Utility of a haemoglobin test of gingival crevicular fluid: A multicentre, observational study.

OBJECTIVE: The purpose of this study was to verify the accuracy and utility of clinical parameters (plaque index, gingival crevicular fluid volume, probing depth, clinical attachment level, bleeding on probing and gingival index) and biochemical parameters (aspartate aminotransferase, protein and haemoglobin) in a longitudinal analysis during the supportive periodontal therapy period. SUBJECTS AND METHODS: A total of 279 test sites of 128 patients were investigated clinically and biochemically. After the first examination of clinical and biochemical parameters, periodontal support treatments were administered immediately and performed once every three months up to the second examination. RESULTS: All of the clinical and biochemical parameters were significantly lower at the second examination than at the first, except for the plaque index and bleeding on probing. Of these parameters, in particular, aspartate aminotransferase and haemoglobin in the gingival crevicular fluid were significantly reduced compared to those of the first examination in both the ≤4 and ≥5 mm probing depth groups, and they clearly suggested that periodontitis tended to recover. CONCLUSION: Adding the haemoglobin test to the bleeding on probing test strongly improves the accuracy of measurement of clinical parameters after periodontal treatment.

Degradation of Incretins and Modulation of Blood Glucose Levels by Periodontopathic Bacterial Dipeptidyl Peptidase 4.

Severe periodontitis is known to aggravate diabetes mellitus, though molecular events related to that link have not been fully elucidated. Porphyromonas gingivalis, a major pathogen of periodontitis, expresses dipeptidyl peptidase 4 (DPP4), which is involved in regulation of blood glucose levels by cleaving incretins in humans. We examined the enzymatic characteristics of DPP4 from P. gingivalis as well as two other periodontopathic bacteria, Tannerella forsythia and Prevotella intermedia, and determined whether it is capable of regulating blood glucose levels. Cell-associated DPP4 activity was found in those microorganisms, which was effectively suppressed by inhibitors of human DPP4, and molecules sized 73 kDa in P. gingivalis, and 71 kDa in T. forsythia and P. intermedia were immunologically detected. The kcat/Km values of recombinant DPP4s ranged from 721 ± 55 to 1,283 ± 23 µM-1s-1 toward Gly-Pro-4-methylcoumaryl-7-amide (MCA), while those were much lower for His-Ala-MCA. Matrix-assisted laser desorption ionization-time of flight mass spectrometry (MALDI-TOF MS) analysis showed His/Tyr-Ala dipeptide release from the N termini of incretins, glucagon-like peptide 1 (GLP-1) and glucose-dependent insulinotropic polypeptide, respectively, with the action of microbial DPP4. Moreover, intravenous injection of DPP4 into mice decreased plasma active GLP-1 and insulin levels, accompanied by a substantial elevation in blood glucose over the control after oral glucose administration. These results are the first to show that periodontopathic bacterial DPP4 is capable of modulating blood glucose levels the same as mammalian DPP4; thus, the incidence of periodontopathic bacteremia may exacerbate diabetes mellitus via molecular events of bacterial DPP4 activities.

Evaluation of bleeding on probing and gingival crevicular fluid enzyme activity for detection of periodontally active sites during supportive periodontal therapy.

This study aimed to analyze the enzyme activity in gingival crevicular fluid (GCF) and its association with clinical parameters, especially bleeding on probing (BOP), and thus reconsider the significance and accuracy of recording BOP. A total of 184 patients who had entered supportive periodontal therapy were selected and GCF was collected from 401 sites before recording the clinical parameters, probing pocket depth (PPD), BOP, clinical attachment level, gingival index and plaque index. The enzyme activity of neutrophil elastase and aspartate aminotransferase and amount of protein in GCF were also analyzed. In the clinical parameters for biochemical data, amount of GCF showed the most correlation. A cut-off value for BOP and PPD were determined by the ROC curve and Youden index. Analysis was performed with all clinical parameters and biochemical data. Of the 401 sites, 51 were less than the cut-off value and were BOP-negative. On the other hand, 29 sites had values more than the cut-off value, with 14 BOP-negative sites and 15 BOP-positive sites. A conclusion is as follows: twenty-nine sites with values more than the cut-off value were diagnosed as sites requiring periodontal management, however, 14 of these were BOP-negative. These results suggest that combining other biochemical tests with examination of BOP and PPD may improve the validity of periodontal disease diagnosis. In future studies, it will be essential to find a marker that can precisely detect periodontal disease activity, and to develop a diagnostic tool for chair-side use.

Improvement of pustulosis palmaris et plantaris by periodontal infection control in a patient with chronic periodontitis.

BACKGROUND: Pustulosis palmaris et plantaris (PPP) is a chronic, inflammatory, autoimmune-type disease characterized by sterile pustules of skin. The skin inflammation is influenced by several factors such as drugs, sunlight, metabolic and psychogenic factors as well as metal allergy. Here, we report a rare case that intensive periodontal treatment might have contributed to the improvement of skin inflammation. RESULTS: Skin inflammation regressed 1 month after intensive periodontal treatment. Both CD4/CD8 ratio and % of B cells in the blood sample were slightly decreased corresponding to the improvement of periodontal inflammation. CONCLUSIONS: Infection control of periodontal lesions might be one of attractive therapeutic targets in management of PPP.

Cathepsin B, D, and L regulation in cyclosporin A-mediated gingival hyperplasia of a patient with sarcoidosis.

BACKGROUND: Cyclosporin A (CsA) is an immunosuppressant with side effects including gingival hyperplasia. Sarcoidosis is a systemic disease characterized by granulomas. Here, we report on a rare case of sarcoidosis with gingival hyperplasia to clarify whether clinical observation corresponds to in vitro results. METHODS: Gingival fibroblasts (HGFs) were isolated from healthy gingiva and cultured with CsA. Total RNA was collected and expression of mRNAs examined using semi-quantitative RT-PCR analysis. Cathepsin B, D, and L expression in overgrown gingiva of the patient was examined by immunohistochemistry. RESULTS: Cathepsin D, L, and vascular endothelial growth factor (VEGF)165 mRNA were markedly suppressed in CsA-treated HGFs, whereas cathepsin B, matrix metalloproteinase-1 (MMP-1) and tissue inhibitor of metalloproteinase-1 (TIMP-1) mRNA were not reduced. Next, the decrease of cathepsin B and L expression in enlarged gingiva was observed, whereas an increase of cathepsin D expression was observed. Clinically, the enlarged gingival lesions were fully resolved by performing oral infection control. CONCLUSIONS: Cathepsins regulation might be an important factor in the development of CsA-mediated gingival hyperplasia.

The Role of Lactic Acid on Wound Healing, Cell Growth, Cell Cycle Kinetics, and Gene Expression of Cultured Junctional Epithelium Cells in the Pathophysiology of Periodontal Disease.

Lactic acid (LA) is short-chain fatty acid, such as butyric acid and propionic acid, that is produced as a metabolite of lactic acid bacteria, including periodontopathic bacteria. These short-chain fatty acids have positive effects on human health but can also have negative effects, such as the promotion of periodontal disease (PD), which is caused by periodontal pathogens present in the gingival sulcus. PD is characterized by apical migration of junctional epithelium, deepening of pockets, and alveolar bone loss. Thus, the junctional epithelial cells that form the bottom of the gingival sulcus are extremely important in investigating the pathophysiology of PD. The aim of this study was to investigate the effect of LA on wound healing, cell growth, cell cycle kinetics, and gene expression of cultured junctional epithelium cells. The results showed that stimulation with 10 mM LA slowed wound healing of the junctional epithelial cell layer and arrested the cell cycle in the G0/G1 (early cell cycle) phase, thereby inhibiting cell growth. However, cell destruction was not observed. LA also enhanced mRNA expression of integrin α5, interleukin (IL)-6, IL-8, intercellular adhesion molecule-1, and receptor activator of nuclear factor kappa-B ligand. The results of this study suggest that stimulation of junctional epithelial cells with high concentrations of LA could exacerbate PD, similarly to butyric acid and propionic acid.

Localization of bromodeoxyuridine-incorporating, p63- and p75(NGFR)- expressing cells in the human gingival epithelium.

The objective of this study was to find the sites with proliferative activity in the human gingival epithelium, where stem cells are likely to exist. Gingival tissues were excised from 16 adult patients and immunohistochemically examined for the presence of bromodeoxyuridine (BrdU)-incorporating, p63- and low affinity nerve growth factor receptor (p75(NGFR))-expressing cells. BrdU-incorporating cells were rarely present in the junctional epithelium. The number of BrdU-incorporating cells in the sulcular and oral gingival epithelia was significantly higher than that found in the junctional epithelium (ANOVA, P < 0.01). A considerable number of p63-positive nuclei were detected in the basal layer to lower spinous layers in the sulcular and oral gingival epithelia, but only few p63-positive cells were present in the junctional epithelium. p75(NGFR)-positive cells were exclusively located in the basal layer in the sulcular and oral gingival epithelia, and in limited basal area in the junctional epithelium neighboring the sulcular epithelium. In the oral gingival epithelium, intense immunostaining of BrdU, p63 and p75(NGFR) was correspondingly observed on the base and side of the rete ridges. These areas probably exhibit high proliferative activity owing to the presence of stem cells.

Prevalence and risk factors for peri-implant diseases in Japanese adult dental patients.

We investigated the prevalences and risk factors for peri-implant diseases in Japanese adult dental patients attending a follow-up visit at dental hospitals or clinics as part of their maintenance program. This cross-sectional multicenter study enrolled patients with dental implants who attended regular check-ups as part of a periodontal maintenance program during the period from October 2012 through September 2013. Patients with implants with at least 3 years of loading time were included in the study. The condition of peri-implant tissue was examined and classified into the following categories: healthy, peri-implant mucositis, and peri-implantitis. Patients were also evaluated for implant risk factors. A total of 267 patients (110 men, 157 women; mean age: 62.5 ± 10.7 years) were analyzed. The prevalence of patient-based peri-implant mucositis was 33.3% (n = 89), and the prevalence of peri-implantitis was 9.7% (n = 26). Poor oral hygiene and a history of periodontitis were strong risk factors for peri-implant disease. The present prevalences were lower than those previously reported. The quality of periodontal therapy before and after implant installation and patient compliance and motivation, as indicated by plaque control level, appear to be important in maintaining peri-implant tissue health.

Correlation Between Gingival Crevicular Fluid Hemoglobin Content and Periodontal Clinical Parameters.

BACKGROUND: Probing depth (PD) and bleeding on probing (BOP) are essential clinical parameters used for periodontal diagnosis. This study investigated whether detection of hemoglobin (Hb) in gingival crevicular fluid (GCF), along with PD and BOP, would improve diagnostic accuracy. METHODS: After plaque index (PI) was measured, GCF was collected from the gingival sulci of 401 anterior teeth in the maxilla and mandible from 184 patients who had entered periodontal maintenance therapy. Clinical parameters (gingival index [GI], PD, clinical attachment level [CAL], and BOP) were recorded. Hb values in GCF were assessed by immunochromatography. Moreover, cutoff values for PI, GI, and CAL based on the degree of PD and amount of GCF were created and analyzed. RESULTS: Hb was detected in 64.8% of GCF samples in 105 BOP-negative (-) sites in the periodontally stable group out of 107 sites that were less than all cutoff values. There were 71 BOP(-) sites in the periodontal-management-required group out of 122 sites that were more than all cutoff values, although no improvement in periodontal disease was observed. Hb was detected in 88.7% of GCF samples from these 71 BOP(-) sites. CONCLUSIONS: Hb was observed in more than 60% of GCF samples in BOP(-) gingival sulci in both periodontally stable and periodontal-management-required groups. These results suggest inspection of Hb derived from microbleeding in gingival sulci may serve as an index for preclinical diagnosis.