Effects of Lactobacillus reuteri-containing lozenges on healing after surgical removal of mandibular third molars: a randomised controlled trial.

We investigated the effect of probiotic supplements on oral wound healing, swelling, pain and discomfort after surgical removal of mandibular third molars. A second aim was to evaluate if the intervention could influence the concentrations of oxytocin in saliva. Sixty-four consecutive volunteers (18-34 years) were enrolled to a double-blind randomised placebo-controlled trial with two parallel arms. Following surgery, the patients were asked to take three lozenges per day containing two strains of Lactobacillus reuteri (DSM 17938 and ATCC PTA 5289) or placebo for two weeks. The clinical healing and extra-oral swelling were scored two weeks post-operatively. Samples of wound exudate were cultivated for the presence of Staphylococcus aureus and ß-haemolytic streptococci. Salivary oxytocin concentrations were analysed from pre- and post-surgery samples using ELISA technique. Compliance and the subjective perception of swelling, pain and discomfort were reported daily through visual analogue scales in a logbook. All patients except three completed the protocol and the postoperative course was uneventful in most cases. Minor extra-oral swellings were noted in five patients, but none required antibiotic treatment. At the 2-week follow-up, there were no significant differences in clinical wound healing index, extra-oral swelling, bacterial growth or salivary oxytocin levels between the groups. The self-reported data unveiled, however, a significantly reduced sense of swelling, in particular during the second week after surgery in the probiotic test group (P<0.05). Likewise, significantly fewer nights with disturbed sleep and fewer days with sick-leave from work were reported among the participants in the test group (P<0.05). No differences were found in the post-operative use of analgesics. In conclusion, we found no significant influence of probiotic supplements on objective wound healing after surgical extraction of impacted mandibular third molars. However, since the patients' perceived significant post-operative ameliorations, further studies are needed to explore the patient's value of the intervention.

Effect of probiotic lozenges containing Lactobacillus reuteri on oral wound healing: a pilot study.

It has been suggested that beneficial bacteria may stimulate wound healing. The aim was to investigate the effect of topical applications of probiotic lactobacilli on the healing of standardised oral wounds. This pilot study employed a randomised, placebo-controlled, double-blind cross-over design. Standardised biopsies were punched in the oral mucosa of 10 healthy volunteers, with and without exposure to two strains of Lactobacilli reuteri administrated as lozenges and topical oil. The healing was scored clinically after 2, 5 and 8 days. The amount of exudate was quantified through filter papers and the levels of selected cytokines and chemokines were determined with multiplex immunoassays. Saliva samples were collected before the biopsy and after healing for determination of oxytocin with ELISA. Subjectively perceived pain and discomfort was reported through a daily logbook. There was a clear tendency of improved healing in test group at the 2-and 5-day check-ups but the difference compared with the placebo intervention was not statistically significant (P=0.08). Higher but non-significant expressions of the tumour necrosis factor (TNF) superfamily ligand members 13 (APRIL) and 13B (BAFF), as well as the chemokine interleukin 8 (IL-8), were displayed in wound exudates from the probiotic group as compared with placebo, particularly after 5 and 8 days. The salivary levels of oxytocin were significantly lower (P<0.05) in the placebo group at the 8-day follow-up. The mean number of days with pain and/or discomfort after the biopsies was similar in both groups. No side-effects were reported. The findings of this pilot study justify a larger clinical trial to elucidate the possible role of probiotic supplements on oral wound healing.

The relationship between matrix metalloproteinases (MMP-3, -8, -9) in serum and peripheral lymphocytes (CD8+ , CD56+ ) in Down syndrome children with gingivitis.

BACKGROUND AND OBJECTIVE: Altered immune response may be a major contributor to periodontal disease in Down syndrome. This study investigated the relationship between peripheral lymphocytes and matrix metalloproteinases (MMPs) in serum in Down syndrome children with gingivitis. MATERIAL AND METHODS: Children with Down syndrome (n = 10) and healthy controls (n = 10) were clinically and radiographically examined during dental treatment under general anaesthesia. Peripheral blood and gingival crevicular fluid were collected from each subject and concentrations were determined: serum MMP-2, -3, -8 and -9; serum tissue inhibitors of metalloproteinases (TIMP) -1, -2 and -3; and gingival crevicular fluid. Leukocytes were isolated from peripheral blood and the relative amounts (%) of the various cell phenotypes were analysed using flow cytometry. In addition, peripheral blood cells were treated with Porphyromonas gingivalis lipopolysaccharide and levels of MMPs and TIMPs measured. RESULTS: Concentrations of MMP-3, MMP-8 and TIMP-1 in serum were significantly higher (p < 0.05) in the Down syndrome group compared to the controls. When peripheral blood leukocytes were cultured in the presence or absence of P. gingivalis lipopolysaccharide, MMP-8 levels were significantly (p < 0.05) higher in the Down syndrome group compared to controls. Children with Down syndrome exhibited significant positive correlations between CD8(+) T cells and MMP-8 (r = 0.630; p = 0.050), between CD8(+) T cells and MMP-9 (r = 0.648; p = 0.043), and between CD56(+) NK cells and MMP-3 (r = 0.828; p = 0.003) compared to controls. CONCLUSIONS: The positive relationship of serum MMP-3, -8 and -9 with immune cells in children with Down syndrome may facilitate migration of CD8(+) T cells and CD56(+) NK cells into the periodontal tissue, which may contribute to the increased degradation of periodontal tissue in individuals with Down syndrome.

Altered relationship between MMP-8 and TIMP-2 in gingival crevicular fluid in adolescents with Down's syndrome.

BACKGROUND AND OBJECTIVE: Periodontitis is more frequently found in subjects with Down's syndrome. The aim was to investigate whether the relationship between MMPs and TIMPs) in the gingival crevicular fluid of subjects with Down's syndrome is altered compared with controls. MATERIAL AND METHODS: Twenty-one adolescents with Down's syndrome and gingivitis (DS-G), 12 subjects with Down's syndrome and periodontitis (DS-P), 26 controls with gingivitis (HC-G) and eight controls with periodontitis (HC-P) were clinically examined. All patients were between 11 and 20 years of age. Gingival crevicular fluid was collected from each subject and the concentrations of MMPs (2, 3, 8, 9 and 13) and TIMPs (1, 2 and 3) (expressed as pg/µL adjusted for volume of gingival crevicular fluid) were determined using multianalyte kits from R&D Systems. RESULTS: The concentrations of MMP-2, MMP-3, MMP-8, MMP-9 and TIMP-2 in gingival crevicular fluid were significantly higher (p < 0.005) in the DS-G group compared with the HC-G group. The correlation coefficient between MMP-8 and TIMP-2 differed significantly (p = 0.006) between the DS-G group and the HC-G group. On the contrary, the correlation coefficients between MMPs and TIMPs did not differ significantly between the DS-P group and the HC-P group. However, the DS-P group exhibited a significantly lower concentration of TIMP-2 in the gingival crevicular fluid compared with the HC-P group. CONCLUSION: Down's syndrome subjects with gingivitis exhibit higher concentrations of MMPs in gingival crevicular fluid with an altered relationship between MMP-8 and TIMP-2, which might impair the periodontal tissue turnover.

Differences in metabolic risk factors between normal weight and overweight children.

BACKGROUND: The effect of overweight on metabolic risk factors and the role of physical activity (PA) in pre-pubertal children is unclear. OBJECTIVE: To study differences in metabolic risk factors between groups of normal weight and overweight children and how these risk factors are associated with objectively measured PA and cardio-respiratory fitness (CRF). DESIGN: A cross-sectional study was conducted with 68 children aged 8?11 years. Children were categorized into normal weight (n = 39) and overweight/obese (n = 24/5). PA and CRF were measured objectively. An oral glucose tolerance test (OGTT) was performed and triglycerides (TG) and HDL-cholesterol (HDL-C) were measured. A metabolic risk score (MRS) was calculated from the standardized values of insulin, glucose, TG, inverted HDL-C and blood pressure. RESULTS: There was a significant (P < 0.05) difference between normal weight and overweight children in clustered metabolic risk, insulin (AUC), fasting insulin and systolic blood pressure. PA and CRF did not differ significantly between groups. In linear regression analysis combining the two groups, PA was negatively associated with insulin (AUC) (? = ?0.25, 95% CI = ?0.50, ?0.002) and CRF was negatively associated with fasting insulin (? = ?0.41, 95% CI = ?0.67, ?0.15). CONCLUSIONS: Metabolic risk factors are elevated in overweight pre-pubertal children compared with normal weight controls. This is not explained by lower PA or CRF in the overweight group although PA and CRF were associated with lower insulin levels in pooled analyses. This highlights the importance of preventing overweight in children from an early age in order to prevent the metabolic syndrome and its associated diseases.

Triclosan reduces microsomal prostaglandin E synthase-1 expression in human gingival fibroblasts.

OBJECTIVE: The effect of triclosan (2,4,4'-trichloro-2'-hydroxydiphenyl ether) on the expression of cyclooxygenase-2 (COX-2) and microsomal prostaglandin E synthase-1 (mPGES-1) and on the translocation of the nuclear factor-kappaB (NF-kappaB) in relation to prostaglandin E2 (PGE2) production was investigated in human gingival fibroblasts challenged with tumor necrosis factor alpha (TNFalpha). METHODS: Fibroblasts were established from gingival biopsies obtained from six children. COX-2 mRNA and protein expression was quantified using mRNA quantitation and enzyme immunometric assay kits. mPGES-1 mRNA was analysed by RT-PCR, mPGES-1 protein and NF-kappaB translocation by immunoblotting. PGE2 was determined by radioimmunoassay. RESULTS: The cytokine TNFalpha enhanced the expression of mRNA as well as the protein levels of both COX-2 and mPGES-1 and subsequently the production of PGE2 in gingival fibroblasts. Treatment of gingival fibroblasts with triclosan (1 microg/ml) significantly reduced the stimulatory effect of TNFalpha (10 ng/ml) on the expression of mPGES-1 at both the mRNA and the protein level by an average of 21% and 43%, respectively, and subsequently the production of PGE2 (p<0.01). Triclosan did not, however, affect the translocation of NF-kappaB or the expression of COX-2 in TNFalpha-stimulated cells. CONCLUSION: The results show that triclosan reduces the augmented biosynthesis of PGE2 by inhibiting the mRNA and the protein expression of mPGES-1 in gingival fibroblasts. This finding may partly explain the anti-inflammatory effect of the agent previously reported in clinical studies.

Matrix metalloproteinase-1 and tissue inhibitor of metalloproteinase-1 production in human gingival fibroblasts: the role of protein kinase C.

BACKGROUND: Matrix metalloproteinase-1 (MMP-1) plays an important role in tissue remodelling and in the pathology of inflammatory diseases including periodontitis. The activity of MMP-1 is firmly controlled by the endogenous tissue inhibitor of metalloproteinase-1 (TIMP-1). OBJECTIVE: The aim of the study was to investigate the production and regulation of MMP-1 and TIMP-1 with special regards to the enzyme protein kinase C (PKC) in human gingival fibroblasts. METHODS: Gingival fibroblasts were treated with substances related to PKC such as phorbol 12-myristate 13-acetate (PMA), interleukin-1beta, Ca2+ -ionophore A231817 and inhibitors of PKC, p38 mitogen-activated protein kinase (p38 MAPK) and tyrosine kinase. RESULTS: The PKC activator PMA stimulated the production of MMP-1 and TIMP-1 at both the transcriptional and the translational level. The production of MMP-1 and TIMP-1 stimulated by PMA was abolished by the PKC inhibitor bisindolylmaleimide. Treatment of the cells with interleukin-1beta or A23187 synergistically increased the stimulatory effect of PMA on MMP-1 production. In contrast, TIMP-1 production was unaffected by interleukin-1beta and reduced by A23187. Tyrosine kinase inhibitor herbimycin A reduced MMP-1 production induced by PMA, whereas the p38 MAPK-inhibitor SB 203580 synergistically increased the stimulatory effect of PMA on both MMP-1 and TIMP-1 production. CONCLUSION: The present study shows that MMP-1 and TIMP-1 production is regulated differently by interleukin-1beta and calcium in human gingival fibroblasts and that this difference is markedly amplified in the presence of the PKC-activator PMA. Taken together, the discrepancy in the production of MMP-1 and TIMP-1 in gingival fibroblasts may contribute to tissue destruction in periodontal diseases.

Herpes simplex virus type 1 infection and glucocorticoid treatment regulate viral yield, glucocorticoid receptor and NF-kappaB levels.

The interplay between the endocrine and immune systems has come into focus in recent years with the insight that endocrine parameters may affect susceptibility to both auto-immune and infectious diseases. Our interest in immunoendocrine regulation led us to investigate the effects of glucocorticoids on Herpes simplex virus type 1 (HSV-1) infections. Glucocorticoids used to treat inflammatory conditions are not yet recommended for HSV-1 therapy, since they have been reported to prolong viral shedding both in vivo and in vitro. Here we report that glucocorticoids did not alter the viral yield in human gingival fibroblast (HGF) cell culture when glucocorticoid treatment and viral infection occured simultaneously, but the viral yield increased when cells were treated with the glucocorticoid dexamethasone (dex) prior to viral infection. We found that viral infection in our primary cell system increased NF-kappaB levels and DNA binding. In addition, the amount of glucocorticoid receptor (GR) increased following viral infection, and HSV-1 infection as such could induce glucocorticoid-driven transcription of a reporter gene in human embryo kidney (HEK) 293 cells stably transfected with GR. Dex treatment did not affect HSV-1-induced binding of p65 to an NF-kappaB element in an electrophoretic mobility shift assay, and acyclovir was still efficient as an anti-viral drug in the presence of dex. Further studies of the observed effects of HSV-1 infection and glucocorticoid treatment on GR and NF-kappaB regulation could give insights into the immunoendocrine mechanisms important for defence and therapy against viral infections.

Enhanced cyclooxygenase-2 mRNA expression in human gingival fibroblasts induced by cell contact with human lymphocytes.

Prostaglandins, especially prostaglandin E2 (PGE2), play a crucial role in the pathogenesis of periodontal disease. We have previously reported that inflammatory mediators interleukin-1 (IL-1) and tumor necrosis factor alpha (TNFalpha) increase the production of PGE2 in human gingival fibroblasts. In this study, we investigated the effect of cell-to-cell interactions between gingival fibroblasts and lymphocytes on PGE2 production by using co-culture technique. Cell-to-cell contact between gingival fibroblasts and lymphocytes synergistically enhanced the production of PGE2 in co-cultures. In contrast to lymphocytes, the cyclooxygenase-2 (COX-2) mRNA expression in gingival fibroblasts was strongly enhanced following cell contact between gingival fibroblasts and lymphocytes. The level of COX-1 mRNA expression, however, was not affected either in gingival fibroblasts or in lymphocytes by the interactions between fibroblasts and lymphocytes. The study demonstrates that cell contact between gingival fibroblasts and lymphocytes strongly stimulates PGE2 production partly due to enhanced COX-2 mRNA expression in gingival fibroblasts. The cell-to-cell contact between gingival fibroblasts and lymphocytes should be considered as an important regulatory aspect for the enhancement of PGE2 in periodontal disease.

Involvement of tyrosine kinases on cyclooxygenase expression and prostaglandin E2 production in human gingival fibroblasts stimulated with interleukin-1beta and epidermal growth factor.

The purpose of the present study was to investigate the involvement of cyclooxygenase-1 (COX-1), cyclooxygenase-2 (COX-2), and tyrosine kinase on prostaglandin E2 (PGE2) production in human gingival fibroblasts stimulated by interleukin-1beta (IL-1beta) and/or epidermal growth factor (EGF). The cytokine IL-1beta and to a lesser extent EGF, enhanced COX-2 mRNA levels in gingival fibroblasts. Simultaneous treatment with EGF and IL-1beta resulted in enhanced COX-2 mRNA levels accompanied by a synergistic stimulation of PGE2 biosynthesis compared to the cells treated with IL-1beta or EGF alone. Neither IL-1beta EGF nor the combination of IL-1beta and EGF enhanced COX-1 mRNA levels in gingival fibroblasts. The tyrosine kinase inhibitors, Herbimycin A and PD 153035 hydrochloride, reduced COX-2 mRNA levels as well as PGE2 production induced by IL-1beta or the combination of IL-1beta and EGF whereas COX-1 mRNA levels were not affected. Furthermore, the COX-2 specific inhibitor, NS-398, abolished the PGE2 production induced by IL-1beta, EGF, or the combination. These results indicate that the synergy between IL-1beta and EGF on PGE2 production is due to an enhanced gene expression of COX-2 and that tyrosine kinase(s) are involved in the signal transduction of COX-2 in gingival fibroblasts.

Effect of an antibacterial dental varnish on the levels of prostanoids, leukotriene B4, and interleukin-1 beta in gingival crevicular fluid.

The aim of this study was to investigate the effects of a chlorhexidine/thymol-containing dental varnish on the levels of prostaglandin E2 (PGE2), prostaglandin I2 (PGI2), leukotriene B4 (LTB4), and interleukin-1 beta (IL-1 beta) in gingival crevicular fluid (GCF). The material consisted of 15 adolescents undergoing treatment with fixed orthodontic appliances. Four buccal sites adjacent to bands or brackets and exhibiting a mild chronic gingival inflammation were selected in the upper quadrants of each patient. According to a split-mouth technique, the first and second quadrants were randomly treated with either a varnish (Cervitec) containing 1% chlorhexidine diacetate and thymol (CHX/thymol) or a placebo varnish without active ingredients. The varnishes were applied immediately after the baseline registration, and follow-up examinations were carried out after 3, 8, and 30 days. GCF was sampled with the aid of a paper strip and the volume was determined using a Periotron 8000. The concentrations of PGE2, PGI2, LTB4, and IL-1 beta in GCF were assessed using radioimmunoassay and ELISA techniques. The results unveiled statistically significant reductions of PGE2, PGI2, and LTB4 levels in GCF following the active varnish treatment when compared to baseline values. A slight drop in IL-1 beta levels was registered after both active and placebo varnish applications, but the differences were not significant. The results suggest that treatment with an antibacterial varnish decreases the levels of inflammatory mediators PGE2, PGI2, and LTB4 in gingival crevicular fluid and further support the concept that topical application of a CHX/thymol-containing varnish is beneficial in patients with chronic gingival inflammation.

Benzydamine reduces prostaglandin production in human gingival fibroblasts challenged with interleukin-1 beta or tumor necrosis factor alpha.

Benzydamine [1-benzyl-3-(3-dimethylamino)propoxy-1H-indazole] is a drug with analgesic, anesthetic, antimicrobial and anti-inflammatory activity. The purpose of the present study was to investigate the effect of benzydamine on prostaglandin production in human gingival fibroblasts. Benzydamine significantly reduced the basal production of both prostaglandin E2 (PGE2) and 6-keto-PGF1 alpha, the stable breakdown product of prostaglandin I2 (PGI2), in unstimulated human gingival fibroblasts. When the cells were treated simultaneously with benzydamine and the cytokines IL-1 beta or TNF alpha, the agent benzydamine reduced (P < 0.05) the stimulatory effect of IL-1 beta and TNF alpha respectively, on PGE2 and PGI2 production in human gingival fibroblasts. Furthermore, benzydamine reduced (P < 0.05) both the basal level and the cytokine-induced 3H-arachidonic acid release 3H-(AA) in gingival fibroblasts. The addition of exogenous arachidonic acid to the cells resulted in enhanced PGE2 production, which was reduced (P < 0.05) in the presence of benzydamine. The study indicates that benzydamine reduces the prostaglandin synthesis in gingival fibroblasts, partly at the level of phospholipase A2, by diminishing the liberation of arachidonic acid (AA) from phospholipids, and partly at the level of cyclooxygenase. The inhibitory effect of benzydamine on prostaglandin production may explain the anti-inflammatory effect of the drug in the management of patients with oral inflammatory conditions.

Signal transduction pathways involved in the synergistic stimulation of prostaglandin production by interleukin-1beta and tumor necrosis factor alpha in human gingival fibroblasts.

Accumulating evidence indicates that prostaglandins play an important role in the pathogenesis of periodontal disease. In this study, the effects and interactions between IL-1beta and TNFalpha on prostaglandin production and its regulation were investigated. The cytokines IL-1beta and TNFalpha stimulated prostaglandin E2 (PGE2) and prostacyclin (PGI2) production in gingival fibroblasts. Simultaneous treatment of the cells with IL-1beta and TNFalpha resulted in a synergistic stimulation of PGE2 and PGI2 formation. IL-1beta and, to a lesser extent, TNFalpha stimulated the release of 3H-arachidonic acid (3H-AA), and simultaneous addition of IL-1beta and TNFalpha further increased the release of 3H-AA from pre-labeled gingival fibroblasts. Furthermore, IL-1beta and, to a lesser extent, TNFalpha induced the expression of cyclooxygenase-2 (COX-2) mRNA. Simultaneous addition of IL-1beta and TNFalpha synergistically enhanced COX-2 mRNA levels, accompanied by a corresponding stimulation of PGE2 synthesis. Neither IL-1beta, TNFalpha, nor the combination of these two cytokines affected COX-1 mRNA levels. PMA, known to activate protein kinase C (PKC), enhanced the stimulatory effect of IL-1beta, TNFalpha, and the combination on COX-2 mRNA levels accompanied by a corresponding increase in PGE2 production. The phospholipase A2 (PLA2) inhibitor, BPB, and the PKC inhibitor, BIS, reduced PGE2 production, whereas dexamethasone, indomethacin, and NS-398 completely abolished PGE2 production induced by IL-1beta, TNFalpha, and the combination. The study indicates that the synergistic stimulation of prostaglandin production by IL-1beta, and TNFalpha is mediated partly at the level of COX-2 and partly at the level of PLA2 and that PKC is involved in the signal transduction of the synergy between the two cytokines. The synergy between IL-1beta and TNFalpha may play an important role in the inflammatory processes in gingival tissue in vivo.

Effect of a chlorhexidine/thymol-containing varnish on prostaglandin E2 levels in gingival crevicular fluid.

The aim was to study the effect of a chlorhexidine/thymol-containing varnish (Cervitec) on the levels of prostaglandin E2 (PGE2) in gingival crevicular fluid (GCF). The material consisted of 25 adolescents and young adults with fixed orthodontic appliances exhibiting gingival inflammation. Four buccal sites, adjacent to bands and brackets, were selected on each patient and randomly treated with either a varnish containing chlorhexidine diacetate (1% w/w) and thymol (1% w/w) or a placebo varnish without active ingredients. After baseline registration, the varnishes were applied twice within 3 d. Follow-up examinations were performed after 3, 8 and 30 d. The gingival inflammation was assessed by bleeding on probing, volume of GCF with a Periotron 8000 and PGE2 level in GCF by using a radioimmuno assay. Compared with baseline, a statistically significant reduction in the volume of GCF was recorded at the chlorhexidine/thymol treated sites in contrast to the placebo. The mean PGE2 levels were significantly reduced after the test varnish treatment compared with baseline and differed significantly from placebo after 8 d. The findings suggest that treatments with the antibacterial varnish result in reduced gingival inflammation and may thus be beneficial for patients with fixed orthodontic appliances.

Bradykinin synergistically stimulates interleukin 6 production in human gingival fibroblasts challenged with interleukin 1 or tumour necrosis factor alpha.

The production of interleukin 6 (IL-6) was studied in human gingival fibroblasts challenged with bradykinin (BK), in the presence or absence of either tumour necrosis factor alpha (TNF-alpha) or interleukin 1 (IL-1). The inflammatory mediator BK as well as the cytokines TNF-alpha and IL-beta dose dependently stimulated IL-6 production in gingival fibroblasts. When the cells were treated simultaneously with BK and either IL-1 beta or TNF-alpha, the inflammatory mediator BK synergistically upregulated IL-6 production in a dose-dependent manner. The BK B1 receptor agonist des-arg9-BK as well as the BK B2 receptor agonist Lys-BK also induced IL-6 production and synergistically enhanced the effect of IL-1 and TNF-alpha on the production of IL-6. The upregulation of IL-6 production induced by BK was abolished by the anti-inflammatory agent dexamethasone (DEX) and the phospholipase A2 (PLA2) inhibitor 4-bromphenacyl bromide (BPB). Treatment of the cells with the cyclooxygenase (COX) inhibitor flurbiprofen resulted in a minor reduction of the stimulatory effect of BK. The results show that BK together with IL-1 or TNF-alpha act in concert to enhance IL-6 production and that the synergy was obtained by both the BK B1 and the BK B2 receptor agonist. The study indicates that the synergy between BK and IL-1 or TNF-alpha on IL-6 production is mediated partly at the level of PLA2 and partly at the level of COX. The synergism between the pro-inflammatory mediator BK and the cytokines IL-1 or TNF-alpha indicates that gingival fibroblasts, by producing cytokines, affect the local immune response in the connective tissue and thereby play a role in the pathogenesis of periodontal disease.

Prostaglandin E2 level in gingival crevicular fluid from patients with Down syndrome.

The levels of prostaglandin E2 (PGE2) and interleukin-1 beta (IL-1 beta) were determined in gingival crevicular fluid (GCF) collected from patients with gingivitis: 15 Down syndrome children and 15 controls. The mean level of PGE2 in GCF was significantly higher (P < 0.05) in the Down syndrome group (10.0 pg/microliters GCF) than in the control group (4.6 pg/microliters GCF). In GCF samples collected from sites characterized as noninflamed, the mean level of PGE2 was significantly higher (P < 0.001) in the Down syndrome group than in the controls. The mean level of PGE2 in samples from inflamed sites, on the other hand, did not differ between the two groups. The mean level of IL-1 beta was not significantly higher in the Down syndrome group than in the controls. This study shows that the level of PGE2 detected in GCF from Down syndrome patients is increased, a fact that may be of importance in the pathogenesis of the periodontal disease frequently seen in these patients.

Effect of phenytoin on interleukin-1 beta production in human gingival fibroblasts challenged to tumor necrosis factor alpha in vitro.

Effects and interaction of tumor necrosis factor alpha (TNF alpha) and the antiepileptic drug phenytoin (PHT) on interleukin-1 beta (IL-1 beta) production as well as on prostaglandin E2 (PGE2) formation were studied in gingival fibroblasts in vitro. TNF alpha, in contrast to PHT, dose-dependently stimulated the production of cell-associated IL-1 beta. The stimulatory effect of TNF alpha on IL-1 beta production was accompanied by enhanced PGE2 formation. When PHT and TNF alpha were added simultaneously, the drug potentiated the stimulatory effect of TNF alpha on both IL-1 beta production and PGE2 formation. The major PHT metabolite, p-HPPH, did not affect IL-1 beta production, either alone or in combination with TNF alpha. The production of IL-1 beta induced by TNF alpha and the combination of TNF alpha and PHT was further enhanced in the presence of the prostaglandin endoperoxide (PGH) synthase inhibitors, indomethacin and flurbiprofen. The PHT-mediated enhancement of TNF alpha-induced IL-1 beta production and PGE2 formation in gingival fibroblasts may be an important link in the pathogenesis of gingival overgrowth induced by PHT.

Interleukin-1 beta induces expression of cyclooxygenase-2 mRNA in human gingival fibroblasts.

The effect of interleukin-1 beta (IL-1 beta) on the expression of cyclooxygenase-1 and -2 (COX-1 and COX-2) mRNA and its relation to prostaglandin E2 (PGE2) biosynthesis in human gingival fibroblasts was studied. IL-1 beta increased levels of mRNA for COX-2 whereas the COX-1 mRNA level was unaffected. The increased COX-2 mRNA levels were accompanied by enhanced PGE2 formation. The phorbol, 12-myristate 13-acetate (PMA), known to stimulate protein kinase C (PKC), also induced expression of COX-2 mRNA. When gingival fibroblasts were treated simultaneously with IL-1 beta and PMA, the cytokine IL-1 beta synergistically increased levels of COX-2 mRNA, accompanied by a corresponding increase in PGE2 biosynthesis. The anti-inflammatory steroid, dexamethasone (DEX) abolished the enhanced expression of COX-2 mRNA as well as PGE2 formation induced by IL-1 beta, PMA or the combination of IL-1 beta and PMA. The study indicates that the IL-1 beta induced PGE2 formation is mediated by an enhanced gene expression of COX-2 in gingival fibroblasts suggesting that the enzyme COX-2 may play an important role in the regulation of prostanoid formation at inflammatory lesions in gingival tissue.

Effects and interactions of tumour necrosis factor alpha and bradykinin on interleukin-1 production in gingival fibroblasts.

Effects of and interactions between tumour necrosis factor alpha (TNF alpha) and bradykinin (BK) on production of interleukin-1 (IL-1 alpha, IL-1 beta) in human gingival fibroblasts were studied. The cytokine TNF alpha induced production of cell-associated IL-1 alpha and IL-1 beta in gingival fibroblasts, with IL-1 beta being most abundant. Addition of BK, in the presence of TNF alpha, for 1 h and 6 h, respectively, synergistically enhanced the TNF alpha induced IL-1 beta production, whereas BK alone did not induce IL-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 TNF alpha induced IL-1 beta 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 TNF alpha induced IL-1 beta production. Similar to BK, the phorbol esters (PMA, PDBu, 13-PA) alone did not induce IL-1 beta production in the gingival fibroblasts. The results indicate that TNF alpha induces production of cell-associated IL-1 in gingival fibroblasts, which can be upregulated by a PKC dependent pathway.

Epidermal growth factor potentiates interleukin 1 and tumor necrosis factor-induced prostaglandin biosynthesis in human gingival fibroblasts.

The effects of and interactions between epidermal growth factor (EGF), transforming growth factor alpha (TGF-alpha) interleukin 1 (IL-1) and tumour necrosis factor alpha (TNF-alpha) on arachidonic acid release and prostaglandin biosynthesis in human gingival fibroblasts were studied. IL-1 alpha, IL-1 beta and TNF-alpha, but not EGF nor TGF-alpha, stimulated prostaglandin E2 (PGE2) formation in the gingival fibroblasts. The effect of IL-1 alpha, IL-1 beta and TNF-alpha on PGE2 formation was significantly potentiated by EGF in a dose-dependent manner. Similarly, TGF-alpha synergistically potentiated IL-1 beta stimulated PGE2 formation. IL-1 beta but not EGF stimulated the release of 3H-arachidonic acid (3H-AA) from prelabelled gingival fibroblasts. In contrast to the effect on PGE2 formation, no synergistic interaction between EGF and IL-1 was seen on arachidonic acid (AA) release. Addition of unlabelled exogenous AA, in the presence of EGF, resulted in a significant increase in PGE2 formation compared to that seen in fibroblasts not exposed to EGF. The results demonstrate that EGF and IL-1 as well as EGF and TNF-alpha act in concert to enhance prostanoid formation in gingival fibroblasts. Data indicates that EGF potentiates the IL-1 and TNF-alpha induced PGE2 formation at the level of prostaglandin endoperoxide synthase (cyclooxygenase). The synergistic effects of inflammatory cytokines and growth factors may be of physiological importance for regulation of regenerative tissue growth during inflammation and repair.