摘要:

Because a general study of activated neutrophils may have relevance to periodontal diseases and accompanying inflammation, we studied a function of mouse polymorphonuclear leukocytes (PMNs) that exude into the peritoneal cavity in response to inflammation caused byi.p.injection of 2% casein. The effects ofE. coli‐lipopolysaccharide (E‐LPS) and a chemotactic factor, N‐for‐myl‐N‐methionyl‐N‐leucyl‐L‐phenylalanine (FMLP), on the level of intracellular calcium ([Ca2+]i) in these PMNs were examined. From analysis made with a laser cytometer (ACAS 570), the PMNs in exudates harvested 3–9 h after the onset of inflammation were shown to undergo [Ca2+]i elevation in response to 10–6M FMLP. The peak concentration of [Ca2+]i elicited by FMLP was highest in exudate cells 6 h after casein injection. In addition, about 65% of the PMNs in the 3‐h exudate were FMLP sensitive displaying an elevated [Ca2+]i, whereas more than 85% of them in 6‐ and 9‐h exudates became FMLP sensitive. Also, the maximum level of [Ca2+]i after FMLP stimulation was potentiated by pretreatment of the cells with E‐LPS (0.2 μg/ml). The present study suggests that PMNs induced by casein injection and appearing in mouse peritoneal exudate at different times possess significantly different ability to undergo [Ca2+]i elevation, and different susceptibility t

ISSN:0022-3484    

DOI:10.1111/j.1600-0765.1995.tb01267.x

出版商:Blackwell Publishing Ltd    

年代:1995

数据来源: WILEY

摘要:

SummaryOur previous studies showed that the expression of CD23 on polymorphonuclear leukocytes (PMNLs) in gingival crevicular fluid (GCF) from adult periodontitis (AP) patients was higher than in autologous peripheral blood (PB). Percentages of eosinophils in GCF PMNLs ranged between 6 and 14%. The purpose of the present studies was to increase understanding of the potential role of eosinophils and their products, including CD23, in periodontal disease. We analysed the eosinophil fraction in GCF and PB by flow cytometry using monoclonal antibodies to CD23b (BB10), eosinophil cationic protein (ECP) in stored and secretory forms (EG1 and EG2), and CD67 (80H3). Simultaneously, we measured IgE and soluble CD23 titer and GCF and serum by ELISA. Flow cytometric analysis of BB10, EG2 and 80H3 binding showed that GCF eosinophils from AP were activated. A large BB10+EG2+cellular fraction was detected in GCF from AP whereas it was very low in autologous serum (9.30±2.460 vs 0.16±0.10, p>0.001). GCF from gingivitis patients exhibited no flow cytometric evidence for the presence of BB10+ EG2+ cells. BB10+ EG1+ cells, or inactivated eosinophils rated lower in GCF than in PB both in gingivitis and periodontitis patients (0.45±0.63 vs 1.83±0.96 and 0.15±0.30 vs 1.30±0.20, p>0.05. respectively). IgE titer in AP patients reached 1208.1 ±421.2 IU/ml in GCF while only 49.1 ±50.4 in sera. Soluble CD23 in GCF reached 236.1 ±81.3 ng/ml in GCF and 5.6±1.8 ng/ml in sera. GCF of gingivitis patients, however, contained no detectable sCD23. Thus. GCF from AP patients displayed a high rate of activated eosinophils secreting ECP. while GCF of gingivitis patients did not. These results suggest that ECP‐secreting activated eosinophils are relevant to the pathology of adult pe

ISSN:0022-3484    

DOI:10.1111/j.1600-0765.1995.tb01268.x

出版商:Blackwell Publishing Ltd    

年代:1995

数据来源: WILEY

摘要:

SummaryMany bacterial and host cells contain large amounts of polyamines that can be released at infection sites as a result of cell lysis. Consequently, the putrescine and spermidine content of gingival fluid from inflamed periodontal pockets (0.1 to 1 mM) is sharply elevated in comparison to peripheral blood. At these levels, polyamines potentiated fMet‐Leu‐Phe‐induced Ca2+signaling in polymorphonuclear leukocytes (PMNs)in vitro. Consistent with the essential role of Ca2+signaling in PMN activation, secondary granule release and superoxide anion production by fMet‐Leu‐Phe‐stimulated PMNs was enhanced in the presence of polyamines. Thus, polyamines may play a local role in modulating the antimicrobial activity of PMNs in periodon

ISSN:0022-3484    

DOI:10.1111/j.1600-0765.1995.tb01269.x

出版商:Blackwell Publishing Ltd    

年代:1995

数据来源: WILEY

摘要:

SummaryUsing two‐dimensional (2D) electrophoresis and western blot assay, we analyzed antigenic proteins inPorphyromonas gingivalisuniquely recognized by antibodies in sera of periodontitis subjects. Proteins in the total membrane fraction of P.gingivalis381 were resolved into at least 70 protein spots by 2D electrophoresis. In the gel stained with silver, the substance around 47 kDa protein on the acidic side (at an isoelectric point of about 4.5) was stained as a smear. Antigenic substances were characterized using purified IgGs from sera of 16 adult periodontitis (AP), 19 rapidly progressive periodontitis (RPP) and 14 periodontally healthy volunteers. Western blots demonstrated that 75 kDa protein reacted with IgGs from 75% of AP patients (p>0.001), the antigenic substance around acidic 47 kDa protein reacted with IgGs from 81.3% of AP (p>0.01) and 68.4% of RPP patients (p>0.01) and the acidic 47 kDa protein reacted with 87.5% of AP (p>0.01) and 78.9% of RPP patients (p>0.01). The reaction frequency was significantly different from that of the healthy volunteers. Also 51 kDa and 41 kDa proteins reacted with 47 and 43 of 49 IgG samples, respectively. The substance around acidic 47 kDa protein reacted with mouse antiserum to P.gingivalis‐LPS. After treatment with pronase or heat, the antigenic reactions disappeared not only from the proteins, but also from the area around the acidic 47 kDa protein. When the fraction was digested with lipase, the antigenic reaction of the area decreased. It appeared that the acidic 47 kDa protein and the LPS around it formed a complex, and that the LPS contained the lipid portion. These results suggested that the acidic 47 kDa protein can be classified among the lipid A‐associated proteins, and that it is the major antigenic protein which is recognized by antibodies in sera of almost periodontal patients in P. gingivali

ISSN:0022-3484    

DOI:10.1111/j.1600-0765.1995.tb01270.x

出版商:Blackwell Publishing Ltd    

年代:1995

数据来源: WILEY

摘要:

SummaryNeuropeptides, including substance P (SP) may play a role in neurogenic inflammation. Although SP‐immunoreactive (SP‐IR) axons are known to be present within the oral mucosa (OM) and salivary glands, the functional significance of SP in oral mucosa and sublingual salivary gland (SLG) is not fully understood. The present experiments were carried out to study the effects of SP infused into the left common carotid artery on vascular permeability in the OM and in the SLG of male rats. Vascular permeability was assessed on the basis of Evans Blue extravasation. Separate groups of animals received histamine (H1) receptor antagonist (chloropyramine, 10 mg kg−1i.v.) or prostaglandin synthesis inhibitor (indomethacin, 4 mg kg−1i.v.) prior to SP infusions. Infusion of SP in doses of 30 and 74 pmol min−1increased the vascular permeability of OM by 162.3±16.3% (n=8, p>0.05) and 482.7±46.7% (n=8, p>0.001), respectively. SP in a dose of 15 pmol min−1did not increase Evans Blue extravasation in OM (38.3±4.0 μ g−1, n=8, compared to the control: 44.0±7.9 μg g−1, n = 8, NS). Although SP increased plasma extravasation in OM, it failed to affect vascular permeability in SLG (15 pmol min−1SP: 46.9±6.9 μg g–1, n = 6, NS; 30 pmol min−1SP: 54.1 ±6.2 μg g−1, n= 11 NS; 74 pmol min−1SP: 49.7±2.3 μg g−1, n = 7, NS; compared to the control: 48.9±5.8 μg g−1, n = 8). After chloropyramine administration the SP effect on vascular permeability decreased in OM by 41.5±5.9% (n=10, p>0.05). Indomethacin pretreatment similarly diminished the effect of SP on the dye extravasation in OM by 43.9±6.1% (n = 8, p>0.01). Our results suggest that the effect of SP on plasma extravasation in the oral mucosa is partly elicited via the release of vasoactive agents (histamine, prostaglandins), and the microvasculature of SLG has lower s

ISSN:0022-3484    

DOI:10.1111/j.1600-0765.1995.tb01271.x

出版商:Blackwell Publishing Ltd    

年代:1995

数据来源: WILEY

摘要:

SummaryEffects of and interactions between tumour necrosis factor α (TNFα) and bradykinin (BK) on production of interleukin‐1 (IL‐lα, IL‐lβ) in human gingival fibroblasts were studied. The cytokine TNFα induced production of cell‐associated IL‐lα and IL‐1β in gingival fibroblasts, with IL‐lβ being most abundant. Addition of BK, in the presence of TNFα, for 1 h and 6 h, respectively, synergistically enhanced the TNFα induced IL‐lβ production, whereas BK alone did not induce 1L‐1 production. Similar to BK, two phorbol esters, phorbol 12,13 dibutyrate (PDBu) and phorbol 12‐myristate‐13‐acetate (PMA) which are known to stimulate protein kinase C (PKC), synergistically enhanced the TNFα induced IL‐lβ production in the gingival fibroblasts. On the contrary, a phorbol ester which does not activate protein kinase C, 13‐phorbolacetate (13‐PA), did not potentiate the TNFα induced IL‐lβ production. Similar to BK, the phorbol esters (PMA, PDBu, 13‐PA) alone did not induce IL‐1β production in the gingival fibroblasts. The results indicate that TNFα induces production of cell‐associated IL‐1 in gingival fibrobla

ISSN:0022-3484    

DOI:10.1111/j.1600-0765.1995.tb01272.x

出版商:Blackwell Publishing Ltd    

年代:1995

数据来源: WILEY

摘要:

SummaryPrevious reports have indicated that certain mouthrinses, even when used as directed can induce oral pain. In order to help determine the causal agent(s), various commercially available mouthrinses, as well as control solutions, were tested in a psychophysical study in which subjects rated categories of pain during and after mouthrinsing. More specifically, the studies tested the effects of ethanol concentration on induced pain. The results show that there is a direct relationship between ethanol content and the amount of induced pain. Furthermore, the amount of pain was found to increase with time of rinsing, and to slowly decrease after cessation of rinsing. Lastly, comparison of ethanol/water controls with a marketed product (Clear Choice®) matched for ethanol content showed that, while ethanol was the key factor in mouthwash‐induced oral pain, other presently unidentified agents can also add to the effe

ISSN:0022-3484    

DOI:10.1111/j.1600-0765.1995.tb01273.x

出版商:Blackwell Publishing Ltd    

年代:1995

数据来源: WILEY

摘要:

SummarySoluble endothelial leukocyte adhesion molecule‐1 (sE‐selectin) levels in peripheral blood (PB) and gingival capillary blood (GCB) of both healthy donors (HD) and patients with adult periodontitis (AP) were assayed by ELISA. Binding of sE‐selectin to polymorphonuclear neutrophils (PMN) from PB, GCB and crevicular fluid (GF), and expression of L‐selectin and sialyl‐Lewisx(sLex) on these cells were analyzed by immunofluorescence and flow cytometry. No significantly enhanced serum levels of sE‐selectin in patients with AP. compared to HD (28±5 ng/ml vs 19±3 ng/ml, respectively), and no differences in the concentration of sE‐selectin in GCB (16±1 ng/ml vs 16±2 ng/ml, respectively) were observed. On PB‐PMN no significant differences in the expression of L‐selectin and sLexwere found and binding of sE‐selectin to PB‐PMN was comparable between HD and patients with AP. Binding of sE‐selectin to GCB‐PMN was significantly higher in patients with AP compared to HD (mean channel fluorescence (MCF)=88.5± 13.2 vs MCF=24.2±5.3, respectively). The expression of sLexon GCB‐PMN did not differ significantly between the two groups. A significant decrease in the expression of the adhesion molecule L‐selectin on GCB‐PMNs compared to PB‐PMN was found in patients with AP but not in HD. CF‐PMN showed decreased expression of both L‐selectin and sLexcompared to PMN from PB and GCB, both in HD and patients with AP. Taken together, these data suggest that PMN from patients with AP had reduced selectin‐mediated adhesive

ISSN:0022-3484    

DOI:10.1111/j.1600-0765.1995.tb01274.x

出版商:Blackwell Publishing Ltd    

年代:1995

数据来源: WILEY

摘要:

SummaryAlthough vital plaque micro‐organisms are part of the natural ecosystem in the oral cavity they are also the key factor in the development of diseases induced by the human dental plaque. In a previous study (9) the portion of vital bacteria related to the total number of plaque micro‐organisms (i.e. the microbial vitality) appeared low in small plaque samples. The objective of this investigation was to determine the exact relationship of microbial vitality and age of supragingival plaque during the early phases of human dental plaque formation. Between intervals of optimal oral hygiene, thirteen participants refrained from all oral hygiene measures for periods of 1, 2, 4, 8, 24 and 72 h. Plaque was completely sampled from a defined area situated on the vestibular surface of the teeth 13, 14, 15, 23, 24 and 25. The pooled plaque from these areas was immediately processed. Total bacterial counts (BC) as enumerated by darkfield microscopy, and colony‐forming units (CFU) were recorded. The microbial vitality was calculated indirectly as plating efficiency (PE=CFU per BC) and directly assessed using a vital fluorescence (VF) technique. In the 1 h old plaque samples the median values of PE and VF were 29% and 18%, respectively. Thereafter, the microbial vitality increased significantly with plaque age. The 24 h old plaque samples yielded values of 77% (PE) and 62% (VF). It was concluded that the microbial vitality of the early dental plaque investigated was considerably lower compared to that of a more mature p

ISSN:0022-3484    

DOI:10.1111/j.1600-0765.1995.tb01275.x

出版商:Blackwell Publishing Ltd    

年代:1995

数据来源: WILEY

摘要:

SummaryA 3‐dimensional gingival epithelial model has been developed and characterized. Oral epithelial cells and connective tissue fibroblasts were isolated from human gingival tissue and used to create anin vitrooral mucosa co‐culture model. Fibroblasts were seeded on a scaffold of nylon mesh, allowed to proliferate and secrete collagen and extracellular matrix proteins to form a stroma capable of supporting the growth of epithelial cells. Epithelial cells were seeded on top of a confluent stromal layer, proliferated and differentiated to form a stratified squamous epithelium. Resident epithelial cells were stimulated, by manipulation of growth medium and culture conditions, to form a multi‐layered oral mucosa‐like tissue. Histologic analyses revealed cellular architecture exhibiting stromal‐epithelial interaction which supports the growth and differentiation of an epithelial layer. lmmunohistochemical analyses confirmed production of types I and III collagen. Immunofluorescence of the stromal layer identified type IV collagen and fibronectin. Fibronectin was also detected on surface epithelium. Differentiation of basal, spinous and granular cells was observed, and the presence of differentiation markers, acidic (K10, 14–16, 19) and basic (Kl–8) cytokeratins were confirmed using broad spectrum cytokeratin antibodies, AE1 and AE3. Development of a discontinuous basal lamina zone, with hemidesmosomes, was observed by electron microscopy. The co‐culture was metabolically active, as measured by the thiazoyl blue (MTT) assay for mitchondrial function and [3H] thymidine incorporation into DNA. The human gingival epithelial co‐culture model was viable up to 35 days post‐epithelial seed. This model may offer opportunities for limited study of periodontal ti

ISSN:0022-3484    

DOI:10.1111/j.1600-0765.1995.tb01276.x

出版商:Blackwell Publishing Ltd    

年代:1995

数据来源: WILEY