ABSTRACT
AIMS: Abundance of connexin 43 (Cx43), a transmembrane protein that forms hemichannels (HCs) and gap junctions (GJs), is dynamically regulated in human gingival fibroblasts (GFBLs) during wound healing. This may be important for fast and scarless gingival wound healing as Cx43 is involved in key cell functions important during this process. Our aim was to uncover the factors that regulate Cx43 expression and abundance in GFBLs. We hypothesized that cytokines and growth factors released during wound healing coordinately regulate Cx43 abundance in GFBLs. RESULTS: TGF-ß1, -ß2, -ß3, PGE2 and IL-1ß significantly upregulated, while TNF-α and IFN-γ downregulated Cx43 in cultured GFBLs. TGF-ß1, -ß2, -ß3, IL-1ß and IFN-γ modulated Cx43 abundance at both mRNA and protein levels, while TNF-α and PGE2 regulated only Cx43 protein abundance, suggesting involvement of distinct transcriptional/post-transcriptional and translational/post-translational mechanisms, respectively. TGF-ß1-induced upregulation of Cx43 was mediated by TGFßRI (ALK5) and SMAD2/3 signaling, and this was potently suppressed by PGE2, IL-1ß, TNF-α and IFN-γ that inhibited SMAD2/3 phosphorylation. CONCLUSION: Regulation of Cx43 abundance in GFBLs involves transcriptional/post-transcriptional and translational/post-translational mechanisms that are distinctly modulated by an interplay between TGF-ß isoforms and PGE2, IL-1ß, TNF-α and IFN-γ.
Subject(s)
Connexin 43/genetics , Fibroblasts/drug effects , Gene Expression Regulation/drug effects , RNA, Messenger/genetics , Receptor, Transforming Growth Factor-beta Type I/genetics , Transforming Growth Factor beta1/pharmacology , Adolescent , Adult , Aged , Biological Assay , Connexin 43/metabolism , Dinoprostone/pharmacology , Female , Fibroblasts/cytology , Fibroblasts/metabolism , Gingiva/cytology , Gingiva/drug effects , Gingiva/metabolism , Humans , Interferon-gamma/pharmacology , Male , Primary Cell Culture , Protein Isoforms/pharmacology , RNA, Messenger/metabolism , Receptor, Transforming Growth Factor-beta Type I/metabolism , Signal Transduction , Smad2 Protein/genetics , Smad2 Protein/metabolism , Smad3 Protein/genetics , Smad3 Protein/metabolism , Tumor Necrosis Factor-alpha/pharmacology , Wound Healing/drug effectsABSTRACT
BACKGROUND AND OBJECTIVE: Elevated levels of matrix metalloproteinase-7 (MMP7) have been observed in serum samples of subjects with type 2 diabetes mellitus (T2DM) and in gingival tissues of subjects with periodontitis. The aim of the present study was to collect in vivo and in silico evidence on the role of MMP7 in the interplay between T2DM and generalized periodontitis (GP). MATERIAL AND METHODS: The extent of MMP7 expression and localization were immunohistochemically analyzed in gingival tissues of patients with GP with T2DM (T2DM/GP, n = 11), systemically healthy patients with GP (n = 7), and systemically and periodontally healthy controls (n = 11). An in silico network model was built to determine the interactions between MMP7 and T2DM pathways. Regulation of neutrophil transmigration by MMP7 was analyzed in a knock-out mice model. RESULTS: In human gingival tissues, the proportion of cells with robust MMP7 expression was elevated in patients with T2DM/GP in comparison to controls (P = .014). According to the in silico analysis, "hydroxyl radical" and "hydrogen peroxide" compounds were among the most central nodes of the network, and were within the shortest paths connecting "glucose" to "MMP7." In MMP7 knock-out mice, an intense accumulation of neutrophils was observed in the gingival epithelium as compared to wild-type mice (P = .0001). CONCLUSION: Elevated MMP7 expression in gingival tissues of patients with T2DM/GP is related to the activation of reactive oxygen species by hyperglycemia. Suppression of MMP7 expression results in impaired neutrophil transmigration in gingiva.
Subject(s)
Diabetes Mellitus, Type 2/metabolism , Matrix Metalloproteinase 7/metabolism , Periodontitis/metabolism , Adult , Aged , Animals , Case-Control Studies , Female , Humans , Immunoenzyme Techniques , Male , Mice , Mice, Knockout , Middle Aged , Periodontitis/diagnostic imaging , Radiography, Panoramic , TurkeyABSTRACT
BACKGROUND AND OBJECTIVE: Periodontitis is a highly prevalent chronic inflammatory disease that causes tooth loss, morbidity and confers an increased risk for systemic disease. Tissue destruction during periodontitis is due in large part to collagen-degrading matrix metalloproteinases (MMPs) released by resident cells of the periodontium in response to proinflammatory cytokines. Platelets are immune-competent blood cells with a newly recognized role in chronic inflammation; however, their role in the pathogenesis of periodontitis is undefined. Consequently, the objective of this study was to assess the effect of platelet factor 4 (PF4), a major platelet-derived cytokine, on MMP-1 (collagenase) expression in human gingival fibroblasts (HGFs). MATERIAL AND METHODS: HGFs were cultured in the presence or absence of recombinant PF4. Pro-MMP-1 secretion was quantified by enzyme-linked immunosorbent assay analysis of the cell culture supernatants. MMP-1 transcription was quantified by real-time polymerase chain reaction. Regulation of MMP-1 production by the p44/42 MAP kinase (MAPK) signaling pathway was examined in the presence or absence of PF4. RESULTS: Exposure to PF4 caused a ~ 2-3-fold increase in MMP-1 transcription and secretion from cultured HGFs. PF4 treatment also enhanced phosphorylation of p44/42 MAPK, which has been previously shown to induce MMP-1 expression in fibroblasts. Blockade of p44/42 MAPK signaling with the cell-permeant inhibitors PD98059 and PD184352 abrogated PF4-induced pro-MMP-1 transcription upregulation and release from cultured HGFs. CONCLUSION: We conclude that PF4 upregulates MMP-1 expression in HGFs in a p44/42 MAPK-dependent manner. These findings point to a previously unidentified role for platelets in the pathogenesis of periodontal diseases.
Subject(s)
Fibroblasts/drug effects , Fibroblasts/enzymology , Gingiva/cytology , Matrix Metalloproteinase 1/metabolism , Platelet Factor 4/pharmacology , Cells, Cultured , Electrophoresis, Polyacrylamide Gel , Enzyme-Linked Immunosorbent Assay , Humans , Immunoblotting , MAP Kinase Signaling System/drug effects , Phosphorylation , Real-Time Polymerase Chain Reaction , Signal Transduction , Up-RegulationABSTRACT
Microvesicles (MVs) are extracellular vesicles secreted by various cell types that are involved in intercellular communication. We hypothesized that in human periodontal disease, the pocket epithelium releases MVs, which then modulate gene expression in the underlying fibroblasts to control periodontal inflammation. MVs were isolated from culture medium of gingival epithelial cells (GECs) treated with oral bacterial biofilm extract or left untreated. Biofilm treatment significantly increased MV release from the GECs. Mass spectrometry of GEC-MVs identified a total of 2,173 proteins, of which about 80% were detected in MVs from both control and biofilm-treated GECs. Among 80 signature genes of human gingival fibroblasts, 20 were significantly regulated (P < 0.05) by MVs from control and biofilm-treated GECs in a similar manner. Matrix metalloproteinase 1 and 3 and interleukin 6 and 8 showed the strongest regulation at the mRNA and protein levels. Several cellular signaling pathways were activated by GEC-MVs in human gingival fibroblasts, including Smad and mitogen-activated protein kinase-associated pathways ERK1/2, JNK, and p38. However, ERK1/2 signaling dominated in the MV-induced gene expression changes. The results demonstrate that GEC-MVs have a strong regulatory effect on the expression of fibroblast genes associated with inflammation and matrix degradation and that bacterial biofilm stimulates the generation of GEC-MVs. This suggests that bacterial biofilms can contribute to the initiation and progression of periodontal disease by promoting a tissue-destructive phenotype in gingival fibroblasts via the enhanced secretion of epithelial MVs.
Subject(s)
Epithelial Cells/metabolism , Extracellular Vesicles/physiology , Fibroblasts/physiology , Gingiva/cytology , Periodontal Diseases/metabolism , Biofilms , Cells, Cultured , Enzyme-Linked Immunosorbent Assay , Gene Expression Regulation , Humans , Inflammation/metabolism , Interleukin-6/metabolism , Interleukin-8/metabolism , Mass Spectrometry , Matrix Metalloproteinase 1/metabolism , Matrix Metalloproteinase 3/metabolism , Microscopy, Electron , Microscopy, Fluorescence , Phenotype , Signal TransductionABSTRACT
Neural crest (NC)-derived stem cells (NCSC) have an exceptionally wide differentiation potential, but their use in regenerative therapy has been hampered by their scarcity in adult tissues and complex isolation protocols. Human oral mucosal gingiva may provide an attractive source of these cells as it contains NC-derived cells, the tissue is easily accessible and wound healing is fast and scarless with very little morbidity. To this end, we first investigated whether NC-derived cells are retained in adult gingiva by examining 8-months-old NC-reporter Wnt1-Cre/R26RYFP mice. We then hypothesised that gingival cell NC-like phenotype can be further enhanced by floating neurosphere cultures generated from standard human gingival fibroblast (GF) and pooled CFU-F (GSC) cultures. Findings showed that NC-derived cells are retained in the gingival connective tissue of aged mice. Human GFs and GSCs expressed NC-related genes nestin, Snai1, Twist1, Pax3, Sox9 and FoxD3, and generated neurospheres. This was mediated via calcium- and connexin 43-dependent cell communication, which is similar to neurospheres formed by neural progenitors. Cells in the spheres showed significantly increased expression of NC-related genes, and down regulation of fibroblast-related type I collagen. Structurally, the neurospheres were polarised with nestin positive cells located on the outer layers underlined with an extracellular matrix rich in molecules typical to embryonic NC. Sphere-derived cells expressed significantly elevated levels of neural markers, and differentiated into Tau, neurofilament-M and GFAP-positive cells suggesting neural differentiation potential. Thus, human GF and GSC cultures may provide an efficient source of NC-derived cells via enrichment by floating sphere cultures.
Subject(s)
Gingiva/cytology , Neural Crest/cytology , Neural Stem Cells/cytology , Spheroids, Cellular/cytology , Adolescent , Animals , Cell Culture Techniques , Cell Differentiation , Cells, Cultured , Connexin 43/genetics , Connexin 43/metabolism , Female , Fibroblasts/cytology , Humans , Male , Mice , RNA Interference , RNA, Messenger/biosynthesis , RNA, Small Interfering/geneticsABSTRACT
The resistance patterns of Campylobacter spp. isolated from retail broiler chicken meat originating either from Estonia, Lithuania or Latvia collected in Estonia were determined. Additionally, in collaboration with the laboratories of several Estonian hospitals, antimicrobial susceptibility patterns were determined for Campylobacter isolates from patients with severe Campylobacter enteric infections. The isolates were identified at the species level by the PCR method. Respectively, 88.8% of the isolates were C. jejuni, and 11.2% were C. coli. In total, 126 Campylobacter isolates of broiler chicken meat and human origin were tested for minimal inhibitory concentrations (MICs) with the broth microdilution VetMIC(TH) method (National Veterinary Institute; Uppsala, Sweden) for a total of six antimicrobials. Resistance to one or more antimicrobials was detected in 62 (63.3%) of Campylobacter broiler chicken meat isolates and in 20 (71.4%) of human-origin isolates. Large proportions of the broiler chicken meat isolates were resistant to ciprofloxacin (60.2%). Multidrug resistance (i.e. to three or more unrelated antimicrobials) was detected in five (5.1%) C. jejuni isolates. Among the human isolates, 20 (71.4%) were resistant to fluoroquinolones, and two (7.1%) C. jejuni isolates exhibited multidrug resistance. The chicken meat isolates of Estonian origin were the most susceptible. However, a high proportion of fluoroquinolone-resistant C. jejuni isolates were found in Latvian and Lithuanian products. The results of this study indicate that the problems caused by the inappropriate use of antimicrobials extend beyond the country in which a food originates; therefore, both domestic and international interventions and agreements are required to implement common policies on antimicrobial usage and to minimize the emergence of Campylobacter drug resistance.
Subject(s)
Anti-Bacterial Agents/pharmacology , Campylobacter Infections/microbiology , Campylobacter coli/drug effects , Campylobacter jejuni/drug effects , Meat/microbiology , Animals , Baltic States , Campylobacter/drug effects , Campylobacter/isolation & purification , Campylobacter Infections/drug therapy , Campylobacter Infections/transmission , Campylobacter coli/isolation & purification , Campylobacter jejuni/isolation & purification , Chickens , Drug Resistance, Multiple, Bacterial/drug effects , Food Contamination , Food Microbiology , Humans , Microbial Sensitivity Tests , Polymerase Chain Reaction , Quinolones/pharmacologyABSTRACT
Adhesion of epithelium to the extracellular matrix is crucial for the maintenance of systemic and oral health. In the oral cavity, teeth or artificial dental implants penetrate the soft tissue of the gingiva. In this interface, gingival soft tissue needs to be well attached via the epithelial seal to the tooth or implant surface to maintain health. After injury or wounding, epithelial tissue rapidly migrates to form the initial epithelial cover to restore the barrier against infection. These events are crucially dependent on deposition of extracellular matrix and proper activation and function of integrin receptors in the epithelial cells. Recent experimental evidence suggests that epithelial integrins also participate in the regulation of periodontal inflammation. In this review, we will discuss the structure and function of epithelial integrins and their extracellular ligands and elaborate on their potential role in disease and repair processes in the oral cavity.
Subject(s)
Epithelial Attachment/physiology , Integrins/physiology , Keratinocytes/physiology , Mouth Mucosa/chemistry , Wound Healing/physiology , Animals , Cell Adhesion , Epithelial Attachment/cytology , Extracellular Matrix Proteins/metabolism , Humans , Integrin alpha6beta4/metabolism , Integrins/chemistry , Keratinocytes/chemistry , Mouth Mucosa/cytology , Protein Binding , Protein Structure, Tertiary , Transforming Growth Factor beta1/metabolismABSTRACT
BACKGROUND: Previous studies suggest that offspring of mothers with psychotic disorders have an almost two-fold higher mortality risk from birth until early adulthood. We investigated predictors of mortality from late adolescence until middle age in offspring of mothers with psychotic disorders. METHOD: The Helsinki High-Risk Study follows up offspring (n=337) of women treated for schizophrenia spectrum disorders in mental hospitals in Helsinki before 1975. Factors related to mortality up to 2005 among offspring of these mothers was investigated with a survival model. Hazard rate ratios (HRR) were calculated using sex, diagnosis of psychotic disorder, childhood socio-economic status, maternal diagnosis, and maternal suicide attempts and aggressive symptoms as explanatory variables. The effect of family was investigated by including a frailty term in the model. We also compared mortality between the high-risk group and the Finnish general population. RESULTS: Within the high-risk group, females had lower all-cause mortality (HRR 0.43, p=0.05) and mortality from unnatural causes (HRR 0.24, p=0.03) than males. Having themselves been diagnosed with a psychotic disorder was associated with higher mortality from unnatural causes (HRR 4.76, p=0.01), while maternal suicide attempts were associated with higher suicide mortality (HRR 8.64, p=0.03). Mortality in the high-risk group was over two-fold higher (HRR 2.44, p<0.0001) than in the general population, and remained significantly higher when high-risk offspring who later developed psychotic disorders were excluded from the study sample (HRR 2.30, p<0.0001). CONCLUSIONS: Offspring of mothers with psychotic disorder are at increased risk of several adverse outcomes, including premature death.
Subject(s)
Child Mortality/trends , Child of Impaired Parents/statistics & numerical data , Mothers/psychology , Mothers/statistics & numerical data , Psychotic Disorders/epidemiology , Psychotic Disorders/psychology , Adolescent , Adult , Child, Preschool , Female , Humans , Infant , Infant, Newborn , Male , PrevalenceABSTRACT
BACKGROUND: Foetal nutrition and growth seem to affect the risk of developing schizophrenia. Exposure to famine during foetal development and low birthweight increase the risk. However, few studies have investigated the association between schizophrenia and adult height and weight or patterns of growth. METHOD: The study population consisted of two subpopulations: families with at least one member with schizophrenia, and families of offspring of mothers with psychotic disorder, and controls. Using a seven-parameter model of height growth curves, we compared the parameters of persons who later developed schizophrenia and their unaffected siblings from the same families. We also studied how growth curve parameters differed in children with genetic risk for schizophrenia and controls, and whether weight, height and body mass index (BMI) at different ages predicted later development of schizophrenia. RESULTS: The predicted growth curves based on a parametric model were nearly identical for persons with schizophrenia and their unaffected siblings. Adult height of daughters of mothers with psychoses was borderline significantly (p=0.0536) lower compared to controls, while no difference was detected among sons (p=0.3283). CONCLUSIONS: No association between growth characteristics and schizophrenia in families with at least one member with schizophrenia was found. Family-related factors should be taken into account as possible confounders in future studies on growth and schizophrenia.
Subject(s)
Growth/physiology , Schizophrenia/epidemiology , Adolescent , Adult , Age Factors , Body Height/genetics , Body Height/physiology , Body Mass Index , Body Weight/genetics , Body Weight/physiology , Child , Child Development/physiology , Child Health Services/statistics & numerical data , Child of Impaired Parents/psychology , Child of Impaired Parents/statistics & numerical data , Confounding Factors, Epidemiologic , Female , Finland/epidemiology , Genetic Predisposition to Disease/epidemiology , Genetic Predisposition to Disease/genetics , Growth/genetics , Humans , Linear Models , Longitudinal Studies , Male , Models, Genetic , Nuclear Family/psychology , Risk Factors , Schizophrenia/diagnosis , Schizophrenia/genetics , Siblings/psychologyABSTRACT
BACKGROUND AND OBJECTIVE: Gingival overgrowth is a side-effect of nifedipine and cyclosporin medications. Integrins are transmembrane glycoproteins that mediate cell adhesion, regulate cell proliferation and participate in the regulation of tissue fibrosis. The aim of this study was to investigate whether expression of epithelial cell integrins is linked to the development of drug-induced gingival overgrowth. MATERIAL AND METHODS: Human gingival biopsies of patients taking nifedipine, cyclosporin, or a combination of both medications, were used. Expression of the alpha5beta1, alphavbeta1 and alphavbeta6 integrins, and of cellular extra domain A of fibronectin, was localized in frozen sections using immunohistochemistry. RESULTS: The activated conformation of the beta1, alpha5beta1 and alphavbeta6 integrins were more frequently expressed in distinct locations in the oral epithelium in the combined drug group. Cellular extra domain A of fibronectin, a ligand for both alpha5beta1 and alphavbeta6 integrins, was expressed within the connective tissue of all groups. It was also expressed around the basal keratinocytes of the control, nifedipine and cyclosporin-induced gingival overgrowth groups, but not in the combined medication group. No relationship between the presence of inflammation and integrin expression was found. CONCLUSION: The results indicate that expression of certain integrins is up-regulated in the epithelium of drug-induced gingival overgrowth where they could participate in controlling the formation of elongated rete ridges and tissue fibrosis.
Subject(s)
Gingival Overgrowth/chemically induced , Gingival Overgrowth/metabolism , Integrins/biosynthesis , Adult , Analysis of Variance , Antigens, Neoplasm/biosynthesis , Calcium Channel Blockers/adverse effects , Cyclosporine/adverse effects , Drug Combinations , Female , Fibronectins/metabolism , Fluorescent Antibody Technique , Gingiva/metabolism , Humans , Immunosuppressive Agents/adverse effects , Integrin alpha5beta1/biosynthesis , Integrin beta1/biosynthesis , Keratinocytes/metabolism , Male , Nifedipine/adverse effects , Statistics, Nonparametric , Up-RegulationABSTRACT
Hereditary gingival fibromatosis (HGF) is a rare condition that can occur as an isolated disease or as part of a syndrome or chromosomal abnormality. In severe cases, the gingival enlargement may cover the crowns of teeth and cause severe functional and esthetic concerns. Histological and cell culture studies have uncovered some of the molecular and cellular changes associated with HGF. However, the pathogenesis of the disease is still largely unknown. Recent studies about the genetic characteristics of HGF have provided novel clues about the potential pathogenic mechanisms. In particular, mutation in the son-of-sevenless (SOS-1) gene has been associated with one form of the disease. However, HGF displays genetic heterogeneity, and mutations in other genes are also likely involved. This review outlines the current knowledge about the histological, cellular, and genetic characteristics of HGF. In addition, the potential role of the SOS-1 molecule and related novel intracellular signaling pathways in the pathogenesis of HGF will be discussed.
Subject(s)
Fibromatosis, Gingival/genetics , Cell Proliferation , Fibroblasts/pathology , Fibromatosis, Gingival/pathology , Genetic Heterogeneity , Humans , Mitogen-Activated Protein Kinases/genetics , Mutation/genetics , Receptors, Transforming Growth Factor beta/genetics , SOS1 Protein/genetics , Transforming Growth Factor beta/geneticsABSTRACT
AIMS: Interactions of cells with the extracellular matrix are important for normal wound healing and may play a role in scar formation. Remarkably, wound healing in human gingiva does not result in scar formation and serves as a model for wound regeneration. Endo180 (CD280) is a cell surface receptor that has novel functions to regulate cell migration and bind and internalize collagens that are key processes in wound healing. The aim of this study was to examine the expression of Endo180 during gingival wound regeneration. METHODS AND RESULTS: Biopsies were collected from normal human gingiva and 1-60 days after wounding and expression of Endo180 was analysed by immunostaining. Expression of Endo180 by cultured fibroblasts and keratinocytes was studied by immunoblotting and semiquantitative reverse transcriptase-polymerase chain reaction. In normal gingiva, Endo180 was expressed by basal epithelial cells, fibroblasts, myofibroblasts, pericytes, macrophages and endothelial cells. In wounds, Endo180 expression was spatiotemporally increased in the migrating and differentiating wound epithelium, in subsets of myofibroblasts, pericytes, macrophages and endothelial cells. Growth factors involved in wound healing up-regulated the expression of Endo180 in keratinocytes and fibroblasts. CONCLUSIONS: The findings suggest that Endo180 plays a role in re-epithelialization and connective tissue remodelling during wound regeneration.
Subject(s)
Gingiva/metabolism , Receptors, Mitogen/metabolism , Wound Healing/physiology , Cells, Cultured , Endothelium, Vascular/metabolism , Endothelium, Vascular/pathology , Epithelial Cells/metabolism , Epithelial Cells/pathology , Fibroblasts/metabolism , Fibroblasts/pathology , Fluorescent Antibody Technique, Indirect , Gene Expression Regulation , Gingiva/injuries , Gingiva/pathology , Humans , Keratinocytes/metabolism , Keratinocytes/pathology , Macrophages/metabolism , Macrophages/pathology , Pericytes/metabolism , Pericytes/pathology , RNA, Messenger/metabolism , Receptors, Mitogen/genetics , Time FactorsABSTRACT
BACKGROUND: Small leucine-rich proteoglycans (SLRPs) decorin, biglycan, fibromodulin and lumican are secreted extracellular matrix molecules that associate with fibrillar collagens and regulate collagen fibrillogenesis. Collagens are the major extracellular matrix components of periodontal connective tissues where they provide mechanical attachment of the tooth to the bone and gingiva and mediate signals that regulate cell functions, including remodeling of the periodontal ligament and bone. Structural organization of collagen may also be important for the defense against periodontal disease, because in certain conditions abnormal collagen fibrils associate with increased susceptibility to periodontal disease. OBJECTIVES: The purpose of this study was to find out the role of SLRPs to regulate collagen fibril and fibril bundle formation in periodontal tissues. METHODS: The localization of SLRPs in human and mouse periodontal tissues was studied using immunohistochemical methods. To assess the function of SLRPs we studied periodontal tissues of mice harboring targeted deletions of decorin, fibromodulin or lumican genes and lumican and fibromodulin double knockout mice using histological and electronmicroscopical methods. RESULTS: The SLRPs were coexpressed in human and mouse gingival and periodontal ligament connective tissues where they colocalized with collagen fibril bundles. Teeth in the knockout animals were fully erupted and showed normal gross morphology. Targeted deletion of decorin, fibromodulin, lumican or both lumican and fibromodulin resulted in abnormal collagen fibril and fibril bundle morphology that was most evident in the periodontal ligament. Each of the gene deletions resulted in a unique fibril and fibril bundle phenotype. CONCLUSIONS: These findings indicate that decorin, fibromodulin and lumican coordinately regulate the fibrillar and suprafibrillar organization of collagen in the periodontal ligament.
Subject(s)
Fibrillar Collagens/chemistry , Fibrillar Collagens/genetics , Periodontal Ligament/chemistry , Proteoglycans/genetics , Proteoglycans/physiology , Adolescent , Adult , Animals , Chondroitin Sulfate Proteoglycans/analysis , Chondroitin Sulfate Proteoglycans/genetics , Chondroitin Sulfate Proteoglycans/physiology , Decorin , Extracellular Matrix Proteins/analysis , Extracellular Matrix Proteins/genetics , Extracellular Matrix Proteins/physiology , Female , Fibromodulin , Gene Deletion , Humans , Immunoenzyme Techniques , Keratan Sulfate/analysis , Keratan Sulfate/genetics , Keratan Sulfate/physiology , Leucine , Lumican , Male , Mice , Mice, Knockout , Periodontal Ligament/physiology , Proteoglycans/analysisABSTRACT
BACKGROUND AND OBJECTIVE: Decorin, biglycan, fibromodulin and lumican are structurally related molecules that belong to the family of small leucine-rich proteoglycans (SLRPs). These SLRPs are secreted extracellular matrix molecules that interact with type I collagen and regulate collagen fibrillogenesis. They may also modulate cell functions that are important in maintenance of connective tissue structure. The aim of this study was to localize decorin, biglycan, fibromodulin and lumican in human gingiva. METHODS: Localization of decorin and its proform (prodecorin), biglycan, fibromodulin and lumican and mature and proform of type I collagen was studied by immunohistochemical staining of frozen tissue sections from healthy human attached gingiva. Double immunostaining with anti-SLRP or anti-type I procollagen antibodies and specific markers for different connective tissue cells was used to study association of these molecules with cells. RESULTS: The mature and proforms of decorin and collagen and biglycan, fibromodulin and lumican showed distinct localization in the extracellular matrix, where they associated with type I collagen fiber bundles. Prodecorin also localized to the epithelial basement membrane zone. Fibroblasts, myofibroblasts, endothelial cells and pericytes showed immunoreactivity for procollagen, prodecorin, biglycan and fibromodulin, whereas lumican associated with fibroblasts and myofibroblasts only. Biglycan and fibromodulin were also associated with macrophages. Basal epithelial cells of the gingival epithelium showed immunoreactivity for biglycan, fibromodulin and lumican. CONCLUSIONS: Decorin, biglycan, fibromodulin and lumican associate with type I collagen and may collaborate to regulate collagen fibrillogenesis in human gingiva. Each of the SLRPs showed a distinct association with different connective tissue cells, suggesting that the cells produce these molecules and/or that the cells interact with them. Localization of biglycan, fibromodulin and lumican at the epithelial cells suggests novel functions for these SLRPs in human gingival epithelium.
Subject(s)
Collagen Type I/analysis , Gingiva/chemistry , Procollagen/analysis , Proteoglycans/analysis , Adult , Biglycan , Chondroitin Sulfate Proteoglycans/analysis , Decorin , Extracellular Matrix/chemistry , Extracellular Matrix Proteins/analysis , Fibromodulin , Humans , Keratan Sulfate/analysis , Lumican , Staining and Labeling/methodsABSTRACT
Squamous cell carcinoma (SCC) of the oral cavity is a highly invasive tumour of stratified squamous epithelium that spreads through degradation of the basement membrane (BM) and extracellular matrix (ECM). There are currently no reliable tissue or serum markers to predict whether the tumour has metastasized at the time of diagnosis. Verrucous carcinoma (VC) of the oral cavity is a rare low-grade variant of oral SCC that penetrates into the subepithelial connective tissue. Many matrix metalloproteinases (MMPs), such as MMP-1, -2, -7, -9, -13, and -14, as well as integrin receptors have been implicated in cancer invasion. Integrin alphavbeta6 is induced in SCC and appears to be involved in up-regulation of MMP-9 expression by oral keratinocytes and promotion of their migration. The aim of this study was to investigate whether the pattern of MMP expression or that of alphavbeta6 integrin contributes to the differences in the biological behaviour of oral SCC and VC. The results show that the less aggressive nature of oral VC may be connected to its MMP expression profile. Typically, VCs were devoid of epithelial MMP-3, -7, -9, -12 and -13 expression, compared with SCCs. MMP-19 was expressed by epithelial keratinocytes in hyperproliferative areas of verrucous hyperplasia, VC, and SCC, but was absent in the invasive cancer cell nests of SCC. MMP-26 was expressed by hyperproliferative keratinocytes in VC as well as by invasive cancer cells in SCCs. MMP-10 was expressed widely in the epithelium of all SCC specimens. alphavbeta6 integrin expression was also detected in some cases of epithelial hyperplasia but was significantly more abundant in cancers at the invasive front. The absence of MMP-7, -9 and -12 from epithelial cells may serve as a good prognostic marker of non-invasive oral carcinoma. Blocking the activity of invasion-specific MMPs or alphavbeta6 integrin might offer novel therapeutic modalities in early-stage oral carcinoma.
Subject(s)
Carcinoma, Squamous Cell/genetics , Carcinoma, Verrucous/genetics , Matrix Metalloproteinase 7/analysis , Matrix Metalloproteinase 9/analysis , Metalloendopeptidases/analysis , Mouth Neoplasms/genetics , Antigens, Neoplasm/analysis , Biomarkers, Tumor/analysis , Carcinoma, Squamous Cell/enzymology , Carcinoma, Squamous Cell/pathology , Carcinoma, Verrucous/enzymology , Carcinoma, Verrucous/pathology , Cell Adhesion Molecules/analysis , Collagenases/analysis , Gene Expression Regulation, Neoplastic/genetics , Humans , Hyperplasia/enzymology , Hyperplasia/genetics , Hyperplasia/pathology , Immunohistochemistry/methods , In Situ Hybridization/methods , Integrins/analysis , Matrix Metalloproteinase 10 , Matrix Metalloproteinase 12 , Matrix Metalloproteinase 13 , Matrix Metalloproteinase 3/analysis , Matrix Metalloproteinases/analysis , Matrix Metalloproteinases, Secreted , Mouth Neoplasms/enzymology , Mouth Neoplasms/pathology , Neoplasm Invasiveness/pathology , Prognosis , KalininABSTRACT
Organotypic cultures have been used to study epithelial cell behavior for many years. The aim of this study was to develop an organotypic culture method that better mimics the three-dimensional morphology of interdigitating rete ridges and connective tissue papillae and that also conserves the basement membrane zone (BMZ). Bovine tongue mucosa connective tissue, separated from epithelium after 1 M NaCl incubation, was used as organotypic culture substratum for different human keratinocyte cell lines. Organotypic cultures were characterized by electron and immunofluorescence microscopy for expression of integrin subunits and extracellular matrix components. While spontaneously immortalized mucosal keratinocytes produced highly irregular stratified organotypic cultures, the normal human epidermal keratinocytes (NHEK) demonstrated culture morphology that resembled in vivo epidermis. However, in this model, the histomorphology, expression of differentiation markers involucrin, keratin 10 and 14, and integrins varied significantly between the cell lines. Some cultures appeared to have an extended survival since they were maintained up to 40 days without histological signs of degeneration. The ultrastructure of the BMZ including hemidesmosomes was similar to the normal dermo-epidermal junction. Extracellular matrix molecules, including tenascin, laminin-1 and -5, were expressed in the cultures demonstrating their secretion solely by keratinocytes. Distribution and expression of integrins in NHEK cultures was similar to that seen in vivo skin with the exception of additional expression of alpha5beta1 and alpha(v)beta6 integrins. Organotypic NHEK cultures show similarities to normal stratified epithelium and are potentially useful for multiple applications for studies on epithelial cell behavior in vitro.
Subject(s)
Keratinocytes/ultrastructure , Mouth Mucosa/cytology , Mouth Mucosa/ultrastructure , Tongue/cytology , Tongue/ultrastructure , Animals , Basement Membrane/ultrastructure , Biomarkers , Cattle , Cell Differentiation/physiology , Cell Line , Cells, Cultured , Extracellular Matrix/metabolism , Extracellular Matrix/ultrastructure , Gingiva/cytology , Gingiva/ultrastructure , Integrins/biosynthesis , Microscopy, Electron , Microscopy, Electron, Scanning , Organ Culture Techniques , Tissue FixationABSTRACT
BACKGROUND: Various approaches to treating the periodontal condition associated with Papillon-Lefèvre syndrome have been reported. These include oral hygiene instruction, use of mouthrinses, frequent debridement, multiple antibiotic regimens, periodontal surgery, extraction of hopeless teeth, and extraction of all deciduous teeth. Because Papillon-Lefèvre syndrome is rare, most publications are case reports, and very few document long-term successful treatment of the periodontal condition. METHODS: In 1986, a 3.5-year-old Indo-Canadian male was diagnosed with Papillon-Lefèvre syndrome and began periodontal treatment. Initial therapy consisted of debridement every 3 weeks, a 0.12% chlorhexidine mouthrinse, 2 regimens of metronidazole, and oral hygiene instruction for his parents. After 10 months it became apparent that the treatment was having little beneficial effect, since the periodontal destruction continued and teeth 51 and 61 exfoliated. At age 4, all remaining deciduous teeth were extracted and complete dentures inserted for the following 2-year edentulous period; then a 3-month maintenance schedule was maintained. RESULTS: The patient is now 17 years old and all his adult teeth are present with the exception of the third molars. His oral hygiene varies between moderate and good, with his most recent plaque score at 80% effectiveness. There are no probing depths greater than 4 mm, with the exception of the distal of the lower second molars where opercula are present. CONCLUSIONS: Extraction of all the deciduous teeth followed by a period of edentulousness may partially explain the fact that there has been no recurrent attachment loss in the permanent teeth up to age 17. Other explanations are discussed as part of the literature review of Papillon-Lefèvre syndrome.
Subject(s)
Papillon-Lefevre Disease/complications , Periodontal Diseases/prevention & control , Adolescent , Anti-Bacterial Agents/therapeutic use , Dental Plaque/prevention & control , Follow-Up Studies , Humans , Male , Mouthwashes/therapeutic use , Oral Hygiene , Patient Education as Topic , Periodontal Pocket/prevention & control , Tooth Eruption , Tooth Extraction , Tooth, Deciduous/surgeryABSTRACT
BACKGROUND: Historically, animal models for the study of periodontal diseases have incorporated surgically created defects, plaque retentive ligatures, as well as soft and high-sucrose diets which may not accurately reflect progression of the natural disease. Spontaneous periodontal disease is seen in a few animal species, but these are often expensive to maintain and are unsuitable for manipulation using advanced molecular biology techniques. Mice are inexpensive, easy to maintain, and are routinely used for transgenic experiments and are therefore an optimal animal for research purposes. However, it is commonly accepted that mice do not spontaneously develop periodontal disease. The purpose of this study was to determine if a mouse population that exhibits periodontal breakdown in the wild could be found, allowing for genetic manipulation of naturally occurring periodontal disease. METHODS: We examined over 2,500 dry skulls of several Peromyscus species from various locations and habitats on the west coast of North America for periodontal bone loss in the molars, using furcation involvement as an indicator of disease severity. Alveolar bone loss was classified as Grade I) horizontal component of bone loss in the furcations; II) through-and-through furcations; and III) through-and-through furcations with alveolar bone loss into the apical third of the root. RESULTS: The proportions of individual mice experiencing bone loss were 3.8% for Class I-III involvement, 1.3% for Class II-III involvement, and 0.5% for Class III alone. Three subspecies of P. keeni and one subspecies of P. maniculatus had periodontal disease prevalences in 7% to 13.5% of their samples. Mice from isolated islands had 1.8- to 4.7-fold higher disease prevalence than those located on the mainland, with even greater prevalence on small islands. No statistically significant differences between genders were found. CONCLUSIONS: It appears that periodontal disease is far more common in this mouse genus than previously believed. Some of the subspecies demonstrated severe periodontal disease at a prevalence comparable to that found in humans.
Subject(s)
Alveolar Bone Loss/veterinary , Peromyscus/classification , Alveolar Bone Loss/classification , Animals , British Columbia , Chi-Square Distribution , Disease Models, Animal , Female , Furcation Defects/classification , Furcation Defects/veterinary , Male , Pacific States , Peromyscus/genetics , Sex FactorsABSTRACT
During wound-healing, cells are required to migrate rapidly into the wound site via a proteolytically generated pathway in the provisional matrix, to produce new extracellular matrix, and, subsequently, to remodel the newly formed tissue matrix during the maturation phase. Two classes of molecules cooperate closely to achieve this goal, namely, the matrix adhesion and signaling receptors, the integrins, and matrix-degrading and -processing enzymes, the matrix metalloproteinases (MMPs). There is now substantial experimental evidence that blocking key molecules of either group will prevent or seriously delay wound-healing. It has been known for some time now that cell adhesion by means of the integrins regulates the expression of MMPs. In addition, certain MMPs can bind to integrins or other receptors on the cell surface involved in enzyme activation, thereby providing a mechanism for localized matrix degradation. By proteolytically modifying the existing matrix molecules, the MMPs can then induce changes in cell behavior and function from a state of rest to migration. During wound repair, the expression of integrins and MMPs is simultaneously up-regulated. This review will focus on those aspects of the extensive knowledge of fibroblast and keratinocyte MMPs and integrins in biological processes that relate to wound-healing.