Ethanol consumption enhances periodontal inflammatory markers in rats

ObjectiveThe aim of this study was to assess the short term effect of ethanol administration on periodontal disease in rats.DesignRats received either ethanol 2 g/kg or water by gastric gavage twice a day. On the fifth day ligatures were tied around the molars of half of the rats to induce periodontitis. After 7 days gingival tissue was removed and assayed for inflammatory markers. Finally, hemi-mandibles were extracted to evaluate bone loss by histomorphometrical techniques.ResultsThe experimental periodontitis increased significantly the mRNA expression (p < 0.001) and activity (p < 0.001) of inducible nitric oxide synthase (iNOS) in the gingival tissue, whilst short time ethanol administration increased iNOS activity (p < 0.05) and produced an additive effect on iNOS mRNA expression augmented by periodontitis (p < 0.01). The short time ethanol administration also potentiated the periodontitis stimulatory effect on the mRNA expression of interleukin (IL)-1β (p < 0.01 and p < 0.001, in semi-quantitative and real time PCR, respectively) and on the height of periodontal ligament (p < 0.05). However, the ligature-induced periodontitis, but not ethanol administration, increased the prostaglandin E₂ content (p < 0.05) and, diminished the alveolar bone volume (p < 0.05), as compared to sham rats.ConclusionThe present results suggest that ethanol consumption could represent a risk indicator for periodontal disease since augments the expression of inflammatory markers, in healthy rats, and increases them, at short term, during the illness. However, scale longitudinal investigation and more case–control studies are needed to confirm this statement.     

Changes of adiponectin and inflammatory cytokines after periodontal intervention in type 2 diabetes patients with periodontitis

ObjectiveTo determine serum adiponectin, C-reactive protein (CRP), TNF-α and IL-6 levels in impaired glucose tolerance (IGT) and type 2 diabetes mellitus (T2DM) patients with periodontitis before and after periodontal intervention, and to investigate the relationship between T2DM and periodontitis.MethodsA total of 50 IGT and 106 T2DM patients with periodontitis were enrolled. The T2DM patients were divided into two groups: T2DM without macrovascular disease (DM1) group and T2DM with macrovascular disease (DM2) group. Each group was randomly divided into two subgroups according to whether they performed periodontal intervention. The normal control group (NC group) consisted of 30 healthy adults. The serum adiponectin, CRP, TNF-α and IL-6 levels were measured at baseline and 3 months after periodontal intervention.ResultsThe serum adiponectin levels at baseline had decreased tendency with significant difference between each two groups, while CRP, TNF-α, and IL-6 levels had increased tendency with significant difference between each two groups among NC, IGT, DM1 and DM2 groups (all P < 0.01). At 3 months after periodontal intervention, the serum adiponectin levels were increased than those without periodontal intervention (all P < 0.01), while CRP, IL-6 and TNF-α significantly decreased (all P < 0.05) in both IGT and DM1 groups. In DM2 group, only CRP levels at 3 months after periodontal intervention were significantly decreased (P < 0.05). Moreover, the HbAlc levels in T2DM patients were improved at 3 months after periodontal invention (P < 0.01).ConclusionPeriodontal intervention is helpful for glucose control, which may be associated with increased serum adiponectin levels and decreased inflammatory cytokine levels.     

Effects of connective tissue growth factor on human periodontal ligament fibroblasts

ObjectiveThe aim of this study was to evaluate the effects of different concentrations of connective tissue growth factor (CTGF) on human periodontal ligament fibroblasts(HPLFs).DesignHPLFs were cultured and identified. Then, different concentrations of CTGF (1, 5, 10, 50, 100 ng/ml) were added to the HPLF culture. Next, CCK-8 assays, alkaline phosphatase (ALP) assays, hydroxyproline determination, alizarin red staining methods, Transwell chambers and real-time PCR methods were applied to observe the effects of CTGF on the proliferation, ALP activity, synthesis of collagen, formation of mineralized nodules and migration. We also studied expression of ALP, fiber link protein (FN), integrin-binding sialoprotein (IBSP), osteocalcin (OC), and integrin beta 1 (ITGB1) mRNA by HPLFs. Statistical significance was assumed if P < 0.05 or P < 0.01.ResultsThe addition of CTGF (1, 5, 10 ng/ml) remarkably promoted the proliferation and collagen synthesis of HPLFs compared with controls. CTGF (1, 5, 10, 50 ng/ml) improved ALP activity of HPLFs, and at all concentrations, CTGF (1, 5, 10, 50, 100 ng/ml) improved the expression of ALP, FN, IBSP and ITGB1 mRNA. In addition, CTGF (1, 5, 10, 50, 100 ng/ml) promoted the migration of HPLFs, which was dose-dependent, with maximal promotion in the 10 ng/ml group (P < 0.05 or P < 0.01).ConclusionsThus, in a certain range of concentrations, CTGF can promote the biological effects, including proliferation, migration and collagen synthesis of HPLFs, to promote the differentiation of HPLFs in the process of osteogenesis.     

Short-term periodontal and microbiological changes following orthognathic surgery

ObjectiveAim of the present study was to evaluate the influence of orthognathic surgery on the development of periodontal and microbiological changes.Materials and methodsFifteen consecutively treated patients with a mean age of 24.9 ± 7.7 years receiving orthognathic surgery were included in the present study. Plaque index (PI) and concentrations of 11 periodonto-pathogenic bacteria were recorded one day prior to surgery (t₀) and one week (t₁) and six weeks (t₂) post-surgery. In addition, a complete periodontal examination including pocket probing depth (PPD), gingival recession (GR), clinical attachment level (CAL), bleeding on probing (BOP) and width of keratinized gingiva (WKG) was conducted at t₀ and t₂. For statistical analysis, general linear model and paired t-test were applied.ResultsA significant increase of PI (t₀–t₁, p = 0.037) was followed by a significant decrease (t₁–t₂, p = 0.017). Apart from Eikenella corrodens (p = 0.036), no significant microbiological changes were recorded. PPD significantly increased on oral sites (p = 0.045) and GR especially on buccal sites (p = 0.001). In the incision area the development of GR was significantly higher on the test (buccal) than on the control sites (oral). Both gingival biotypes were affected by GR.ConclusionsOrthognathic surgery causes statistically significant changes of periodontal parameters, but these changes do not necessarily impair the aesthetic appearance of the gingival margin.     

The effect of initial periodontal treatment on plasma,gingival crevicular fluid and salivary levels of 8-hydroxy-deoxyguanosine in obesity

ObjectiveRecent studies have shown adverse effects on the periodontium from the increased production of reactive oxygen species (ROS) in obesity. The purpose of this study was to investigate the effects of obesity on 8-hydroxy-deoxyguanosine (8-OHdG) levels in the bodily fluids of patients with and without periodontal disease and to evaluate changes after initial periodontal treatment.DesignForty-five obese individuals and 45 normal-weight individuals were included in this study. Obese and normal-weight groups were classified into three sub-groups: chronic periodontitis (CP), gingivitis (G) and periodontally healthy controls (CTRL). Gingival crevicular fluid (GCF), plasma, saliva samples and clinical measurements were obtained at baseline and a month after initial periodontal treatment. Levels of 8-OHdG were analysed by ELISA.ResultsWhile plasma 8-OHdG levels were significantly higher at baseline in the obese patients with periodontal disease than in the normal-weight individuals (P < 0.05), no significant differences in GCF and saliva 8-OHdG levels were found (P ˃ 0.05). GCF and salivary 8-OHdG levels in obese patients with G and CP were significantly higher than in CTRL groups at baseline (P < 0.05). After treatment, 8-OHdG levels were decreased in all groups with periodontal disease (P < 0.01). Statistically significant positive correlations were observed between GCF 8-OHdG levels and GI in all the groups (P < 0.001).ConclusionsThe significant increase of plasma 8-OHdG levels in obese patients did not correlate with saliva and GCF 8-OHdG levels when compared to normal-weight individuals. Periodontal treatment had a positive effect on the periodontal parameters and 8-OHdG levels of both obese and normal-weight individuals.     

GuttaFlow Bioseal promotes spontaneous differentiation of human periodontal ligament stem cells into cementoblast-like cells

Objectives

To evaluate in vitro the cementogenic potential and the biological effects of GuttaFlow Bioseal, GuttaFlow 2, MTA Fillapex and AH Plus on human periodontal ligament stem cells (hPDLSCs).

Frequency of periodontal pathogens in equivalent peri-implant and periodontal clinical statuses

ObjectivesThis study tested the hypotheses that there is: (1) higher bacterial frequency in peri-implantitis/periodontitis, followed by mucositis/gingivitis and peri-implant/periodontal health; (2) similar bacterial frequency between comparable peri-implant and periodontal clinical statuses.Design of studyThe presence of Porphyromonas gingivalis, Tannerella forsythia, Campylobacter rectus, Prevotella intermedia, Treponema denticola and Aggregatibacter actinomycetemcomitans was evaluated in peri-implant (n = 53) and periodontal (n = 53) health; mucositis (n = 50), gingivitis (n = 50), peri-implantitis (n = 50) and periodontitis (n = 50).ResultsThe pattern of peri-implant bacterial frequency was not as expected (peri-implantitis > mucositis > health). Except for P. intermedia (p > 0.05), bacterial frequency was higher in peri-implantitis than health (p < 0.05). The frequency of P.gingivalis and red complex species were higher in peri-implantitis than mucositis (p < 0.05). In periodontal samples, T. forsythia and T. denticola showed the expected pattern of frequency (periodontitis > gingivitis > health). The frequencies of C. rectus and T. forsythia were higher in healthy teeth/gingivitis than healthy implants/mucositis, respectively (p < 0.05). The frequency of P. gingivalis and A. actinomycetemcomitans were similar between periodontitis and peri-implantitis (p > 0.05) while all other species occurrences were higher in periodontitis than peri-implantitis (p < 0.05).ConclusionsBacterial frequency increased from peri-implant/periodontal health to peri-implantitis/periodontitis but not from mucositis/gingivitis to peri-implantitis/periodontitis. There was a trend towards higher bacterial frequency in teeth than implants.     

Effects of ovariectomy on periodontal tissues following tooth replantation

ObjectivesThe aim of the study was to analyze the effects of ovariectomy on periodontal tissues following immediate tooth replantation by histomorphometric, immunohistochemistry, and μCT analysis.Materials and methodsEighty wistar rats (Rattus norvegicus albinos) with normal estrous cycles were randomly divided into two groups: ovariectomized (OVX) and Sham. Two months after surgery, the rats’ upper right incisor was extracted followed by immediate reimplantation. The animals were sacrificed after 28, 45, and 60 days healing time. Histomorphometric and immunohistochemical analysis were performed by evaluation of PCNA and TRAP straining.ResultsThe periodontal ligament was reinserted into the bone and cementum in the both groups. The immunohistochemical analysis revealed PCNA positive cells on the periodontal ligament in both groups at 28 days. Root resorption was noted at 45 days with immunoreactive cells for TRAP present in bone and tooth surface however no statistical differences between the groups were noticed. Histomorphometric analysis showed significant difference between groups in the periodontal ligament and root resorption parameters for the sub-items: intensity of chronic inflammatory infiltrate at 60 days (p < 0.01), the organization of the periodontal ligament at 28 days (p < 0.05), depth of root resorption at 45 days (p < 0.05) and at 60 days (p < 0.001). The μCT analysis showed multiple areas of bone resorption in association with OVX at 28 and 60 days with no significant differences between times in vivo.ConclusionThe ovariectomy did not have significant influence in periodontal tissue parameters following tooth reimplantation.     

Immunohistochemical analysis of bone resorption regulators (RANKL and OPG), angiogenic index, and myofibroblasts in syndrome and non-syndrome odontogenic keratocysts

ObjectiveThe aim of this study was to immunohistochemically analyse bone resorption regulators (receptor activator of nuclear factor kappa B ligand [RANKL] and osteoprotegerin [OPG]), angiogenic index, and myofibroblasts in Gorlin syndrome-related odontogenic keratocysts (SOKCs) and non-syndrome odontogenic keratocysts (NSOKCs).Study designTwenty-two SOKCs, 22 primary NSOKCs, and eight recurrent NSOKCs were evaluated by immunohistochemistry using anti-RANKL and anti-OPG antibodies. The angiogenic index was determined by microvessel count (MVC) using anti-CD34 antibody. Anti-α-smooth muscle actin (α-SMA) antibody was used for the identification of myofibroblasts.ResultsAnalysis of the expression of RANKL and OPG in the epithelial lining and fibrous capsule did not reveal significant differences between groups (P > 0.05). In the epithelial lining, the RANKL/OPG ratio was RANKL < OPG and RANKL = OPG in most primary NSOCKs (54.5%) and SOKCs (59.1%), respectively (P > 0.05). In the fibrous capsule, the ratio was RANKL = OPG in most primary (81.8%) and recurrent NSOKCs (75.0%) and in most SOKCs (45.5%) (P > 0.05). No significant differences in the angiogenic index or number of myofibroblasts were observed between primary NSOKCs, recurrent NSOKCs, and SOKCs (P > 0.05).ConclusionsThe present results suggest that differences in the biological behaviour of SOKCs and NSOKCs may not be related to the expression of RANKL and OPG, to the RANKL/OPG ratio, to the angiogenic index, or to the number of myofibroblasts in these lesions.     

Effects of prostaglandin E2 on clonogenicity,proliferation and expression of pluripotent markers in human periodontal ligament cells

Background and objectiveBased on our earlier work on the response of periodontal ligament (PDL) cells to mechanical stress by induction of cyclooxygenase expression and production of prostaglandin PGE₂ that could regulate mineralization of PDL cells, it was hypothesized that PGE₂ had potential effects on PDL stemness. In this study, we aimed to investigate clonogenicity, proliferation and expression of certain pluripotent markers, considered to be characteristics of PDL stemness, in response to treatment with exogenously-added PGE₂.Material and methodsHuman PDL cells were cultured and treated with various doses of PGE₂, and the aforementioned characteristics of PDL stemness were analyzed.ResultsThe clonogenicity and proliferation were significantly enhanced by PGE₂ at low concentrations (0.01, 0.1 and 1 ng/ml; P < 0.05), but only the proliferation was significantly diminished by PGE₂ at a high concentration (100 ng/ml; P < 0.05). Expression of NANOG and OCT4 mRNA and protein was increased by PGE₂ treatment at 0.1 and 1 ng/ml. Consistently, expression of stage-specific embryonic antigen 4, a putative stem cell marker, was significantly augmented by PGE₂ treatment at 1 ng/ml (P < 0.05).ConclusionOur findings suggest that although a high dose of PGE₂ (100 ng/ml) inhibits proliferation of PDL cells, PGE₂ at low doses appears to play a role in the maintenance of PDL stemness.     

Matrix metalloproteinase 8 (MMP8) gene polymorphisms in chronic periodontitis

ObjectivePrevious studies have suggested that some functional polymorphisms in the matrix metalloproteinase (MMPs) genes are associated with the risk of periodontal disease. However, to date no study has investigated MMP8 gene variants in relation to chronic periodontitis (CP). The aim of this study was to analyse polymorphisms in the MMP8 gene and their associations with microbial composition and clinical manifestation of CP.DesignA total of 619 unrelated Czech subjects were included in the present study. Two polymorphisms [?799C/T (rs11225395) and +17C/G (rs2155052)] in the MMP8 gene were studied in 341 patients with CP and 278 unrelated non-periodontitis controls. Both polymorphisms were detected using the polymerase chain reaction–restriction fragment length polymorphism (PCR–RFLP) methods. Subgingival bacterial colonisation (occurrence of bacteria in subgingival pockets and gingival sulci) was investigated by a commercial semiquantitative kit in selected subjects (N = 169).ResultsOur results showed no differences in the allele and genotype frequencies of the MMP8 ?799C/T and +17C/G polymorphisms between patients with CP and controls (p > 0.05). Nevertheless, the haplotype T(?799)/C(+17) was significantly more frequent in patients with CP than in controls [43.7% vs. 37.6%, p < 0.05, OR = 1.273 (95% CI: 1.013–1.601)]. Despite significant differences determined in the occurrence of periodontal bacteria between patients with CP and non-periodontitis controls (from p < 0.000001 to p < 0.05), no significant relationships between periodontal pathogens, MMP8 polymorphisms and CP were found (p > 0.05).ConclusionsAlthough none of the investigated SNPs in the MMP8 gene was individually associated with periodontitis, specific haplotype showed association with clinical manifestation of chronic periodontitis in a Czech population.     

Effectiveness of periodontal treatment on the improvement of inflammatory markers in children

AimIt is known that atherosclerosis begins in childhood, a behaviour towards oral health care and metabolic control, since an early age, is essential for patients with cardiovascular disease. The aim of this research was to evaluate the effectiveness of periodontal treatment full-mouth scaling and root planning (FMSRP), applied to children without systemic diseases, correlating with periodontal clinical and blood parameters (lipid profile and inflammatory markers).Materials and methodsThe 29 patients were divided into two groups, group 1 (14) – scaling and rot planning (SRP), group 2 (15) – FMSRP and the follow-up was conducted among 180 days.ResultsThe results showed a significant improvement in clinical periodontal parameters (p < 0.05) in both groups. In the analyzed blood parameters there was a greater evidence, with a significant improvement (p < 0.05), in total cholesterol (TC), triglycerides (TGs), fibrinogen (FGN), and interleukin-6 (IL-6)ConclusionsThus, we suggest that both periodontal treatments were effective in children without any systemic diseases.     

Effects of boric acid on experimental periodontitis and alveolar bone loss in rats

ObjectiveThe goal of the present study was to evaluate the histopathologic and morphometric effects of systemic boric acid in a rat periodontitis model.DesignTwenty-four Wistar rats were divided into three groups of eight animals each: non-ligated (NL), ligature only (LO), and ligature and treated with boric acid (BA) (3 mg/kg per day for 11 days). A 4/0 silk suture was placed in a subgingival position around the mandibular first molars; after 11 days the rats were sacrificed, and changes in alveolar bone levels were measured clinically and tissues were histopathologically examined to assess the differences amongst the study groups.ResultsThe ratio of presence of inflammatory cell infiltration (ICI) and osteoclast number in the LO group was significantly higher than that of the NL and BA groups (p < 0.05). The ratio of presence of osteoblastic activity in the LO group was significantly lower than that of the NL and BA groups (p < 0.05). Alveolar bone loss was also significantly higher in the LO group compared to the BA and NL groups (p < 005).ConclusionsThis study has demonstrated that systemic administration of boric acid reduced periodontal inflammation and alveolar bone loss in periodontal disease in rats.     

Effects of biglycan on physico-chemical properties of ligament-mineralized tissue attachment sites

Matrix proteoglycans define matrix structure, mineralization, and resulting biomechanics of tissues and their attachment sites.ObjectiveWe therefore investigated physical and (bio)chemical differences in enamel and periodontal tissues/attachment sites from mice that lack a specific nanoscale small leucine-rich proteoglycan (SLRPs) named biglycan (BGN).DesignExperimental groups consisted of N = 4, biglycan knockout (BGNKO) and N = 5 wildtype (WT) 8-week-old, male C3H mice. Morphology, histochemical and mechanical analyses were performed through micro X-ray computed tomography (Micro XCT?), immunohistochemistry, and microindentation. Unless mentioned otherwise, all differences between BGNKO and WT were demonstrated to be statistically significant through Student's t-tests with a 95% confidence interval (P ≤ 0.05).ResultsHistomorphometry performed by using Micro XCT? images indicated significantly higher BGNKO-enamel (0.46 ± 0.03 mm³) and BGNKO-root (1.81 ± 0.10 mm³) volumes compared to WT-enamel (0.37 ± 0.02 mm³) and WT-root (1.65 ± 0.07 mm³). BGNKO tooth size was relatively larger than WT mice, with no significant difference between skull sizes. Immunohistochemistry indicated BGN expression in the periodontal ligament (PDL), alveolar bone (AB), at the bone–PDL and cementum–PDL attachment sites in WT mice. Deeper AB resorption pits within interdental region of BGNKO specimens compared to WT resulting in significant differences in PDL-space of BGNKO (93 ± 13 μm) and WT (74 ± 11 μm) were observed. Microhardness of BGNKO-enamel (2.46 ± 0.60 GPa) and BGNKO-AB (0.52 ± 0.10 GPa) was significantly lower than WT-enamel (2.67 ± 0.60 GPa) and WT-AB (0.54 ± 0.10 GPa).ConclusionResults indicate that BGNKO-mice exhibit significant differences in tissue properties compared to WT-mice.     

Study of the participation of MMP-7, EMMPRIN and cyclophilin A in the pathogenesis of periodontal disease

Background and objectivePeriodontal disease is an infectious disease resulting from the immunoinflammatory response of the host to microorganisms present in the dental biofilm which causes tissue destruction. The objective of this study was to evaluate the immunohistochemical expression of matrix metalloproteinase 7 (MMP-7), extracellular matrix metalloproteinase inducer (EMMPRIN) and cyclophilin A (CypA) in periodontal disease.DesignGingival tissue samples were divided as follows: clinically healthy gingiva (n = 32), biofilm-induced gingivitis (n = 28), and chronic periodontitis (n = 30). Histological sections of 3 μm were submitted to immunoperoxidase method and undergone quantitative analysis. The results were analyzed statistically by the Mann-Whitney and Spearman correlation tests, with the level of significance set at 0.05 (α = 0.05).ResultsImmunopositivity for MMP-7, EMMPRIN and CypA differed significantly between the three groups, with higher percentages of staining in chronic periodontitis specimens, followed by chronic gingivitis and healthy gingiva specimens (p < 0.05). Immunoexpression of CypA and MMP-7 was higher in the intense inflammatory infiltrate observed mainly in cases of periodontitis (p < 0.05). CypA expression was positively correlated with MMP-7 (r = 0.831; p < 0.001) and EMMPRIN (r = 0.289; p = 0.006). In addition, there was a significant positive correlation between probing depth and expression of MMP-7 (r = 0.726; p < 0.001), EMMPRIN (r = 0.345; p = 0.001), and CypA (r = 0.803; p < 0.001).ConclusionThese results suggest that MMP-7, EMMPRIN and CypA are associated with the pathogenesis and progression of periodontal disease.     

Cytotoxicity and potential anti-inflammatory activity of velutin on RAW 264.7 cell line differentiation: Implications in periodontal bone loss

ObjectivesHypoxia-inducible factor-1α (HIF-1α) has been implicated in periodontal tissue inflammation and possibly in osteoclast differentiation, while polyphenols are known to be anti-inflammatory natural compounds that are capable of regulating the NF-κB protein complex pathway. The objective of this study was to investigate cytotoxicity and HIF-1α expression through the NF-κB pathway by polyphenol velutin (Euterpe oleracea Mart.), found in the pulp of acai fruit, during inflammatory RAW 264.7 differentiation.DesignRAW 264.7 mouse monocyte macrophage cells were stimulated with RANKL (30 ng/mL) and Porphyromonas gingivalis lipopolysaccharide (1 μg/mL). Cells were treated with various concentrations of velutin (0.5–2 μM) to check for viability, morphology, osteoclast differentiation, and HIF-1α expression (Western blot).ResultsAlamar blue cell viability assay showed no toxicity to RAW cells with the use of velutin in all concentrations tested (p > 0.05). Velutin did not induce cell apoptosis based on caspase 3/7 assay (p > 0.05). Fluorescence images stained by DAPI showed no alteration in the morphology of RAW cell nuclei (p > 0.05) treated with velutin. TRAP assays demonstrated a dose-dependent reduction in osteoclast formation by velutin when compared with control (p < 0.05). Velutin showed a reduction in HIF-1α expression related to IκB phosphorylation when compared with control (p < 0.001).ConclusionsAt the tested concentrations, velutin was not cytotoxic to RAW 264.7 and differentiated cells. Velutin reduced osteoclast differentiation and downregulated HIF-1α through the NF-κB pathway.     

Expression of human beta defensins (HBDs) 1, 2 and 3 in gingival crevicular fluid of patients affected by localized aggressive periodontitis

AimTo study the effect of nonsurgical periodontal therapy on the expression frequencies of human beta-defensin (HBD)-1, -2, and -3 in the gingival crevicular fluid (GCF) of patients affected by localized aggressive periodontitis.Materials and methodsTwenty patients affected by localized aggressive periodontitis (age range, 20–35 years) and 20 healthy subjects (age range, 21–37 years) were examined with clinical periodontal parameters and radiographic examination with the long-cone parallel technique. All periodontitis patients underwent nonsurgical periodontal therapy combined with doxycycline treatment and a maintenance program (including brushing with regular toothpaste). GCF samples were collected from patients and healthy control subjects at baseline as well as 3 months after periodontal therapy for the patient group.ResultsIn the patient group, the expression frequencies of HBD-1, -2, and -3 mRNA at baseline were 30%, 85%, and 35%, respectively, which changed after periodontal therapy to 80%, 45%, and 85%, respectively (all P < 0.001). In the healthy control subjects, the expression frequencies were 95%, 40%, and 95% for HBD-1, -2, and -3, respectively, which were different from those of diseased patients at baseline (all P < 0.001).ConclusionsThe appropriate expression of HBD peptides in health and disease may contribute to the maintenance of periodontal homeostasis, possibly through its antimicrobial effects and the promotion of adaptive immune responses.     

Porphyromonas gingivalis antigens differently participate in the proliferation and cell death of human PBMC

ObjectiveModulation of cell-mediated immunity by microorganisms in periodontal diseases has been widely studied; however, the proliferative activity and/or programmed death of mononuclear cells under periodontopathogenic stimuli are not yet well understood. The aim of this study was to investigate in vitro proliferation and death of peripheral blood mononuclear cells (PBMC) upon stimulation with Porphyromonas gingivalis (Pg) antigens.DesignIn 19 patients with chronic periodontitis (CP) and 16 controls without periodontitis (NP) the following clinical parameters were evaluated: bleeding on probing, probing depth, and clinical attachment level. PBMC were cultured under Pg stimuli and apoptosis/necrosis and proliferation assays were carried out for 18 and 48 h, respectively. Fluorescence of labelled cells was determined using flow cytometry.ResultsPBMC of CP and NP subjects exhibited a lower proliferative response to Pg LPS (p < 0.05) and HmuY protein (p < 0.001) compared with non-stimulated cells. Early apoptosis was induced by Pg LPS (p < 0.01) and Pg extract (p < 0.05), whilst all antigens induced late apoptosis (Pg LPS: p < 0.001; Pg extract: p < 0.001; HmuY: p < 0.01) and necrosis (Pg LPS: p < 0.01; Pg extract: p < 0.001; HmuY: p < 0.001). Pg LPS induced higher late apoptosis than HmuY (p < 0.05). Only Pg LPS-induced necrosis tended to be higher in CP compared with NP.ConclusionsThe inhibitory effect of cell proliferation caused by Pg LPS and HmuY protein is not observed when these antigens comprise Pg extract. Despite induced apoptosis, some still unknown mechanism determines the inflammatory outcome in cell death stimulated by HmuY.