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)

This study was designed to examine developing acellular cementum in rat molars by immunohistochemistry, to elucidate (1) how Hertwig's epithelial root sheath disintegrates and (2) whether epithelial sheath cells transform into cementoblasts through epithelial-mesenchymal transition (EMT). Initial acellular cementogenesis was divided into three developmental stages, which can be seen in three different portions of the root: portion 1, where the epithelial sheath is intact; portion 2, where the epithelial sheath becomes fragmented; and portion 3, where acellular cementogenesis begins. Antibodies against three kinds of matrix proteinases, which degrade epithelial sheath-maintaining factors, including basement membrane and desmosomes, were used to investigate proteolytic activity of the epithelial sheath. Tissue non-specific alkaline phosphatase (TNALP) and keratin were used to investigate EMT. Epithelial sheath cells showed immunoreactivity for all three enzymes at fragmentation, which suggests that epithelial sheath disintegration is enzymatically mediated. Dental follicle cells and cementoblasts showed intense immunoreactivity for TNALP, and from portion 1 through to 3, the reaction extended from the alveolar bone-related zone to the root-related zone. Cells possessing keratin/TNALP double immunoreactivity were virtually absent. Keratin-positive epithelial sheath cells showed negligible immunoreactivity for TNALP, and epithelial cells did not appear to migrate to the dental follicle. Together, these findings suggest that a transition phenotype between epithelial cells and cementoblasts does not exist in the developing dental follicle and hence that epithelial sheath cells do not undergo EMT during initial acellular cementogenesis. In brief, this study supports the notion that cementoblasts derive from the dental follicle.
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PMID:Hertwig's epithelial root sheath cell behavior during initial acellular cementogenesis in rat molars. 2485 38

Several techniques have been employed to detect proteins on membranes. These include the use of quantum dot luminescent labels, oxyblot immunochemical detection, polymer immunocomplexes, "coupled" probing approach, in situ renaturation of proteins for detecting enzyme activities in crude or purified preparations, immunochromatographic assay, western-phosphatase assay, and use of Congo red dye (a cosmetic color named Alta), Pro-Q Emerald 488 dye, or amine-reactive dye in combination with alkaline phosphatase- and horseradish peroxidase-antibody conjugates for the simultaneous trichromatic fluorescence detection of proteins. Several methods have been used to improve the detection of proteins on membranes, including glutaraldehyde treatment of nitrocellulose blots, elimination of keratin artifacts in immunoblots probed with polyclonal antibodies, and washing of immunoblots with excessive water and manipulation of Tween-20 in wash buffer. These methods are briefly reviewed in this chapter.
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PMID:Other Notable Methods of Membrane Protein Detection: A Brief Review. 2613 83

To elucidate the fate of the epithelial root sheath during initial cellular cementogenesis, we examined developing maxillary first molars of rats by immunohistochemistry for keratin, vimentin, and tissue non-specific alkaline phosphatase (TNALP) and by TdT-mediated dUTP nick end labeling (TUNEL). The advancing root end was divided into three sections, which follow three distinct stages of initial cellular cementogenesis: section 1, where the epithelial sheath is intact; section 2, where the epithelial sheath becomes fragmented; and section 3, where initial cellular cementogenesis begins. After fragmentation of the epithelial sheath, many keratin-positive epithelial sheath cells were embedded in the rapidly growing cellular cementum. A few unembedded epithelial cells located on the cementum surface. Dental follicle cells, precementoblasts, and cementoblasts showed immunoreactivity for vimentin and TNALP. In all three sections, there were virtually no cells possessing double immunoreactivity for vimentin-keratin or TNALP-keratin and only embedded epithelial cells showed TUNEL reactivity. Taken together, these findings suggest that: (1) epithelial sheath cells divide into two groups; one group is embedded in the cementum and thereafter dies by apoptosis, and the other survives on the cementum surface as epithelial cell rests of Malassez; and (2) epithelial sheath cells do not undergo epithelial-mesenchymal transition during initial cellular cementogenesis.
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PMID:Hertwig's Epithelial Root Sheath Fate during Initial Cellular Cementogenesis in Rat Molars. 2616 Sep 88

The tooth enamel organ (EO) is a complex epithelial cell assembly involved in multiple aspects of tooth development, including amelogenesis. The present study focuses on the role of the nonameloblast layers of the EO, the stratum intermedium, the stellate reticulum, and the outer enamel epithelium (OEE). The secretory stage stratum intermedium was distinguished by p63-positive epithelial stem cell marks, highly specific alkaline phosphatase labeling, as well as multiple desmosomes and gap junctions. At the location of the presecretory stage stellate reticulum, the pre-eruption EO prominently featured the papillary layer (PL) as a keratin immunopositive network of epithelial strands between tooth crowns and oral epithelium. PL cell strands contained numerous p63-positive epithelial stem cells, while BrdU proliferative cells were detected at the outer boundaries of the PL, suggesting that the stellate reticulum/PL epithelial cell sheath proliferated to facilitate an epithelial seal during tooth eruption. Comparative histology studies demonstrated continuity between the OEE and the general lamina of continuous tooth replacement in reptiles, and the outer layer of Hertwig's epithelial root sheath in humans, implicating the OEE as the formative layer for continuous tooth replacement and tooth root extension. Cell fate studies in organ culture verified that the cervical portion of the mouse molar EO gave rise to Malassez rest-like cell islands. Together, these studies indicate that the nonameloblast layers of the EO play multiple roles during odontogenesis, including the maintenance of several p63-positive stem cell reservoirs, a role during tooth root morphogenesis and tooth succession, a stabilizing function for the ameloblast layer, the facilitation of ion transport from the EO capillaries to the enamel layer, as well as safe and seamless tooth eruption.
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PMID:Daughters of the Enamel Organ: Development, Fate, and Function of the Stratum Intermedium, Stellate Reticulum, and Outer Enamel Epithelium. 2761 44

Effects of bone morphogenetic protein (BMP)-4 on proliferation and differentiation capacities of dental pulp cells through BMP-4 acting on human dental pulp cells cultured in vitro were investigated. Dental pulp tissues of lesion-free teeth extracted from patients due to orthodontics were taken, and human dental pulp cells were cultured in vitro using the tissue explant method. Immunocytochemical staining was used for the identification of vimentin and keratin. The dental pulp cells were divided into groups A and B. A total of 100 ng/ml BMP-4 was added into group A, while no inducer was added into group B as the control group. The cell growth curves at day 1, 2, 3, 5 and 7 after culture were drawn. At day 7, the cell count, alkaline phosphatase (ALP) activity, number of calcified nodules, and expression levels of dentin sialophosphoprotein (DSPP), dentin matrix protein-1 (DMP-1) and each gene related to dentinogenesis in each group were detected, respectively. Human dental pulp cells were conformed to the biological characteristics of dental pulp cells according to the identification of vimentin and keratin via immunocytochemical staining. With the prolongation of culture time, the number of cells in both groups was gradually increased, reaching the peak at day 5 and began to decline at day 7. The number of cells in group A was significantly greater than that in group B (p<0.05). According to the results of reverse transcription-quantitative polymerase chain reaction (RT-qPCR), the relative messenger ribonucleic acid (mRNA) expression levels of ALP, DSPP and DMP-1 in group A were significantly higher than those in group B (p<0.05). BMP-4 can promote the growth of dental pulp cells and remarkably enhance the differentiation of dental pulp cells into odontoblasts.
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PMID:Expression and influence of BMP-4 in human dental pulp cells cultured in vitro. 3054 66

Keratin has intrinsic biocompatibility and contains several peptide-binding motifs that support the attachment of a wide variety of cell types. We have previously shown that keratin extracted from human hair can promote cell adhesion and proliferation of 3T3 fibroblasts, MG63 osteoblasts, and human adipose stem cells (hASCs). Despite its bioactivity advantages, keratin possesses fragile mechanical properties that introduce challenges for tissue engineering. To remedy this, we examined the results of combining keratin with chitosan, a combination facilitated via induction of an azide functional group, which acted as a photocrosslinker, to improve mechanical strength. Analysis of the keratin/chitosan composite showed that films of this material demonstrated good adhesion and promoted the proliferation of human adipose stem cells. Most importantly, this biomaterial was shown to promote the osteogenic differentiation of hASCs, in terms of up-regulations in type I collagen, runt-related transcription factor 2, and alkaline phosphatase gene expression. We further demonstrated that lyophilizing the keratin/chitosan forms highly interconnected and porous scaffolds that might provide an ideal environment for tissue culture. We believe that keratin/chitosan composite biomaterials can be used in bioactive surface modification, and the crosslinkable properties can produce natural polymer 3D scaffolds for the application of tissue engineering research.
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PMID:Keratin/chitosan UV-crosslinked composites promote the osteogenic differentiation of human adipose derived stem cells. 3226 4


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