Salivary chromogranin A and myeloid-related protein-8/14 from periodontitis patients with type 2 diabetes.

OBJECTIVES: The bidirectional relationship between diabetes mellitus and chronic periodontitis is well known from clinical trials. Periodontitis in diabetic patients is characterized by severe inflammation and tissue destruction. The purpose of this study was to investigate the levels of chromogranin A (CgA), a stress marker, and myeloid-related protein (MRP)-8/14, an inflammatory marker, in saliva from patients with periodontitis and diabetes mellitus, and to investigate the relationship between CgA and MRP-8/14 in all individuals and in the three groups separately. METHOD AND MATERIALS: Stimulated saliva was collected from 20 diabetic patients with chronic periodontitis, 16 patients with chronic periodontitis, and 21 healthy individuals. Salivary CgA and MRP-8/14 were determined with enzyme-linked immunosorbent assay. Salivary CgA and MRP-8/14 levels were assessed in the saliva of diabetic periodontitis and periodontitis patients, and the relationship with periodontal disease severity was investigated. RESULTS: CgA values in saliva samples from chronic periodontitis patients and diabetic patients with chronic periodontitis were significantly higher than those of the control group. MRP-8/14 values in saliva from chronic periodontitis patients and diabetic patients with chronic periodontitis was significantly higher than that in the control group. Salivary CgA level was positively correlated to MRP-8/14 in all individuals, but there was no significant correlation within the chronic periodontitis patient group, diabetic patients with chronic periodontitis group, and the healthy patient group. No significant correlation between salivary CgA/MRP-8/14 and clinical parameters of periodontitis was found in the three groups. CONCLUSIONS: The results suggest that salivary CgA and MRP-8/14 could be related to the pathogenesis of periodontitis and diabetes. CgA concentration in saliva was positively associated with increased MRP-8/14 in all individuals.

Effect of Plasma Oxidation-Treated TiOx Film on Early Osseointegration.

PURPOSE: To develop a TiOx film with high wettability using plasma oxidation methodology and to evaluate the effect of such surface modification on new bone formation and early osseointegration. MATERIALS AND METHODS: Twenty sandblasted and acid-etched (SLA) titanium implants layered with a TiOx film layer were modified by plasma oxidation using a Radio Frequency Plasma Enhanced Chemical Vapor Deposition (RFPECVD) system. Twenty SLA implants without any treatment were employed as the control group. Scanning electron microscopy was used to evaluate surface morphology of the coating. X-ray photoelectron spectroscopy (XPS) was utilized to identify the chemical composition of the implant surface, and the wettability was tested with the help of the contact angle calculation. All implants were randomly inserted into the proximal tibia of 20 rats with a split-plot design. Four weeks after implantation, early osseointegration of the two groups was analyzed by a removal torque test and histologic analysis. RESULTS: The surface characteristics showed that both SLA and plasma oxidization-treated SLA (PO-SLA) surfaces displayed similar typical isotropic irregular indentations. As revealed by the XPS analyses, both TiO2 and Ti2O3 were coexistent in the PO-SLA surface. The contact angle measurement revealed that the super-hydrophilic surface was created out in the PO-SLA surface. Four weeks after implantation, a higher removal torque value was observed in the group of implants with the PO-SLA surface compared with the control group (12.68 ± 1.07 vs 9.95 ± 1.42 Ncm, P < .05), and a higher rate of bone-to-implant contact was also detected in the same group of implants with the PO-SLA surface (47.79% ± 9.59% vs 39.41% ± 9.00%, P < .05). The bone area ratio was also higher in the PO-SLA surface group than the control group (39.10% ± 10.01% vs 29.01% ± 7.24%, P < .05). CONCLUSION: It was indicated that the PO-SLA surface has combined the effect of high wettability and micro/nano-structures to notably increase early bone apposition.

A novel IRF6 mutation causing non-syndromic cleft lip with or without cleft palate in a pedigree.

Non-syndromic cleft lip with or without cleft palate (NSCLP) is the most common congenital craniofacial malformation, and its harmful influence on affected individuals is apparent. Despite many studies, the causative genes and their mechanisms are not completely clear. We recruited a Han Chinese NSCLP family and explored the causative variant in this pedigree. We performed whole-exome sequencing on two patients. Bioinformatics screening and analysis were used to identify the mutation. We also performed species conservation analysis, mutation function predictions, and homology protein modelling to evaluate the influence of the mutation. We identified a rare mutation in interferon regulatory factor 6 (IRF6) (c.26G>A; p.Arg9Gln) as a candidate of causative mutation. This mutation was predicted to be deleterious. The codon is conserved in many species. The residue change caused by this mutation would affect the structure of IRF6 to a degree. Our study suggested that the rare IRF6 variant is probably the pathogenic mutation in this family. Our result adds evidence that IRF6 variants play a role in the aetiology of orofacial clefts.

Effect of tyrosine-rich amelogenin peptide on behavior and differentiation of endothelial cells.

BACKGROUND: Enamel matrix derivative (EMD) is an effective biomaterial for periodontal tissue regeneration and might stimulate angiogenesis. Tyrosine-rich amelogenin peptide (TRAP) is present in EMD and is thought to contribute in its biological activity. In the present study, we investigated the effect of chemically synthesized TRAP on proliferation, migration, angiogenic structure formation, and differentiation of human umbilical vein endothelial cells (HUVECs) in vitro. MATERIAL AND METHODS: The effects of TRAP isolated from EMD and chemically synthesized TRAP on proliferation/viability, migration, and angiogenic structure formation were investigated. Expression of angiopoietin-2 (ang-2), von Willebrand factor (vWF), E-selectin, intracellular adhesion molecules 1 (ICAM-1), vascular endothelial growth factor (VEGF) receptors FMS-like tyrosine kinase 1 (FLT-1), and kinase insert domain receptor (KDR) was measured on both messenger RNA (mRNA) and protein levels. RESULTS: The proliferation/viability of HUVECs was inhibited by TRAP at concentration of 100 µg/ml and slightly stimulated by EMD at similar concentration. Both EMD and TRAP stimulated endothelial cell migration in microchemotaxis chamber. The effect of both TRAP preparations on the migration was significantly higher than that of EMD. All substances stimulated formation of angiogenic structure in vitro. The expression of ICAM-1, E-selectin, FLT-1, KDR, and vWF was significantly increased by both TRAP and EMD at a concentration 50 µg/ml. The expression of ang-2 was not affected by TRAP but was significantly increased by EMD. CONCLUSION: Our in vitro study shows that TRAP confer the most effects of EMD on the endothelial cells. CLINICAL RELEVANCE: TRAP might be used as a basis for development of new approaches for periodontal regeneration.

[Comparative analysis of 6 kinds of bacteria in the subgingival plaque in different types of patients with periodontal diseases].

PURPOSE: To detect the existence of Aa,Pg,Tf,Cr,Ec and Pn in the subgingival plaque, and determine their relationships among different types of periodontal diseases. METHODS: Dental plaques from 120 subjects were sampled, including 40 volunteers with health periodontal status(Group A) , forty patients with dental plaque-induced gingival diseases(Group B) and 40 patients with moderate or severe chronic periodontitis (Group C) . These samples were detected based on bacterial composition using the terminal restriction fragment length polymorphism of 16S rRNA genes by multiple-polymerase chain reaction. The data was analysed with SPSS 13.0 software package for Chi-square test. RESULTS: The detection rate of Pn, Cr and Pg had significant differences between group A and B. The detection rate of Ec, Cr, Pg, Aa and Tf had significant differences between group C and B. The detection rate of Ec, Pn, Cr, Pg, Aa and Tf had significant differences between group A and C. CONCLUSIONS: The rate of Ec, Pn, Cr, Pg and Tf detected in moderate or patients with moderate or severe chronic periodontitis are significantly higher than that in healthy subjects, indicating that these bacteria have certain correlation with chronic periodontitis. The rate of Ec, Cr, Pg and Tf detected in severe chronic periodontitis are significantly higher than that in dental-induced gingivitis, suggesting their close relationship with the progress of periodontal disease.

Total Antioxidant Capacity and Total Oxidant Status in Saliva of Periodontitis Patients in Relation to Bacterial Load.

The detection of salivary biomarkers has a potential application in early diagnosis and monitoring of periodontal inflammation. However, searching sensitive salivary biomarkers for periodontitis is still ongoing. Oxidative stress is supposed to play an important role in periodontitis progression and tissue destruction. In this cross-sectional study, we investigated total antioxidant capacity (TAC) and total oxidant status (TOS) in saliva of periodontitis patients compared to healthy controls and their relationship with periodontopathic bacteria and periodontal disease severity. Unstimulated saliva was collected from 45 patients with generalized severe periodontitis and 37 healthy individuals and the TAC/TOS were measured. In addition, salivary levels of Aggregatibacter actinomycetemcomitans, Porphyromonas gingivalis, Tannerella forsythia, Treponema denticola, and Fusobacterium nucleatum in saliva were measured. Salivary TAC was lower in periodontitis patients compared to healthy controls. Moreover, a significant negative correlation of salivary TAC with clinical attachment loss was observed in periodontitis patients. No significant difference in the salivary TOS was observed between periodontitis patients and healthy controls. Bacterial load was enhanced in periodontitis patients and exhibited correlation with periodontal disease severity but not with salivary TAC/TOS. Our data suggest that changes in antioxidant capacity in periodontitis patients are not associated with increased bacterial load and are probably due to a dysregulated immune response.

Escin inhibits lipopolysaccharide-induced inflammation in human periodontal ligament cells.

Periodontitis is a chronic inflammatory disease associated with gram-negative subgingival microflora infection. Accumulating experimental evidence suggests that escin exerts anti-inflammatory and anti-edematous effects. This study was designed to investigate the in vitro effects of escin on the inflammatory reaction of human periodontal ligament cells (hPDLs). hPDLs were stimulated with lipopolysaccharide (LPS). The cells were treated with various concentrations of escin. The viability of hPDLs was evaluated using the MTT method. The expression of Toll-like receptor 2 (TLR2) in hPDLs and the levels of IL-1ß, TNF-α and IL-6 in the supernatant were measured. Escin significantly attenuated LPS-induced cytotoxicity in a concentration-dependent manner in hPDLs. Treatment with escin partly blocked the expression of TLR2. Escin also lowered the increase of pro-inflammatory cytokines (IL-1ß, TNF-α and IL-6) induced by LPS. The present findings show that escin exerts a protective effect against LPS-induced inflammation in hPDLs. It was also shown that escin is a promising medicine for the treatment of periodontitis.