Anti-inflammatory effects of hesperidin on human gingival fibroblasts stimulated by lipopolysaccharide of Porphyromonas gingivalis in vitro.

Periodontal disease is an inflammatory disease caused by periodontopathogenic bacteria, the inflammatory response generated against them, and host factors. Furthermore, environmental factors can lead to disease progression. Using lipopolysaccharide (LPS)-stimulated human gingival fibroblast (HGF), this study investigated the bioactivity of HGF after exposure to hesperidin (Hesp) and the anti-inflammatory activity of Hesp against early periodontitis. HGF were cultured in Dulbecco's modified Eagle's medium containing 15% fetal bovine serum. They were exposed to LPS for 6 h, followed by Hesp (1, 10, 30, and 50 µM) exposure for 4 h. Cell proliferation was evaluated using reduction staining with alamerBlue™. Inflammatory cytokines [interleukin (IL)-6 and IL-8] and Toll-like receptor 4 (TLR4) levels were assessed using reverse transcription quantitative polymerase chain reaction. Hesp 50 µM + LPS inhibited cell proliferation. The Hesp exposure group inhibited the expression of IL-8 and IL-6. No significant difference in TLR4 expression was observed. Hesp significantly suppressed IL-6 and IL-8 expression by inhibiting downstream signaling without inhibiting TLR4 activation.

Immunogenic effects of enamel matrix derivative on human alveolar ridge mucosa-derived vascular endothelial cells under lipopolysaccharide stimulation.

Early peri-implant disease detection remains difficult. Enamel matrix derivative (EMD), which is used for periodontal tissue regeneration, promotes leukocyte chemotactic factor and adhesion molecule expression in vascular endothelial cells. We hypothesized that stimulating vascular endothelial cells with EMD would induce an inflammatory response in the peri-implant mucosa, enabling early peri-implant infection detection. To verify this hypothesis, we assessed the intercellular adhesion between human alveolar ridge mucosa-derived vascular endothelial cells (ARMEC) stimulated with lipopolysaccharide (LPS) and EMD and human periodontal ligament-derived vascular endothelial cells (PDLEC). Leukocyte chemotactic factors and cell adhesion molecules were investigated and we established an experimental model of peri-implant disease by stimulating ARMEC (representing the peri-implant mucosa) with Porphyromonas gingivalis-derived LPS. ARMEC and PDLEC were obtained from patients (n = 6) who visited the Nippon Dental University Niigata Hospital. The cells were divided into four subcategories, each cultured with: LPS (1 µg/mL), EMD (100 µg/mL), LPS + EMD, and pure medium. Cell viability, leukocyte chemotactic factor (interleukin-8: IL-8), adhesion molecules (intercellular adhesion molecule-1: ICAM-1), tight junction protein gene expression (zonula occludens-1: ZO-1 and Occludin), and transendothelial electrical resistance (TEER) was then determined. LPS reduced ARMEC viability, whereas simultaneous stimulation with EMD improved it. LPS and EMD stimulation enhanced IL-8 and ICAM-1 gene expression, suppressed TEER, and decreased ZO-1 and Occludin expression levels compared to that with stimulation with LPS alone. EMD stimulates leukocyte migration, increase vascular permeability, and trigger an immune response in the peri-implant mucosa, thus facilitating the early detection and treatment of peri-implant disease.

Effects of nicotine and lipopolysaccharide stimulation on adhesion molecules in human gingival endothelial cells.

Smoking is a risk factor for periodontitis, and the immune response of periodontal tissues in patients with periodontitis may be strongly affected by smoking. The purpose of this study was to elucidate the bioactivity and signal transduction of human gingival endothelial cells (HGECs) due to nicotinic stimulation using a cultured medium supplemented with lipopolysaccharide (LPS) as a model of periodontitis. HGECs were cultured in medium supplemented with LPS, nicotine, nicotine + LPS, and medium supplemented without nicotine or LPS (control). Cell proliferation was assessed using Alamar blue. Cytotoxicity was assessed by lactate dehydrogenase leakage. The expression of adhesion molecule-1 (ICAM-1, VCAM-1) was assessed by reverse transcription quantitative polymerase chain reaction (RT-qPCR) and enzyme-linked immunosorbent assay. The expression of nicotinic acetylcholine receptor (nAChR) subunits (α3, α5, α7, ß2 and ß4) was evaluated by RT-PCR. The involvement of p38 mitogen-activated protein kinase (p38MAPK) and protein kinase C (PKC) cell signaling pathways in ICAM-1 and VCAM-1 expression was investigated by RT-qPCR with specific inhibitors. HGECs stimulated with LPS, nicotine and nicotine + LPS showed inhibition of cell proliferation, increase of cell death, and increase of gene and protein expression of ICAM-1. Moreover, HGECs showed the presence of α5 and α7 nAChR subunits. The expression of ICAM-1 in HGECs stimulated with LPS, nicotine, and nicotine + LPS was significantly suppressed by p38MAPK inhibitor, but not by a PKC inhibitor. The nAChR subunits of HGECs are α5 and α7, and that HGECs stimulated with nicotine and LPS express ICAM-1 via p38MAPK pathway.

Dental implant care and trouble among dependent patients based on the questionnaire survey among Japanese dental practitioners.

BACKGROUND: Self-care and professional care of implants may prove difficult for elderly people who require nursing care. However, the actual state of care and problems remains unknown. In this study, we investigated the actual state of implant problems in elderly people living in their own home or in a nursing home who received visiting dental treatment. METHODS: We mailed questionnaire survey forms to 2339 representatives or specialists who were members of the Japanese Society of Oral Implantology, the Japanese Society of Gerodontology or the Japan Prosthodontic Society. We narrowed down the respondents to those who provided visiting dental treatment, and analyzed the actual state of implants observed during visiting dental treatment (type, care, problems, countermeasures, etc.). RESULTS: Of the 924 dentists who responded to the questionnaire survey, 291 (22%) provided visiting dental treatment. While the majority of implant types encountered in the previous 12 months were root-form implants, there were still a certain number of blade and subperiosteal implants. Daily implant care involved mostly cleaning with a toothbrush + auxiliary tools. The most frequent implant problems encountered in the past were difficulty in cleaning and peri-implantitis. Medication and antiphlogistic treatment were most frequently adopted as countermeasures to implant problems, followed by observation. When we classified the results into those for the dentists who provided implant treatment and those for the dentists who did not, we found that many of the dentists who did not provide implant treatment opted for observation or medication, while those who provided implant treatment also implemented removal of superstructure, retightening of screws, repair and so forth. CONCLUSIONS: We found that many of the implant troubles encountered by dentists who provided visiting dental care were difficulty in cleaning or peri-implantitis, and that the actions taken against these troubles varied depending on the experience of the dentist performing the implant treatment. Our study also revealed that dentists who provide visiting dental care need to acquire knowledge and skills of implant treatment, to have actions prepared in case they encounter such cases, or to closely coordinate with dentists who specialize in implants.

Effect of high-glucose conditions on human periodontal ligament endothelial cells: in vitro analysis.

Endothelial cells participate in key aspects of vascular biology, such as maintenance of capillary permeability and regulation of inflammation. According to previous reports, endothelial cells have revealed highly specific characteristics depending on the organs and tissues. In particular, periodontal endothelial cells have a higher permeability than vascular endothelial cells of other types of tissue. Periodontal disease is not only a chronic disease in oral, but also affect the entire body. Diabetes and periodontal disease are closely related, with periodontal disease even been referred to as the sixth complication of disease. However, no reports have investigated the pathophysiology of microvascular in periodontal tissue once diabetes has developed. Therefore, the aim of the present study is to investigate changes in the properties of human periodontal endothelial cells (HPDLECs) that were cultured under high-glucose conditions. We isolated HPDLECs from human periodontal ligament cells. HPDLECs were cultured under high-glucose (5.5, 11.0, 22.0 mM) and investigated proliferation, apoptosis, tube formation and the expression of cell adhesion molecules. A 5.5 mM (100 mg/dl) control was used in this study. HPDLECs stimulated with high glucose concentration exhibited suppression of cell proliferation and an increased percentage of apoptosis-positive cells. This results suggested that apoptosis was caused by TNF-α expression. The expression levels cell adhesion molecules increased. These results suggest that when HPDLECs are stimulated with a high glucose concentrations, PKC in the intracellular cell substrate is activated, increasing the expression of intercellular and vascular adhesion molecules. Thus, the results of this study demonstrate that diabetes exacerbates periodontal disease.

The perivascular phenotype and behaviors of dedifferentiated cells derived from human mature adipocytes.

Derived from mature adipocytes, dedifferentiated fat (DFAT) cells represent a special group of multipotent cells. However, their phenotype and cellular nature remain unclear. Our study found that human DFAT cells adopted perivascular characteristics and behaviors. Flow cytometry and immunofluorescent staining revealed that human DFAT cells positively expressed markers highly related to perivascular cell lineages, such as CD140b, NG2 and desmin, but were negative for common endothelial markers, including CD31, CD34, and CD309. Furthermore, DFAT cells displayed vascular network formation ability in Matrigel, and they noticeably promoted and stabilized the vessel structures formed by human umbilical vascular endothelial cells (HUVECs) in vitro. These results provide novel evidence on the pericyte nature of human DFAT cells, further supporting the recent model for the perivascular origin of adult stem cells, in which tissue-specific progenitor cells in mesenchymal tissues associate with blood vessels, exhibiting perivascular characteristics and functions.

The phenotype and tissue-specific nature of multipotent cells derived from human mature adipocytes.

Dedifferentiated fat (DFAT) cells derived from mature adipocytes have been considered to be a homogeneous group of multipotent cells, which present to be an alternative source of adult stem cells for regenerative medicine. However, many aspects of the cellular nature about DFAT cells remained unclarified. This study aimed to elucidate the basic characteristics of DFAT cells underlying their functions and differentiation potentials. By modified ceiling culture technique, DFAT cells were converted from human mature adipocytes from the human buccal fat pads. Flow cytometry analysis revealed that those derived cells were a homogeneous population of CD13(+) CD29(+) CD105(+) CD44(+) CD31(-) CD34(-) CD309(-) α-SMA(-) cells. DFAT cells in this study demonstrated tissue-specific differentiation properties with strong adipogenic but much weaker osteogenic capacity. Neither did they express endothelial markers under angiogenic induction.

Characterization and Streptococcus mutans adhesion on air polishing dentin.

Air polishing is known as an effective and time saving tooth cleaning method. However, this method increased surface roughness and bacterial adhesion on dentin surface. The aim of this study was to characterize and examine Streptococcus mutans adhesion on dentin surface after air polishing as compared to the conventional method. The dentin blocks (4 × 4 × 1 mm) were polished by a rubber cup with polishing material (Polishing) and air-polished by 25 µm glycine (G25), 65 µm glycine (G65), and 65 µm sodium bicarbonate (NHC65) microparticles. Surface roughness (Ra) was measured by a laser electron microscope. The amount of adhered S. mutans was quantified using a resazurin reduction assay (alamarBlue(®)). The Ra of G25 and G65 was significantly (p < 0.01) smaller than that of NHC65 and greater than that of Polishing. However, there was no significant difference in S. mutans adhesion among Polishing, G25, and G65, while NHC65 showed significantly (p < 0.01) higher S. mutans adhesion. Within the limitations of this in vitro study, air polishing using glycine microparticles conditioned S. mutans adhesion on dentin surface in a similar fashion than the conventional method, and less than air polishing using sodium bicarbonate microparticles.

Effects of air polishing on the resin composite-dentin interface.

The aim of this study was to examine defect depths and volumes at the resin composite-dentin (R/D) interface after air polishing with different particles and spray angles. Samples were 54 dentin specimens that were formed in saucer-shaped cavities filled with resin composite. Each specimen was air polished with either sodium bicarbonate (NaHCO3) or one of two glycine (Gly) powders. The air polisher was set at angles of 90° to the interface and at 45° to the interface from both the dentin and resin composite sides. Air polishing with Gly powder produced defects with less depth and volume than NaHCO3 powder (p < 0.05). Air polishing with a spray angle of 45° to the interface from the resin composite side produced fewer defects (p < 0.05) than polishing from the dentin side. Air polishing to the R/D interface from the resin composite side produced fewer defects to the interface because the hardness of the resin composite was higher than that of dentin.

In vitro analysis of mesenchymal stem cells derived from human teeth and bone marrow.

Mesenchymal stem cells derived from human teeth and bone marrow have been characterized by many research groups, but demonstrate inconsistent cellular phenotypes or functions, partly because of differences in culture methodology. Therefore, our aims were to resolve these inconsistencies and discuss the potential uses of these cells in research/clinical applications. We isolated and characterized dental stem cells (DSCs) from the dental pulp, periodontal ligament, apical papilla (APSCs) and dental follicle (DFSCs) of mature and immature teeth, along with bone marrow-derived stem cells (BMSCs) from the iliac crest. We compared the clonogenic and proliferative potentials of these cells in terms of colony-forming efficiency, proliferation potential, population doubling time and cell cycle. All DSCs, particularly APSCs and DFSCs, possessed greater proliferative potential than BMSCs. All stem cells expressed typical mesenchymal and embryonic markers, and developed alizarin red-positive mineralization nodules and Oil red O-positive lipid droplets when cultured in osteogenic and adipogenic media, respectively. Immunocytochemistry revealed that all stem cells developed neuronal markers when cultured in a control medium without neural inductive supplements. After 7 days of neurogenic culture, the differentiated cells showed a transition from fibroblast-like to neuron-like cell bodies with long processes, suggesting that the stem cells differentiated into mature neurons. Karyotyping confirmed that the stem cells maintained a normal karyotype and were chromosomally stable. Our results provide new insights into the physiological properties of stem cells with a normal karyotype and indicate that DSCs are appropriate for basic research and clinical applications.

The characterization of dentin defects produced by air polishing.

The objective of this study is to characterize the defects in the dentin surface after air polishing for three types of polishing powders and five different nozzle distances. Human teeth were embedded in acrylic resin and then polished until the dentin surface became exposed. The nozzle of the polisher was fixed at a specified distance (2, 3, 4, 5, or 6 mm) and orientation (45°) with respect to the dentin surface. The three powders were CLASSIC (NaHCO(3), 65 µm diameter), PERIO (glycine, 25 µm diameter), and SOFT (glycine, 65 µm diameter). With respect to nozzle distance, we find a significant difference in the mean defect depth only at 6 mm. The spray distance of 6 mm produced the shallowest defect depths. This variable had no effect on the defect volume. SOFT powder had significantly less depths and volumes of defects than the other two powders. The contact angle of the dentin increased significantly in all polishing tests, compared to an unpolished dentin surface (control). We find that spray distance of 6 mm from the nozzle of the polisher and orienting on 45° angle produced less dentin surface defects than any other distance in all powder systems used. At this distance, SOFT powder produced statistically significant smaller depth and volume defects than the other two powder groups.

Estimation of the Periodontal Inflamed Surface Area by Simple Oral Examination.

The periodontal inflamed surface area (PISA) is a useful index for clinical and epidemiological assessments, since it can represent the inflammation status of patients in one contentious variable. However, calculation of the PISA is difficult, requiring six point probing depth measurements with or without bleeding on probing on 28 teeth, followed by data input in a calculation program. More simple methods are essential for screening periodontal disease or in epidemiological studies. In this study, we tried to establish a convenient partial examination method to estimate PISA. Cross-sectional data of 254 subjects who completed active periodontal therapy were analyzed. Teeth that represent the PISA value were selected by an item response theory approach. The maxillary second molar, first premolar, and lateral incisor and the mandibular second molar and lateral incisor were selected. The sum of the PISAs of these teeth was significantly correlated with the patient's PISA (R2 = 0.938). More simply, the sum of the maximum values of probing pocket depth with bleeding for these teeth were also significantly correlated with the patient's PISA (R2 = 0.6457). The simple model presented in this study may be useful to estimate PISA.

Prospective Longitudinal Changes in the Periodontal Inflamed Surface Area Following Active Periodontal Treatment for Chronic Periodontitis.

Periodontal disease is a chronic inflammatory disease of the periodontal tissue. The periodontal inflamed surface area (PISA) is a proposed index for quantifying the inflammatory burden resulting from periodontitis lesions. This study aimed to investigate longitudinal changes in the periodontal status as evaluated by the PISA following the active periodontal treatment. To elucidate the prognostic factors of PISA, mixed-effect modeling was performed for clinical parameters, tooth-type, and levels of periodontal pathogens as independent variables. One-hundred-twenty-five patients with chronic periodontitis who completed the active periodontal treatment were followed-up for 24 months, with evaluations conducted at 6-month intervals. Five-times repeated measures of mean PISA values were 130+/-173, 161+/-276, 184+/-320, 175+/-417, and 209+/-469 mm2. Changes in clinical parameters and salivary and subgingival periodontal pathogens were analyzed by mixed-effect modeling. Plaque index, clinical attachment level, and salivary levels of Porphyromonas gingivalis were associated with changes in PISA at the patient- and tooth-level. Subgingival levels of P. gingivalis and Prevotella intermedia were associated with changes in PISA at the sample site. For most patients, changes in PISA were within 10% of baseline during the 24-month follow-up. However, an increase in the number of bleeding sites in a tooth with a deep periodontal pocket increased the PISA value exponentially.

Effect of particle diameter on air polishing of dentin surfaces.

We examined the abrasiveness of glycine powders with particle diameters of 63 and 100 mum by measuring the depth and volume of defects produced during air polishing of human dentin. A total of 36 extracted human teeth were embedded in acrylic resin. The resin blocks were polished until the dentin surfaces were exposed. The nozzle of an air polisher was mounted 4 mm from the dentin surface, and the dentin surface was treated for 5 s at one of two angles of incidence (45 degrees or 90 degrees). Three materials were used in the polishing process: NaHCO(3) powder with a mean particle diameter of 100 microm (Handy Jet Powder), glycine powder with a mean particle diameter of 63 microm (Handy Jet Powder PMTC), and glycine powder with a mean particle diameter of 100 microm (Handy Jet Powder Recall). The defect depth at both angles was significantly deeper after treatment with Handy Jet Powder or Handy Jet Powder PMTC. The defect volume was the greatest with Handy Jet Powder, followed by Handy Jet Powder PMTC, and Handy Jet Powder Recall. The larger diameter glycine powder resulted in less damage to the dentin.

The relationship between physiologic halitosis and periodontopathic bacteria of the tongue and gingival sulcus.

To determine the influence of oral status on halitosis, the relationship between halitosis and periodontopathic bacteria present in plaque on the tongue and the subgingival sulcus was examined in 62 periodontally healthy adults. Halitosis indicators used were the organoleptic score; gas chromatography results [total volatile sulfur compounds (VSCs) = H(2)S + CH(3)SH + (CH(3))(2)S]; Halimeter values; and the results of three clinical tests, plaque control record (PlCR), plaque index (PlI), and tongue coat status. Significant correlations with organoleptic scores was observed for PlCR, PlI, tongue coat status, VSC amounts, and Halimeter values, indicating that halitosis in periodontally healthy subjects tended to originate from tongue plaque deposits. Polymerase chain reaction analysis was used to detect six periodontopathic bacteria (Aggregatibacter actinomycetemcomitans, Fusobacterium nucleatum, Porphyromonas gingivalis, Prevotella intermedia, Tannerella forsythensis, and Treponema denticola) from the tongue and subgingival plaque. Significant effects on the organoleptic scores, tongue coat status, total VSC, H(2)S and CH(3)SH amounts, and Halimeter values were observed only for T. denticola and F. nucleatum and only in the tongue plaque, not in the subgingival plaque. Thus, therapies developed to inhibit the growth of these bacteria may lead to future treatments of halitosis.

Characterization and comparative DNA methylation profiling of four adipogenic genes in adipose-derived stem cells and dedifferentiated fat cells from aging subjects.

Adipose-derived stem cells (ASCs) and dedifferentiated fat (DFAT) cells are alternative cell sources in tissue engineering and regeneration because they are easily obtained and exhibit multilineage differentiation. However, aging may attenuate their regenerative potential and metabolic functions. Reports characterizing DFAT cells derived from aging donors are rare, and comparisons of DNA methylation profiles between aging ASCs and DFAT cells are poorly understood. Therefore, this study aimed to characterize DFAT cells relative to ASCs derived from aging subjects and compare the DNA methylation profiles of four adipogenic genes in these cells. ASCs and DFAT cells from aging donors exhibited characteristics similar to those of stem cells, including colony formation, proliferation, and multilineage differentiation abilities. However, compared with ASCs, DFAT cells exhibited increased proliferation, smooth muscle actin alpha (SMA-α) expression and decreased cellular senescence. DNA methylation profiling of ASCs and DFAT cells by combined bisulfite restriction analysis (COBRA) demonstrated hypermethylation patterns in three potent adipogenic genes-peroxisome proliferator-activated receptor gamma 2 (PPARγ2), fatty acid-binding protein 4 (FABP4), and lipoprotein lipase (LPL)-but hypomethylation of CCAAT/enhancer binding protein alpha (C/EBPα) in the aging group. Statistically significant differences were observed between the aging group and the young group. Epigenetic regulation maintains the stability of ASCs and DFAT cells in an age-dependent manner. Our findings suggested that although the DNA methylation patterns of three adipogenic genes correlated with hypermethylation and aging, ASCs and DFAT cells exhibited cellular stability and several stem cell characteristics, offering further opportunities for personalized regeneration and energy maintenance by adipogenesis during aging.

A multicenter prospective cohort study on the effect of smoking cessation on periodontal therapies in Japan.

Few prospective studies have reported the effects of periodontal therapy on patients who attempted to quit smoking. This study aimed to assess how smoking cessation affects periodontal therapy. Twenty-five smokers with periodontitis were investigated by dividing them into two groups, a smoking cessation support group and a continued smoking group. Those in the support group received counseling and nicotine replacement therapy, followed by periodontal treatment conducted by dentists who had completed an e-learning course on smoking cessation. Clinical parameters were measured at baseline, 3, and 6 months. Most clinical parameters improved for those in the smoking cessation support group. There were no significant improvements in bleeding on probing (BOP) or the number of severe periodontal disease sites in the continued smoking group. Probing pocket depth (PPD) and clinical attachment levels (CAL) at sites that received scaling and root planing (SRP) significantly improved in all subjects. BOP did not improve at reevaluation in the smoking relapse subgroup. Patients in the smoking cessation support program led by dental professionals showed more improvement in BOP than those in the continued smoking group.

Optimal Examination Sites for Periodontal Disease Evaluation: Applying the Item Response Theory Graded Response Model.

Periodontal examination data have a complex structure. For epidemiological studies, mass screenings, and public health use, a simple index that represents the periodontal condition is necessary. Periodontal indices for partial examination of selected teeth have been developed. However, the selected teeth vary between indices, and a justification for the selection of examination teeth has not been presented. We applied a graded response model based on the item response theory to select optimal examination teeth and sites that represent periodontal conditions. Data were obtained from 254 patients who participated in a multicenter follow-up study. Baseline data were obtained from initial follow-up. Optimal examination sites were selected using item information calculated by graded response modeling. Twelve sites-maxillary 2nd premolar (palatal-medial), 1st premolar (palatal-distal), canine (palatal-medial), lateral incisor (palatal-central), central incisor (palatal-distal) and mandibular 1st premolar (lingual, medial)-were selected. Mean values for clinical attachment level, probing pocket depth, and bleeding on probing by full mouth examinations were used for objective variables. Measuring the clinical parameters of these sites can predict the results of full mouth examination. For calculating the periodontal index by partial oral examination, a justification for the selection of examination sites is essential. This study presents an evidence-based partial examination methodology and its modeling.

A preliminary report on dental implant condition among dependent elderly based on the survey among Japanese dental practitioners.

BACKGROUND: The objective of this study was to ascertain the situation relevant to implants, the status of oral self-care, the status of aftercare provided by the dentist who placed the implant, and the usage status of the implant card, in homebound or institutionalized older adults who are receiving home-visit dental care due to the inability to visit a dental clinic on their own. METHODS: A survey questionnaire was sent by post mail to 2339 people who are representative members or dental specialists belonging to any of the following three academic societies: Japanese Society of Oral Implantology, Japanese Society of Gerodontology, and Japan Prosthodontic Society. The survey questions asked were about provision/no provision of implant treatment, provision/no provision of home-visit dental care, the situation of patients after implant treatment, the situation of implants in the context of home-visit dental care, and the usage status and recognition of the implant card. RESULTS: No less than 30% of the dentists had patients who were admitted to the hospital or became homebound after receiving implant treatment at their clinic. Twenty-two percent of the dentists had been consulted about the implants. Dentists who continued to provide post-operative implant care through home-visit dental care accounted for approximately 80%. On the other hand, however, 40% of the dentists did not know the post-implantation status of their implant patients. Of the patients receiving home-visit dental care, approximately 3% had implants (identified mainly by visual inspection). It was found that more than 50% of the dentists offering implant treatment did not use the implant card, and even in cases where it was used, most of the cards were not in the standardized format. CONCLUSIONS: Within the limitation of low response rate to the questionnaire in this preliminary study, we concluded that many of practitioners including specialists indicated the need of universal record of implant for dependent elderly cares.

Establishment and characterization of novel epithelial-like cell lines derived from human periodontal ligament tissue in vitro.

In this study, novel human-derived epithelial-like cells (hEPLCs) lines were established from periodontal ligament (PDL) tissues, which were composed of a variety of cell types and exhibited complex cellular activities. To elucidate the putative features distinguishing these from epithelial rest of Malassez (ERM), we characterized hEPLCs based on cell lineage markers and tight junction protein expression. The aim of this study was, therefore, to establish and characterize hEPLCs lines from PDL tissues. The hEPLCs were isolated from PDL of third molar teeth. Cellular morphology and cell organelles were observed thoroughly. The characteristics of epithelial-endothelial-mesenchymal-like cells were compared in several markers by gene expression and immunofluorescence, to ERM and human umbilical-vein endothelial cells (HUVECs). The resistance between cellular junctions was assessed by transepithelial electron resistance, and inflammatory cytokines were detected by ELISA after infecting hEPLCs with periodontopathic bacteria. The hEPLCs developed into small epithelial-like cells in pavement appearance similar to ERM. However, gene expression patterns and immunofluorescence results were different from ERM and HUVECs, especially in tight junction markers (Claudin, ZO-1, and Occludins), and endothelial markers (vWF, CD34). The transepithelial electron resistance indicated higher resistance in hEPLCs, as compared to ERM. Periodontopathic bacteria were phagocytosed with upregulation of inflammatory cytokine secretion within 24 h. In conclusion, hEPLCs that were derived using the single cell isolation method formed tight multilayers colonies, as well as strongly expressed tight junction markers in gene expression and immunofluorescence. Novel hEPLCs lines exhibited differently from ERM, which might provide some specific functions such as metabolic exchange and defense mechanism against bacterial invasion in periodontal tissue.