Gene/Protein Disease Symptom Drug Enzyme Compound
Pivot Concepts:   Target Concepts:
Query: EC:3.1.3.1 (alkaline phosphatase)
47,916 document(s) hit in 31,850,051 MEDLINE articles (0.00 seconds)

Oncostatin M (OSM) has been described as a bone-remodeling factor either stimulating osteoblast activity or osteoclast formation in vitro. To elucidate the in vivo effect of OSM on bone remodeling, we injected an adenoviral vector encoding murine OSM in knee joints of mice. OSM strongly induced interleukin (IL)-6 gene expression, a known mediator of osteoclast development. We investigated the OSM effect in wild-type and IL-6-deficient mice and found a similar degree of OSM-induced joint inflammation. Within the first week of inflammation, the periosteum along the femur and tibia increased in cell number and stained positive for the osteoblast marker alkaline phosphatase. At these sites bone apposition occurred in both strains as demonstrated by Goldner and Von Kossa staining. In vitro OSM enhanced the effect of bone morphogenetic protein-2 on osteoblast differentiation. Immunohistochemistry demonstrated expression of receptor activator of nuclear factor-kappa B ligand (RANKL) and its receptor, receptor activator of nuclear factor-kappa B (RANK), in the periosteum but osteoclasts were not detected at sites of bone apposition. Induced mRNA expression for the receptor activator of nuclear factor-kappa B ligand inhibitor osteoprotegerin probably controlled osteoclast development during OSM overexpression. Our results show that OSM favors bone apposition at periosteal sites instead of resorption in vivo. This effect was not dependent on or inhibited by IL-6.
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PMID:Adenoviral transfer of murine oncostatin M elicits periosteal bone apposition in knee joints of mice, despite synovial inflammation and up-regulated expression of interleukin-6 and receptor activator of nuclear factor-kappa B ligand. 1200 Jul 25

Bone replacement materials for reconstruction of bone defects must be biocompatible and biodegradable and must have osteoconductive or even osteogenic potential. Ideally, their shape should also be adaptable to the defect and they should possess long-term adaptability to the biomechanical situation at the implantation site. Human mesenchymal stem cells of the cambium layer of the periosteum were cultivated, placed in a fibrin suspension on a preformed carrier structure (PGLA polymer + beta-TCP), and cultivated under conditions of osteogenic differentiation. After 10, 20, 30, and 40 days, histological examination was performed, alkaline phosphatase activity and levels of osteocalcin, DNA, and collagen were determined, and the influence of addition of TGF-beta1 at a concentration of 5 ng/ml to the culture medium was investigated. Demonstration of bone-specific marker proteins indicated that the in vitro combination of mesenchymal stem cells, PGLA polymer, beta-TCP, and fibrin resulted in de-novo synthesis of human preosseous tissue, while addition of TGF-beta1 resulted in greater new bone formation with significantly higher concentrations of marker proteins. Histological examination showed the presence of newly formed bone at the surface of the implant. As compared with the use of structured TCP or hydroxyapatite implants as in earlier works, use of a combination of autologous cell material, PGLA polymer, and beta-TCP results in a malleable, vital implant that is adaptable to the bone defect. This combination thus may represent a new option for the treatment of bone defects.
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PMID:In vitro-cultivation of human periosteum derived cells in bioresorbable polymer-TCP-composites. 1201 77

Deer antlers are the only mammalian organs that can be repeatedly regenerated; each year, these complex structures are shed and then regrow to be used for display and fighting. To date, the molecular mechanisms controlling antler regeneration are not well understood. Vitamin A and its derivatives, retinoic acids, play important roles in embryonic skeletal development. Here, we provide several lines of evidence consistent with retinoids playing a functional role in controlling cellular differentiation during bone formation in the regenerating antler. Three receptors (alpha, beta, gamma) for both the retinoic acid receptor (RAR) and retinoid X receptor (RXR) families show distinct patterns of expression in the growing antler tip, the site of endochondral ossification. RAR alpha and RXR beta are expressed in skin ("velvet") and the underlying perichondrium. In cartilage, which is vascularised, RXR beta is specifically expressed in chondrocytes, which express type II collagen, and RAR alpha in perivascular cells, which also express type I collagen, a marker of the osteoblast phenotype. High-performance liquid chromatography analysis shows significant amounts of Vitamin A (retinol) in antler tissues at all stages of differentiation. The metabolites all-trans-RA and 4-oxo-RA are found in skin, perichondrium, cartilage, bone, and periosteum. The RXR ligand, 9-cis-RA, is found in perichondrium, mineralised cartilage, and bone. To further define sites of RA synthesis in antler, we immunolocalised retinaldehyde dehydrogenase type 2 (RALDH-2), a major retinoic acid-generating enzyme. RALDH-2 is expressed in the skin and perichondrium and in perivascular cells in cartilage, although chondroprogenitors and chondrocytes express very low levels. At sites of bone formation, differentiated osteoblasts which express the bone-specific protein osteocalcin express high levels of RALDH2. The effect of RA on antler cell differentiation was studied in vitro; all-trans-RA inhibits expression of the chondrocyte phenotype, an effect that is blocked by addition of the RAR antagonist Ro41-5253. In monolayer cultures of mesenchymal progenitor cells, all-trans-RA increases the expression of alkaline phosphatase, a marker of the osteoblastic phenotype. In summary, this study has shown that antler tissues contain endogenous retinoids, including 9-cis RA, and the enzyme RALDH2 that generates RA. Sites of RA synthesis in antler correspond closely with the localisation of cells which express receptors for these ligands and which respond to the effects of RA.
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PMID:A role for retinoic acid in regulating the regeneration of deer antlers. 1243 67

Age-related expansion of the external surface of the femoral neck in order to offset generalized bone loss is potentially an important mechanism whereby hip strength and hence resistance to hip fracture is maintained. However, it has been widely assumed that bone formation is precluded from this external interface due to the presence of a synovial membrane associated with the hip joint. In this study we have demonstrated histologically that bone formation does indeed occur on the outer "periosteal" surface of the proximal femoral neck. It was therefore hypothesized that an impairment or reduction in periosteal bone formation might be seen in cases of femoral neck fracture compared with age-matched controls. Qualitative analysis of whole femoral neck samples from female subjects and age- and sex-matched post-mortem controls demonstrated that these groups expressed similar distributions of the bone formation marker, alkaline phosphatase (AP), at the periosteal surface [whole biopsy mean % periosteal AP-positive surface: control=16.0 (range=0.5-43.0), fracture=13.4 (range=1.0-34.6), p=0.44]. In conclusion, despite a wide intersubject variation, bone formation at the femoral neck periosteum is a feature of elderly women even if they have had a hip fracture.
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PMID:Evidence for bone formation on the external "periosteal" surface of the femoral neck: a comparison of intracapsular hip fracture cases and controls. 1273 Jul 75

To investigate the effect of basic fibroblast growth factor(bFGF) gene transfection on the proliferation and differentiation of mesenchymal stem cells (MSCs) and to provide basis for accelerating bone defect repairing using gene-enhanced tissue engineering technology, Rabbit periosteum-derived MSCs were transfected with the full-length rat bFGF cDNA in vitro. The transient and stable gene expression of bFGF were determined by immunohistochemistry. The proliferation and the synthesis alkaline phosphatase (ALP) and osteocalcin(OC) of the transfected MSCs were also examined. The results showed that bFGF cDNA could be transferred into osteoblasts and expressed stably at least 4 weeks. The proliferation and OC content of genetically modified MSCs were increased significantly, whereas the ALP activity remained no change. In conclusion, transfer of gene encoding bFGF to MSCs increases its proliferation and osteogenesis property. Based on the successful conjunction of the existing techniques of tissue engineering with the novel possibilities offered by modern gene transfer technology, an innovative concept, molecular tissue engineering, was put forward for the first time. As a new branch of tissue engineering, it represents both a new area and an important trend in tissue engineering research.
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PMID:[Gene-enhanced tissue engineering: applications in osteoinduction using cultured mesenchymal stem cells transduced with the bFGF gene]. 1456 9

The effect of low intensity pulsed ultrasound on human periosteal cells was investigated. Normal human periosteum was obtained to culture the periosteal cells. After characterization, cultures of periosteal cells at Days 2 and 4 were treated with ultrasound for 5, 10, and 20 minutes respectively. Assessments were done to assess total number of viable cells, cell proliferation, alkaline phosphatase activity, osteocalcin secretion, vascular endothelial growth factor expression, and calcium nodule formation. With the cells not treated with ultrasound as the control, the results showed that ultrasound did not affect the total number of viable cells. It stimulated cell proliferation at the early phase of cell culture. The activity of alkaline phosphatase was increased significantly in the culture at Day 4. A similar effect was seen with osteocalcin secretion and the responses were dose-dependent. The vascular endothelial growth factor secretion increased in Day 2 and Day 4 cultures with the dose-dependent effect. Formation of calcium nodules was significantly higher with ultrasound treatment. We think that low intensity pulsed ultrasound stimulated periosteal cell proliferation and differentiation toward osteogenic lineage. The dose-dependent effect on osteogenic activities may modify the existing treatment regimen. Ultrasound treatment should be started from the beginning of fracture healing.
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PMID:Low intensity pulsed ultrasound stimulates osteogenic activity of human periosteal cells. 1504 27

Several inorganic materials have been shown previously to hold some osteogenic capacity. The purpose of this study is to compare the bone-forming abilities of hydroxyapatite ceramic, high-density porous polyethylene, and bone collagen within the periosteal island flap of rabbit tibia using histological and biochemical analysis. With this goal, four discrete experimental groups were formed, each comprising 22 New Zealand male rabbits. A sac was created on each rabbit tibial periosteum flap in each of the groups, and each of the previously mentioned materials was placed within this sac separately. One of these groups was thought as a control group without any material being placed inside the periosteal sac. Biopsies were taken at weeks 1, 2, 4, and 8 for biochemical analysis and at weeks 2 and 8 for histological evaluation. Neo-osteogenesis was evaluated quantitatively by determination of alkaline phosphatase and osteocalcin levels biochemically as well as by the percentage of new bone formation inside the periosteal sac histologically. Results show statistically that the osteogenic effect of high-density porous polyethylene is greater than that of the other materials used in this study (P < 0.05).
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PMID:Comparing the osteogenic capacities of bone substitutes: hydroxyapatite, high-density porous polyethylene, and bone collagen: a biochemical and histological analysis. 1521 35

In adult animals, bone marrow is the major site of blood cell production, which is controlled by interactions between the local stroma and blood cell progenitors. The endosteal/subendosteal environment comprises bone-lining and adjacent reticular cells and sustains haemopoietic stem cell (HSC) self-renewal, proliferation and differentiation. We have questioned the specific role of each of these stroma cells in controlling HSC fate. We have isolated two distinct stroma-cell populations containing subendosteal reticulocytes (F-RET) and osteoblasts (F-OST) from periosteum-free fragments of murine femurs by a two-step collagenase-digestion procedure. Both populations produce similar extracellular matrix (collagen I, laminin, fibronectin, decorin), except for collagen IV, which is low in F-OST. They also express osteogenic markers: osteopontin, osteonectin, bone sialoprotein and alkaline phosphatase (ALP). The quantity and activity of ALP are however higher in F-OST. When co-cultured with bone marrow mononuclear cells or lineage-negative haemopoietic progenitors, F-OST stroma induces low proliferation and high maintenance of early haemopoietic progenitors, whereas F-RET stroma induces high short-term proliferation and differentiation. Analysis by reverse transcription/polymerase chain reaction has revealed higher levels of Jagged-1 expression by F-OST cells than by the F-RET population. Thus, two adjacent stroma cells (subendosteal and endosteal) play distinct roles in controlling the stem-cell capacity and fate of HSC and probably contribute distinctly to HSC niche formation.
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PMID:Bone marrow subendosteal microenvironment harbours functionally distinct haemosupportive stromal cell populations. 1557 25

Mineralized extracellular matrix formation is representative for the osteoinductive capacity of biomaterials and is often tested in vitro. Characteristics of in vitro mineralization of primary rat osteoblastic cells (bone marrow, calvaria, periosteum, fetal and adult long bone) and UMR-106 cells were compared by von Kossa staining, FTIR, X-ray diffractometry, TEM and related to parameters of early (ALP and collagen I formation) and late (osteocalcin secretion) osteoblast expression. All cultures expressed high alkaline phosphatase activity and were able to form bone apatite. However, a nodular versus diffuse mineralization pattern was observed. Bone marrow, calvaria and periosteum (early passage) derived cells mineralized restrictively on the three-dimensional area of a nodule. The extracellular matrix consisted of collagen I fibers, among matrix vesicles loaded with needle-like crystals. Long bone, late passage periosteum derived and UMR-106 cells exhibited a diffuse mineralization pattern. Needle-like crystals were observed between the cells but collagen fibers and matrix vesicles could not be detected. Secretion of osteocalcin was detected in cultures derived from bone marrow and absent in UMR-106 and long bone derived cell cultures. The present study demonstrates that dystrophic calcification can not be distinguished from cell-mediated calcification with von Kossa, FTIR and X-ray diffractometry. Primary osteoblastic cells capable of forming nodules are recommended to evaluate the osteoinductive properties of biomaterials.
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PMID:Calcification as an indicator of osteoinductive capacity of biomaterials in osteoblastic cell cultures. 1576 32

The ability to generate new bone for reconstructive surgery use is a major clinical need. Tissue engineering with osteoprogenitor cells isolated from the patient's periosteum and seeded into bioresorbable scaffolds offers a promising approach to the generation of skeletal tissue. To our knowledge, there is no description about the expression of Ets2 in tissue engineered "bone neotissue." The aim of our study was to manufacture cell-seeded three-dimensional bone constructs with human periosteal cells on poly (lactic-co-glycolic acid) polymer fleeces to describe the expression pattern of Ets2 and its target genes osteocalcin and osteopontin; expression analysis of type I collagen, core-binding factor-1, alkaline phosphatase, and osteonectin; the ability of matrix mineralization and ALP enzymatic activity showed the osteogenic character of the constructs. A significant correlation between the expression of Ets2 and osteopontin mRNA (r = -0.70; p < 0.05) could be shown. A 1.35-fold increase of Ets2 expression from days 1 to 9 was detected, followed by a slight decrease from days 11 to 15. Until the end of the culture period, the expression of Ets2 reached a comparable high level as detected on day 9. In contrast, the expression level of osteopontin mRNA reached a maximum at day 7, followed by a progressive 3.04-fold decrease until day 21. This study shows for the first time that Ets2 gene and its transcriptional target genes are expressed in tissue-engineered bone constructs. These findings have the potential to provide much-needed information about the role and function of Ets2 in human osteogenesis processes and creation of "bone neotissue."
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PMID:Expression pattern of the chromosome 21 transcription factor Ets2 in cell-seeded three-dimensional bone constructs. 1590 Jun 11


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