Biomedical subjects

H Shimauchi

Publications and source records attributed to H Shimauchi.

At least 19 recordsLinked to original sources

PubMed2005

Cleavage of PDGF receptor on periodontal ligament cells by elastase.

Human leukocyte elastase, a neutrophil serine protease, is considered to be a potential immunoregulatory protease. Since the PDGF receptor (PDGFR) on periodontal ligament (PDL) cells is a crucial element for various functions, such as wound healing in periodontal tissue, we investigated the effect of elastase on the expression of PDGFR on PDL cells by flow cytometry and Western blotting. We found that PDGFR-alpha disappeared with an increasing dose of elastase, and PDGFR-beta was degraded into several fragments. Elastase degraded both receptors on fixed cells, indicating that the degradation resulted from direct proteolysis on the cell surface. Elastase also then disturbed the phosphorylation of ERK1/2, JNK/SARK, and p38, triggered by PDGF-AA and PDGF-BB, suggesting that elastase inhibited PDGFR-dependent cell activation in PDL cells. These results suggest that elastase may modulate the PDGF-mediated activity of PDL cells during periodontal wound healing.

Adolescent↗

PubMed2004

Detection frequency of periodontitis-associated bacteria by polymerase chain reaction in subgingival and supragingival plaque of periodontitis and healthy subjects.

The aim of this study was to compare the detection frequencies of 25 bacterial species in subgingival and supragingival plaque of 18 untreated periodontitis subjects and 12 periodontally healthy subjects. Genomic DNA was extracted from subgingival and supragingival plaque samples, and bacterial detection was performed by polymerase chain reaction of the 16S rRNA genes. Fourteen bacteria showed no relationship with periodontitis, and 11 of these 14 species were frequently detected (> or =50%) in subgingival plaque in both periodontitis and healthy subjects. Nine bacteria such as Eubacterium saphenum, Prevotella intermedia, and Treponema denticola seemed to be related to periodontitis; their detection frequencies in subgingival plaque samples were higher in periodontitis than in healthy subjects, but these differences were not statistically significant by multiple comparisons (0.002< or =P<0.05). Two species (Mogibacterium timidum and Porphyromonas gingivalis) were detected significantly more frequently in subgingival plaque of periodontitis subjects than of healthy subjects (P<0.002), with P. gingivalis being detected only in periodontitis subjects, suggesting that these two species are closely related to periodontitis. There were no significant differences in the detection frequencies of the 25 bacteria between subgingival and supragingival plaque, suggesting that the bacterial flora of supragingival plaque reflects that of subgingival plaque.

Adult↗

PubMed2003

Proteolysis of ICAM-1 on human oral epithelial cells by gingipains.

Cysteine proteinases (gingipains) from Porphyromonas gingivalis are considered key virulence factors of severe periodontitis and host immune evasion. Since expression of intercellular adhesion molecule-1 (ICAM-1) on gingival epithelium is indispensable in polymorphonuclear leukocyte (PMN) migration at the site of periodontitis, we examined the effects of gingipains on the expression of ICAM-1 on human oral epithelial cell lines (KB and HSC-2) by flow cytometry and Western blotting. We found that three purified forms of gingipains efficiently reduced ICAM-1 expression on the cells in a time- and dose-dependent manner. Gingipains reduced the expression on fixed cells and degraded the ICAM-1 in the cell membranes, indicating that the reduction resulted from direct proteolysis. They then disturbed the ICAM-1-dependent adhesion of PMNs to the cells. These results indicate that gingipains cleave ICAM-1 on oral epithelial cells, consequently disrupting PMN-oral epithelial cell interaction, and are involved in immune evasion by the bacterium in periodontal tissues.

Adhesins, Bacterial↗

PubMed2002

In vivo platelet response to lipopolysaccharide in mice: proposed method for evaluating new antiplatelet drugs.

We previously found evidence (based on the use of 5HT as a marker) that i.v. injection of a lipopolysaccharide (LPS) into mice induces a rapid accumulation of platelets in liver and lung. Our previous studies lacked measurement of the platelet count itself, but we have now compared the LPS-induced changes in 5HT levels with the change in platelet count. We also examined the effects on the platelet response of some drugs that act on platelets. In mice, sublethal doses of LPS induced parallel decreases in platelets and 5HT in the blood. The 5HT lost from the blood accounted well for the 5HT accumulated in liver and lung. Soon after this accumulation, the levels of platelets and 5HT in the blood recovered in parallel, and these recoveries corresponded well with the decreases in 5HT occurring in liver and lung. Aspirin and dexamethasone were effective at both reducing pulmonary platelet-accumulation and promoting their return to the circulation. By contrast, oestrogen tended to reduce the return of platelets from lung to circulation. Heparin did not inhibit pulmonary platelet-accumulation but it did decrease their return to the circulation. These results suggest that (i) in response to sublethal doses of LPS, platelets translocate into the liver and lung, then return to the circulation; (ii) this platelet response involves mechanisms that can be modified by drugs; and (iii) the use of this platelet response as a tool for drug evaluation might help identify new drugs with therapeutic potential.

Animals↗

PubMed2002

Expression of MMP-8 and MMP-13 mRNAs in rat periodontium during tooth eruption.

The present study was designed to investigate mRNA expression of matrix metalloproteinase-8 (MMP-8) and MMP-13 in forming periodontium during tooth eruption in the rat. RT-PCR for the decalcified paraffin sections indicated expression of MMP-8 and MMP-13 in the periodontal tissues. In situ hydridization demonstrated expression of MMP-8 in osteoblasts, osteocytes, periodontal ligament cells, cementoblasts, and cementocytes along with collagen types I and III. In contrast, transcripts of MMP-13 were confined to a small population of osteoblasts and osteocytes in alveolar bone. The results suggested that MMP-8 may be involved in remodeling the periodontium during tooth eruption, and its expression may be coordinated with that of collagen types I and III, whereas the participation of MMP-13 may be rather limited.

Alveolar Process↗

PubMed2001

Osteoblastic differentiation of periosteum-derived cells is promoted by the physical contact with the bone matrix in vivo.

The periosteum contains osteoprogenitors that differentiate to osteoblasts in bone growth or repair. Our previous studies suggested the hypothesis that the physical contact of the periosteum with the bone matrix is requisite for the differentiation of osteoblasts. To test the hypothesis, the present study was designed to investigate how the contact between the periosteum and the bone matrix influences the osteoblastic differentiation of periosteal cells with establishing a new experimental model in vivo. Differentiation of osteoblasts was assessed by gene expression of type I collagen, osteocalcin and bone sialoprotein using in situ hybridization. A barrier was designed to prevent periosteal cells from contacting the bone matrix using the membrane filter. The membrane filter was inserted surgically between the surface of rat parietal bone and the periosteum after being punched out with pin holes. Periosteal cells were allowed to contact with the bone surface only through the pin holes. The pin hole was filled with cells derived from the periosteum 1 week after inserting the filter. Differentiation of osteoblasts in week 2 and noticeable bone formation in week 3 were identified on the bone surface only under the pin hole but not under the filter. The present study demonstrated that the physical contact with the bone matrix promotes osteoblastic differentiation of periosteum-derived cells in vivo.

Animals↗

PubMed2001

The effect of short-term tooth intrusion on human pulpal blood flow measured by laser Doppler flowmetry.

The purpose was to examine the effect of brief intrusive forces on human pulpal blood flow (PBF). Laser Doppler flowmetric measurements were made from 17 vital upper left central incisors of 17 participants who had clinically healthy tooth crowns and periodontal tissues. Brief intrusive forces (0.5,1,5 N; duration 20 s) were applied to the incisal edges of the examined teeth, and apical displacement of the teeth and the PBF were measured simultaneously. Recordings were made with and without an opaque rubber dam applied to the examined teeth. Intrusive force significantly reduced PBF flux both with and without the dam (P<0.05, Friedman analysis). The results indicate that transient apical displacement can reduce PBF.

Adult↗

PubMed2001

Porphyromonas gingivalis-specific IgG subclass antibody levels as immunological risk indicators of periodontal bone loss.

BACKGROUND: It has been well demonstrated a positive association between the magnitude of host antibody response and periodontal disease status. Previous studies also reported that Porphyromonas gingivalis-specific IgG subclass antibodies were elevated in sera from adult periodontitis patients. However, the rôle and the association of these IgG subclass antibodies to the development of periodontal diseases are poorly understood. AIM: The aim of present investigation was to examine the relation of serum IgG subclass antibody levels and alveolar bone loss in treated and untreated periodontitis patients. METHODS: Serum samples were taken from 20 treated and maintained periodontitis patients (SPT patients), 30 untreated patients and 19 periodontally healthy subjects. We determined the IgG subclass antibody titers to P. gingivalis whole cells using an enzyme-linked immunosorbent assay (ELISA). Subgingival plaque samples were taken from the mesio-buccal surface of 6 randomly selected teeth of SPT patients and evaluated for the presence of P. gingivalis by immunofluorescence microscopy. Clinical measurements were also taken including full mouth intraoral radiographs to measure interproximal alveolar bone loss at baseline (BLS1) and at a 5-year recall visit in the SPT patients (BLS2). RESULTS: Our results indicated that both patient groups had detectable levels of IgG1, IgG2, and IgG4. Significantly higher IgG1 was observed in both patient groups compared to the healthy subjects. The untreated patients also exhibited significantly elevated IgG2 response (p<0.05). The mean IgG4 level of the SPT patients was significantly higher compared to the other subject group (p<0.05). A statistically significant positive correlation between IgG2 levels and changes in bone levels (DeltaBLS: BLS2-BLS1) was seen in the SPT patients (p<0.001). SPT patients with high IgG2 and low IgG4 showed greater bone loss than those with low IgG2 and high IgG4 (p<0.05), although the mean prevalence of P. gingivalis in the 2 groups did not differ significantly. CONCLUSION: Our data suggest that the prolonged IgG2 response after periodontal treatment may be indicative of recurrent or persistent periodontal destruction.

Adult↗

PubMed2001

Production of interleukin-8 and nitric oxide in human periapical lesions.

Bacterial infection of the pulp and root canal system leads to the recruitment of immunocompetent cells in the periapex and stimulates inflammatory cell responses to produce a variety of inflammatory mediators. Cytokines, reactive oxygen intermediates, and reactive nitrogen intermediates are frequently found at sites of acute inflammation. In this study, we measured the levels of interleukin (IL)-8 and nitric oxide (NO) in the periapical exudate (PE) from human periapical lesions and investigated the association of these mediators with the clinical symptoms of periapical periodontitis. PE samples were collected from root canals during routine endodontic treatments. The IL-8 concentration was measured by the enzyme-linked immunosorbent assay, and the NO level was measured as nitrite/nitrate concentration assayed by the Griess reaction. Detectable levels of IL-8 and nitrite/nitrate were found in 24 and 19 of 27 PE-samples, respectively. Although PE-IL-8 and nitrite/nitrate concentration showed a broad range, a significantly positive correlation was found between both mediators. Also, significantly higher IL-8 levels were found in PE from lesions that had painful symptoms at the sampling visit. However, there was no relationship between elevated NO levels and clinical symptoms. These results suggest that the up-regulation of IL-8 may have a critical role in the development of the symptoms of periapical disease.

Analysis of Variance↗

PubMed2000

Cleavage of CD14 on human gingival fibroblasts cocultured with activated neutrophils is mediated by human leukocyte elastase resulting in down-regulation of lipopolysaccharide-induced IL-8 production.

Activated polymorphonuclear leukocytes (PMNs) release various types of proteases and express them on the cell surface. The proteases play important roles in PMN-mediated events. In the present study, flow cytometric analysis revealed that CD14 expression on human gingival fibroblasts (HGF) was markedly reduced by PMA-activated PMNs in a coculture system. We found that this reduction was caused by both secreted and cell surface proteases produced by activated PMNs. A protease responsible for the reduction was found to be human leukocyte elastase (HLE) secreted from the activated PMNs by use of various protease inhibitors, although HLE was only partially involved in CD14 reduction caused by cell-bound molecule(s) on fixed PMNs. Analysis with purified HLE revealed a time- and dose-dependent reduction of CD14 on HGF, and complete reduction was observed by 20 microg/ml HLE treatment for 30-60 min, but the other molecules such as CD26, CD59, CD157, and MHC class I on HGF were only slightly reduced. This reduction of CD14 resulted from direct proteolysis by HLE on the cell surface, because HLE reduced CD14 on fixed HGF and also on purified cell membranes. As a result of CD14 proteolysis, IL-8 production by HGF was suppressed when triggered by 10 ng/ml LPS, but not by IL-1alpha, indicating that HLE inhibited a CD14-dependent cell activation. These findings suggested that activated PMNs have a potential negative feedback mechanism for HGF function at the inflammatory site, particularly in periodontal tissues.

Adolescent↗

PubMed2000

Immunobiological activities of a chemically synthesized lipid A of Porphyromonas gingivalis.

A synthetic lipid A of Porphyromonas gingivalis strain 381 (compound PG-381), which is similar to its natural lipid A, demonstrated no or very low endotoxic activities as compared to Escherichia coli-type synthetic lipid A (compound 506). On the other hand, compound PG-381 had stronger hemagglutinating activities on rabbit erythrocytes than compound 506. Compound PG-381 also induced mitogenic responses in spleen cells from lipopolysaccharide (LPS)-hyporesponsive C3H/HeJ mice, as well as LPS-responsive C3H/HeN mice. The addition of polymyxin B resulted in the inhibition of mitogenic activities, however, compound 506 did not show these capacities. Additionally, compound PG-381 showed a lower level of activity in inducing cytokine production in peritoneal macrophages and gingival fibroblasts from C3H/HeN mice, but not C3H/HeJ mice, in comparison to compound 506. Thus, this study demonstrates that the chemical synthesis of lipid A, mimicking the natural lipid A portion of LPS from P. gingivalis, confirms its low endotoxic potency and immunobiological activity.

Animals↗

PubMed1999

Nasopharyngeal-associated lymphoreticular tissue (NALT) immunity: fimbriae-specific Th1 and Th2 cell-regulated IgA responses for the inhibition of bacterial attachment to epithelial cells and subsequent inflammatory cytokine production.

To investigate the antibacterial activity of mucosal Th1 and Th2 immune responses induced nasally and orally, mice were immunized with mucosal vaccine containing fimbrial protein of Porphyromonas gingivalis, a causative agent for a destructive chronic inflammation in the periodontium, and cholera toxin (CT) as mucosal adjuvant. Nasal vaccine containing low doses of fimbriae (10 micrograms) and CT (1 microgram) induced Ag-specific Th1/Th2-type response in CD4+ T cells in mucosal effector tissues, including nasal passage and submandibular glands, which accounted for the generation of Ag-specific IgA-producing cells. In contrast, oral immunization required higher amounts of fimbriae and CT for the induction of Ag-specific IgA responses. Fimbriae-specific IgA mAbs generated from submandibular glands of nasally immunized mice inhibited P. gingivalis attachment to and reduced subsequent inflammatory cytokine production from epithelial cells. These findings suggest that nasal vaccination is an effective immunization regimen for the induction of Ag-specific Th1 and Th2 cell-driven IgA immune responses that possess the ability to inhibit bacterial attachment to epithelial cells and subsequent inflammatory cytokine production.

Animals↗

PubMed1999

Autoregulatory effect of interleukin-10 on proinflammatory cytokine production by Porphyromonas gingivalis lipopolysaccharide-tolerant human monocytes.

Pretreatment of human peripheral blood monocytes with a very low concentration (0.1 ng/ml) of Porphyromonas gingivalis lipopolysaccharides (LPS) resulted in a significant decrease of interleukin-6 (IL-6) production, but not IL-8 production, by restimulation of a high concentration (1 microg/ml) of the same LPS. In contrast, the same pretreatment with Escherichia coli LPS resulted in the enhanced production of both IL-6 and IL-8 after restimulation. The selective induction by P. gingivalis LPS tolerance of IL-6 production developed in a time-dependent manner during the primary culture. P. gingivalis LPS-pretreated cells were also refractory to a high-dose E. coli LPS restimulation in terms of IL-6 production. The expression of IL-6 mRNA decreased 10 h after restimulation of P. gingivalis LPS-pretreated monocytes. Furthermore, an up-regulation of anti-inflammatory cytokine IL-10 upon a second high-dose LPS rechallenge occurred at the same time point in the pretreated cells. We studied the role of IL-10 in the process of IL-6 down-regulation. Neutralization by an anti-IL-10 polyclonal antibody prevented IL-6 down-regulation in P. gingivalis LPS-pretreated monocytes, whereas IL-8 production was not affected. Addition of exogenous IL-10 during the high-dose LPS stimulation of untreated cells substituted for the LPS pretreatment and resulted in the inhibition of IL-6 production in a dose-dependent manner. A higher dose of IL-10 was required to suppress IL-8 synthesis from monocytes. Our data suggest that IL-10 mediates IL-6 down-regulation in P. gingivalis LPS-tolerant monocytes in an autocrine manner.

Base Sequence↗

PubMed1998

Balance of interleukin-1 beta and interleukin-1 receptor antagonist in human periapical lesions.

Interleukin-1 beta (IL-1 beta) has been considered as a major potent mediator of bone resorption and implicated in the development of human periapical lesions. Among naturally occurring interleukin-1 (IL-1) inhibitors, IL-1 receptor antagonist (IL-1ra) is a 22 kDa protein that shares homology with IL-1 beta and IL-1 alpha, binds to IL-1 receptor with similar affinity to IL-1, and has no known agonist properties. In this study, we measured the periapical exudate (PE) levels of IL-1 beta and IL-1ra from human periapical lesions. PE samples were collected from root canals during routine endodontic treatments, and the enzyme-linked immunosorbent assay was used to measure PE-IL-1 beta and IL-1ra. Detectable levels of both IL-1 beta and IL-1ra were found in 25 of 29 clinical samples. Relatively high levels of IL-1ra compared with IL-1 beta (mean IL-1ra:IL-1 beta ratio = 128:7; range: 0.9 to 495.4), and significantly positive correlation between IL-1ra and IL-1 beta levels was found. The PE-IL-1ra:IL-1 beta ratios obtained from symptomatic lesions were significantly lower than those from asymptomatic lesions. These results suggest that IL-1ra-mediated IL-1 antagonism occurred to block locally produced IL-1 activity, and the balance of IL-1 to IL-1ra production may be crucial in the development of periapical lesions.

Analysis of Variance↗

PubMed1998

Antigen direction of specific T-cell clones into gingival tissues.

This study was performed to investigate T-cell traffic to periodontal tissues during infection with a periodontal pathogen Actinobacillus actinomycetemcomitans (Aa). Rowett rat T-cell clones, A3 (CD4+ CD8-, alpha beta TCR+, NKRP-1-, specific to Aa) and G2 (CD4- CD8-, alpha beta TCR+, NKRP-1+, which reacts to Aa, Gram-negative and -positive bacteria), both expressed the same prominent adhesion molecules (LFA-1, VLA-4) to the same extent. Binding of both T-cell clones to rat endothelial cells in vitro was blocked by antibody to VLA-4. Rowett rats were infected with Aa and infused with Aa-stimulated, isogenic T-clone lymphocytes that had been labelled in vitro with 125IUdR. Radioactivity associated with recovery of clone A3, but not G2, was significantly elevated in the gingivae of infected rats, suggesting migration to infected animals' gingival tissues. Migration of radioactive Aa-specific A3 clone cells traced by autoradiography reached a maximum at 24 hr (1.2% of total lymphocytes as radiolabelled cells in infected gingiva versus 0.6% in noninfected), indicating an apparent antigen-directed retention in infected rats' gingival tissues. The G2 clone was not retained in the gingival tissues (0.20% of total lymphocytes as radiolabelled cells in infected gingiva versus 0.26% in non-infected). However, the possibility of A3 retention directed by inflammation or tissue-selective homing could not be excluded. In further experiments, other adoptively transferred T-clone lymphocytes [clones G23 (Th1) and F13 (Th2)] with specificity for the 29,000 MW outer membrane protein of Aa with the same prominent adhesion molecules could be recovered from rat gingivae previously challenged with this antigen. However, transferred T-clone lymphocytes [clone G26 (Th1)] with specificity for a different Aa antigen were not recovered. Therefore, the dynamics of cell entry into periodontal lesions vary for activated T lymphocytes with different antigenic specificities, indicating the significance of antigen in lymphocyte traffic to periodontal tissues.

Actinobacillus Infections↗

PubMed1997

The change of periapical exudate prostaglandin E2 levels during root canal treatment.

We have previously demonstrated that detectable levels of prostaglandin E2 exist in periapical exudates (PE-PGE2) from root canals of periapical periodontitis. Our data from a cross-sectional study also suggested the association of PE-PGE2 concentrations with the active phase of periapical lesions. The objective of this study was to investigate the longitudinal changes of PE-PGE2 levels during root canal treatment. Periapical exudate specimens were quantitatively sampled from root canals of 20 nonvital teeth at consecutive treatment visits to measure PGE2 concentrations. The mean PE-PGE2 levels significantly decreased following the endodontic therapy (158.0 +/- 54.1 pg/microliter in the first samples versus 48.4 +/- 37.9 pg/microliter in the second samples). A significant correlation was also found between the changes in PE-PGE2 levels and the clinical disease expression of periapical periodontitis (p < 0.01). Decreased PGE2 production is considered to reflect the remission of disease activity. These results suggest the central roles of PGE2 in regulatory mechanisms for the expression of periapical disease.

Dental Pulp Cavity↗

PubMed1997

Hyporesponsiveness of inflamed human gingival fibroblasts from patients with chronic periodontal diseases against cell surface components of Porphyromonas gingivalis.

Inflamed human gingival fibroblasts (HGF) of patients with chronic periodontal diseases have less active interleukin-8 (IL-8) production compared with normal HGF of volunteers with healthy gingival tissues, after stimulation with Porphyromonas gingivalis surface components such as fimbriae, lipopolysaccharide (LPS) and its lipid A, but not LPS or lipid A from other bacterial species. A decrease in number of specific binding sites for P. gingivalis fimbrial molecules in inflamed HGF is also observed by Scatchard plot analysis. A short exposure (6 h) to P. gingivalis LPS resulted in significant potentiation of the LPS-dependent IL-8 production in normal HGF, whereas a long exposure (48 h) to the LPS significantly reduced IL-8 production. Tyrosine phosphorylation of proteins of 127 kDa and 186 kDa in inflamed HGF stimulated with P. gingivalis fimbriae or its LPS was observed by immunoblotting, and these two phosphoproteins were termed tolerance-induced protein, TIP. Protein bands of 45 kDa which bound to radioiodinated P. gingivalis fimbriae in the presence and absence of fetal bovine serum (FBS), and major 73-kDa and minor 30-kDa and 45-kDa bands which bound to radioiodinated P. gingivalis LPS in the presence of FBS in normal and inflamed HGF were observed by using photocrosslinking. These findings suggest that the hyporesponsiveness of HGF induced by a prolonged exposure to P. gingivalis may emerge because of HGF damage or result from host defense in chronic periodontal lesions.

Adult↗

PubMed1996

Splenic B-cell activation in lipopolysaccharide-non-responsive C3H/HeJ mice by lipopolysaccharide of Porphyromonas gingivalis.

Porphyromonas gingivalis 381 lipopolysaccharide (LPS) definitely exhibited mitogenic activity in purified B-cells, separated from spleens of LPS-responsive C3H/HeN mice and LPS-non-responsive C3H/HeJ mice by using a magnetic cell sorting system. The mitogenic activity induced by P. gingivalis LPS was incompletely inhibited by polymyxin B. P. gingivalis LPS also induced a higher production of interleukin-6 (IL-6) in splenic B-cells of C3H/HeN mice as compared with Escherichia coli LPS. Furthermore, P. gingivalis LPS, but not E. coli LPS, induced definite IL-6 production in C3H/HeJ mice. P. gingivalis LPS increased tyrosine, serine/threonine phosphorylation of proteins with various major induced bands in splenic B-cells of both C3H/HeN and C3H/HeJ mice. Additionally, radioiodinated P. gingivalis LPS, similarly to E. coli LPS, bound to a 73-kDa protein on C3H/HeJ as well as C3H/HeN B-cells. Thus P. gingivalis LPS may activate B-cells of C3H/HeJ as well as C3H/HeN mice via the LPS-specific binding protein on the cells.

Acute-Phase Proteins↗