Gene/Protein Disease Symptom Drug Enzyme Compound
Pivot Concepts: Gene/Protein Disease Symptom Drug Enzyme Compound   Target Concepts: Gene/Protein Disease Symptom Drug Enzyme Compound

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Query: EC:3.1.3.1 (alkaline phosphatase)
47,916 document(s) hit in 31,850,051 MEDLINE articles (0.00 seconds)

The presence of many types of polypeptide growth factors in the mineralized extracellular matrix of bone is now well established. These factors are generally referred to as bone-derived growth factors (BDGFs), and are similar, or possibly identical, to the following species; platelet-derived growth factor (PDGF); acidic and basic forms of fibroblast growth factor (aFGF, bFGF); transforming growth factor beta (TGF-beta); and insulin-like growth factor 1 (IGF-1). Several osteoinductive factors, such as bone morphogenetic protein (BMP) and osteogenin, a skeletal growth factor (SGF), and osteoblast-derived BDGFs, have also been identified. Complete description of the biological functions of these BDGFs which are relevant to bone will ultimately require specific bioassays involving specific cell types in vitro, as well as in vivo animal implant models. Studies with primary rat osteoblast-like cells exposed either to mixed BDGFs, pure TGF-beta, or heparin-purified PDGF, aFGF, or bFGF from bovine bone have shown a general dose-dependent mitogenic effect. Phenotypic changes which accompany the BDGF-induced wave of proliferation include: decreased osteocalcin secretion and a reduction in 1,25-(OH)2 vitamin D3-stimulated osteocalcin synthesis; reduced alkaline phosphatase specific activity; decreased cyclic AMP responsiveness to parathyroid hormone (PTH); and increased collagen synthesis. Bone exhibits the most complex spectrum of growth factor activities of any tissue yet described. In bovine bone powder free of blood and cartilage contamination, the volume concentration of mitogens is up to 20 times greater than that in serum. Bone cells and other indigenous cell types must be considered as possible sources of the BDGFs, in addition to sequestration from blood. Mechanisms for the unmasking or release of BDGFs from the mineralized matrix that result in local action on osteoblasts, endothelial cells, and other target cells are undoubtedly important for the development and maintenance of bone tissue.
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PMID:Polypeptide growth factors in bone matrix. 306 10

Periostea were dissected from 17-day-old chicken embryo calvariae, placed on millipore filters, and cultured on fluid media containing serum or on serum and plasma containing "plasma clots," in three ways: 1) with the osteogenic layer facing the filter, 2) with the osteogenic layer away from the filter, 3) folded such that the osteogenic layer was in apposition with itself within the fold. The cultures were studied histologically as well as biochemically. Periostea that were cultured folded showed differentiation of osteoblastlike cells after 2 days, and production of osteoid at day 4. Tissues cultured with the osteogenic layer away from the filter demonstrated similar osteoblastic differentiation and osteoid production. Both types of cultures exhibited an increase in histochemically detectable alkaline phosphatase activity over the 4 day culture period that was associated with osteoblasts and the osteogenic area. Periostea cultured with the osteogenic layer facing the filter produced no osteoid. In these cultures, histochemically detectable alkaline phosphatase activity decreased and virtually disappeared over the 4 day culture period. The possibility that the creation of a suitable micro-environment is required for osteodifferentiation in this culture system is discussed.
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PMID:Differentiation of osteoid-producing cells in vitro: possible evidence for the requirement of a microenvironment. 308 98

These studies were intended to assess the osteogenic activity of monofluorophosphate (MFP) in vitro, and to identify the enzyme(s) responsible for MFP hydrolysis-alkaline phosphatase (ALP) and/or acid phosphatase (AcP). ALP and AcP activities were determined by hydrolysis of p-nitrophenylphosphate (PNPP) at pH greater than 8 and pH 5.5, respectively, and MFP hydrolysis was determined, between pH 5.5 and pH 9.0, from measurements of [fluoride ion], using an ion-specific electrode. We found (1) that MFP was an alternative substrate for purified ALP, but not for AcP; (2) that MFPase activity in the embryonic chick resembled ALP, but not AcP, with respect to pH-dependent hydrolysis, sensitivity to effectors (r = 0.98, P less than .001), and tissue distribution (r = 0.96, P less than .001); and (3) that intestinal MFPase activity in the embryonic chick co-purified with ALP activity (r = 0.93, P less than .01) and resembled ALP, but not AcP, in its distribution along the small intestine, being highest in the duodenum and lowest in the distal ileum (r = 0.96, P less than .001). We also found that in vitro exposure to MFP increased (1) the proliferation rate of embryonic chick calvarial cells in serum-free monolayer cultures (i.e., 3[H]-thymidine incorporation into DNA, P less than .001); (2) ALP activity in calvarial cells (P less than .005) and in intact calvaria (P less than .05); and (3) collagen production by intact calvaria (i.e., 3[H]-proline incorporation as 3[H]-hydroxyproline, P less than .05).(ABSTRACT TRUNCATED AT 250 WORDS)
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PMID:Monofluorophosphate is hydrolyzed by alkaline phosphatase and mimics the actions of NaF on skeletal tissues, in vitro. 310 98

A new culture system was developed to clarify the biocompatibility of implant materials with bone tissue using the MC3T3-E1 osteogenic cell line. The cells were inoculated onto specimens such as aluminium oxide, titanium, dental casting silver-palladium alloy (PD), and a plastic coverslip. To study the effects of these materials on cell growth, differentiation, and calcification, DNA and protein content, alkaline phosphatase activity, and calcium content, respectively, were determined. The results from biochemical analysis suggest titanium and aluminum oxide to have adequate biocompatibility, while PD has an irritant effect on cell metabolism. It is clear that an objective view of the differentiation and calcification processes of osteogenic cells can be understood through such analysis. From the results of this study, our culture system appears suitable for evaluating the biocompatibility of implant materials with bone tissue.
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PMID:Development of a new system for evaluating the biocompatibility of implant materials using an osteogenic cell line (MC3T3-E1). 316 81

To examine an inhibitory mechanism of Cd on bone formation, the effects of Cd on calcification were investigated in a culture of a clonal osteogenic cell line, MC3T3-E1. At 3 days after inoculation, Cd was added to the medium containing 7 mM beta-glycerophosphate, and culture was continued for 8 days. Cd at 1.78 microM and above caused a significant decrease in 45Ca accumulation. The decrease in mineralization by Cd was similar to that in collagen content or alkaline phosphatase (ALP) activity. Histologically, the cell density and the mineralization degree were lower than those of the controls. Ultrastructurally, degenerated cells were observed with undifferentiated cells which had fewer rough-surfaced endoplasmic reticulum and many mitochondria. This suggests that Cd may inhibit the differentiation into osteoblasts as well as the cell function. On the other hand, calcification of cells at 8 days after inoculation was inhibited by Cd at 1.78 microM and above. The decrease in collagen content and ALP activity by Cd was much lower than that in calcification. Cd-treated cells were well differentiated into osteoblasts morphologically, but the mineralization degree was lower than that of the controls. Ultrastructurally, cell damage was not recognized so strongly compared with long-term Cd treatment. The mineralization of osteoblasts was also inhibited by Zn levels which left both collagen content and ALP activity unaffected. From these results, it was suggested that the inhibitory effect of Cd on in vitro calcification of MC3T3-E1 cells may be due to both a depression of cell-mediated calcification and a decrease in physiochemical mineral deposition.
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PMID:Inhibitory effects of cadmium on in vitro calcification of a clonal osteogenic cell, MC3T3-E1. 318 26

The anterior part of the mammalian nasal septum (NS) persists throughout the life span as hyaline cartilage, in contrast to cartilage in most parts of the body, which is gradually replaced by bone during development. In this study, we have cultured differentiating rat NS under various experimental conditions in an attempt to gain some insight into the osteogenic potential, if any, of the NS and its surrounding connective tissue. Differentiating NS from E15 and E19 rat embryos were dissected and grown under the following conditions: 1) organ cultured in Waymouth's medium or modified Eagle's medium, with or without serum; 2) cultured on chick chorioallantoic membrane (CAM); 3) implanted under rat kidney capsule (KC). Bone-like substance (BLS) never developed in organ cultures, but was observed in CAM cultures and KC implants after 7 days. The BLS was located external to the perichondrium of the NS and was stained red by the van Gieson's technique, indicating the presence of mature collagen. Further evidence of its bone-like characteristics was demonstrated by the presence of alkaline phosphatase and type I collagen. The CAM and KC represent two experimental conditions under which progenitor cells in the nasal septum area may be induced to synthesize BLS.
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PMID:Development of bone-like substance in cartilaginous rat nasal septum under experimental conditions. 318 74

Murine adult bone marrow exhibits mineralizing capacity in vitro as is demonstrated by the new in vitro assay we report here. In less than 2 weeks after the onset of the cultures, mineralization is obtained in more than 80% of the marrow cultures. Moreover, morphological studies reveal that during incubation phenotypic changes related to osteogenic differentiation occur at the extracellular matrix as well within cell populations. Well banded collagen is synthesized. Matrix vesicles and needles of hydroxy-apatite crystals are observed via transmission electron microscopy. Osteoblast-like cells are present with membrane-associated alkaline phosphatase activity. The mineralization is specific for cultured bone marrow and is not observed in cultured spleen fragments as is shown via 85Sr uptake, calcein uptake and histomorphology. No inducing agent is added to the tissue culture medium except for 10% fetal calf serum, beta-glycerophosphate (10(-2) M) and ascorbic acid. However, the prerequisite for obtaining mineralization is the three-dimensional structure of the marrow in culture. The in vitro organ culture we developed may provide the opportunity to identify which marrow cells have osteogenic potential and to investigate the mechanisms triggering differentiation towards osteogenesis.
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PMID:Mineralization of adult mouse bone marrow in vitro. 324 57

New aspects of the distribution and developmental appearance of the 44-kDa bone phosphoprotein (44K BPP, also called sialoprotein I or osteopontin) and bone gamma-carboxyglutamic acid (Gla)-containing protein (BGP, also called osteocalcin) during osteogenesis and dentinogenesis were investigated with immunocytochemical techniques using monospecific, affinity-purified polyclonal antibodies. Sections from newborn rat incisors and from various bone anlagen of newborn animals and fetuses were processed for detection of 44K BPP or BGP antigenicity. In addition, histochemical reactions for detection of alkaline phosphatase or calcium salts were performed on a number of the sections. The 44K BPP appears to be synthesized and secreted by chondrocytes only in the areas of cartilage-to-bone transition; these cells could participate indirectly in the process of bone formation by providing a suitable scaffold onto which primary marrow osteoblasts attach and spread. During osteogenesis, 44K BPP is found in bone-forming cells almost concomitantly with the appearance of alkaline phosphatase and before osteoid deposition, whereas BGP is still absent during early stages of mineralization. We hypothesize that this dramatic difference between the developmental appearance of 44K BPP and BGP reflects the delayed expression of the BGP gene relative to that of 44K BPP. In long-term cultures of bone marrow from adult mice, some fibroblastic cells expressed the 44K BPP phenotype; these cells could represent early osteogenic progenitor cells. Some experiments also suggested that, as with BGP, 44K BPP or an immunologically related protein is synthesized by some odontoblasts and secreted into predentin, prior to the onset of mineralization.
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PMID:Developmental expression of 44-kDa bone phosphoprotein (osteopontin) and bone gamma-carboxyglutamic acid (Gla)-containing protein (osteocalcin) in calcifying tissues of rat. 326 Aug 80

Telangiectatic osteosarcoma (TOS) which occurred in the metaphysis of the right femoral bone in a 13-year-old female was reported. It showed osteolytic and cystic lesion without sclerotic change on roentgenogram and consisted histologically of various sized blood-filled spaces lined by layers of round to oval tumor cells in the thin fibrous septa. In some solid areas, a proliferation of atypical tumor cells with large prominent nucleoli was evident, embedded in the lace-like osteoid tissue. Mitotic cells were easily encountered. A large population of tumor cells revealed high alkaline phosphatase activity as well as 5'-nucleotidase activity, indicative of osteogenic cell origin. Ultrastructurally, they showed osteogenic characteristics of well-developed rough endoplasmic reticula, cytoplasmic microfibrils, and dense bodies, but not for those of endothelial cells. In this report, we suggest that alkaline phosphatase activity in biopsy and surgical specimens is useful for distinguishing TOS from other osteolytic bone tumors, with regard to its ontogenic discussion.
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PMID:High alkaline phosphatase activity of telangiectatic osteosarcoma (TOS) and its diagnostic significance. 347 63

The purpose of this study was to correlate the histologic and biochemical responses of the interparietal suture to a range of tensile forces. Stainless steel spring implants, calibrated to generate expansive forces from 50 to 250 g, were placed across the interparietal suture in 85 female Sprague-Dawley rats. After experimental periods from 2 hours to 14 days, the interparietal sutures were evaluated by radiography, histology, and biochemistry. An in vivo/in vitro system was used for the biochemical analysis; total protein, proline incorporated, percent collagen, and alkaline phosphatase activity were measured. The radiographs and histology showed that in vivo suture expansion was achievable with 50 to 70 g of force, but the heavier forces showed greater sutural opening, more cellular proliferation, and more bone formation. This increased biologic response by the heavier forces was substantiated by an increase in sutural protein and alkaline phosphatase activity but not in percent collagen. It was concluded that changes in the total protein content of the suture were not primarily caused by proliferation of osteogenic cells and fibroblasts but due to an influx of transudate. In contrast, the increase in incorporation of 3H-proline and alkaline phosphatase activity correlated with the observance of bone formation. This study indicated a positive correlation between the magnitude of tensile forces and osteogenic response.
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PMID:The morphologic and biochemical effects of tensile force application to the interparietal suture of the Sprague-Dawley rat. 347 67


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