High glucose levels contribute to vascular fibrosis via the activation of the endothelial-to-mesenchymal transition in periodontitis.

OBJECTIVE: To determine the changes of Porphyromonas gingivalis (P. gingivalis) growth and metabolism and identify whether the vascular epithelium change could be induced in diabetic periodontitis. BACKGROUND: Maintaining favourable vascular function is a precondition for periodontal regeneration. In diabetic periodontitis, high glucose levels could enhance the metabolism of pathogens, and a complex condition involving inflammation and high glucose levels would disrupt homeostasis of the epithelium and promote fibrosis by endothelial-to-mesenchymal transition (EndMT). METHODS: Porphyromonas gingivalis was cultured with glucose to judge its metabolic activity. Human umbilical vein endothelial cells (HUVECs) were treated with P. gingivalis-lipopolysaccharide (LPS) (10 µg/ml) and/or high glucose concentrations (25 mM), and transforming growth factor (TGF)-ß inhibitor was used to block EndMT. Inflammation level was assessed by flow cytometry. Multiple biological functions including EndMT, angiopoiesis, and cell migration were analysed. Additionally, gene expressions and protein levels were determined with qPCR and western blot, respectively. Finally, blood vessels were cultured ex vivo, and EndMT and fibrosis markers were detected by immunohistochemistry. RESULTS: Glucose could promote P. gingivalis growth and biofilm formation as well as the expression of virulence factor genes including FimA, RgpA, RgpB, and Kgp. P. gingivalis-LPS and glucose could increase intracellular reactive oxygen species (ROS) and promote fibrosis via EndMT in HUVECs, along with attenuating angiopoiesis and cell migration, which could be resumed by blocking EndMT with TGF-ß inhibitor. Vascular fibrosis was observed after the addition of glucose via EndMT regulation. CONCLUSION: Glucose augmented the growth and metabolism of P. gingivalis and promoted fibrosis by the activation of EndMT, as well as the inhibition of angiopoiesis and cell migration.

[Clinical and genetic analysis of a pedigree affected with hereditary dentinogenesis imperfecta type II].

OBJECTIVE: To explore the clinical and genetic characteristics of a Chinese pedigree affected with hereditary dentinogenesis imperfecta (DGI) type II. METHODS: Clinical data of the pedigree members were collected. Genomic DNA was extracted from peripheral blood samples and subjected to whole exome sequencing. RESULTS: Clinical characteristics of the affected family members have included amber teeth along with significant attrition, constricted roots and dentine hypertrophy leading to pulpal obliteration, which were suggestive of DGI type II. All of the affected members were found to have harbored a novel heterozygous c.2837delA (p.Asp946Valfs*368) variant of the DSPP gene which was predicted to be likely pathogenic. CONCLUSION: The c.2837delA variant of the DSPP gene probably underlay the disease in this pedigree. Above finding has expanded the variant spectrum of DSPP gene and provided a basis for molecular diagnosis and genetic counseling for this pedigree.

Th17/Treg balance and indoleamine 2,3 dioxygenase activity in periodontitis-associated atherosclerotic patients.

OBJECTIVE: This study investigated the peripheral Th17/Treg balance and its potential controlling factor indoleamine 2,3 dioxygenase (IDO) in patients with periodontitis and atherosclerosis (AS), as well as its correlation with Porphyromonas gingivalis infection. METHODS: In this retrospective study, P. gingivalis-infected atherosclerotic patients (Pg-AS), atherosclerotic patients (AS), P. gingivalis-infected periodontitis patients (Pg), and healthy controls (HCs) were selected after clinical examination, subgingival plaque examination, and plasma anti-P. gingivalis antibody analysis. Treg and Th17 cell percentages, related transcription factors, and functional cytokines in peripheral blood were analysed. Plasma tryptophan (Trp) and kynurenine (Kyn) were measured to determine IDO activity. RESULTS: Atherosclerotic patients (Pg-AS and AS groups) had significantly lower IDO activity and higher Th17/Treg ratio than those in the Pg and HC groups. The Th17/Treg ratio was higher and IDO activity was lower in the Pg-AS group compared with the AS group. Transcription factors and cytokines exhibited the same trend as the Th17 and Treg cells. Additionally, IDO activity was negatively correlated with the plasma anti-P. gingivalis antibody titre and the Th17/Treg ratio in the atherosclerotic group. CONCLUSIONS: P. gingivalis may reduce IDO activity and further promote Th17/Treg imbalance to facilitate AS development. IDO may be a novel molecular marker to predict periodontitis-associated AS.

Remodeling the periodontitis microenvironment for osteogenesis by using a reactive oxygen species-cleavable nanoplatform.

Modestly removing the excessive reactive oxygen species (ROS) plays a crucial role in regulating the microenvironment of periodontitis and provides favorable conditions for osteogenesis. However, the current strategy for scavenging ROS is not controllable, substantially limiting the outcomes in periodontitis. Herein, we introduced a controllable ROS-scavenging nanoplatform by encasing N-acetylcysteine (NAC, (a well-known ROS scavenger) into tailor-made ROS-cleavable amphiphilic polymer nanoparticles (PEG-ss-PCL NPs) as an intracellular delivery carrier. The existing ROS in the inflammatory microenvironment facilitated polymer degradation via breakage of thioketal bonds, and then led to encapsulated NAC release. NAC eliminated all ROS induced by lipopolysaccharide (LPS), while PssL-NAC adjusted the ROS level slightly higher than that of the control group. The percentage of apoptotic cells cultured with NAC and PssL-NAC decreased observably compared with that of cells cultured with 10 µg/ml LPS. The microenvironment regulated by PssL-NAC was highly suitable for osteogenic differentiation based on PCR and Western blot results, which showed higher expression levels of BMP2, Runx2, and PKA. Analysis of ALP activity and Alizarin red S staining showed consistent results. Additionally, the injection of PssL-NAC into the periodontitis area could alleviate the tissue destruction induced by ligation of the maxillary second molar. PssL-NAC showed a better ability to decrease osteoclast activity and inflammation, consequently improving the restoration of destroyed tissue. Our study suggests that ROS-responsive polymer nanoparticles loaded with NAC (PssL-NAC) can be new promising materials for the treatment of periodontitis. STATEMENT OF SIGNIFICANCE: More and more studies indicate that periodontal tissue damage is closely related to the high reactive oxygen species (ROS) environment. Excessive ROS will aggravate periodontal tissue damage and is not conducive to tissue repair. However, as an essential signal molecule in human physiological activities, ROS absence is also useless for tissue repair. In this study, we proposed to improve ROS imbalance in the environment of periodontitis as a strategy to promote periodontal regeneration and successfully synthesized a smart drug-releasing nanoplatform that can respond to ROS. Besides, we validated its ability to regulate the ROS environment and promote osteogenesis through experimental data in vivo and in vitro.

RANKL expression in periodontal disease: where does RANKL come from?

Periodontitis is an inflammatory disease characterized by periodontal pocket formation and alveolar bone resorption. Periodontal bone resorption is induced by osteoclasts and receptor activator of nuclear factor-κB ligand (RANKL) which is an essential and central regulator of osteoclast development and osteoclast function. Therefore, RANKL plays a critical role in periodontal bone resorption. In this review, we have summarized the sources of RANKL in periodontal disease and explored which factors may regulate RANKL expression in this disease.

[The clinical effect of root amputation in the treatment of periodontal/alveolar abscess].

OBJECTIVE: To study the clinical effect of root amputation in the treatment of periodontal/alveolar abscess teeth with one severe lesion root. METHODS: 30 periodontal/alveolar abscess teeth with one severe lesion root were chosen in the study. After root canal treatment, supragingival scaling, subgingival scaling and root planning, occlusal adjustment were done. Then the teeth were treated by root amputation. The clinical effect was evaluated 3 months, 6 months and 1 year after surgery. RESULTS: One year after surgery, 27 of 30 teeth were successful, 1 mandibular molar occurred root fracture, 1 mandibular molar was removed because of tooth loosening secondary to periodontal damage. 1 patient lost. CONCLUSION: Root amputation is an effective solution of periodontal/alveolar abscess.

[Experimental study on the potential role of BME-10X collagen/hydroxyapatite bone graft in periodontal tissue regeneration of beagle].

OBJECTIVE: To evaluate the potential role of BME-10X collagen/hydroxyapatite (HA) bone graft in periodontal tissue regeneration. METHODS: Four 18 months old male beagles in the experiment were treated with non-surgical periodontal therapy. At the sites of mandibular 3rd and 4th premolars at the time of dogs were treated with non-surgical periodontal therapy one week later, the teeth with the same name in the same jaw were selected to the experimental group (T group) or the control group (C group) at random. The defects in T group were filled with BME-10X collagen/HA bone graft while the defects in C group were filled with nothing. The dogs were sacrificed in twelve weeks and analyzed by histopathology. RESULTS: The defects in T group got more tissue regeneration compared with the defects in C group. The height of new bone (NB) was (3.01 +/- 0.14) mm in T group versus (1.32 +/- 0.11) mm in C group (P < 0.05). The height of new periodontal ligament (NP) was (3.12 +/- 0.19) mm in T group versus (1.35 +/- 0.12) mm in C group (P < 0.05). The height of new cementum (NC) was (3.30 +/- 0.15) mm in T group versus (2.70 +/- 0.12) mm in C group (P > 0.05). The new tissue guided by the bone graft was the same as the normal tissue in histopathology analysis. CONCLUSION: The results of the study suggest that BME-10X collagen/HA bone graft is a good bone graft for periodontal tissue regeneration.

[Evaluation of mandibule bone loss of periodontitis with standard digital panoramic tomogram].

OBJECTIVE: To compare the mandibular indices differences between periodontitis and non-periodontitis subjects by digital panoramic tomograms in order to find a simple and effective method to evaluate the density of alveolar bone and study the relativity between local bone loss and systemic bone loss. METHODS: Standard digital panoramic tomograms of sixty periodontitis subjects and sixty non-periodontitis subjects were taken. Density and height cortical width (CW) and panoramic mandibular index (PMI) of alveolar bone were measured on the digital panoramic tomograms. SPSS 12.0 was used for statistics analyses. RESULTS: The periodontitis subjects had lower height of alveolar bone (8.76 mm vs. 11.85 mm, P=0.000), lower density of alveolar bone (106.08 vs. 113.33, P=0.034), thinner cortical width (3.80 mm vs. 4.27 mm, P=0.008), while the difference of PMI between the two groups was not significant (P>0.05). The relativity between both density and height of alveolar bone with CW was significant (P<0.05), but not significant with PMI (P>0.05). CONCLUSION: The optical method is a simple and effective method to measure the density of the alveolar bone. Compared with the non-periodontitis subjects, the periodontitis subjects not only have alveolar bone changed significantly but also have thinner cortical width.