PubMed Health⌕ Search

Biomedical subjects

H Larjava

Publications and source records attributed to H Larjava.

At least 73 records · Page 4Linked to original sources
PubMed1990

Differential localization of type I and type III procollagen messenger ribonucleic acids in inflamed periodontal and periapical connective tissues by in situ hybridization.

Inflammatory lesions of periodontal and periapical connective tissue were studied by in situ hybridization to detect cells responsible for type I and type III collagen production. Formalin-fixed and paraffin-embedded tissue specimens from patients with oral lesions of various stages of inflammation were hybridized with cDNA probes specific for human pro alpha 1(I) and pro alpha 1(III) collagen mRNAs, and with bacteriophage lambda DNA as a control probe. This technique permitted us to localize fibroblasts active in type I collagen synthesis in the vicinity of inflammatory infiltrates in all the samples studied. Cells containing high levels of type III collagen mRNA were seen in early abscess formation and they were particularly abundant in pyogenic granuloma and irritation fibroma. Type I collagen mRNA was prominent in gingival fibrosis. In the infrabony lesions with active inflammatory infiltrations the production of collagen was confined mostly to the periphery of the lesions. These findings give indirect evidence that cytokines liberated during the early stages of the inflammatory process stimulate expression of the type III collagen gene by fibroblasts. In chronic lesions a gradual switch from type III to type I collagen gene expression occurs. The change in collagen types appears to underlie the observed isolation of the inflammation by a collagenous capsule. In all the samples studied fibroblasts exhibited marked variation in their levels of procollagen mRNAs, supporting previous views about their heterogeneity in connective tissues. The approach presented here offers new possibilities to study cellular interactions and metabolic activities in inflammatory lesions.

Adolescent↗
PubMed1989

Gingival crevicular fluid fibronectin degradation in periodontal health and disease.

The molecular forms of fibronectin (FN) in gingival crevicular fluid of five subjects with at least two sites exhibiting clinical signs of inflammation and pockets of at least 4 mm (test group) and five subjects with clinically healthy periodontium (control group) were investigated. Samples were collected with standard filter paper strips. In the test group samples from both diseased and healthy sites were collected. After collection the test group received one episode of periodontal treatment (scaling and root planning). The sampling and clinical recording were repeated for the diseased sites after about 2 wk. The crevicular fluid FN was analyzed using sodium dodecyl sulphate gel electrophoresis followed by western blotting with polyclonal antibodies against FN. Both intact FN and FN fragments were found in all samples. A larger proportion of FN was in degraded form in the diseased sites than in the healthy or the treated sites. FN was also degraded into smaller peptide fragments in the diseased than in the treated sites. These results suggest that crevicular fluid FN is partially degraded both in periodontal health and disease and that the degree of degradation of FN increases with periodontal inflammation and decreases with periodontal treatment.

Adult↗
PubMed1989

Altered distribution of type I collagen mRNA in periodontal disease.

Earlier studies on collagen metabolism in the periodontium have suggested that periodontal disease is associated with reduced amounts of type I collagen in the pocket walls, and with an overall increase in collagen synthesis. In this study we analyzed gingival biopsies of patients suffering from adult type chronic periodontal disease by in situ hybridization, which permitted localization of fibroblasts containing different amounts of proa(I) collagen mRNA. The results showed that expression of type I collagen mRNA is reduced in the vicinity of periodontal pockets and dental plaque. Deeper in the connective tissue, high levels of proa(I) collagen mRNA were observed particularly around inflammatory cell infiltrates which may contribute to the fibrotic reaction observed. In all areas studied the levels of type I collagen mRNA in periodontal fibroblasts varied considerably, supporting previous views about their heterogeneity.

Collagen↗
PubMed1989

A protease of Bacteroides gingivalis degrades cell surface and matrix glycoproteins of cultured gingival fibroblasts and induces secretion of collagenase and plasminogen activator.

To assess the direct effects of Bacteroides gingivalis on periodontal cells, human gingival fibroblasts were cultured in the presence of B. gingivalis extracts or a trypsinlike enzyme partially purified from the bacteria by chromatography on benzamidine-Sepharose and Sephacryl S-200. Analysis of cell surface glycoproteins by the periodate-[3H]borohydride labeling technique combined with sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE)-fluorography demonstrated that fibronectin and some other high-molecular-weight cell surface glycoproteins were degraded by a 35,000-Mr(35K) B. gingivalis protease. Immunostaining of the fibroblast cultures showed degradation of intercellular matrix fibronectin by the 35K protease. The pattern of fibronectin degradation was monitored by examining the reaction products with the SDS-PAGE-immunoblotting technique. The protease degraded fibronectin rapidly and more extensively than did corresponding amounts of pancreatic trypsin. Collagenase secretion by the fibroblasts was assayed by incubating cell culture medium with soluble type I [3H]collagen at 25 degrees C followed by SDS-PAGE-fluorography analysis of the reaction products. The medium was also assayed for plasminogen activator activity by using a casein-agarose diffusion plate assay. The fibroblasts cultured with the 35K protease secreted increased amounts of collagenase and plasminogen activator into the medium. The results suggest that periodontal infection by B. gingivalis causes proteolytic damage of the host cell surface structures. Concomitantly, B. gingivalis may induce the cells to degrade their pericellular matrix.

Bacteroides↗
PubMed1989

Localization of integrin receptors for fibronectin, collagen, and laminin in human skin. Variable expression in basal and squamous cell carcinomas.

VLA integrins in human skin were examined by indirect immunofluorescence utilizing antibodies recognizing the beta 1, alpha 2, alpha 3, or alpha 5 subunits. Staining of fetal, newborn, or adult skin with antibodies to beta 1, alpha 2, or alpha 3 subunits gave essentially similar staining patterns: intense staining was associated with the basal layer of the epidermis, hair follicles, and blood vessel walls. The alpha 5 subunit could be detected only in epidermis and the inner root sheath of hair follicles in fetal skin. In epidermis, the staining reaction for the beta 1 subunit was not only found in sites interfacing with the basement membrane zone, but also around the entire periphery of these cells. We speculate that these receptors might have previously unrecognized functions in cell-cell interactions or that these findings may suggest the presence of previously unrecognized ligands in the intercellular spaces of keratinocytes. Examination of nine nodular basal cell carcinomas revealed a prominent staining reaction with anti-beta 1 and anti-alpha 3 antibodies at the periphery of the tumor islands. In contrast, staining of five squamous cell carcinomas revealed either the absence of integrins or altered and variable expression. Thus, matrix components and their receptors may participate in modulation of growth, development, and organization of human skin.

Adult↗
PubMed1989

Characterization of one phenotype of human periodontal granulation-tissue fibroblasts.

Human granulation-tissue fibroblasts were cultured from oral chronic inflammatory lesions and compared with fibroblasts of healthy gingival connective tissue with respect to cell-surface sialoglycoproteins, and the synthesis of extracellular matrix components. Granulation-tissue fibroblasts exhibited a slower growth rate and larger size than their controls. Their cell-surface sialoglycoproteins resembled those of the control cells, except that the relative amount of glycoproteins in the 140-kd region was lower. The ratio of mRNAs for pro alpha l (I) and pro alpha l (III) collagen chains was decreased in granulation-tissue fibroblasts, although electrophoretic fractionation of the proteins did not reveal consistent differences in type I/type III collagen ratio. Granulation-tissue fibroblasts secreted into the culture medium a dermatan sulfate proteoglycan with a lower molecular weight. After digestion with chondroitinase ABC, the molecular weight of the core protein appeared to be identical with that of the control fibroblasts, suggesting a difference in the glycosylation of the core protein. These results support the theory that granulation-tissue fibroblasts represent a distinct phenotype of fibroblastic cells.

Adolescent↗
PubMed1988

The small dermatan sulphate proteoglycans synthesized by fibroblasts derived from skin, synovium and gingiva show tissue-related heterogeneity.

Dermatan sulphate proteoglycans (DSPGs) synthesized in the presence of 35SO4 were characterized in culture media of fibroblast lines obtained from skin, synovium, and gingiva. The molecular mass of DSPG varied from 95-130 kDa as estimated by SDS/polyacrylamide-gel electrophoresis. Gingival fibroblasts constantly produced larger DSPGs than skin fibroblasts. This was due to the larger dermatan sulphate (DS) chains, which also showed tissue-related heterogeneity in the distribution of 4- and 6-sulphated disaccharide units. The N-glycosylated cores (44 and 47 kDa) obtained following chondroitinase ABC treatment were of identical size in all tissues. The cores from the different tissues were also of the same size (38 kDa) when addition of the N-linked oligosaccharides was inhibited by tunicamycin or when they were removed by N-glycanase treatment. No evidence for low-molecular-mass sulphated oligosaccharides was found. All tissues contained two mRNA species (1.6 and 1.9 kb) for the DSPG core protein. These data suggest that the pattern of transferase activities involved in the construction of DS chains differs from one tissue to another. This variation may modulate the functions of DSPG in the extracellular matrix.

Blotting, Northern↗
PubMed1988

Attachment and spreading of human gingival fibroblasts on potentially bioactive glasses in vitro.

The attachment and spreading of cultured fibroblasts on potentially bioactive glasses (bioglasses) of ten different compositions were studied. Human gingival fibroblasts were allowed to attach and spread on bio-glasses for 1-72 h. Unreactive silica glass and cell culture polystyrene served as controls. The attachment and spreading of cells were examined by 3H-thymidine labeling of cells, planimetric analysis, cytological staining, immunocytochemistry, and scanning electron microscopy. The cell attachment to bioglasses and silica glass and the cell spreading on bioglasses were slower and cell morphology more elongated compared to control plastic. In spite of great differences in bioglass compositions no great differences in cell behavior on these surfaces were detected. Thus the initial events in the tissue-implant interface might be independent on the bioglass composition, and furthermore the differences in the organization of the tissue-implant interface in vivo might depend on the nature of the surrounding tissues and subsequent changes of the implant surface and the extracellular environment.

Adult↗
PubMed1988

Effect of citric acid treatment on the migration of epithelium on root surfaces in vitro.

Explants of bovine gingival mucosa were cultured for four days on scaled and citric acid-conditioned root surfaces. Demineralization of the hard tissue with citric acid exposed the collagenous matrix of the root. Undemineralized islands were frequently seen among the collagen fibers of the treated roots. When cultured on scaled, control root surfaces, the epithelium migrated inwards between the connective tissue of the explant and the root surface. On citric acid-treated roots, epithelial migration in this direction was rare although it was possible. Citric acid treatment of the substratum directed the epithelium to migrate outwards from the explant. Only a few of the controls showed epithelial migration to the outward direction. The findings indicate that demineralization of the root surface has an influence on the direction in which the epithelium initially starts to migrate. An induced delay of epithelial migration between the gingival connective tissue and the hard tissue may be favorable for connective tissue attachment to the root surface.

Animals↗
PubMed1987

Fibronectin fragmentation induced by dental plaque and Bacteroides gingivalis.

Degradation of fibronectin (FN) by subgingival and supragingival plaque and Bacteroides gingivalis (Bg) was studied in vitro. The degradation of FN by both types of plaque was relatively rapid, continuous but incomplete. Some differences were found between supra- and subgingival samples. Supragingival plaque extracts produced several FN fragments of 110-180 kd during short incubations of 15-60 min. The predominant fragment after overnight incubation was a 110 kd polypeptide. With subgingival plaque extract a more extensive degradation of FN was noted. The main degradation product was a 120 kd fragment after overnight incubation. Several peptide fragments were released from fibronectin by Bg extracts. Their molecular size was different from those produced by trypsin, elastase or dental plaque. When cell extracts of Bg were fractionated by high performance liquid chromatography, three separate peaks of fibronectin degrading activity were obtained. Two of those peaks also contained trypsin-like enzyme activity. The degradation of fibronectin and the subsequent formation of biologically active peptides may have many effects in periodontal pockets. These may include modifying effects on plaque growth and wound healing.

Bacteroides↗
PubMed1987

Inhibition of gingival fibroblast growth by Bacteroides gingivalis.

Human gingival fibroblasts were exposed in culture to cell extracts of different black-pigmented Bacteroides species, and their growth was monitored by determining thymidine uptake and counting cells. Of the Bacteroides species tested (B. gingivalis, B. asaccharolyticus, and B. intermedius), B. gingivalis gave the extract with the strongest inhibitory effect on fibroblast thymidine uptake. Linear inhibition reaching 80% of the control level was obtained with a dose of 100 micrograms of B. gingivalis extract protein per ml. The effect of B. asaccharolyticus resembled that of B. gingivalis, but even at the highest dose tested B. intermedius had only a slight inhibitory effect. When fibroblasts were counted after 2- and 4-day exposures to B. gingivalis extracts, a clear depression in the number of fibroblasts was found. The effects of extracts obtained from early and late growth phases of B. gingivalis cultures were similar. A fraction of B. gingivalis consisting essentially of lipopolysaccharides (LPSs) was obtained by degrading the extract proteins with proteinase K. Silver staining of polyacrylamide gels revealed a LPS pattern with a molecular mass ranging from 37 to 60 kilodaltons. This LPS-rich fraction caused inhibition of thymidine uptake by gingival fibroblasts similar to that caused by the native extract alone. Thus, the inhibition of gingival fibroblast growth by B. gingivalis appeared to be LPS mediated. This inhibitory effect of B. gingivalis on oral fibroblast growth may be a virulence factor of this bacterium.

Bacteroides↗
PubMed1987

Abnormal dermal proteoglycan in aspartylglycosaminuria: a possible mechanism for ultrastructural changes of collagen fibrils in a glycoprotein storage disorder.

Changes in the structure and organization of collagen fibrils were recently described in the skin of aspartylglycosaminuria patients. The skin of the patients contained a normal amount and distribution of glycosaminoglycans, but the dermatan sulfate of aspartylglycosaminuria skin was more sensitive to chondroitinase AC digestion, resulting in unsaturated 4-sulfated disaccharides which were not detected in controls. Isolated dermatan sulfate chains as well as the chains present in the intact core protein synthesized by skin fibroblasts from an aspartylglycosaminuria patient were also digestible with chondroitinase AC, while those of a control fibroblast culture could be digested with chondroitinase ABC only. This is indirect evidence for abnormal epimerization of dermatan sulfate in the skin of aspartylglycosaminuria patients, which may be associated with the changes in collagen fibril formation.

Acetylglucosamine↗
PubMed1987

Alterations in scleroderma fibroblast surface glycoproteins associated with increased collagen synthesis.

Fibroblasts were cultured from affected and unaffected skin sites of 6 patients with localized scleroderma. As a parameter of fibroblast activation, collagen synthesis and cellular pro alpha 1(I)collagen mRNA levels were measured. Cell surface glycoproteins were labelled with the periodate/borohydride method and fractionated electrophoretically. A distinct reduction in the relative amount of surface glycoproteins in the 120 kDa region was observed in two affected cell lines producing increased amounts of collagen and in one affected cell line with normal collagen production when compared to unaffected fibroblast lines. Other, non-systematic alterations in the surface glycoproteins of the affected cell lines were also detected. Compared to unaffected and healthy control fibroblasts no alterations in the surface protein profiles were seen in the other three affected cell lines. These cells did not show an increase in collagen production either. The results suggest that the activation of collagen synthesis found in scleroderma fibroblasts might be connected with alterations in the normal cell surface glycoprotein pattern.

Adult↗
PubMed1986

Changes in the expression of cell surface sialoglycoproteins during transition of human monocytes into macrophages.

Cell surface sialoglycoproteins of human mononuclear phagocytes in different maturation stages were labelled by the periodate/borohydride method and separated by SDS-polyacrylamide gel electrophoresis. The main surface glycoproteins of peripheral blood monocytes had molecular masses of 115 and 95 kDa. During in vitro transition into adherent macrophages, the monocyte-characteristic surface glycoproteins disappeared. Most of the changes in the surface glycoprotein pattern occurred during the first 24 h and after 96 h the changes were completed. The major sialoglycoproteins of the macrophage cell surface had molecular masses of 130 and 55 kDa. The macrophage cell surface showed further changes when cultured in the presence of synovial fluid (10%). These results may reflect the in vivo maturation of monocytes into tissue macrophages. In synovium, tissue-derived factors may also take part in differentiation.

Cell Membrane↗
PubMed1986

Oral health of patients with insulin-dependent diabetes mellitus.

Oral health, the amount of salivary Streptococcus mutans and lactobacilli, and the flow rate, pH and buffer capacity of paraffin-stimulated whole saliva were analyzed in 35 adult diabetic patients and their age- and sex-matched non-diabetic, clinically healthy controls. All patients had insulin-dependent diabetes (IDDM) with a mean (+/- SD) duration of 14.0 +/- 9.1 yr. The prevalence of dental caries was as high in the diabetic group as in the controls but the past caries experience was remarkably lower in those individuals whose diabetes had started at a very early age (less than or equal to 7 yr). In agreement with the clinical data, the salivary levels of cariogenic microorganisms were of the same order of magnitude in both study groups. However, the relative proportion of S. mutans from the total cultivable aerobic microflora was significantly higher (P less than 0.01) in diabetics compared to the controls. The other studied salivary parameters did not differ between the groups. Remarkable individual differences were observed in the correlation between glucose levels of blood and whole saliva among diabetics. In spite of the noncariogenic dietary habits, the adult diabetic patients seem to be at least as susceptible to dental caries as non-diabetics, probably due to the leakage of glucose from blood into the oral cavity.

Adolescent↗
PubMed1986

Collagens in neurofibromas and neurofibroma cell cultures.

Neurofibromas contain approximately 30-50% collagen of their lipid-free dry weight, which is about half of the value of skin but approximately twice that described for peripheral nerve endoneurium. Immunohistochemical stainings indicate that neurofibromas contain types I, III, IV, and V collagens and fibronectin. Most of the neurofibroma cells are type IV collagen and S-100 protein positive, which provides immunohistochemical evidence that neurofibromas are mostly composed of Schwann cell-like cells. The proteoglycan/collagen ratio is 4 to 10 times higher in the neurofibromas than in the surrounding dermal tissue. This would explain the typical soft consistency of the neurofibromas and may contribute to a favorable milieu for tumor growth. Pure fibroblastic cell cultures are obtained from neurofibromas after repeated passages. The cultured cells synthesized type I and III collagens and fibronectin, indicating that these cells are important in the production of the fibrous connective tissue proteins in neurofibromas.

Cells, Cultured↗