• diabetes mellitus
• peri‐implantitis
• streptozotocin‐induced diabetes

• Animals
• Peri-Implantitis/pathology
• Peri-Implantitis/diagnostic imaging
• Peri-Implantitis/etiology
• Male
• X-Ray Microtomography
• Mice
• Mice, Inbred C57BL
• Diabetes Mellitus, Experimental/complications
• Disease Models, Animal
• Alveolar Bone Loss/diagnostic imaging
• Alveolar Bone Loss/pathology
• Alveolar Bone Loss/etiology
• Inflammation
• Ligation

• R21 DE031431 NIDCR NIH HHS
• NIH/NIDCR DE031431 NIDCR NIH HHS

• Gingipain
• Hyperglycemia
• Inflammation
• RT‒PCR
• Zebrafish

• Marmara University Scientific Research and Project Commission

• Dental pulp stem cell
• Glycogen synthase kinase-3β inhibitor
• Inflammation
• Lipopolysaccharide
• NLRP3 inflammasome

• Low-level laser therapy
• knee joint
• prednisolone
• rheumatoid arthritis
• therapeutic ultrasound.

• diabetes
• injectable and thermosensitive hydrogel
• interleukin 1 receptor antagonist protein
• periodontitis

• cardiovascular disease
• diabetes
• metabolic syndrome
• microbiome and inflammation
• periodontitis

• Animals
• Cytokines
• Diabetes Mellitus, Type 2/complications
• Humans
• Lipids
• Metabolic Syndrome/complications
• Microbiota
• Periodontitis/complications

• R01 DE025225 NIDCR NIH HHS

Interaction of C. albicans with oral bacteria is crucial for its persistence, but also plays a potential role in the infection process. In the oral cavity, it grows as part of dental plaque biofilms. Even though growth and interaction of C. albicans with certain bacterial species has been studied, little is known about its biofilm growth in vitro in the simultaneous presence of Gram-negative and Gram-positive bacteria. The aim was to evaluate the growth of C. albicans in polymicrobial biofilms comprising oral Gram-negative and Gram-positive bacteria. Further, we also aimed to assess the potential of C. albicans in the Candida-bacteria polymicrobial biofilm to elicit cytokine gene expression and cytokine production from human blood cells.

C. albicans cell counts increased significantly up to 48 h in polymicrobial biofilms (p < 0.05), while the bacterial counts in the same biofilms increased only marginally as revealed by qPCR absolute quantification. However, the presence of bacteria in the biofilm did not seem to affect the growth of C. albicans. Expression of IL-8 gene was significantly (p < 0.05) higher upon stimulation from biofilm-supernatants than from biofilms in polymicrobial setting. On the contrary, TNF-α expression was significantly higher in biofilms than in supernatants but was very low (1–4 folds) in the monospecies biofilm of C. albicans. ELISA cytokine quantification data was in agreement with mRNA expression results.

Persistence and enhanced growth of C. albicans in polymicrobial biofilms may imply that previously reported antagonistic effect of A. actinomycetemcomitans was negated. Increased cytokine gene expression and cytokine production induced by Candida-bacteria polymicrobial biofilms and biofilm supernatants suggest that together they possibly exert an enhanced stimulatory effect on IL-8 and TNF-α production from the host.

• Candida albicans
• Cytokines
• Gram-negative bacteria
• Gram-positive bacteria
• Polymicrobial biofilms
• qPCR

• Diabetes
• Estrogen deficiency
• Periodontium
• Rats

• Alveolar Bone Loss/metabolism
• Animals
• Bone Density/physiology
• Diabetes Mellitus, Experimental/metabolism
• Estrogens/deficiency
• Female
• Mast Cells/metabolism
• Osteoclasts/metabolism
• Osteocytes/metabolism
• Ovariectomy/methods
• Periodontal Ligament/metabolism
• Periodontium/metabolism
• Rats
• Rats, Wistar
• Streptozocin/pharmacology

Severe periodontitis is a risk factor for poor glycemic control. The appropriate medical treatment and plaque control of periodontitis positively affects blood-sugar control in diabetes patients. We aimed to identify the factors associated with glycemic control and examine the periodontal treatment (PT) experience and oral health-related quality of life (OHQoL) for patients with poor glycemic control in type 2 diabetes mellitus (T2DM). This multicenter case–control study recruited 242 patients with poor glycemic control and 198 patients with good glycemic control. We collected patients’ information through face-to-face interviews using a structured questionnaire. The Oral Health Impact Profile-14 (OHIP-14) was used to measure OHQoL. Based on PT status, the patients were classified into three groups: a non-periodontal disease group, a PT group, and a non-PT (NPT) group. Regression models were used to analyze the data. No interdental cleaning (adjusted odds ratio (aOR) = 1.78) and positive attitudes toward periodontal health (aOR = 1.11) were significantly more likely to be associated with poor glycemic control in patients with T2DM. The PT group had a significantly lower OHIP-14 score than the NPT group (6.05 vs. 9.02, p < 0.001), indicating a better OHQoL among patients with poorly controlled T2DM. However, the OHQoL did not differ significantly in patients with well-controlled T2DM between the PT and NPT groups. This suggested that diabetic patients with poor glycemic control must improve periodontal care practices and receive proper PT, if necessary, to improve their OHQoL.

• Glycemic control
• oral health-related quality of life
• periodontal care behavior
• periodontal treatment experience

Rheumatoid arthritis (RA) and periodontitis share risk factors and inflammatory pathways that could be related to cytokines, such as interleukin (IL)-6, IL-8, IL-17A, and tumour necrosis factor-α (TNF-α). The aim of this study was to compare periodontal status and salivary levels of selected cytokines between patients diagnosed with RA and periodontitis. RA patients were assessed for the potential influence of anti-rheumatic therapy.

One hundred and six patients were enrolled in a cross-sectional study. Medical assessment and periodontal examination were performed in 35 patients with chronic periodontitis, in 35 patients with RA and chronic periodontitis, and in 36 controls. Unstimulated whole saliva samples were analysed for IL-6, IL-8, IL-17A, and TNF-α.

Significant differences in biomarkers and periodontal parameters were found among groups. Study groups exhibited higher mean pocket depth (PD), number of PD > 4 mm, and mean clinical attachment loss, when compared with controls. The RA group had lower bleeding on probing index and PD, but higher values of plaque indices than the periodontitis group. Concentration of evaluated cytokines were higher in the RA and periodontitis groups, compared with controls. The periodontitis group showed also higher levels of IL-6, IL-17A, and TNF-α in comparison to RA. RA patients were treated with disease-modifying anti-rheumatic drugs (DMARDs) and glucocorticosteroids.

Salivary levels of IL-6, IL-8, IL-17A, and TNF-α can be affected by periodontitis, RA, and presumably DMARDs. DMARD therapy appears to reduce destructive and inflammatory processes in periodontal tissues because lower values of PD, BOP, and salivary levels of IL-6, IL-17A, and TNF-α were found in RA.

• interleukin 17A
• interleukin 6
• interleukin 8
• periodontitis
• rheumatoid arthritis
• salivary diagnostics
• tumour necrosis factor α

Patients with diabetes tend to have an increased incidence of osteoporosis, which may be associated with hyperglycemia; however, the pathogenic mechanisms governing this interaction remain unknown. The present study sought to investigate whether elevated extracellular glucose levels of bone mesenchymal stem cells (BMSCs) could influence osteoblastic differentiation and whether the intracellular Sonic hedgehog (Shh) pathway could adjust the effects. Furthermore, to verify the results in vivo, a rat tooth extraction model was constructed. BMSCs were incubated in eight types of culture medium, including low glucose (LG), LG + lentivirus (Lenti), LG + Lenti-small interfering RNA (Lenti-siRNA), LG + Lenti-Shh, high glucose (HG), HG + Lenti, HG + Lenti-siRNA and HG + Lenti-Shh. The lentiviral transfection efficiency was observed using a fluorescence microscope; protein and mRNA expression was detected by western blotting and reverse transcription-quantitative polymerase chain reaction (RT-qPCR). The matrix mineralization and alkaline phosphatase (ALP) activity of BMSCs were examined by Alizarin red staining and ALP activity assays, respectively. The expression of osteogenesis-related genes in BMSCs were quantified by RT-qPCR. The alveolar ridge reduction was measured and histological sections were used to evaluate new bone formation in the tooth socket. With high concentrations of glucose, Shh expression, matrix mineralization nodules formation, ALP activity and the levels of bone morphogenic protein 4 (BMP4), bone sialoprotein (BSP) and osteopontin (OPN) expression were greatly reduced compared with LG and corresponding control groups. Whereas activated Shh signaling via Lenti-Shh could increase the number of matrix mineralization nodules, ALP activity, and the expression levels of BMP4, BSP and OPN in BMSCs. Additionally, in vivo assays demonstrated that Lenti-Shh induced additional bone formation. Collectively, the results of the present study indicated that HG inhibited the Shh pathway in osteoblasts and resulted in patterning defects during osteoblastic differentiation and bone formation, while the activation of Shh signaling could suppress these deleterious effects.

Alcohol intake is associated with oral diseases and bone changes including alveolar bone loss in humans and in experimental animals. The main aim of the present study is to assess the effect of long-term alcohol intake, at different frequencies, on periodontal bone loss (PBL) in adult rats.

Thirty-six (36) rats were divided into 3 groups: Control (daily water intake, n=12), daily alcohol intake (20% ethanol, n=12), and social alcohol intake (20% ethanol twice a week, n=12). The rats were sacrificed after 90 days and their right maxillae were removed. Initially, a random portion from each group was analyzed through SEM (scanning electron microscope) to assess surface topography. Next, all pieces were dissected and stained with methylene blue 1% and photographed in stereomicroscope at 10x magnification. The PBL was assessed by measuring the distance between cement-enamel junction and alveolar bone crest.

Results showed higher (p=0.0368) alcohol solution amount in the daily intake group than in the twice week intake one. The SEM showed qualitatively flat bone surface in the control group, the social intake group presented surface with few minor hollows, and the daily intake group evidenced increased number and diameter of wells. The comparison between groups showed higher bone loss (p<0.05) in both frequencies than in the control, but the bone loss was lower (p<0.05) in the social alcohol intake group than in the daily intake one.

Alcohol intake may cause alveolar bone loss in periodontitis-free rats depending on the frequency.

Key words:Alcohol intake, alveolar bone loss, alcohol-induced periodontitis, alcoholic rats.

Curcumin exhibits anti-inflammatory effects and has been suggested as a treatment for inflammatory diseases. The aim of this study was to investigate the effects of curcumin on the lipopolysaccharide induced inflammatory response in rat gingival fibroblasts in vitro and ligation-induced experimental periodontitis in vivo, and to speculate the possible anti-inflammatory mechanism of curcumin.

The gingival fibroblasts were incubated with different concentrations of curcumin in the absence or presence of lipopolysaccharide (LPS). Concentrations of interleukin-1β(IL-1β), tumor necrosis factor-α (TNF-α), osteoprotegerin (OPG) and soluble receptor activator of nuclear factor kappa-B ligand (RANKL) culture supernatants of rat gingival fibroblasts were determined by enzyme linked immunosorbent assay. The nuclear fraction of rat gingival fibroblasts was extracted and nuclear factor kappa-B (NF-κB) activation was assessed by western blotting to elucidate related mechanisms. Curcumin was given every two days by oral gavage. The gingival inflammation and alveolar bone loss between the first and second molars were observed by hematoxylin and eosin staining. Collagen fibers were observed by picro-sirius red staining. Alveolar bone loss was assessed by micro-CT analysis.

Curcumin attenuated the production of IL-1β and TNF-α in rat gingival fibroblasts stimulated by LPS, and inhibited the LPS-induced decrease in OPG/sRANKL ratio and NF-κB activation. Curcumin significantly reduced gingival inflammation and modulated collagen fiber and alveolar bone loss in vivo.

curcumin modulates inflammatory activity in rat periodontitis by inhibiting NF-κB activation and decreasing the OPG/sRANKL ratio induced by LPS.

• Curcumin
• Micro-CT
• NF-κB
• OPG/RANKL
• Periodontitis

Type 1 diabetes with periodontitis shows elevated TNF-α expression. Tumor necrosis factor (TNF)-α stimulates the expression of receptor activator of nuclear factor-κB ligand (RANKL) and sclerostin. The objective of this study was to determine the effect of TNF-α expression of osteocytic RANKL and sclerostin in type 1 diabetes rats with periodontitis using infliximab (IFX), a TNF-α antagonist. Rats were divided into two timepoint groups: day 3 and day 20. Each timepoint group was then divided into four subgroups: 1) control (C, n = 6 for each time point); 2) periodontitis (P, n = 6 for each time point); 3) diabetes with periodontitis (DP, n = 8 for each time point); and 4) diabetes with periodontitis treated with IFX (DP+IFX, n = 8 for each time point). To induce type 1 diabetes, rats were injected with streptozotocin (50 mg/kg dissolved in 0.1 M citrate buffer). Periodontitis was then induced by ligature of the mandibular first molars at day 7 after STZ injection (day 0). IFX was administered once for the 3 day group (on day 0) and twice for the 20 day group (on days 7 and 14). The DP group showed greater alveolar bone loss than the P group on day 20 (P = 0.020). On day 3, higher osteoclast formation and RANKL-positive osteocytes in P group (P = 0.000 and P = 0.011, respectively) and DP group (P = 0.006 and P = 0.017, respectively) than those in C group were observed. However, there was no significant difference in osteoclast formation or RANKL-positive osteocytes between P and DP groups. The DP+IFX group exhibited lower alveolar bone loss (P = 0.041), osteoclast formation (P = 0.019), and RANKL-positive osteocytes (P = 0.009) than that of the DP group. On day 20, DP group showed a lower osteoid area (P = 0.001) and more sclerostin-positive osteocytes (P = 0.000) than P group. On days 3 and 20, the DP+IFX group showed more osteoid area (P = 0.048 and 0.040, respectively) but lower sclerostin-positive osteocytes (both P = 0.000) than DP group. Taken together, these results suggest that TNF-α antagonist can diminish osteocytic RANKL/sclerostin expression and osteoclast formation, eventually recovering osteoid formation. Therefore, TNF-α might mediate alveolar bone loss via inducing expression of osteocytic RANKL and sclerostin in type 1 diabetes rats with periodontitis.

• Alveolar Bone Loss
• Animals
• Bone Morphogenetic Proteins/metabolism
• Diabetes Mellitus, Experimental/complications
• Diabetes Mellitus, Experimental/metabolism
• Genetic Markers
• Immunohistochemistry
• Infliximab/pharmacology
• Male
• Osteocytes/drug effects
• Osteocytes/metabolism
• Periodontitis/complications
• Periodontitis/metabolism
• RANK Ligand/metabolism
• Rats
• Rats, Inbred F344
• Tumor Necrosis Factor-alpha/antagonists & inhibitors

• Basic Science Research Program through the National Research Foundation of Korea (NRF) funded by the Ministry of Education
• Basic Science Research Program through the National Research Foundation of Korea (NRF) funded by the Ministry of Science, ICT and Future Planning
• Yonsei University College of Dentistry Fund

• S‐PRG filler
• bone loss
• inflammation
• periodontal disease

• periodontal diseases
• proteasome
• ubiquitination

EphrinA2‐EphA2 and ephrinB2‐EphB4 critically engage in bidirectional signalling to modulate alveolar bone remodelling. The present study aimed to investigate the effects of lipopolysaccharides (LPS) derived from Porphyromonas gingivalis on ephrin/Eph signalling in periodontal ligament fibroblasts (PDLFs).

The primary cultured PDLFs were incubated in the absence (as a control) or presence of P. gingivalisLPS at 0.001‐10 μg/mL for 24 hours. The PDLFs were then stimulated with P. gingivalisLPS at the optimal concentration (0.1 μg/mL) for different periods (6‐48 hours). The expression of ephrinA2, ephrinB2, EphA2 and EphB4 was assessed by quantitative reverse‐transcription real‐time polymerase chain reaction and western blotting. The osteoblastic markers alkaline phosphatase, osteocalcin and Runt‐related transcription factor 2 (Runx2), and the osteoclastogenesis‐related factors receptor activator of nuclear factor kappa‐B ligand (RANKL) and osteoprotegerin were also evaluated.

The ephrinA2 and EphA2 expression was upregulated and EphB4 expression was downregulated by stimulation of P. gingivalisLPS. EphrinA2 mRNA expression in the PDLFs was significantly upregulated from 12 to 48 hours (P<.05), whereas EphA2 exhibited no change for the first 24 hours, after which there was a significant increase at 48 hours (P<.05). EphB4 exhibited lower mRNA expression at 12 and 24 hours than did the control (P<.05), but the change was insignificant at 48 hours. In contrast, the expression of ephrinB2 remained unchanged. The expressions of ephrinA2, EphA2, ephrinB2 and EphB4 at the protein level showed a similar pattern to that at the mRNA level. The expression of Runx2 and osteocalcin significantly decreased, whereas that of RANKL/osteoprotegerin increased.

The present study suggest that P. gingivalisLPS would contribute to a dysregulation of bone remodelling, whereby ephrinA2/EphA2 expression is stimulated and EphB4 expression is inhibited.

• ephrin/Eph
• lipopolysaccharide
• periodontal ligament fibroblasts

Diabetes mellitus, characterized by abnormally high blood glucose levels, gives rise to impaired bone remodeling. In response to high glucose (HG), the attenuated osteogenic differentiation capacity of human periodontal ligament stem cells (hPDLSCs) is associated with the loss of alveolar bone. Recently, DNA methylation was reported to affect osteogenic differentiation of stem cells in pathological states. However, the intrinsic mechanism linking DNA methylation to osteogenic differentiation ability in the presence of HG is still unclear. In this study, we found that diabetic rats with increased DNA methylation levels in periodontal ligaments exhibited reduced bone mass and density. In vitro application of 5-aza-2′-deoxycytidine (5-aza-dC), a DNA methyltransferase inhibitor, to decrease DNA methylation levels in hPDLSCs, rescued the osteogenic differentiation capacity of hPDLSCs under HG conditions. Moreover, we demonstrated that the canonical Wnt signaling pathway was activated during this process and, under HG circumstances, the 5-aza-dC-rescued osteogenic differentiation capacity was blocked by Dickkopf-1, an effective antagonist of the canonical Wnt signaling pathway. Taken together, these results demonstrate for the first time that suppression of DNA methylation is able to facilitate the osteogenic differentiation capacity of hPDLSCs exposed to HG, through activation of the canonical Wnt signaling pathway.

Periodontitis is the commonest oral disease affecting population worldwide. This disease is notorious for the devastation of tooth supporting structures, ensuing in the loss of dentition. The etiology for this disease is bacterial biofilm, which accumulates on the teeth as dental plaque. In addition to the biofilm microorganisms, other factors such as environmental, systemic and genetic are also responsible in progression of periodontitis. Diabetes mellitus (DM) is metabolic disorder which has an impact on the global health. DM plays a crucial role in the pathogenesis of periodontitis. Periodontitis is declared as the “sixth” major complication of DM. Evidence based literature has depicted an enhanced incidence and severity of periodontitis in subjects with DM. A “two way” relationship has been purported between periodontitis and DM. Mutual management of both conditions is necessary. Periodontal therapy (PT) may assist to diminish the progression of DM and improve glycemic control. Various advanced technological facilities may be utilized for the purpose of patient education and disease management. The present paper clarifies the etio-pathogenesis of periodontitis, establishing it as a complication of DM and elaborating the various mechanisms involved in the pathogenesis. The role of PT in amelioration of DM and application of digital communication will be discussed. Overall, it is judicious to create an increased patient cognizance of the periodontitis-DM relationship. Conjunctive efforts must be undertaken by the medical and oral health care professionals for the management of periodontitis affected DM patients.

• Cost-effectiveness
• Advanced glycation end products
• Complications
• Glycated hemoglobin
• Inflammation
• Mobile health
• Periodontitis
• Periodontal therapy
• Scaling and root planing
• Type 2 diabetes mellitus

Osteoblasts produce various types of cytokines under pathological conditions and control osteoclast differentiation. Tumor necrosis factor-α (TNF-α) has been demonstrated to exert complex effects in osteoblasts under local inflammatory conditions, including in periodontal and periapical diseases. Interleukin-34 (IL-34) has been recently identified as a novel regulatory factor for the differentiation and function of osteoclasts. The present study provides the first evidence, to the best of our knowledge, that the expression of IL-34 is induced by TNF-α through nuclear factor-κB (NF-κB) activation in MC3T3-E1 osteoblastic cells. TNF-α induced IL-34 expression in a dose- and time-dependent manner. Immunocytochemistry with an NF-κB antibody demonstrated that NF-κB was mainly localized in the cytoplasm of the untreated MC3T3-E1 cells. Rapid translocation of NF-κB from the cytoplasm to the nucleus was observed in the cells treated with TNF-α for 15 min. Translocation and transcriptional activity of NF-κB were also determined by western blotting and a luciferase reporter assay, respectively. Pretreatment with 100 μM CAPE, an inhibitor of NF-κB, significantly inhibited TNF-α-induced IL-34 expression. These results indicate that TNF-α induces IL-34 expression via NF-κB in osteoblasts.

• Adaptive Immunity
• Alveolar Bone Loss/immunology
• Alveolar Bone Loss/microbiology
• Bacteria/immunology
• Bacteria/metabolism
• Bacteria/pathogenicity
• Humans
• Immunity, Innate
• Inflammation/microbiology
• Inflammation/pathology
• Interleukin-1/metabolism
• Lymphocytes/immunology
• Lymphocytes/pathology
• Macrophages/metabolism
• Macrophages/pathology
• Osteoclasts/metabolism
• Periodontal Diseases/immunology
• Periodontal Diseases/microbiology
• Tumor Necrosis Factor-alpha/immunology