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Biomedical subjects

I Olsen

Publications and source records attributed to I Olsen.

At least 73 records · Page 4Linked to original sources
PubMed1999

Retroviral transduction of human periodontal cells with a temperature-sensitive SV40 large T antigen.

The periodontal ligament (PDL) is considered to contain subpopulations of cells responsible for the development, repair and regeneration of the periodontium. Cell cultures have been used as model systems in order to understand the complex cellular and biochemical events underlying these processes. In order to obtain long-term cultures of these cells that can be cloned and characterized, primary cultures of PDL and gingival cells were infected with an amphotropic retroviral construct encoding a temperature-sensitive SV40 large T antigen (tsT). After selection for drug resistance, the cells expressed the T antigen and proliferated at 34 degrees C for more than 40 passages. However, when the T antigen was inactivated by incubation at 39 degrees C, the cultures became growth-arrested and the granularity of the cells increased, possibly as a result of differentiation. Reverse transcribed-polymerase chain reaction and flow cytometry showed that the tsT-transduced cells expressed a number of soft and hard connective-tissue antigens, including osteocalcin, osteonectin, osteopontin, collagen type I and alkaline phosphatase. Moreover, incubation of the transduced PDL cells at 39 degrees C was found to upregulate the expression of osteocalcin, osteopontin and collagen type I, but downregulate osteonectin. At this temperature, the presence of the dexamethasone downregulated type I collagen, while vitamin D3 had no effect on the expression of any of the antigens examined. Under all culture conditions, antigen expression was far higher in the transduced PDL cells than the gingival cells. The findings thus show that growth of the tsT-transduced PDL and gingival cells is temperature-dependent and that the presence of the T antigen increases their lifespan but does not ablate the expression of certain of their characteristic phenotypic and functional features.

Alkaline Phosphatase↗
PubMed1999

Severe, rapidly progressing peri-implantitis.

The replacement of teeth by means of titanium implants is a safe and predictable procedure in most patients. Some studies show that a small number of patients lose the majority of their implants. Unfortunately, it is impossible to identify these patients prior to surgery. It is important to report such cases so that we may, in the future, be able to identify them before surgery. The present case showed a severe and rapid exfoliation of titanium implants. Out of 8 implants inserted in the anterior lower jaw of this patient, only two maintained osseointegration over a 2-year period.

CD4-Positive T-Lymphocytes↗
PubMed1999

LFA-3 (CD58) mediates T-lymphocyte adhesion in chronic inflammatory infiltrates.

Previous studies have suggested that LFA-3 has an important role in a number of chronic inflammatory pathologies, although an active role for LFA-3 within in vivo inflammatory reactions has not previously been directly observed in humans. To assess the importance of LFA-3 in this process, this study used an adaptation of the Stamper-Woodruff lymphocyte adhesion assay to measure the binding of exogenous activated lymphocytes to the T-cell-dominated chronic inflammatory infiltrate of oral lichen planus. Antibody blockade experiments showed that anti-LFA-3 monoclonal antibody reduced lymphocyte adhesion by approximately 29%, while anti-ICAM-1 produced a reduction of 26%. These results thus suggest that both LFA-3 and ICAM-1 are likely to mediate cell-cell interactions within lesional tissues in vivo. Moreover, these findings are also the first to directly demonstrate that LFA-3-mediated adhesion, like that of ICAM-1, is functionally important in the molecular pathology of inflammatory mucosal disease.

Adult↗
PubMed1999

Localized adhesion molecule expression and circulating LFA-3 levels in adult and early onset forms of periodontitis.

Because of their importance in mediating cellular interactions in chronic inflammatory diseases, this study has examined the expression of a number of adhesion molecules in adult (n=11), generalized early onset (n=5) and localized early onset (n=2) forms of periodontitis. In comparison with immunostaining profiles of cryostat sections of healthy gingival tissue (n=7), the beta 1 integrins VLA-1, VLA-2 and VLA-4 were found to be up-regulated in periodontitis, with VLA-6 being markedly elevated. Although only small differences were observed in ICAM-1 and LFA-3 expression in the gingival epithelium, there was particularly notable up-regulation of these adhesion molecules within the inflammatory infiltrates of the diseased tissues. However, there were no statistically significant differences between the serum levels of a soluble form of LFA-3 in periodontitis patients (n=47) compared with healthy control subjects (n=40), although the generalized early onset and adult periodontitis groups exhibited wider ranges of circulating LFA-3. These findings show that there is localized modulation of adhesion molecule expression in the chronic inflammatory periodontal diseases studied, but that the levels of LFA-3 in the circulation nevertheless remain unaffected.

Adolescent↗
PubMed1999

Alkaline phosphatase activity is upregulated in regenerating human periodontal cells.

The activity of alkaline phosphatase (ALP) is considered to indicate the presence of osteoblast cells and the formation of new bone. In the present study this enzyme was investigated in cells obtained from retrieved polytetrafluoroethylene membranes (M cells) of periodontal disease patients treated by guided tissue regeneration (GTR) and from the regenerated tissue underlying the membrane (RT cells). Normal periodontal ligament (PL) and gingival cells were also grown from the corresponding healthy tissues of human subjects. ALP activity was measured colourimetrically, using paranitrophenyl phosphate as the substrate, after 4 and 7 d of culture in the absence and presence of dexamethasone (DEX), a synthetic glucocorticoid which induces osteoblast differentiation. The results showed that basal levels of ALP activity were expressed by all the cells and that DEX upregulated ALP levels in the M, RT and PL cells but not in the gingival cells. Moreover, both the basal and DEX-induced ALP activities were statistically significantly higher in the RT cells than in any of the other cells. Our results suggest that both the GTR-associated and normal PL cells express osteoblast-like characteristics and, furthermore, that the RT cultures in particular contain a high proportion of osteoprogenitor cells.

Adolescent↗
PubMed1999

Brain abscesses caused by oral infection.

Brain abscesses are rare but can be life-threatening infections. Recent progress in microbiological classification and identification has indicated that they are sometimes caused by oral infection and dental treatment. It has been postulated that oral microorganisms may enter the cranium by several pathways: 1) by direct extension, 2) by hematogenous spread, 3) by local lymphatics, and 4) indirectly, by extraoral odontogenic infection. In the direct extension, oral infections spread along the fascial planes. Hematogenous spreading occurs along the facial, angular, ophthalmic, or other veins which lack valves, through the cavernous sinus and into the cranium. Another hematogenous pathway is through the general circulation. Oral bacteria may cause systemic infections, e.g., endocarditis, and then indirectly initiate brain abscess. Microbiota, complications, and the prevention and management of odontogenic brain abscesses are also discussed in this review.

Anti-Bacterial Agents↗
PubMed1998

Diffusion blotting for rapid production of multiple identical imprints from sodium dodecyl sulfate polyacrylamide gel electrophoresis on a solid support.

A simple and fast method for diffusion blotting of proteins from precast SDS-PAGE gels (0.5 mm) was developed. The efficiency of protein transfer was evaluated using 14C labelled proteins. Diffusion blotting for three minutes, gave a transfer of 10% compared to electroblotting. By doubling the transfer time for each subsequent imprint, four imprints were made from the same lane with similar amounts of protein transferred onto each imprint. With a transfer time of three minutes each, it was possible to obtain at least ten imprints with all the proteins visible in all the imprints. There was no detectable loss in resolution as compared to electroblotting. The method also works well with an immuno-detecting system. The number of imprints which can be obtained, is dependent on the sensitivity of the detection system and the amount of protein applied. The greatest advantage of diffusion blotting compared to electroblotting is that several imprints can be made from each lane, and different antisera can be tested on identical imprints. The gel remains on its plastic support which prevents it from stretching and compression; this ensures identical imprints and facilitates more reliable molecular mass determination. If only a few imprints are made, sufficient protein remains within the gel for general protein staining. These advantages make diffusion blotting the method of choice when quantitative protein transfer is not required.

Animals↗
PubMed1998

Flow cytometry for assessing biocompatibility.

Flow cytometry (FCM) was examined as a possible procedure for measuring in vitro the biocompatibility of implant materials for orthopedic and dental surgery. The human osteoblast-like cell line MG63 was grown on hydroxyapatite (HA) and P2O5 glass-reinforced HA composite discs and compared with the same cells grown on polystyrene culture dishes. While morphological observation at the light and electron microscopic levels showed no major deleterious effects, FCM indicated that cell size was somewhat reduced, particularly by growth on the HA composite. Morever, this material also appeared to delay the progression of the cells from the G0/G1 into the S phase of the cell cycle. In addition to this low level of inhibition of cell growth relative to control cultures, FCM analysis also demonstrated that the glass-reinforced HA caused some down-regulation of the expression of osteocalcin and fibronectin, two antigens which play a vital part in the integrity and function of bone and soft connective tissue, respectively. These results thus show, first, that although HA and the HA composite used in these experiments were generally biocompatible, they nevertheless had certain suboptimal effects on the cells; and second, that FCM could be a highly useful procedure for effectively screening and evaluating important biological responses to implant materials.

Biocompatible Materials↗
PubMed1998

IL-6 expression by oral fibroblasts is regulated by androgen.

Interleukin 6 (IL-6) is a multi-functional cytokine which has a major role in tissue damage. It is secreted by many types of cell, including oral fibroblasts, and has been implicated in the pathogenesis of periodontal diseases, particularly those associated with sex hormones. In the present study we investigated whether the androgen dihydrotestosterone (DHT) affects the expression and regulation of IL-6 in gingival fibroblasts. Using a 'capture' ELISA assay, it was found that increasing DHT concentrations progressively reduced IL-6 production by gingival cells from normal individuals and from patients with gingival inflammation and gingival hyperplasia. In contrast, cells from periodontal ligament tissue produced only barely detectable levels of IL-6. The anti-androgen cyproterone acetate acted as an androgen analogue in the gingival fibroblasts, potently inhibiting IL-6 production, and did not reverse the DHT-mediated downregulation of the cytokine. Flutamide also failed to abrogate DHT inhibition of IL-6 production, and it had no effect on IL-6 production in the absence of DHT. Moreover, semi-quantitative RT-PCR showed that DHT acted at the level of transcription of the IL-6 gene, causing a marked reduction in the relative level of IL-6 mRNA in the gingival cells.

Androgen Antagonists↗
PubMed1998

Control of the human cell cycle by a bacterial protein, gapstatin.

The oral gram-negative bacterium Actinobacillus actinomycetemcomitans is a major pathogen in human periodontal disease. Saline extraction releases a range of surface-associated components from this bacterium, including one which exhibits potent anti-proliferative activity as assessed by its capacity to inhibit DNA synthesis by human and other mammalian cells. Cultures incubated with this bacterial fraction for a prolonged period comprise a high proportion of cells containing a 4n level of DNA. Studies using hydroxyurea-synchronized cultures showed that cells treated with the surface-associated fraction were arrested in the G2 phase of the cell cycle and did not enter mitosis. This G2/M blockade was observed only when the bacterial fraction was added to the cells during early S phase. Our data also suggest that the active bacterial component binds to surface receptors expressed by the human cells and may act by a novel mechanism which involves down-regulation of cyclin B1 expression. The anti-proliferative activity of the bacterial fraction, purified by a combination of ammonium sulphate precipitation, HPLC anion exchange and gel filtration, has been shown to be an 8 kDa protein, which we have called gapstatin. Purified gapstatin was shown to be responsible for the the inhibitory effects of the surface-associated fraction on mammalian cells.

Aggregatibacter actinomycetemcomitans↗
PubMed1998

Immunohistochemical and serological comparison of idiopathic and hepatitis C virus-associated forms of oral lichen planus.

The oral form of the inflammatory disease lichen planus occurs spontaneously due to unknown aetiological factors. However, it has recently been observed to occur with increased frequency in patients infected with the hepatitis C virus. Because of the prominent role of adhesion molecules in immune cell interactions, we have compared the expression of these antigens in the hepatitis C virus-associated and idiopathic forms of the disease. The results show similar patterns of expression of very late activation antigen-4, lymphocyte function-associated antigen-3 and intercellular adhesion molecule-1, but relatively elevated levels of these antigens in oral lichen planus patients with no hepatitis C virus infection. In addition to differences in Langerhans cell distribution, serum levels of "soluble" intercellular adhesion molecule-1 as well as immunoglobulin G were significantly increased in the hepatitis C virus-associated group. These findings show that there are some differences in the lesional and systemic immune reactivities of the two types of oral lichen planus which may be related to possibly distinct pathogenic mechanisms.

Antigens↗
PubMed1998

Palmar plantar keratosis and unusual periodontal findings. Observations from a family of 4 members.

The connection between palmar plantar hyperkeratosis and severe periodontal disease was first reported in 1924 by Papillon and Lefevre. The 2 major components of this syndrome (PLS) can also occur as distinct entities. The literature describes a number of cases which do not fit the classical disease descriptions. In this paper, we report on a family with an atypical PLS. The father had marked palmar plantar hyperkeratosis but a very late onset of destructive marginal periodontitis. The son also had palmar plantar hyperkeratosis, but despite the fact that he initially harboured Actinobacillus actinomycetemcomitans, Prevotella intermedia and Porphyromonas gingivalis, he did not develop periodontal disease over a seven-year observation period when improved oral hygiene and professional tooth cleaning were instituted.

Adult↗
PubMed1998

Surface structure, hydrophobicity, phagocytosis, and adherence to matrix proteins of Bacillus cereus cells with and without the crystalline surface protein layer.

Nonopsonic phagocytosis of Bacillus cereus by human polymorphonuclear leukocytes (PMNs) with particular attention to bacterial surface properties and structure was studied. Two reference strains (ATCC 14579(T) and ATCC 4342) and two clinical isolates (OH599 and OH600) from periodontal and endodontic infections were assessed for adherence to matrix proteins, such as type I collagen, fibronectin, laminin, and fibrinogen. One-day-old cultures of strains OH599 and OH600 were readily ingested by PMNs in the absence of opsonins, while cells from 6-day-old cultures were resistant. Both young and old cultures of the reference strains of B. cereus were resistant to PMN ingestion. Preincubation of PMNs with the phagocytosis-resistant strains of B. cereus did not affect the phagocytosis of the sensitive strain. Negatively stained cells of OH599 and OH600 studied by electron microscopy had a crystalline protein layer on the cell surface. In thin-sectioned cells of older cultures (3 to 6 days old), the S-layer was observed to peel off from the cells. No S-layer was detected on the reference strains. Extraction of cells with detergent followed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis revealed a major 97-kDa protein from the strains OH599 and OH600 but only a weak 97-kDa band from the reference strain ATCC 4342. One-day-old cultures of the clinical strains (hydrophobicity, 5.9 to 6.0%) showed strong binding to type I collagen, laminin, and fibronectin. In contrast, reference strains (hydrophobicity, -1.0 to 4.2%) as well as 6-day-old cultures of clinical strains (hydrophobicity, 19.0 to 53.0%) bound in only low numbers to the proteins. Gold-labelled biotinylated fibronectin was localized on the S-layer on the cell surface as well as on fragments of S-layer peeling off the cells of a 6-day-old culture of B. cereus OH599. Lactose, fibronectin, laminin, and antibodies against the S-protein reduced binding to laminin but not to fibronectin. Heating the cells at 84 degreesC totally abolished binding to both proteins. Benzamidine, a noncompetitive serine protease inhibitor, strongly inhibited binding to fibronectin whereas binding to laminin was increased. Overall, the results indicate that changes in the surface structure, evidently involving the S-layer, during growth of the clinical strains of B. cereus cause a shift from susceptibility to PMN ingestion and strong binding to matrix and basement membrane proteins. Furthermore, it seems that binding to laminin is mediated by the S-protein while binding to fibronectin is dependent on active protease evidently attached to the S-layer.

Bacillus cereus↗
PubMed1998

Flow cytometry analysis of gingival and periodontal ligament cells.

Gingival and periodontal ligament (PDL) fibroblasts are the major cellular components of periodontal soft connective tissues, but the precise differences between these cells are not yet known. In the present study, we have therefore examined the phenotypic and functional features of the cells obtained from gingival and PDL biopsy samples. Spindle-shaped cells characteristic of fibroblasts were the main cell type observed in vitro, although epithelial cells were also present in primary gingival cell cultures. Flow cytometry was used to measure the size and granularity of the cultured cells, and showed that the gingival fibroblasts were smaller and less granular compared with the PDL cells. The expression of certain key extracellular matrix (ECM) proteins, fibronectin, collagen type I, and tenascin was measured by flow cytometry. Analysis of the fluorescence profiles of these cultures showed that the majority of cells expressed fibronectin and that the average fluorescence intensity of this antigen in the PDL cells was higher than that in the gingival fibroblasts. Moreover, the fibronectin-positive PDL cells apparently comprised two subpopulations which expressed fibronectin at different levels, suggesting that the cells in the PDL cultures were functionally heterogeneous. The level of collagen type I was also found to be up-regulated in the PDL compared with the gingival cells and, as with fibronectin, was expressed at two different levels by subsets of the PDL cells. In contrast, tenascin was expressed at very similar levels by both the gingival fibroblasts and PDL cells. In addition, measurement of alkaline phosphatase, a marker enzyme for mineralized tissue-forming cells, showed that the PDL cells had higher activity than the gingival fibroblasts and that the alkaline phosphatase activity in the PDL cells was far more markedly up-regulated by dexamethasone. Our findings demonstrate that, despite their similar spindle-shaped appearance, fibroblasts derived from gingival and PDL tissues appear to display distinct functional activities which are likely to play a vital part in the maintenance of tissue integrity and regenerative processes.

Adolescent↗
PubMed1998

Anaerobic bacteremia and fungemia in patients undergoing endodontic therapy: an overview.

Oral focal infection, a concept neglected for several decades, is a subject of controversy. Recent progress in classification and identification of oral microorganisms has renewed interest in focal infection. The aim of this study was to use phenotypic and genetic methods to trace microorganisms released into the bloodstream during and after endodontic treatment back to their presumed source--the root canal. Microbiological samples were taken from the root canals of 26 patients with asymptomatic apical periodontitis of single-rooted teeth. The blood of the patients was drawn during and 10 minutes after endodontic therapy. Microorganisms in blood were collected after anaerobic lysis filtration and cultured anaerobically on blood agar plates. The phenotypic methods used for characterization and tracing of microorganisms in blood and root canals were: biochemical and antimicrobial susceptibility test, SDS-PAGE of whole-cell soluble proteins, and gas chromatography of cellular fatty acids. Phenotypic data were verified by DNA restriction patterns and corresponding ribotypes of the root canal and blood isolates by using a computer-assisted system fro gel analysis. All root canals contained anaerobic bacteria. The frequency of bacteremia varied from 31% to 54%. The microorganisms from the root canal and blood presented identical phenotype and genetic characteristics within the patients examined. These characteristics differed between patients. The present study demonstrated that endodontic treatment can be the cause of anaerobic bacteremia and fungemia. The phenotypic and genetic methods used appeared valuable for tracing microorganisms in the blood back to their origin.

Bacteremia↗
PubMed1997

Distinction of Prevotella intermedia and Prevotella nigrescens from endodontic bacteremia through their fatty acid contents.

Prevotella nigrescens has recently been recognized as a new species distinct from Prevotella intermedia. The distinction is based largely on DNA-DNA hybridization, electrophoretic migration of malate and glutamate dehydrogenase, and peptidase and lipase activities of type strains. Gas chromatography of cellular fatty acids can be a useful adjunct for characterization and identification of bacterial species. In the present study, cellular fatty acid profiles were determined for seven strains of P. intermedia and six strains of P. nigrescens. Six of these 13 strains were isolated from the root canal and blood of three patients during endodontic therapy of teeth with Asymptomatic apical periodontitis. The bacteria were cultivated anaerobically in 10 mL prereduced anaerobically sterilized peptone-yeast extract-glucose broth for 24 h. Dried cells of each isolate were methanolysed and their fatty acid contents determined by the Microbial Identification System software package by MIDI. The data were treated by principal component analysis, which distinguished P. nigrescensfromP. intermedia. Cellular fatty acid profiles of these strains of the species in blood matched the profiles of their respective root canal isolates, as demonstrated by Euclidean Distance Square assessment. This suggested that the organisms in the root canal had spread to the bloodstream during endodontic treatment.

Journal Article↗
PubMed1997

Ribotyping of bacteria from root canal and blood of patients receiving endodontic therapy.

DNA restriction patterns and corresponding ribotypes of 26 bacterial isolates (Propionibacterium acnes, Peptostreptococcus prevotii, Fusobacterium nucleatum, Prevotella intermedia, Prevotella nigrescens, Actinomyces israelii, Streptococcus intermedius, and Streptococcus sanguis) recovered from patients with infected root canals and their peripheral blood, collected during and after endodontic therapy, were examined. Eleven additional reference strains including type strains were also examined. Purified DNA was digested with BglI,EcoRI, and HindIII. Hybridization was carried out with a digoxigenin-labeled cDNA probe obtained by reverse transcription of Escherichia coli 16S + 23S rRNA. Ribotypes of the bacteria recovered from root canal and blood showed identical characteristics within the patients, and differed between the patients. The results were confirmed when the similarity coefficient (S(AB)) of the ribotypes from the isolates were assessed by the Dendron computer-assisted system. These results suggested that the bacteria isolated from the blood originated from the root canal.

Journal Article↗