Subject(s)
Chlorides/adverse effects , Dental Restoration, Permanent/adverse effects , Dermatitis, Allergic Contact/diagnosis , Dermatitis, Allergic Contact/etiology , Drug Eruptions/diagnosis , Drug Eruptions/etiology , Zinc Compounds/adverse effects , Adrenal Cortex Hormones/therapeutic use , Adult , Chlorides/blood , Dermatitis, Allergic Contact/drug therapy , Disease Progression , Drug Eruptions/drug therapy , Eosinophilia/diagnosis , Eosinophilia/drug therapy , Eosinophilia/etiology , Humans , Male , Patch Tests , Zinc Compounds/bloodABSTRACT
Coexpression of desmosomal proteins and vimentin has been reported in a specific mesenchymal phenotype. This study investigated the expression of vimentin-binding desmosomal proteins in human dental pulp fibroblasts (DPF) and odontoblasts. The dental pulp has no cells expressing desmocollin (DSC) 1-3, desmoglein (DSG) 1-3, junction plakoglobin (JUP), or desmoplakin (DPK) 1 and 2 except for odontoblasts expressing DPK. A confocal image by laser-scanning microscopy demonstrated the diffuse distribution of DPK in the cytoplasm throughout the odontoblast processes. In culture, the mRNA expression of JUP and DPK1, but not DSC1-3 and DSG1-3, was detected in all DPF clones tested and also in odontoblast-like cells (OB) expressing osteocalcin and dentin sialophosphoprotein mRNAs established in the differentiation medium. The DPF having the potential to differentiate into OB expressed vimentin, but not DPK before culturing in the differentiation medium, whereas OB expressed vimentin-binding DPK1. These results suggest that DPF usually expresses DPK1 mRNA, and that the DPK1 production and the bonding of vimentin to DPK1 occur in DPF with the differentiation into odontoblasts.
Subject(s)
Cytoskeletal Proteins/metabolism , Odontoblasts/metabolism , Adolescent , Adult , Cells, Cultured , Dental Pulp/metabolism , Dental Pulp/ultrastructure , Desmocollins , Desmoglein 1 , Desmogleins , Desmoplakins , Desmosomes/metabolism , Humans , Immunohistochemistry , Microscopy, Confocal , Odontoblasts/ultrastructure , Reverse Transcriptase Polymerase Chain Reaction , gamma CateninABSTRACT
The cys-cys (C-C) chemokine ligand 21 is a member of the C-C chemokines that constitute a group of heparin-binding cytokines with a pattern of four or six conserved cysteines. The CCL21 is known to be expressed in secondary lymphoid tissues, however it has rarely been reported for the expression on peripheral lymphatic vessels in somatic tissue. Here we investigated the expression of CCL21 on lymphatic vessels identified by anti-desmoplakin in uninflamed and inflamed human gingiva. In uninflamed tissue the expression of CCL21 was detected on lymphatic vessels in gingiva. In uninflamed gingiva the expression of CCL21 was detected on all lymphatic capillaries of the mucosal connective tissue papillae. There were two types of collecting lymphatic vessels in the lamina propria mucosae expressing CCL21 strongly or very weakly. In inflamed gingiva no expression of CCL21 was detected on lymphatic vessels. In all tissue sections no blood vessels expressing CCL21 were observed. These results may suggest that the expression of CCL21 is predominantly induced in the peripheral lymphatic endothelium of the uninflamed mucosal microcirculation, and that under inflamed conditions a reduction of CCL21 occurs in lymphatic endothelium.
Subject(s)
Angiogenesis Inhibitors/biosynthesis , Chemokines, CC/biosynthesis , Gingiva/metabolism , Lymphatic Vessels/metabolism , Lymphatic Vessels/pathology , Periodontitis/metabolism , Adolescent , Adult , Chemokine CCL21 , Cytoskeletal Proteins/biosynthesis , Desmoplakins , Endothelium, Lymphatic/metabolism , Endothelium, Lymphatic/pathology , Endothelium, Vascular/metabolism , Endothelium, Vascular/pathology , Female , Gene Expression Regulation/physiology , Gingiva/pathology , Humans , Male , Middle Aged , Mouth Mucosa/metabolism , Mouth Mucosa/pathology , Periodontitis/pathologyABSTRACT
The Toll-like receptor (TLR) is a part of the innate immune system sensing pathogen-associated molecular patterns (PAMPs). Recently, TLRs 2 and 4 have been demonstrated for the ligand engagements, which result in the induction of cytokines. Here we investigated the expression of TLRs 2 and 4 on lymphatic vessels producing cys-cys chemokine ligand 21 (CCL21) in the human small intestine. The specificity of antibodies to TLRs was tested on a human monocyte leukemia cell line, umbilical vein endothelial cells (HUVEC), and periodontal ligament fibroblasts (PDLF) with the examination for the TLR gene expression by the reverse transcription-polymerase chain reaction (RT-PCR), and lymphatic vessels were identified by antibodies for platelet-endothelial cell adhesion molecule-1 (PECAM-1) and desmoplakin. The expression of CCL21 was not clearly detected on collecting lymphatic vessels in the submucosa while it was generally observed on the central lacteals of villi and lymphatic capillaries in the lamina propria mucosae. The reaction of antibodies to TLRs 2 and 4 was also not clearly detected on collecting lymphatic vessels in the submucosa and central lacteals of villi, but generally observed on lymphatic capillaries expressing CCL21 in the lamina propria mucosae of tissue where the expression of CCL21 and TLRs was not clearly observed in blood vessels. These may suggest that the expression of CCL21, and TLRs 2 and 4 is predominantly induced in the peripheral lymphatic endothelium of the small intestinal microcirculation. The lymphatic endothelium may contribute to allow dendritic cells to home into secondary lymphoid tissue through the expression of TLRs, the ligand engagements of which result in the induction of chemokines.
Subject(s)
Endothelium, Lymphatic/metabolism , Intestinal Mucosa/metabolism , Membrane Glycoproteins/biosynthesis , Receptors, Cell Surface/biosynthesis , Aged , Cell Line, Tumor , Cells, Cultured , Chemokine CCL21 , Chemokines, CC/biosynthesis , Gene Expression Regulation, Neoplastic , Humans , Lipids/chemistry , Lymphatic Vessels/metabolism , Middle Aged , Platelet Endothelial Cell Adhesion Molecule-1/metabolism , Reverse Transcriptase Polymerase Chain Reaction , Toll-Like ReceptorsABSTRACT
The osteogenic cell type of human periodontal ligament fibroblasts (PDLF) undergoes senescence at finite population doubling numbers unrelated to donor ages. This study investigated telomere lengths of osteogenic PDLF from differently aged donors and alterations of the osteoblast-like properties in the aged PDLF with short telomeres. Telomere lengths of osteogenic PDLF were biased towards long or short among all 15- to 51-year-old individuals, and did not show a normal distribution by Pearson's test or a correlation to donor age by simple regression analysis. In osteogenic PDLF, senescence-associated beta-galactosidase was expressed in 78.5% of cells in the clones with short telomeres (mean 3.02 kbp), and in 9.4% of cells in the clones with long telomeres (mean 13.06 kbp). These results suggest that human periodontium comprises aged osteogenic PDLF without correlation to age. Osteogenic PDLF with long telomeres strongly expressed alkaline phosphatase (ALPase) activity whereas cells with short telomeres expressed ALPase activity to a weaker extent. Total activity of ALPase in the clones of osteogenic PDLF with long telomeres was significantly higher than that in the clones with short telomeres. The produced amounts of both osteopontin and osteocalcin in the clones of osteogenic PDLF with long telomeres were slightly but statistically significantly smaller than those in the clones with short telomeres. These findings suggest that aged osteogenic PDLF reduce the expression of ALPase activity but that there is not a critical alteration in bone-associated protein production. Aged osteogenic PDLF may impair the ability to induce ALPase-dependent calcification.
