SPARC is a decoy counterpart for c­Fos and is associated with osteoblastic differentiation of bone marrow stromal cells by inhibiting adipogenesis.

Secreted protein acidic and rich in cysteine (SPARC), also called basement­membrane protein 40 or osteonectin, is a matricellular protein that is abundant not only in bone tissue as a non­collagenous protein but is also ubiquitously expressed in non­calcified tissue. SPARC is located intracellularly and disruption of the Sparc gene has been reported to reduce bone formation and increase fat tissue; however, the mechanism by which SPARC inhibits adipogenesis remains unclear. The present study evaluated the intracellular function of SPARC in adipogenesis using the bone marrow stromal cell line ST2. When ST2 cells with low SPARC production were cloned, intrinsic activator protein­1 (AP­1) activity was markedly higher, mineralized nodule formation was significantly lower and lipid accumulation was significantly increased compared with in the parental ST2 cells. Forced expression of secreted SPARC with the signal peptide­coding sequences of wild­type Sparc or preprotrypsin in SPARC­low ST2 cells significantly reduced AP­1 transcription activity; however, these reductions were not observed in the absence of signal peptide sequences. Recombinant SPARC, produced using Brevibacillus brevis, specifically bound to c­Fos but not c­Jun and inhibited the binding of c­Fos/c­Jun to a TPA­response element sequence. These data suggested that SPARC was incorporated into the cells from the extracellular spaces and serves an intracellular role as a decoy counterpart for c­Fos, as well as being associated with osteoblastogenesis through the inhibition of adipogenesis. These findings may provide new insights into regenerative medicine.

Personalized Medicine Based on the Pathogenesis and Risk Assessment of Endodontic-Periodontal Lesions.

Endodontic-periodontal lesions (EPLs) are chronic inflammatory lesions in the mouth caused by multiple factors. Both periapical and marginal periodontitis are characterized by infection and inflammation around the affected teeth, suggesting that the theory of complex systems might describe the progression of EPL. The diagnosis and treatment of EPLs are complicated by variations of this condition and difficulties distinguishing EPLs from other diseases. Technological advances in diagnostic and treatment methods, including cone beam computed tomography, microscopy, mineral trioxide aggregates, and periodontal regenerative treatment, have improved outcomes, even in untreatable teeth. However, treating EPLs with iatrogenic problems and/or severe periodontitis remains challenging. Assessing the risk of each EPL based on the possible pathogenesis of each EPL is essential for determining individualized treatment and optimizing personalized medicine for individual patients.

The Stages and Grades of Periodontitis Are Risk Indicators for Peri-Implant Diseases-A Long-Term Retrospective Study.

The aim of this study is to evaluate the factors of implant failure in patients with periodontitis and their impact on the prognosis of having a peri-implant disease and/or implant failure. Data regarding 325 implants among 84 patients with periodontitis were retrospectively examined. Patients were classified by Stage (I, II, III, IV) and Grade (A, B, C), implant failures for peri-implant disease and lack of osseointegration. Clinical data, including implant- and patient-related variables were evaluated by principal components analysis (PCA) and two-step cluster analysis (CA). Survival and success rates were 96.3% and 87.1%, respectively. Prevalence of peri-implant disease was significantly higher in Stage IV patients (p < 0.05), and incidence of lost implant due to peri-implantitis was significantly higher in patients with bone augmentation (BA) (p < 0.05). PCA and CA revealed five of eleven variables and four clusters at patient level, and six of fourteen variables and three clusters at implant level. Stage and Grade are useful indicators for the development of peri-implant diseases in which BA and the number of implants are involved.

Proposal for a Paradigm Shift in Personalized Medicine for Patients with a Maxillary Edentulous Jaw by ENT Specialist and Dentist Cooperation.

With the spread of oral implant therapy, serious medical complications related to implant surgery are becoming a social problem. Although the number of complications after implant surgery in the edentulous jaw is decreasing in Japan, maxillary-sinus-related complications (MSRCs) have reached the highest number since 2012. It is essential to identify and eliminate possible predisposing risk factors for MSRCs at an early stage to prevent MSRCs. In this review article, we highlight the causal factors of postoperative complications with or without sinus augmentation for the maxillary molar region to achieve optimal treatment outcomes and reduce complications. In particular, we focus on anatomical variations that can cause the impairment of maxillary sinus drainage. Furthermore, we emphasize that the paradigm for personalized medicine for patients with a maxillary edentulous jaw by ENT specialist and dentist cooperation is shifting from the traditional assessment of maxillary sinus pathologies alone to the new assessment of anatomic variations that can cause the impairment of maxillary sinus drainage in addition to maxillary sinus pathologies.

The importance of early detection for postoperative maxillary cyst before dental implantation: A case report.

INTRODUCTION AND IMPORTANCE: It is difficult that doctors other than otorhinolaryngologist/radiologist find early postoperative maxillary cyst (POMC) because it tends to expand gradually with no symptoms over a period of years. CASE PRESENTATION: A 60-year-old Japanese male who had previously undergone a bilateral Caldwell-Luc operation for the treatment of chronic sinusitis, experienced maxillary sinus floor elevation and implant placement. Eleven years after the implant placement, we discovered that the left POMC existed close to dental implants. Fortunately, dental implants still displayed proper osseointegration. Thus, the patient has been successfully treated for POMC, which had not been proper diagnosed before the implantation, by a marsupialization using nasal endoscopy and successfully preserved dental implant. CLINICAL DISCUSSION: Because the expanding POMC might result in dental implant failure after several years, we think that marsupialization is useful as a risk management for possible failure of dental implant close to POMC when it had not been found before maxillary sinus floor elevation and insertion of dental implant. CONCLUSION: Doctors should recognize that patients will have the risk of the dental implant failure after several years due to the expanding cyst when early POMC had not been diagnosed and treated properly before the implantation.

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.

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.

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 Successful Treatment Regimen for the Prevention of Sinusitis after Maxillary Sinus Floor Elevation Surgery in a High-Risk Case.

Maxillary sinus floor elevation (sinus lift) is a widely recognized dental-surgical approach for dental implant placement. However, for an otorhinolaryngological high-risk patient with severe anatomic-structural impairments of the maxillary sinus drainage pathway, surgical intervention is recommended before sinus lift to avoid postsinus lift maxillary sinusitis. Here, we show a case that postsinus lift maxillary sinusitis in such a high-risk patient was noninvasively prevented by the collaboration of otorhinolaryngologist and dentist. A 48-year-old Japanese male intended to undergo a sinus lift for dental implant placement by periodontist. Otorhinolaryngologist found septal deviation, concha bullosa, the presence of Haller cell, and nasal mucosal swelling by the nasal allergy, while no sinusitis and diagnosed him as a "high-risk case" for postsinus lift maxillary sinusitis. The patient was administered preoperative topical steroid and leukotriene receptor antagonist in addition to perioperative antibiotic prophylaxis so that his complication was noninvasively prevented. Thus, this case suggested that consultation from dentist to otorhinolaryngologist provides benefit to the patients who have been diagnosed as "high-risk case" for postsinus lift maxillary sinusitis.

Hidden noise in immunologic parameters might explain rapid progression in early-onset periodontitis.

To investigate in datasets of immunologic parameters from early-onset and late-onset periodontitis patients (EOP and LOP), the existence of hidden random fluctuations (anomalies or noise), which may be the source for increased frequencies and longer periods of exacerbation, resulting in rapid progression in EOP. Principal component analysis (PCA) was applied on a dataset of 28 immunologic parameters and serum IgG titers against periodontal pathogens derived from 68 EOP and 43 LOP patients. After excluding the PCA parameters that explain the majority of variance in the datasets, i.e. the overall aberrant immune function, the remaining parameters of the residual subspace were analyzed by computing their sample entropy to detect possible anomalies. The performance of entropy anomaly detection was tested by using unsupervised clustering based on a log-likelihood distance yielding parameters with anomalies. An aggregate local outlier factor score (LOF) was used for a supervised classification of EOP and LOP. Entropy values on data for neutrophil chemotaxis, CD4, CD8, CD20 counts and serum IgG titer against Aggregatibacter actinomycetemcomitans indicated the existence of possible anomalies. Unsupervised clustering confirmed that the above parameters are possible sources of anomalies. LOF presented 94% sensitivity and 83% specificity in identifying EOP (87% sensitivity and 83% specificity in 10-fold cross-validation). Any generalization of the result should be performed with caution due to a relatively high false positive rate (17%). Random fluctuations in immunologic parameters from a sample of EOP and LOP patients were detected, suggesting that their existence may cause more frequently periods of disease activity, where the aberrant immune response in EOP patients result in the phenotype "rapid progression".

Site-level progression of periodontal disease during a follow-up period.

Periodontal disease is assessed and its progression is determined via observations on a site-by-site basis. Periodontal data are complex and structured in multiple levels; thus, applying a summary statistical approach (i.e., the mean) for site-level evaluations results in loss of information. Previous studies have shown the availability of mixed effects modeling. However, clinically beneficial information on the progression of periodontal disease during the follow-up period is not available. We conducted a multicenter prospective cohort study. Using mixed effects modeling, we analyzed 18,834 sites distributed on 3,139 teeth in 124 patients, and data were collected 5 times over a 24-month follow-up period. The change in the clinical attachment level (CAL) was used as the outcome variable. The CAL at baseline was an important determinant of the CAL changes, which varied widely according to the tooth surface. The salivary levels of periodontal pathogens, such as Porphyromonas gingivalis and Aggregatibacter actinomycetemcomitans, were affected by CAL progression. "Linear"- and "burst"-type patterns of CAL progression occurred simultaneously within the same patient. More than half of the teeth that presented burst-type progression sites also presented linear-type progression sites, and most of the progressions were of the linear type. Maxillary premolars and anterior teeth tended to show burst-type progression. The parameters identified in this study may guide practitioners in determining the type and extent of treatment needed at the site and patient levels. In addition, these results show that prior hypotheses concerning "burst" and "linear" theories are not valid.

Artificial neural networks for the diagnosis of aggressive periodontitis trained by immunologic parameters.

There is neither a single clinical, microbiological, histopathological or genetic test, nor combinations of them, to discriminate aggressive periodontitis (AgP) from chronic periodontitis (CP) patients. We aimed to estimate probability density functions of clinical and immunologic datasets derived from periodontitis patients and construct artificial neural networks (ANNs) to correctly classify patients into AgP or CP class. The fit of probability distributions on the datasets was tested by the Akaike information criterion (AIC). ANNs were trained by cross entropy (CE) values estimated between probabilities of showing certain levels of immunologic parameters and a reference mode probability proposed by kernel density estimation (KDE). The weight decay regularization parameter of the ANNs was determined by 10-fold cross-validation. Possible evidence for 2 clusters of patients on cross-sectional and longitudinal bone loss measurements were revealed by KDE. Two to 7 clusters were shown on datasets of CD4/CD8 ratio, CD3, monocyte, eosinophil, neutrophil and lymphocyte counts, IL-1, IL-2, IL-4, INF-γ and TNF-α level from monocytes, antibody levels against A. actinomycetemcomitans (A.a.) and P.gingivalis (P.g.). ANNs gave 90%-98% accuracy in classifying patients into either AgP or CP. The best overall prediction was given by an ANN with CE of monocyte, eosinophil, neutrophil counts and CD4/CD8 ratio as inputs. ANNs can be powerful in classifying periodontitis patients into AgP or CP, when fed by CE values based on KDE. Therefore ANNs can be employed for accurate diagnosis of AgP or CP by using relatively simple and conveniently obtained parameters, like leukocyte counts in peripheral blood. This will allow clinicians to better adapt specific treatment protocols for their AgP and CP patients.

Re-evaluation of anti-inflammatory activity of mastic using activated macrophages.

Mastic is a resinous exudate obtained from the stem and the main leaves of Pistacia lentiscus. Mastic has shown several beneficial pharmaceutical properties such as antibacterial and apoptosis-modulating activities. The aim of this study was to investigate whether mastic affects the function of activated macrophages. Both solid and liquid types of mastic inhibited the production of pro-inflammatory substances such as nitric oxide (NO) and prostaglandin (PG)E(2) by lipopolysaccharide (LPS)-activated mouse macrophage-like RAW264.7 cells. This was accompanied by the decline of viable cell number. Western blot and RT-PCR analyses showed that mastic inhibited the expression of inducible NO synthase (iNOS) and cyclooxygenase (COX)-2 at both protein and mRNA levels. ESR spectroscopy revealed that mastic scavenged NO and superoxide radicals very poorly, in contrast to its potent hydroxyl radical scavenging activity. These data demonstrate that mastic inhibits the production of both NO and PGE(2) by activated macrophages mostly via its cytotoxic action. The narrow range of effective concentration of mastic due to its cytotoxicity may limit its potential application as an anti-inflammatory agent.

Selective antibacterial and apoptosis-modulating activities of mastic.

Mastic is a resinous exudate obtained from the stem and the main leaves of Pistacia lentiscus. We have reported the antiplaque effect of mastic-containing chewing gum on the oral cavity. We hypothesize that mastic may be a multifunctional food which has some beneficial pharmaceutical properties. The aim of this study was to assess the biological activity of solid and liquid types of mastic by cytotoxicity against fibroblasts, radical-scavenging activities and inhibitory effect on cell death of oral polymorphonuclear leukocytes (OPMNs). Mastic showed selective antibacterial action against Porphyromonas gingivalis and Prevotella melaninogenica, but no anti-HIV activity. Among a total of thirteen human cell types, promyelocytic leukemia HL-60 was the most sensitive to the cytotoxicity of mastic, followed by myeloblastic leukemia (ML-1, KG-1), erythroleukemia (K-562), oral squamous cell carcinoma (HSC-2, HSC-3, HSC-4), hepatocellular carcinoma (HepG2), glioblastoma (T98G, U87MG) and normal oral cells (gingival fibroblast, pulp cell, periodontal ligament fibroblast, most resistant). Mastic did not induce the differentiation of myelogenous leukemic cells into maturing cells with higher nitroblue tetrazolium-reducing activity, but induced apoptotic cell death, characterized by internucleosomal DNA fragmentation, caspase-3 activation and a decline in the intracellular concentration of putrescine. The cytotoxicity of mastic against leukemic cells did not diminish during its storage. On the other hand, mastic inhibited the spontaneous apoptosis of OPMNs. Mastic showed hydroxyl radical-scavenging activity. The selective antibacterial and apoptosis-modulating activity of mastic suggests its possible beneficial effects on oral health.

Glutathione can efficiently prevent direct current-induced cytotoxicity.

We have reported that direct current (DC) with antibacterial agents used in iontophoresis for root canal disinfection induced host cell necrotic cytotoxicity, and this DC-induced cytotoxicity may be because of generated free radicals and metal ions eluted from metal electrodes. Iontophoresis is still used in some cases, and thus it is necessary to consider how we may prevent DC-induced cytotoxicity of host cells of periapical lesions. Thus, we compared the protective effects of various antioxidants on the DC-induced cytotoxicity against host cells. N-acetyl-L-cysteine and glutathione (GSH) efficiently prevented DC-induced cytotoxicity against human polymorphonuclear cells (PMNs) (p < 0.01). The DC-induced cytotoxicity against PMNs was significantly enhanced by buthionine sulfoximine (p < 0.05), an inhibitor of GSH synthesis, and its effect was rescued by adding the exogenous GSH (p < 0.01). In addition, DC treatment reduced the intracellular GSH levels in a time-dependent manner (p < 0.05). Transmission electron microscopy showed that the DC induced the intense vacuolization and accumulation of cellular debris in autophagolysosomes, and these morphological changes were blocked by adding exogenous GSH. These results suggest that GSH, a thiol antioxidant, effectively prevents the DC-induced cytotoxicity.

Anti-stress, anti-HIV and vitamin C-synergized radical scavenging activity of mulberry juice fractions.

Anti-stress and anti-HIV activity of mulberry juice were separated by centrifugation. The anti-stress activity was enriched in the supernatant fraction whereas the anti-HIV activity in the precipitate fraction. Oral administration of the supernatant fraction significantly reduced the elevated plasma level of lipid peroxide in mice loaded with water immersion restraint stress. The kinetic study revealed that the anti-stress activity was maintained for 4 hours after cessation of the administration of mulberry juice. The lignin fraction in the precipitate fraction scavenged superoxide and hydroxyl radicals more efficiently than other fractions, in a synergistic fashion with sodium ascorbate. Anti-HIV activity of mulberry juice was concentrated in the lignin fraction, whereas blueberry juice, which has no precipitating fibrous materials, did not show anti-HIV activity. The present study suggests the functionality of mulberry juice as an alternative medicine.

Anti-stress activity of mulberry juice in mice.

The possible anti-stress activity of mulberry juice was investigated in mice. When mice were subjected to water immersion restraint stress at 25 degrees C for 8 h, the plasma lipid peroxide level, determined by the d-ROMs test performed 12 h thereafter, was almost doubled. After administration of mulberry juice one or two weeks before the stress loading, the lipid peroxidation was completely blocked. Administration of mulberry juice after the stress loading, without pre-administration, was also protective. ESR spectroscopy revealed that mulberry juice scavenged superoxide anion (generated by hypoxanthine and xanthine oxidase reaction), hydroxyl radical (produced by the Fenton reaction) and NO radical (generated by a NO donor) at approximately 50% efficiency of blueberry juice. Mulberry juice produced smaller amounts of radical at neutral to alkaline pH. The cytotoxic and anti-HIV activities of mulberry juice were 18% and >4-fold those of blueberry juice, respectively. These data suggest that the anti-stress activity of mulberry juice in vivo may be derived from its radical scavenging activity.

Role of free radicals and metal ions in direct current-induced cytotoxicity.

The purpose of this study was to investigate the mechanism of direct current (DC)-induced cytotoxicity. To test the working hypothesis that electrolysis products are responsible for the DC-induced cytotoxicity, the cytotoxic effects between the direct and indirect DC treatment against human polymorphonuclear cells (PMNs) was compared. The indirect DC treatment (treatment with the culture medium exposed to DC) was comparable in cytotoxicity to the direct DC treatment, suggesting that electrolysis products have an important role in DC-induced cytotoxicity. Metal ions released from different electrodes into the culture medium were quantified by the inductively coupled plasma-mass spectroscopy. Higher concentrations of Ag, Zn, and Ni and chromium were released from Ag, Zn, and stainless steel (St) electrodes, respectively, whereas much lower concentrations of Ni and Ti were released from Ni-Ti electrode. Further, electron spin resonance spectroscopy with spin-trapping agent showed that the direct current with the following metal electrodes generated alkoxyl radical (St and Ni-Ti electrodes), hydrogen radical (Ag and Au electrodes), and both carbon and alkoxyl radicals (Zn electrode), respectively. These results suggest that free radicals and metal ions released from electrodes contribute to the cytotoxicity of DC treatment used for iontophoresis.

Enhancement of cytotoxic activity of sodium fluoride against human periodontal ligament fibroblasts by water pressure.

Whether or not water pressure enhances the cytotoxic activity of sodium fluoride (NaF) against human periodontal ligament fibroblast (HPLF) was investigated. Loading with water pressure (up to 5 g) alone did not affect the cell proliferation, but significantly enhanced the cytotoxic activity of millimolar concentrations of NaF. Cytotoxic activity of NaF was reduced by supplementation with Ca2+, whereas it was enhanced by removal of Ca2+ from the culture medium. However, the enhancement of cytotoxicity of NaF under water pressure was observed even in the Ca2+ -free medium. NaF failed to induce apoptosis markers, such as the caspase-3, -8 and -9 activation, the intemucleosomal DNA fragmentation, the loss of cell surface microvilli and the changes in intracellular concentration of polyamines. Transmission electron microscopy demonstrated that the combination of NaF and water pressure slightly increased the incidence of the formation of autophagosomes engulfing organella, suggesting the induction of non-apoptotic cell death in HPLF cells. The present study suggests that the external pressure is an additional factor that enhances the cytotoxicity of NaF against HPLF cells.