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PubMedDate not supplied

Systemic candidiasis in mice. II.--Main role of polymorphonuclear leukocytes in resistance to infection.

Cyclophosphamide (CY) increased whereas the talc embedded in a calcium phosphate gel (TCP) decreased the susceptibility of mice to systemic candidiasis estimated by measuring mean survival time and "renal infectivity" 12 h after challenge. Transfers of plasma from CY- and TCP-treated mice did not modify cnadidiasis susceptibility of recipient mice. Granulopenia and granulocytosis induced respectively by CY and TCP were significantly correlated with susceptibility or resistance to candidiasis. Nevertheless, TCP produced significant reticuloendothelial stimulation which could be also correlated with TCP protection. Reticuloendothelial stimulation with associated granulopenia in TCP-CY-treated mice gave protection against Listeria monocytogenes challenge but not against Candida albicans. Thus, blood polymorphonuclear leukocytes seem to play the main role in natural resistance of mice to candidiasis. This was corroborated after injection of immunostimulants; a good correlation was found between C. albicans resistance and the induced granulocytosis.

Animals↗
PubMed1995

Sigma ligands indirectly modulate the NMDA receptor-ion channel complex on intact neuronal cells via sigma 1 site.

To investigate the modulatory effects of sigma ligands on the N-methyl-D-aspartate (NMDA) receptor-ion channel complex in vivo, we examined the intact cell binding of 3H-N-[1-(2-thienyl)cyclohexyl]piperidine (3H-TCP) to cultured neuronal cells prepared from fetal rat telencephalon. The 3H-TCP binding was saturable, reversible, and inhibited by a selective NMDA receptor antagonist, D-amino-5-phosphonovaleric acid. MII-limolar Mg2+ inhibited 3H-TCP binding both in the absence and presence of L-glutamate. 5-Methyl-10,11-dihydro-5H-dibenzo [a,d]cyclohepten-5,10-imine maleate (MK801) inhibited 3H-TCP intact cell binding in a competitive manner, while haloperidol inhibited it in a noncompetitive manner. The effect of the test drugs to inhibit 3H-TCP intact cell binding was in the order of dextromethorphan, haloperidol > (+/-)MK 801 > (+)pentazocine > (-)pentazocine > DTG > PCP > (+)-N-allylnormetazocine [(+)SKF 10047] > (+)3-(3-hydroxyphenyl)-N- (1-propyl)piperidine [(+)3-PPP] > (-)SKF 10047 > (-)3-PPP. The IC50 values of the six sigma ligands for 3H-TCP binding were closely correlated with the Ki values of the corresponding drugs for DTG site 1 in the guinea pig brain reported by Rothman et al. (1991). These findings suggest that the sigma ligand indirectly modulates the NMDA receptor ion channel complex, presumably through sigma 1 sites in vivo as well as in vitro.

2-Amino-5-phosphonovalerate↗
PubMed1994

Development of a ceramic matrix system for continuous delivery of azidothymidine.

A beta-tricalcium phosphate (TCP) ceramic system was designed to deliver azidothymidine (AZT) in an aqueous medium (phosphate buffered saline pH 7.4) at 37 degrees C. Since AZT is an analog of thymidine, initial studies were conducted with thymidine. Ceramic capsules were made by pressing one gram of < 38 microns beta-TCP particles with or without the desired amount of thymidine or azidothymidine in a 10 mm die at a load of 4000 lbs in an electric hydraulic press. The amount thymidine released from TCP ceramic capsules was directly dependent on the amount of the drug loaded in the ceramic. Treatment of TCP ceramic particles with either sesame seed oil or wheat germ oil (vitamin E) delayed the release of thymidine and azidothymidine from TCP ceramic capsules. Incorporation of thymidine or azidothymidine in the form of a compressed pellet (5/16" diameter at a load of 4000 lbs) further inhibited the release of thymidine and azidothymidine from TCP ceramic capsules fabricated with oil-treated ceramic particles. Thus, ceramic drug delivery systems can be designed to deliver continuously desired therapeutic levels of AZT in AIDS patients.

Calcium Phosphates↗
PubMed1993

[Reductive dechlorination of chlorophenols by an anaerobic mixed culture].

In this study, we investigated the potential for reductive dechlorination of chlorophenols by municipal sewage sludge acclimated to 2,4-dichlorophenol (2,4-DCP) and 3,4-dichlorophenol (3,4-DCP). The optimal temperature and pH for dechlorination of 2,4,6-trichlorophenol (2,4,6-TCP) were 30 degrees C and 7.2. Dechlorination of 2,4,6-TCP was inhibited by sulfate, nitrate, or ferric (III), but was increased by manganese (IV) except that product of 2,4-DCP was accumulated. Organic substrates such as pyruvate or acetate did not influence dechlorination of 2,4,6-TCP, but increased dechlorination of 2,4-DCP; nevertheless lactate showed no influence on dechlorination. When glucose was added as substrate, only 20% of 2,4,6-TCP dechlorination was found. Addition of methanogenesis inhibitor 2-bromoethane sulfonate did not influence dechlorination, but addition of eubacteria inhibitor vancomycin showed no dechlorination activity at the first few days, and finally complete transformation and accumulation of 2,4-DCP occurred. Because mixed culture was from Di-Hwa Wastewater Treatment Plant, adding the same water as culture medium was found to increase the dechlorination of 2,4,6-TCP and its product 2,4-DCP. Dechlorination by adding wastewater of Petroleum Corporation was also increased, especially for 2,4-DCP product, but only 7% of 2,4,6-TCP dechlorination was found after incubation with Hou-Chin river water for 14 days.

Bacteria, Anaerobic↗
PubMed1997

Tri-calcium phosphate ceramics and allografts as bone substitutes for spinal fusion in idiopathic scoliosis as bone substitutes for spinal fusion in idiopathic scoliosis: comparative clinical results at four years.

The authors present the results of a comparative study of two series of posterolateral arthrodeses for scoliosis performed using COTREL DUBOUSSET instrumentation. Fifty-four consecutive patients underwent surgery for idiopathic scoliosis using the same technique. Thirty received a graft consisting of a mixture of corticocancellous autologous and allogenic bone frozen at -80 degrees, and 24 patients were grafted with a mixture of cortico-cancellous autologous bone and sticks of tricalcium phosphate (TCP, Biosorb, SBM, Lourdes, France). All patients were seen at three, six and twelve months, then once a year for at least four years with clinical and radiological evaluation at each visit. At the final follow up visit, no radiologic signs of pseudoarthrosis were found in either group with a minimum follow-up of 4 years. The appearance of bone callus was considered satisfactory at 6 months in all cases; moreover callus seemed to be more important in the TCP series, although this assessment was subjective. TCP resorption was total after 2 years, while allograft fragments were visible on x-rays after 2 years. Minor mechanical complications occurred but did not influence the results. Loss of correction was 8% of that initially obtained in the allograft group and 2% in the TCP group. Loss of correction did not progress after 6 months in the TCP group and after 2 years in the allograft group. Based upon this experience, the use of synthetic bone substitutes such as TCP would appear to be a valuable alternative to allografts in posterolateral spinal arthrodesis for idiopathic scoliosis, and it would eliminate the risk of viral contamination inherent to allograft implantation. To our knowledge, there have been no previous comparative studies concerning the use of tricalcium phosphate versus allograft in the literature.

Adolescent↗
PubMed2001

A constitutive 70 kDa heat-shock protein is localized on the fibres of spindles and asters at metaphase in an ATP-dependent manner: a new chaperone role is proposed.

In the present study, double immunofluorescence and immunoblot analysis have been used to show that centrosomes, isolated from Paracentrotus lividus sea urchin embryos at the first mitotic metaphase, contain the constitutive chaperone, heat-shock protein (HSP) 70. More specifically, we demonstrate that centrosomes contain only the HSP70-d isoform, which is one of the four isoforms identified in P. lividus. We also provide evidence that p34(cell division control kinase-2) and t complex polypeptide-1 (TCP-1) alpha, a subunit of the TCP-1 complex, are localized on the centrosomes. Furthermore, inhibition of TCP-1 in vivo, via microinjecting an anti-(TCP-1 alpha) antibody into P. lividus eggs before fertilization, either impaired mitosis or induced severe malformations in more than 50% of embryos. In addition, we have isolated the whole mitotic apparatus and shown that HSP70 localizes on the fibres of spindles and asters, and binds them in an ATP-dependent manner. These observations suggest that HSP70 has a chaperone role in assisting the TCP-1 complex in tubulin folding, when localized on centrosomes, and during the assembling and disassembling of the mitotic apparatus, when localized on the fibres of spindles and asters.

Adenosine Triphosphate↗
PubMed1993

Eukaryotic cytosolic chaperonin contains t-complex polypeptide 1 and seven related subunits.

We have characterized the cytosolic chaperonin from both rabbit reticulocyte lysate and bovine testis. The heteromeric complex contains eight subunits. Partial amino acid sequence data reveal that one of these is t-complex polypeptide 1 (TCP-1), while the other seven are TCP-1-related polypeptides, implicating the existence of a multigene family of TCP-1 homologues. We provide evidence that TCP-1 ring complex from bovine testis can facilitate the folding of both actin and tubulin, although, as in the case of chaperonin from reticulocyte lysate, two cofactors are required for the generation of properly folded tubulin. An additional molecule of TCP-1 may associate with the chaperonin depending on the purification procedure used. We propose that a highly conserved region in these polypeptides and in other chaperonins of the cpn60 chaperone family participates in ATP binding.

Actins↗
PubMed1996

Cucumber T-complex protein. Molecular cloning, bacterial expression and characterization within a 22-S cytosolic complex in cotyledons and hypocotyls.

T-complex protein (TCP) found in mammalian cells and yeast has been proposed as cytosolic folding machinery. We report here the cloning and initial characterization of a plant TCP cDNA. CSTCP-1 cDNA prepared from mRNA of cotyledons of germinating cucumber seeds encodes a polypeptide composed of 535 amino acid residues. The 59157-Da protein exhibits only 28% identity to both TCP-1p from yeast or and its homolog in Arabidopsis thaliana. Antibodies raised against the bacterially expressed plant protein were used to analyze the intracellular localization of TCP in two different plant tissues: fat-degrading non-dividing cotyledons and meristematic hypocotyls during seed germination. Cell fractionations included differential centrifugation and sedimentation of large complexes at 23000O x g for 4h. The latter fraction was further fractionated by sedimentation velocity centrifugation. This enrichment was required to detect by Western blotting cytosolic 59-kDa species as constituents of 22-S particles. From hypocotyls, a preparation of T-complex was obtained which consisted almost exclusively of proteins in the molecular range of 57-62 kDa. Likewise, the radioactive Cucumis sativus TCP-1 synthesized from CSTCP-1 mRNA in vitro using reticulocyte lysate was shown to migrate as a 61-kDa species.

Amino Acid Sequence↗
PubMed1996

Analysis of osteoblast activity at biomaterial-bone interfaces by in situ hybridization.

To investigate the effects of bioactive materials on bone formation in vivo, a new experimental model using in situ hybridization has been developed. A hole was drilled bilaterally in the distal epiphysis of rabbit femurs with subsequent implantations of beta-tricalcium phosphate (beta-TCP) cylinders in a press-fit manner. Specimens were collected at 3, 7, 14, and 28 days after operation. Femurs with empty drilling holes, and normal distal femurs without operation were used as controls. All specimens were decalcified and hybridized with a procollagen alpha 1(I) complementary RNA probe labeled with digoxygenin. In normal-bone sections, procollagen alpha 1(I) RNA was clearly demonstrated in periosteal osteoblasts, in osteoblasts in the mineralizing zone adjacent to growth plates, and in osteoblasts lining remodeling canals. As for beta-TCP, labeled osteoblasts around the material were not found at day 3, whereas they were most intensively observed at day 7 and a little less at day 14, in accordance with new-bone formation around the material. Weaker signals were also detected in fibroblasts at day 7. At day 28, osteoblasts lining the surface of newly formed bone were mainly negative, whereas those adjacent to the resorption sites of the beta-TCP showed positive signals, demonstrating an active remodeling at the material surface. The temporal expression of procollagen alpha 1(I) RNA in the beta-TCP specimens was fundamentally the same as that in the empty-hole specimens, suggesting no remarkable acceleration or suppression of bone-forming activity of osteoblasts by beta-TCP, which is consistent with osteoconductive bone formation. This in situ hybridization method was suggested to be a powerful tool in analyzing the biological effects of bioactive materials.

Animals↗
PubMed1997

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↗
PubMed1998

Glass-reinforced hydroxyapatite: a comprehensive study of the effect of glass composition on the crystallography of the composite.

Glass-reinforced HA composites were produced using phosphate-based glasses, and a structure refinement was carried out to determine the effect of the glass on the structure of the residual HA. Quantitative phase analysis showed that the glass causes some of the HA to decompose to beta-TCP and, at higher temperatures, to alpha-TCP. It also was indicated that when three phases were present, the formation of the alpha-TCP arose from decomposition of the beta-TCP and not from further decomposition of HA to alpha-TCP. The unit cell dimensions showed a decrease in the a axis and an increase in the c axis, giving an overall unit cell decrease in volume. There also was a significant effect based on the amount of glass added. The changes found in the composite containing the 4 wt% glass were attributed to the loss of carbonate and loss of hydroxyl. This was expected to cause shrinkage in the unit cell; however this was not seen, and therefore the major changes in the unit cell were attributed to the ions from the glass taking an interstitial role in the HA structure, thus not allowing the unit cell to shrink as much as expected.

Ceramics↗
PubMed1999

Characterization of acrylic bone cement using dynamic mechanical analysis.

Dynamic mechanical analysis (DMA) was used to characterize the properties of acrylic bone cement with the addition of tricalcium phosphate (TCP), hydroxyethyl methacrylate (HEMA), and ethylene glycol dimethacrylate (EGDMA). The glass transition temperature of acrylic bone cement is >100 degrees C; the cement has a flat modulus response near human body temperature. The height of the damping peak decreases and becomes broader with increasing TCP content. Thus, TCP is incompatibile with acrylic bone cement. When the frequency is changed from high to low, the damping peak shifts to low temperature. The shift in damping peak with frequency indicates that this relaxation is time-dependent. When acrylic bone cement contains TCP with HEMA and EGDMA, the incompatibility between acrylic bone cement and TCP can be ameliorated.

Biocompatible Materials↗
PubMed1999

Biocompatibility and degradation of collagen bone anchors in a rabbit model.

Bone anchors are used to fasten tendons and ligaments to bone during reconstructive surgery. Although metal anchors are often used, an anchor that could resorb and permit normal bone regeneration would be advantageous. The objective of the study was to evaluate the biocompatibility and degradation of bone anchors that consist of collagen-based bodies, ceramic washers, and polyester sutures. Eighteen rabbits underwent bilateral implantations in the distal femoral condyles. Nine animals received glutaraldehyde-crosslinked fibrillar collagen bone anchors (FC) and nine received glutaraldehyde-crosslinked fibrillar collagen bone anchors containing tricalcium phosphate (FC-TCP). Three animals per group were sacrificed at postimplantation weeks 1, 6, and 12. One femur from each rabbit was evaluated histologically, and the contralateral side underwent biomechanical pull-out testing. Histological evaluation of the implant site indicated that the FC and FC-TCP bone anchors were both biocompatible. The FC-TCP formulation degraded earlier than the FC formulation, and FC-TCP showed significant degradation at 6 weeks; the FC and FC-TCP formulations both showed similar amounts of degradation at 12 weeks. The degrading anchor bodies appeared to be osteoconductive as evidenced by new bone ingrowth into the degrading collagen matrices without a fibrous interface. These results suggest that collagen-based bone anchors have potential as bioresorbable orthopedic implants.

Animals↗
PubMed2000

Calcium-deficient hydroxyapatite-PLGA composites: mechanical and microstructural investigation.

The microstructural and mechanical properties of composites composed of calcium deficient hydroxyapatite (CDHAp) and poly(lactide-co-glycolide) (PLGA) have been investigated. The composites were formed by hydrolysis of alpha-tricalcium phosphate (alpha-TCP) to CDHAp in pressed precomposite compacts of alpha-TCP-PLGA-NaCl. The differences in hydrolysis of alpha-TCP-PLGA-NaCl for two compositions of 80:10:10 wt % and 60:20:20 wt %. were monitored by isothermal calorimetry and X-ray diffraction. The microstructural evolution and variance in final composite microstructure after hydrolysis at 37 degrees C, 45 degrees C, and 56 degrees C were examined by scanning electron microscopy. HAp-PLGA composite formed from the alpha-TCP-PLGA-NaCl (80:10:10) precomposites at 37 degrees C developed a tensile strength of 13.3 +/- 0.9 MPa, a flexural strength of 24.8 +/- 1.7 MPa, and Young's modulus of 2.8 +/- 0.3 GPa. These values were 12.00 +/- 0.2 MPa, 36.1 +/- 2.1 MPa, and 5.5 +/- 0.8 GPa for the precomposite composition 60:20:20. All these mechanical properties showed a variation with hydrolysis temperature and composition. The differences in mechanical properties were related to the final microstructures of the composites, which are governed by the morphological changes in the polymer structure at its glass transition temperature and the extent of cement-type formation of CDHAp by hydrolysis of alpha-TCP.

Biomechanical Phenomena↗
PubMed2001

Molded porous poly (L-lactide) membranes for guided bone regeneration with enhanced effects by controlled growth factor release.

The aim of this study was to develop platelet-derived growth factor (PDGF-BB) loaded moldable porous poly (L-lactide) (PLLA)-tricalcium phosphate (TCP) membranes for guided bone regeneration (GBR) therapy. The membranes were designed to fit various types of bone defect sites. PDGF-BB-dissolved PLLA-TCP in methylene chloride-ethyl acetate solution was cast on a dome shaped metallic mold to fabricate a model membrane. The release rate of PDGF-BB, the osteoblast attachment test, and guided bone regeneration potential were evaluated with PDGF-BB-loaded PLLA-TCP membranes. Regular pores were generated throughout the membrane mainly due to phase inversion of PLLA-methylene chloride-ethyl acetate solution. A therapeutic amount of PDGF-BB was released from the membrane. The release rate could be controlled by varying the initial loading content of PDGF-BB. A significant amount of cells attached onto the PDGF-BB-loaded membrane rather than onto the unloaded membrane. Dome shaped bone formation was achieved in rabbit calvaria at 4 weeks. This indicated that restoration of bone defects to the bone's original shape can be made possible by using molded membranes, which guide bone regeneration along with providing sufficient spaces. Bone forming efficiency was increased remarkably due to PDGF-BB release from PLLA-TCP membranes. These results suggested that the PDGF-BB releasing molded PLLA-TCP membrane may potentially improve GBR efficiency in various types of bone defects.

Animals↗
PubMed1991

Cardiopathic effect of 1,2,3-trichloropropane after subacute and subchronic exposure in rats.

1,2,3-Trichloropropane (1,2,3-TCP) is an industrial water contaminant with potential for human exposure by the oral route. The systemic toxicology of 1,2,3-TCP was evaluated after subacute or subchronic exposure in male and female Sprague-Dawley rats. Animals were treated with 0.01, 0.05, 0.20 and 0.80 mmol kg-1 day-1 for 10 days and 0.01, 0.05, 0.10 and 0.40 mmol kg-1 day-1 for 90 days. Chemical exposure was by oral gavage in corn oil. Lethality did not occur in either study. Toxicity was observed primarily in the high dose group of subacute and subchronically treated rats of both sexes. Weight gain suppression occurred at a dose of 0.8 mmol kg-1 (118 mg kg-1) after 10 days. After 90 days of exposure to 0.40 mmol kg-1, the final body weights were 81% and 86% of control values for males and females, respectively. When major organ weights were normalized by body weight, liver and kidney values were generally increased relative to control in the two highest dose groups after 10- and 90-day chemical exposure. Serum chemistries and histopathology indicated a mild hepatotoxic response to 1,2,3-TCP in the high dose group of each study but did not support any renal toxicity. Thymic weight reduction due to atrophy occurred at 10 days of exposure in high dose groups but was normal in all groups after the 90-day treatment. The primary histological finding in this study was an inflammation-associated cardiopathy produced by 1,2,3-TCP. Myocardial necrosis and degeneration occurred in a diffuse pattern with marked eosinophilia of affected cells. Male and female animals showed a cardiopathic response only at a dose of 0.8 mmol kg-1 1,2,3-TCP after the 10-day exposure.(ABSTRACT TRUNCATED AT 250 WORDS)

Administration, Oral↗
PubMed2002

Characterization of a novel calcium phosphate/sulphate bone cement.

Apatitic cements have shown excellent biocompatibility and adequate mechanical properties but have slow resorption in the human body. To assure that new bone tissue grows into the bone defect, a certain porosity is necessary although hard to achieve in injectable cements with suitable mechanical properties. An attempt was made by mixing alpha-tricalcium phosphate (alpha-TCP), calcium sulphate hemihydrate (CSH) and an aqueous solution containing 2.5 wt% of Na(2)HPO(4). The aim was to obtain a material containing two phases: a) one apatitic phase (calcium-deficient hydroxyapatite; CDHA) and b) one resorbable phase (calcium sulphate dihydrate; CSD). alpha-TCP and CSH mixtures were produced at relative intervals of 20 wt%. The liquid-to-powder (L/P) ratio to obtain a paste was 0.32 mLg(-1). The highest compressive strength (34 MPa) was obtained for the pure alpha-TCP sample. The strength was, in a first approximation, directly correlated to the weight proportions of the powders. X-ray diffraction analysis showed that the relative intensity for CDHA increased linearly, and the one for CSD decreased exponentially, when the amount of alpha-TCP increased. Thus, CSH ceased to transform to CSD when the amount of alpha-TCP increased. Observations in environmental scanning electron microscopy confirmed the X-ray diffraction results. CSH-crystals (100 microm) were embedded in the HA-matrix permitting gradual porosity in the material when resorbed.

Biocompatible Materials↗
PubMed2002

Matrix engineering for osteogenic differentiation of rabbit periosteal cells using alpha-tricalcium phosphate particles in a three-dimensional fibrin culture.

Tissue engineering using periosteal cells is a promising approach for bioactive bone repair. Of central importance in tissue engineering is the cell-matrix interaction. In the present study we tested in vitro the influence of alpha-tricalcium phosphate (alpha-TCP) particles on the expression of osteogenic markers in rabbit periosteal cells embedded in specially manufactured fibrin beads. After cell isolation from tibial periosteum of New Zealand White rabbits, and following monolayer culture, cells were embedded in alginate-fibrin beads containing 7.5% alpha-TCP particles and, as a control group, in beads without particles. The alginate was extracted immediately after polymerization. The beads were cultivated for at least 53 days. The DNA content, alkaline phosphatase activity, and osteocalcin level were determined. In monolayer culture the number of cells increased 6.5-fold. DNA content increased in both three-dimensional culture groups but was significantly higher in the beads containing alpha-TCP. Alkaline phosphatase activity increased in both groups without significant differences. Osteocalcin content was significantly higher in the beads containing alpha-TCP than it was in those without alpha-TCP. These observations indicate that matrix engineering using inorganic particles in fibrin culture can influence the osteogenic differentiation of mesenchymal cells. The three-dimensional culture system presented here facilitates the preparation of grafts for bone reconstruction.

Alkaline Phosphatase↗