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Biomedical subjects

H Larjava

Publications and source records attributed to H Larjava.

At least 73 records · Page 4Linked to original sources
PubMed1991

Differential regulation of extracellular matrix proteoglycan (PG) gene expression. Transforming growth factor-beta 1 up-regulates biglycan (PGI), and versican (large fibroblast PG) but down-regulates decorin (PGII) mRNA levels in human fibroblasts in culture.

Proteoglycans (PGs) comprise a group of extracellular matrix macromolecules which play an important role in matrix biology. In this study, normal human skin and gingival fibroblast cultures were incubated with transforming growth factor-beta 1 (TGF-beta 1), and the expression of three PGs, viz. biglycan (PGI), decorin (PGII), and versican (a large fibroblast proteoglycan) was examined. The results indicate that TGF-beta 1 (5 ng/ml) markedly increased the expression of biglycan (up to 24-fold) and versican (up to 6-fold) mRNAs and the enhancement of biglycan expression was coordinate with elevated type I procollagen gene expression in the same cultures. In contrast, the expression of decorin mRNA was markedly (up to approximately 70%) inhibited by TGF-beta 1. The response to TGF-beta 1 was similar in both skin and gingival fibroblasts, although the gingival cells were clearly more responsive to stimulation by TGF-beta 1 with respect to biglycan gene expression. Analysis of 35S-labeled proteoglycans in the culture media of skin and gingival fibroblasts also revealed stimulation of biglycan and versican production, and reduction in decorin production. Quantitation of both [35S]sulfate and [3H]leucine-labeled decorin in cell culture media by immunoprecipitation revealed a 50% reduction in decorin production in cell cultures treated with TGF-beta 1. This TGF-beta 1-elicited reduction was accompanied by an apparent increase in the size of the decorin molecules, although the size of the core protein was not altered, as judged by Western immunoblotting following chondroitinase ABC digestion. Analysis of the proteoglycans in the matrix and membrane fractions also revealed increased amounts of versican in cultures treated with TGF-beta 1. These results indicate differential regulation of PG gene expression in fibroblasts by TGF-beta 1, and these observations emphasize the role of PGs in the extracellular matrix biology and pathology.

Biglycan↗
PubMed1991

Expression of beta 1 integrins in normal human keratinocytes.

The majority of cell adhesive events to the extracellular matrix are mediated by cell surface receptors, beta 1 integrins. Keratinocytes express at least six different polypeptides of beta 1 integrin class, namely beta 1, alpha 2, alpha 3, alpha 5, and alpha 6 (alpha 6 is mainly associated with beta 4 polypeptide). These epithelial cells use alpha 2 beta 1 as a collagen receptor and alpha 3 beta 1 as a fibronectin receptor, while alpha 6 beta 4 is the major basement membrane receptor. Expression of alpha 5 beta 1 complex is low. Processing of beta 1 integrins is fast in keratinocytes; half-maximal maturation takes only 3 hours. In addition to their function in cell-matrix interactions, beta 1 integrins (alpha 2 beta 1 and alpha 3 beta 1) have also a role in maintaining keratinocyte cell-cell interactions. It is possible that resting basal keratinocytes use beta 1 integrins as cell-cell adhesion receptors, and during activation, like in wound healing, these receptors relocalize to mediate events involving cell-matrix interactions.

Basal Cell Carcinoma↗
PubMed1991

Extracellular matrix molecules and their receptors: an overview with special emphasis on periodontal tissues.

Knowledge of extracellular matrix molecules and their cell receptors has increased exponentially during the last 2 decades. It is now known that the structure and function of each tissue is based on specific combinations of matrix molecules. The major constituents of the extracellular matrix are collagens, proteoglycans, and adhesive glycoproteins. The rapid development of biochemical, molecular biological, and immunological research has revealed a lot of interesting details pertaining to these molecules. Several new collagen types have been discovered. In addition to being responsible for the strength and form of tissues, each collagen type has specific sequences providing them with special features such as flexibility and the ability to interact with other matrix molecules and cells. Proteoglycans are another large group of matrix molecules with a variety of functions. Proteoglycans play an important role in tissue resilience and filtering. Some proteoglycans have a capacity to specifically bind other matrix molecules and growth factors, while others act as matrix receptors on the cell surface. An important part of regulation of the cell behavior is played by adhesive glycoproteins belonging to the fibronectin and laminin families. Several isoforms of fibronectin and laminin that result from alternative RNA splicing serve specific functions such as controlling the attachment, migration, and synthetic activity of cells. A major group of cell receptors for cell-matrix and cell-cell interactions is termed integrins. The integrins are cell surface proteins composed of two polypeptides whose structure dictates the specificity of each receptor. The cytoplasmic domain of the integrins interacts with cytoskeletal elements within the cell, and thereby relays the information from the extracellular space into the protein synthesis machinery. The expression of the integrins is controlled by the extracellular matrix and growth factors, most notably TGF beta. During periodontal diseases several aspects of the cell-matrix interactions may be disturbed. Therefore, an understanding of the special features of the extracellular matrix and their receptors in periodontal tissues is a prerequisite for developing new approaches to the prevention and treatment of periodontal diseases.

Extracellular Matrix Proteins↗
PubMed1991

Analyses of periodontal glycosaminoglycans and proteoglycans. Regulation by microbial, chemical and inflammatory factors.

During the last two decades one of the main lines of research in the Department of Periodontology in Turku has related to connective tissue glycosaminoglycans and proteoglycans. A general outline of current knowledge about proteoglycans is presented, with a review of most published and unpublished results on periodontal glycosaminoglycans and proteoglycans obtained in our Department.

Bacteria↗
PubMed1990

Differences in the biosynthesis and localization of the fibronectin receptor in normal and transformed cultured human cells.

We examined the biosynthesis and localization of the fibronectin receptor integrin from normal and transformed cultured human cells. Normal cells required a minimum of 20 h for the biosynthesis of completely mature fibronectin-receptor beta-subunit, while transformed cells required only 6-8 h. There was a correspondingly major decrease in the amount of the intracellular beta-chain precursor in the transformants. Immunostaining of normal fibroblastic cells with monoclonal antibodies indicated that both alpha- and beta-polypeptides of the fibronectin receptor are localized in cell surface streaks and focal contact areas. In contrast, both subunits lacked this clustering and had a more diffuse distribution on the surfaces of transformed cells, even though quantitative immunofluorescence experiments indicated that similar or larger amounts of each subunit were present on a per cell basis. Both immunostaining and biochemical analyses also indicated the presence of a relatively large intracellular pool of beta-polypeptides in normal fibroblasts that is not present in transformed cells. There was no major transformation-dependent change in total quantities of mature fibronectin receptor subunit expressed and inserted into the plasma membrane, when normalized to total protein synthesis. Our results indicate that malignant transformation of cultured human cells results in altered localization and processing of the fibronectin receptor. Such changes involving pathways of crucial cell surface molecules may contribute to alterations in their interactions with extracellular macromolecules, including during the process of cellular invasion.

Cell Transformation, Neoplastic↗
PubMed1990

Histochemical localization of hyaluronate in human oral epithelium using a specific hyaluronate-binding probe.

Biochemical data suggest that gingival epithelium contains hyaluronate, but there is little histochemical information about its localization. Hyaluronate was here visualized in gingival and buccal mucosa using a specific probe derived from the hyaluronate binding region of cartilage proteoglycan. Hyaluronate was found both in the gingival and buccal epithelium, but its localization was correlated with the type of keratinization. In the keratinized epithelium of gingiva, whether ortho- or parakeratotic, the intercellular spaces from basal to upper spinous layers displayed strong staining, most intense in the middle spinous cell layer. The uppermost vital cell layers as well as the cornified cell layer remained unstained. In the non-keratinized epithelium of buccal mucosa and the local non-keratinized areas of gingiva, only the basal cells and the lowermost spinous cell layers stained for hyaluronate, whereas the majority of the upper epithelium was negative. Electron microscopic examination of the basal and spinous cell layers displayed hyaluronate, both associated with the cell surface and free in the intercellular space. The subepithelial connective tissue showed positive but diffuse staining in all specimens.

Adolescent↗
PubMed1990

Novel function for beta 1 integrins in keratinocyte cell-cell interactions.

We have examined the expression, localization, and function of beta 1 integrins on cultured human epidermal keratinocytes using polyclonal and monoclonal antibodies against the beta 1, alpha 2, alpha 3, and alpha 5 integrin subunits. The beta 1 polypeptide, common to all class 1 integrins, was localized primarily in areas of cell-cell contacts of cultured keratinocytes, as were alpha 2 and alpha 3 polypeptides, suggesting a possible role in cell-cell adhesion for these integrin polypeptides. In contrast, the fibronectin receptor alpha 5 subunit showed no such accumulations in regions of cell-cell contact but was more diffusely distributed in the keratinocyte plasma membrane, consistent with the absence of fibronectin at cell-cell contact sites. Colonies of cultured keratinocytes could be dissociated by treatment with monoclonal antibody specific to the beta 1 polypeptide. Such dissociation of cell-cell contacts also occurred under conditions where the monoclonal antibody had no effect on cell-substrate adhesion. Therefore, beta 1 integrin-dependent cell-cell adhesion can be inhibited without affecting other cell-adhesive interactions. Antibody treatment of keratinocytes maintained in either low (0.15 mM) or high (1.2 mM) CaCl2 also resulted in the loss of organization of intracellular F-actin filaments and beta 1 integrins, even when the anti-beta 1 monoclonal antibody had no dissociating effect on keratinocyte colonies at the higher calcium concentration. Our results indicate that beta 1 integrins play roles in the maintenance of cell-cell contacts between keratinocytes and in the organization of intracellular microfilaments. They suggest that in epithelial cells integrins can function in cell-cell interactions as well as in cell-substrate adhesion.

Actins↗
PubMed1990

In vivo analyses of integrin beta 1 subunit function in fibronectin matrix assembly.

Early development of the urodele amphibian Pleurodeles waltl is accompanied by a process of progressive fibronectin (FN) fibrillogenesis. FN begins to assemble into fibrils on the inner surface of the blastocoele roof at the early blastula stage and progressively forms a complex extracellular matrix. We have analyzed the mechanisms of FN-fibril formation under normal and experimental conditions in vivo with the following probes: iodinated FN, fluorescein-labeled FN, synthetic peptides containing the Arg-Gly-Asp (RGD) cell surface recognition sequence of FN, and polyclonal antibodies against both beta 1 subunit of the amphibian FN receptor and the cytoplasmic domain of beta 1 subunit. We report that in living embryos, exogenous labeled mammalian FN injected into the amphibian blastocoele undergoes FN-fibril formation in spatiotemporal patterns similar to those of endogenous FN. This indicates regulation of fibrillogenesis by the cell surface rather than by changes in the type of FN. Fibrillogenesis is inhibited in a dose-dependent manner both by the GRGDS peptide and monospecific antibodies to amphibian integrin beta 1 subunit. Furthermore, when injected intracellularly into uncleaved embryos or into selected blastomeres, antibodies to the cytoplasmic domain of integrin beta 1 subunit produce a reversible inhibition of FN-fibril formation that follows early cell lineages and cause delays in development. Together, these data indicate that in vivo, the integrin beta 1 subunit and the RGD recognition signal are essential for the proper assembly of FN fibrils in early amphibian development.

Amino Acid Sequence↗
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↗