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)

For bone regeneration constructs using human jaw periosteal cells (JPC) the extent of osteoinductive ability of different three-dimensional scaffolds is not yet established. We analyzed open-cell polylactic acid (OPLA) scaffolds for their suitability as bone engineering constructs using human JPC. Cell adhesion and spreading was visualized on the surface of scaffolds by scanning electron microscopy. JPC proliferation within OPLA scaffolds was compared with proliferation within collagen and calcium phosphate scaffolds. We found a significant increase of proliferation rates in OPLA scaffolds versus Coll/CaP scaffolds at three time points. Live-measurements of oxygen consumption within the cell-seeded scaffolds indicate that the in vitro culturing time should not exceed 12-15 days. OPLA scaffolds, which were turned out to be the most beneficial for JPC growth, were chosen for osteogenic differentiation experiments with or without BMP-2. Gene expression analyses demonstrated induction of several osteogenic genes (alkaline phosphatase, osterix, Runx-2 and insulin-like growth factor) within the 3D-scaffolds after 12 days of in vitro culturing. Element analysis by EDX spectrometry of arising nodules during osteogenesis demonstrated that JPC growing within OPLA scaffolds are able to form CaP particles. We conclude that OPLA scaffolds provide a promising environment for bone substitutes using human JPC.
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PMID:Analysis of OPLA scaffolds for bone engineering constructs using human jaw periosteal cells. 1815 13

Platelet-rich plasma (PRP) represents an autologous source of growth factors essential for bone regeneration. The clinical efficacy of PRP is, however, unpredictable, and this is likely due to the inefficient and inconsistent delivery of PRP-derived growth factors. Previous investigations have shown that current methods of PRP preparation result in a premature release of the relevant bone stimulatory factors. As successful bone regeneration requires multiple factors presented in a physiologic temporal and spatial cascade, the objective of this study is to control the bioavailability of PRP-derived growth factors using a hydrogel carrier system. Specifically, the release of platelet-derived growth factor, transforming growth factor beta-1, and insulin-like growth factor from two types of alginate carriers was compared over time. The effects of the released factors on the growth and alkaline phosphatase (ALP) activity of human osteoblast-like cells were also evaluated. It was found that factor release profiles varied as function of carrier type, and binding of growth factors to the alginate matrix also modulated their release. The bioactivity of released factors was maintained in vitro and they promoted cell proliferation and ALP activity. These results demonstrate the potential of this autologous multifactor delivery system for controlling the bioavailability of PRP-derived factors. Future studies will focus on optimizing this system to increase the clinical efficacy of PRP by matching the distribution and temporal sequencing of PRP-derived factors to the bone healing cascade.
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PMID:Controlled delivery of platelet-rich plasma-derived growth factors for bone formation. 1818 Nov 9

Nutritional and metabolic exposure during critical periods of prenatal and neonatal development in animals and humans may induce long-term effects on health status in later life. The aim of this study was to investigate the effects of neonatal treatment with beta-hydroxy-beta-methylbutyrate (HMB) during the first 3 weeks of life on programming of skeletal development in sheep. The study was performed on 12 male sheep divided into two groups. While the control group was treated with placebo, the lambs in the HMB group received the calcium salt of beta-hydroxy-beta-methylbutyrate at a daily dose of 0.1 g (kg body weight)(-1). The assessment of growth hormone (GH), insulin-like growth factor-1 (IGF-1) and biochemical bone turnover markers in serum was performed in 21- and 130-day-old animals. Post mortem, volumetric bone mineral density, morphological and mechanical properties were determined in femur and lumbar vertebrae. Neonatal treatment with HMB increased serum concentrations of bone-specific alkaline phosphatase, osteocalcin, GH and IGF-1 in 21-day-old lambs by 125.2, 93.8, 71.8 and 70.9%, respectively (P < or = 0.05). The C-terminal telopeptide of type I collagen (CTX) concentration was increased in 130-day-old animals from the HMB group by 33.1% (P = 0.03). Furthermore, HMB administration improved bone weight, volumetric bone mineral density and bone morphological and mechanical properties of femur and lumbar spine. In conclusion, this study showed long-term beneficial effects of neonatal treatment with HMB on programming of peripheral and axial skeleton properties that were mediated by a transient improvement of somatotrophic axis function and acceleration of bone metabolism.
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PMID:Neonatal programming of skeletal development in sheep is mediated by somatotrophic axis function. 1826 56

Controlled release of growth factors or drugs provides great therapeutic advantages for bone defects which do not heal with normal therapeutic treatments. We have accelerated the deposition of bone-like mineral (BLM) on the surface of three-dimensional (3D) poly(lactic-co-glycolic acid) (PLGA) porous scaffolds to 36-48 h by modifying the biomimetic process parameters and applying surface treatments onto PLGA scaffolds. We used simulated body fluid containing insulin-like growth factor-1 (IGF-1; 1 microg/ml) to mineralize the PLGA scaffolds for 48 h. IGF-1 was co-precipitated with mineral on the surface of the PLGA scaffolds. IGF-1-incorporated mineralized scaffolds demonstrated slow controlled release over a 30 day period when they were incubated in phosphate-buffered saline (PBS) at 37 degrees C. Bone marrow stromal cell (BMSC) function on three different types of scaffolds, such as control (non-mineralized) scaffolds, mineralized scaffolds, and IGF-1-incorporated mineralized scaffolds was also investigated. BMSC attachment and proliferation was enhanced for IGF-1-incorporated mineralized scaffolds compared with controls during the culture period. BMSC differentiation was not changed during the culture period among the three groups of scaffolds, as assessed by alkaline phosphatase activity and osteocalcin assay. According to findings from this study, BLM has great potential to be used as a carrier for biological molecules for localized release applications as well as bone tissue-engineering applications.
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PMID:Controlled release of insulin-like growth factor-1 and bone marrow stromal cell function of bone-like mineral layer-coated poly(lactic-co-glycolic acid) scaffolds. 1836 82

This study investigated the effect of exogenous insulin-like growth factor (IGF)-1 on the proliferation and differentiation of osteoblastic cells from Chinese Holstein cattle and the resultant bone nodule formation and mineralisation in vitro. The osteoblastic cells were isolated and cultured, then identified using Giemsa and alkaline phosphatase (ALP) staining methods. The effect of different concentrations of IGF-1 on cell growth was assessed by MTT assay. The ALP activity and osteocalcin (OC) concentration in the osteoblastic cells were measured by a colorimetric assay and a radioimmmunoassay, respectively. Calcium nodules were observed using alizarin red S stain, while the content of matrix calcium was determined by atomic absorption spectrophotometry. Cell proliferation in the cultures was stimulated by IGF-1 at concentrations ranging from 1 to 200ng/mL, with the maximum effect observed at 100ng/mL. This effect was observed from day 1 and peaked at day 5, but decreased at day 7. At concentrations of 10ng/mL and 100ng/mL, IGF-1 significantly induced ALP activity, OC level, matrix calcium content, and nodule formation of the osteoblastic cells by 20-180% (P<0.05 or P<0.01), compared to controls. The results suggested that IGF-1 is an anabolic agent for the proliferation, differentiation, mineralisation and calcium content of dairy cow osteoblasts, and could therefore act as a potential treatment for the metabolic bone diseases in these animals.
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PMID:The stimulatory effect of insulin-like growth factor-1 on the proliferation, differentiation, and mineralisation of osteoblastic cells from Holstein cattle. 1840 40

Interleukin (IL)-6 has been considered as an osteolytic factor involved in periodontal disease. However, the function of IL-6 in osteoblastic differentiation of periodontal ligament cells is not clear. We examined the effects of IL-6 and its soluble receptor (sIL-6R) on osteoblastic differentiation of periodontal ligament cells. Osteoblastic differentiation was induced by ascorbic acid. Osteoblast markers, including alkaline phosphatase activity and Runx2 gene expression, were examined. The mechanism of action of IL-6 on osteoblastic differentiation was evaluated by insulin-like growth factor (IGF)-I production and specific inhibitors for the IL-6-signaling molecule. IL-6/sIL-6R enhanced alkaline phosphatase activity and Runx2. Alkaline phosphatase activity was reduced by anti-IGF-I antibody. Mitogen-activated protein kinase and Janus protein tyrosine kinase inhibitors diminished alkaline phosphatase induced by IL-6/sIL-6R. We conclude that IL-6/sIL-6R increases ascorbic-acid-induced alkaline phosphatase activity through IGF-I production, implying that IL-6 acts not only as an osteolytic factor, but also as a mediator of osteoblastic differentiation in periodontal ligament cells.
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PMID:IL-6 induces osteoblastic differentiation of periodontal ligament cells. 1880 47

Dermal papilla (DP) at the hair follicle base is important for hair growth. Recent studies demonstrated that mouse vibrissa DP cells can be cultured in the presence of fibroblast growth factor-2 (FGF-2), but lose expression of versican and their follicle-inducing activity during the culture, and that activation of the Wnt signal, which is inhibited by glycogen synthase kinase-3 (GSK-3), in the DP cells promotes hair growth activity. We therefore investigated the influence of a GSK-3 inhibitor, (2'Z,3'E)-6-bromoindirubin-3'-oxime (BIO), on the growth of human DP cells and mouse vibrissa follicles in culture. We first demonstrated that, similarly to mouse DP cells, human DP cells were able to be cultured up to 15 passages in the presence of FGF-2, and lost the expression of alkaline phosphatase (ALP). When human DP cells later than ten passages were treated with BIO, the expression of ALP as well as insulin-like growth factor-1 (IGF-1), another DP marker, was significantly elevated. Nuclear and perinuclear translocation of beta-catenin was also observed. We then cultured mouse vibrissa follicles. In the presence of BIO, the follicles could be maintained for at least 3 days without detectable regression of the hair bulbs. The morphology and ALP expression were well preserved. BIO successfully retrieved the expression of DP marker molecules, such as ALP and IGF-1 in cultured human DP cells, and maintained mouse hair bulbs. Thus, treatment with BIO may be useful to prepare DP cells with hair follicle-inducing activity.
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PMID:Inhibition of glycogen synthase kinase-3 enhances the expression of alkaline phosphatase and insulin-like growth factor-1 in human primary dermal papilla cell culture and maintains mouse hair bulbs in organ culture. 1923 12

Recent studies indicate that neonatal spermatogonial stem cells (SSCs) possess pluripotency. However, the mechanisms that regulate the pluripotent differentiation capacity of SSCs remain unclear. Here, we describe a new method to clonally derive pluripotent SSCs from neonatal mouse testis. By coculturing with testicular stromal cells, SSCs can be maintained and expanded in serum-free conditions. Unlike endogenous SSCs, these in vitro expanded SSCs showed strong alkaline phosphatase (AP) activity and displayed characteristics of embryonic stem cells and primordial germ cells, which were therefore designated as AP(+) germline stem cells (AP(+)GSCs). The pluripotency of AP(+)GSCs was confirmed by in vitro differentiation toward hepatic and neuronal lineages and formation of embryonic chimeras after injection into blastocysts. Further investigation revealed that insulin-like growth factor-1 (IGF-1) secreted from Leydig cells was a key factor involved in maintaining the pluripotency of AP(+)GSCs. The blockage of IGF-1 receptor phosphorylation and its downstream PI3K pathway by PPP or LY294002 dramatically reduced their AP activity and expression of pluripotent genes, such as Oct-4, Blimp1, and Nanog. In conclusion, the present study demonstrated that IGF-1 secreted by testicular Leydig cells plays an important role in maintaining the pluripotency of SSCs in culture, which provides an insight into the molecular mechanism underlying germ cell pluripotency.
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PMID:Pluripotency of mouse spermatogonial stem cells maintained by IGF-1- dependent pathway. 1924 85

The purpose of this study was to determine the effects of short-term supplementation with the active compounds in green tea on serum biomarkers in patients with prostate cancer. Twenty-six men with positive prostate biopsies and scheduled for radical prostatectomy were given daily doses of Polyphenon E, which contained 800 mg of (-)-epigallocatechin-3-gallate (EGCG) and lesser amounts of (-)-epicatechin, (-)-epigallocatechin, and (-)-epicatechin-3-gallate (a total of 1.3 g of tea polyphenols), until time of radical prostatectomy. Serum was collected before initiation of the drug study and on the day of prostatectomy. Serum biomarkers hepatocyte growth factor (HGF), vascular endothelial growth factor (VEGF), insulin-like growth factor (IGF)-I, IGF binding protein-3 (IGFBP-3), and prostate-specific antigen (PSA) were analyzed by ELISA. Toxicity was monitored primarily through liver function enzymes. Changes in serum components were analyzed statistically using the Wilcoxon signed rank test. Cancer-associated fibroblasts were treated with EGCG, and HGF and VEGF protein and mRNA levels were measured. HGF, VEGF, PSA, IGF-I, IGFBP-3, and the IGF-I/IGFBP-3 ratio decreased significantly during the study. All of the liver function tests also decreased, five of them significantly: total protein, albumin, aspartate aminotransferase, alkaline phosphatase, and amylase. The decrease in HGF and VEGF was confirmed in prostate cancer-associated fibroblasts in vitro. Our results show a significant reduction in serum levels of PSA, HGF, and VEGF in men with prostate cancer after brief treatment with EGCG (Polyphenon E), with no elevation of liver enzymes. These findings support a potential role for Polyphenon E in the treatment or prevention of prostate cancer.
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PMID:Tea polyphenols decrease serum levels of prostate-specific antigen, hepatocyte growth factor, and vascular endothelial growth factor in prostate cancer patients and inhibit production of hepatocyte growth factor and vascular endothelial growth factor in vitro. 1954 90

Growth factors play an important role in the complex cascade of tissue events in periodontal regeneration, although optimal methods of delivery remain to be identified. We hypothesize that multiple delivery of growth factors, particularly via a microparticle-containing scaffold, will enhance cellular events leading to periodontal regeneration. In this study, cellular responses of periodontal ligament fibroblasts (PDLFs) in scaffolds containing microparticles (MPs) loaded with either bone morphogenetic protein (BMP)-2, insulin-like growth factor (IGF)-1, or a mixture of both MPs were evaluated, and the dual-MP-containing scaffold exhibited the release of different proteins in a sustained and independent fashion. When PDLF-seeded scaffolds were cultured in a flow perfusion bioreactor, cell metabolism and proliferation of PDLFs were significantly increased within 3 days in all IGF-1-containing scaffolds compared with those in groups lacking IGF-1 and particulate delivery enhanced these effects between 3 and 7 days. The dual-MP-containing group showed the most positive results. Both the BMP-2-in-MP and IGF-1-in-MP groups showed greater effects of alkaline phosphatase activity, more osteocalcin and osteopontin production, and more calcium deposition compared with matched GF-adsorbed groups. All osteoblastic markers were at their highest in the dual-MP-containing group at all detected time points. The combined results suggest that our dual-MP-containing scaffold can be used as a cell vehicle to positively affect cell behavior, thus exhibiting the potential to be a candidate scaffold for future periodontal tissue engineering.
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PMID:In vitro cellular responses to scaffolds containing two microencapulated growth factors. 1956 Aug 14


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