interface consistently occurred within the bone, even in the case of the poorly crystallized HA coatings, which had almost completely resorbed on rough implants. These results suggest that once early osteointegration is achieved biodegradation of a bioactive coating should not be detrimental to the bone/coating/implant fixation.     

Calcitriol modulates in vivo and in vitro cytokine production: a role for intracellular calcium

Calcitriol modulates in vivoand in vitro cytokine production: A role for intracellular calcium. Background. Several immunomodulatory properties of calcitriol are currently known, however, only little information is available regarding the in vivo and in vitro effects of calcitriol on cytokine production in chronic renal failure. Methods. To study the in vitro effect of calcitriol on lipopolysaccharide (LPS)-induced cytokine production, peripheral blood mononuclear cells (PBMC, 2.5 ml/ml) from 12 chronic dialytic (HD), 15 undialyzed chronic renal failure (CRF) patients and 10 normal subjects (N) were incubated at 37 degrees for 12 hours with 100 ng of LPS (E. coli and P. maltofilia). Increasing doses of calcitriol from 10-10 to 10-9 M were added and cell associated TNF-alpha and IL-1beta were determined by immunoreactive tests after three freeze-thaw cycles. The intradialytic TNF-alpha and IL-1beta production were evaluated in vivo in 12 HD patients before and after three months of intravenous calcitriol treatment (6 microgram/week). Intracellular calcium [Ca++]i was determined on PBMC with a cytofluorimetric assay using FLUO-3 AM as the indicator. Results. In vitro, TNF-alpha increased from 3.6 +/- 1.9 pg/cell to 1797 +/- 337 in N, from 4.5 +/- 1.7 to 1724 +/- 232 in CRF and from 3.4 +/- 2.3 to 1244 +/- 553 in HD after the LPS stimulus. The production of TNF-alpha was inhibited by calcitriol in a dose-dependent manner [LPS + Vit.D3 100 ng, 2.9 +/- 2.1 in N, 3.7 +/- 1.9 in CRF and 3.4 +/- 1.7 in HD; LPS + Vit.D3 50 ng, 263 +/- 296 (N), 6.73 +/- 11 (CRF), 38 +/- 28 (HD); LPS + Vit.D3 25 ng = 873 +/- 583 (N), 325 +/- 483 (CRF), 588 +/- 507 (HD); LPS + Vit.D3 12.5 ng, 954 +/- 483 (N), 912 +/- 510 (CRF), 875 +/- 527 (HD)]. Comparable data were observed on IL-1beta production. In vivo, the intradialytic TNF-alpha increase (from 8.5 +/- 2.3 to 19 +/- 5.6 pg/2.5 x 106 cell) during hemodialysis was markedly reduced after calcitriol therapy (from 6.6 +/- 3.1 to 11 +/- 4.7). [Ca++]i decreased from 105 +/- 25 to 72 +/- 18 nM (P < 0.05) and a positive correlation between cytokine levels and [Ca++]i was found (r = 0.79; P < 0.001). Conclusions. The in vitro increase of cell-associated cytokine after LPS challenge was inhibited by calcitriol in a dose-dependent manner. These data suggest a possible in vivo modulatory effect of calcitriol therapy on cytokine production in hemodialysis.     

磷酸钙骨水泥生物材料用磷酸四钙的制备

采用CaCO3和CaHPO4的混合粉分别在空气和真空2种气氛条件下制备磷酸四钙。结果表明:在空气中较难制得磷酸四钙,这主要是由于潮湿的空气中含有较多水分和制备工艺中采取随炉缓慢冷却,对反应产生了不利影响;在真空条件下,采用n(Ca)∶n(P)=2∶1,1.8∶1和1.5∶1的3种混合粉均容易制得磷酸四钙,但同时都含有其它杂质相,对其纯度有影响,其中以n(Ca)∶n(P)=2∶1的混合粉制备的磷酸四钙纯度最高,其产物中仅含少量CaO杂质相,这种磷酸四钙可用于磷酸钙骨水泥。     

The relationship between osmotic stress and calcium elevation: in vitro and in vivo rat lens models

Recent evaluations by the U.S. General Accounting Office and the National Alliance for the Mentally Ill of reemployment efforts of the federal-state vocational rehabilitation program found that services offered by state vocational rehabilitation agencies do not produce long-term earnings for clients with emotional or physical disabilities. This paper examines reasons for these poor outcomes and the implications of recent policy reform recommendations. Congress must decide whether to take action at the federal level to upgrade programs affecting persons with severe mental illnesses or to continue to rely on state decision making. The federal-state program largely wastes an estimated $490 million annually on time-limited services to consumers with mental illnesses. Rechanneled into a variety of innovative and more appropriate integrated services models, the money could buy stable annual vocational rehabilitation funding for 62,000 to 90,000 consumers with severe mental illnesses. Larger macrosystem problems involve the dynamics of the labor market that limit job opportunities and the powerful work disincentives for consumers with severe disabilities now inherent in Social Security Disability Insurance, Supplemental Security Income, Medicare, and Medicaid.     

In vivo, in vitro, and computational analysis of dendritic calcium currents in thalamic reticular neurons

Thalamic reticular (RE) neurons are involved in the genesis of synchronized thalamocortical oscillations, which depend in part on their complex bursting properties. We have investigated the intrinsic properties of RE cells using computational models based on morphological and electrophysiological data. Simulations of a reconstructed RE cells were compared directly with recordings from the same cell to obtain precise values for the passive parameters. In a first series of experiments, the low-threshold calcium current (I(Ts)) was studied via voltage clamp in acutely dissociated RE cells that lack most of their dendrites. Simulations based on a cell with truncated dendrites and Hodgkin-Huxley kinetics reproduced these recordings with a relatively low density of I(Ts). In a second series of experiments, voltage-clamp recordings obtained in intact RE cells in slices showed a higher amplitude and slower kinetics of I(Ts). These properties could be reproduced from the reconstructed cell model assuming higher densities of I(Ts) in distal dendrites. In a third series of experiments, current-clamp recordings were obtained on RE cells in vivo. The marked differences with in vitro recordings could be reconciled by simulating synaptic bombardment in the dendrites of RE cells, but only if they contained high distal densities of I(Ts). In addition, simpler models with as few as three compartments could reproduce the same behavior assuming dendritic I(Ts). These models and experiments show how intrinsic bursting properties of RE cells, as recorded in vivo and in vitro, may be explained by dendritic calcium currents.     

Evolution of the local calcium content around irradiated beta-tricalcium phosphate ceramic implants: in vivo study in the rabbit

To evaluate whether dissolved calcium from tricalcium phosphate implants contributes to osseous wound healing in bone defects, the authors used nuclear radioactivated materials. Six months after irradiation, the calcium was still radioactive. Samples of the material were prepared and placed in rabbit condyles for 1, 3 and 9 months. Over time the condyles were retrieved and treated for histology or radiocounting. Measurements of the radioactivity of the slices and histomorphometry of the implants and surrounding tissues were performed. The authors observed that the radioactivity decreased regularly. Connective tissue had penetrated the pores and totally invaded the implants, first at the periphery of the implants, then inside the pores. Comparison of the results of radioactivity and histomorphometry suggest that part of the calcium from the implants was re-used specifically in the new osseous tissue.