Seoul National University Dental Hospital

Mesenchymal stem cell-mediated tissue regeneration is a promising approach for regenerative medicine for a wide range of applications. Here we report a new population of stem cells isolated from the root apical papilla of human teeth (SCAP, stem cells from apical papilla). Using a minipig model, we transplanted both human SCAP and periodontal ligament stem cells (PDLSCs) to generate a root/periodontal complex capable of supporting a porcelain crown, resulting in normal tooth function. This work integrates a stem cell-mediated tissue regeneration strategy, engineered materials for structure, and current dental crown technologies. This hybridized tissue engineering approach led to recovery of tooth strength and appearance.

Aberrantly enhanced vascular endothelial growth factor (VEGF) gene expression is associated with increased tumor growth and metastatic spread of solid malignancies, including human renal carcinomas. Persistent activation of STAT3 is linked to tumor-associated angiogenesis, but underlying mechanisms remain unclear. Therefore, we examined whether STAT3 modulates the stability and activity of hypoxia-inducible factor-1alpha (HIF-1alpha), and in turn enhances VEGF expression. We found that STAT3 was activated in ischemic rat kidneys and hypoxic human renal carcinoma cells. We also found that hypoxia-induced activation of STAT3 transactivated the VEGF promoter and increased the expression of VEGF transcripts. Consistent with these findings, STAT3 inhibition attenuated the hypoxic induction of VEGF. Interestingly, activated STAT3 increased HIF-1alpha protein levels due to the HIF-1alpha stability by blocking HIF-1alpha degradation and accelerated its de novo synthesis. The novel interaction of STAT3 with HIF-1alpha was identified in hypoxic renal carcinoma cells. Furthermore, hypoxia recruited STAT3, HIF-1alpha, and p300 to the VEGF promoter and induced histone H3 acetylation. Therefore, these findings provide compelling evidence that a causal relationship exists between STAT3 activation and HIF-1-dependent angiogenesis and suggest that therapeutic modalities designed to disrupt STAT3 signaling hold considerable promise for the blocking tumor growth and enhancing apoptosis of cancer cells and tissues.

Using a bacterial pathogen-induced acute inflammation model in the skin, we defined the roles of local lymphatic vessels and draining lymph nodes (DLNs) in antigen clearance and inflammation resolution. At the peak day of inflammation, robust expansion of lymphatic vessels and profound infiltration of CD11b+/Gr-1+ macrophages into the inflamed skin and DLN were observed. Moreover, lymph flow and inflammatory cell migration from the inflamed skin to DLNs were enhanced. Concomitantly, the expression of lymphangiogenic growth factors such as vascular endothelial growth factor C (VEGF-C), VEGF-D, and VEGF-A were significantly up-regulated in the inflamed skin, DLNs, and particularly in enriched CD11b+ macrophages from the DLNs. Depletion of macrophages, or blockade of VEGF-C/D or VEGF-A, largely attenuated these phenomena, and produced notably delayed antigen clearance and inflammation resolution. Conversely, keratin 14 (K14)-VEGF-C transgenic mice, which have dense and enlarged lymphatic vessels in the skin dermis, exhibited accelerated migration of inflammatory cells from the inflamed skin to the DLNs and faster antigen clearance and inflammation resolution. Taken together, these results indicate that VEGF-C, -D, and -A derived from the CD11b+/Gr-1+ macrophages and local inflamed tissues play a critical role in promoting antigen clearance and inflammation resolution.

BACKGROUND: Titanium is a commonly used inert bio-implant material within the medical and dental fields. Although the use of titanium is thought to be safe with a high success rate, in some cases, there are rare reports of problems caused by titanium. In most of these problematic reports, only individual reports are dominant and comprehensive reporting has not been performed. This comprehensive article has been prepared to review the toxicity of titanium materials within the medical and dental fields. METHODS: We used online searching tools including MEDLINE (PubMed), Embase, Cochrane Library, and Google Scholar by combining keywords such as "titanium implant toxicity," "titanium implant corrosion," "titanium implant allergy," and "yellow nail syndrome." Recently updated data has been collected and compiled into one of four categories: "the toxicity of titanium," "the toxicity of titanium alloys," "the toxicity of titanium implants," and "diseases related to titanium." RESULTS: Recent studies with regard to titanium toxicity have been increasing and have now expanded to the medical field in addition to the fields of environmental research and basic science. Problems that may arise in titanium-based dental implants include the generation of titanium and titanium alloy particles and ions deposited into surrounding tissues due to the corrosion and wear of implants, resulting in bone loss due to inflammatory reactions, which may lead to osseointegration failure of the dental implant. These titanium ions and particles are systemically deposited and can lead to toxic reactions in other tissues such as yellow nail syndrome. Additionally, implant failure and allergic reactions can occur due to hypersensitivity reactions. Zirconia implants can be considered as an alternative; however, limitations still exist due to a lack of long-term clinical data. CONCLUSIONS: Clinicians should pay attention to the use of titanium dental implants and need to be aware of the problems that may arise from the use of titanium implants and should be able to diagnose them, in spite of very rare occurrence. Within the limitation of this study, it was suggested that we should be aware the rare problems of titanium toxicity.

It was reported that postnatal stem cells are present in adult tissues such as bone marrow, liver, muscle, dental pulp, and periodontal ligament. We isolated postnatal stem cells from human dental tissues such as dental pulp (DPSC), periodontal ligament (PDLSC), periapical follicle (PAFSC), and the surrounding mandibular bone marrow (MBMSC) to ascertain their properties. Immunocytochemistry proved the existence of stem cells in these cell populations using STRO-1 as a stem cell marker. These cells also expressed the mesenchymal stem cell (MSC) markers CD29 and CD44. The isolated cells showed self-renewal capabilities and colony-forming efficiency. Almost all of the dental stem cells showed optimal growth when they were cultured in alpha modification of Eagle's medium (alpha-MEM) supplemented with 10% fetal calf serum (FCS) and 100 microM ascorbic acid. Only the PAFSC showed increased proliferation in 20% FCS and 50 microM ascorbic acid. All of the dental stem cells were capable of differentiating into adipocytes and mineral nodule forming cells. MBMSC, in particular, showed much better mineralization compared to the others. These results indicate that MSCs exist in various tissues of the teeth and can differentiate into osteoblasts, adipocytes, and other kinds of cells with varying efficiency.

Although CRISPR/Cas9 genome editing has provided numerous opportunities to interrogate the functional significance of any given genomic site, there is a paucity of data on the extent of molecular scars inflicted on the mouse genome. Here we interrogate the molecular consequences of CRISPR/Cas9-mediated deletions at 17 sites in four loci of the mouse genome. We sequence targeted sites in 632 founder mice and analyse 54 established lines. While the median deletion size using single sgRNAs is 9 bp, we also obtain large deletions of up to 600 bp. Furthermore, we show unreported asymmetric deletions and large insertions of middle repetitive sequences. Simultaneous targeting of distant loci results in the removal of the intervening sequences. Reliable deletion of juxtaposed sites is only achieved through two-step targeting. Our findings also demonstrate that an extended analysis of F1 genotypes is required to obtain conclusive information on the exact molecular consequences of targeting events.

PURPOSE: Artificial intelligence (AI), represented by deep learning, can be used for real-life problems and is applied across all sectors of society including medical and dental field. The purpose of this study is to review articles about deep learning that were applied to the field of oral and maxillofacial radiology. MATERIALS AND METHODS: A systematic review was performed using Pubmed, Scopus, and IEEE explore databases to identify articles using deep learning in English literature. The variables from 25 articles included network architecture, number of training data, evaluation result, pros and cons, study object and imaging modality. RESULTS: Convolutional Neural network (CNN) was used as a main network component. The number of published paper and training datasets tended to increase, dealing with various field of dentistry. CONCLUSION: Dental public datasets need to be constructed and data standardization is necessary for clinical application of deep learning in dental field.

We developed an automatic method for staging periodontitis on dental panoramic radiographs using the deep learning hybrid method. A novel hybrid framework was proposed to automatically detect and classify the periodontal bone loss of each individual tooth. The framework is a hybrid of deep learning architecture for detection and conventional CAD processing for classification. Deep learning was used to detect the radiographic bone level (or the CEJ level) as a simple structure for the whole jaw on panoramic radiographs. Next, the percentage rate analysis of the radiographic bone loss combined the tooth long-axis with the periodontal bone and CEJ levels. Using the percentage rate, we could automatically classify the periodontal bone loss. This classification was used for periodontitis staging according to the new criteria proposed at the 2017 World Workshop on the Classification of Periodontal and Peri-Implant Diseases and Conditions. The Pearson correlation coefficient of the automatic method with the diagnoses by radiologists was 0.73 overall for the whole jaw (p < 0.01), and the intraclass correlation value 0.91 overall for the whole jaw (p < 0.01). The novel hybrid framework that combined deep learning architecture and the conventional CAD approach demonstrated high accuracy and excellent reliability in the automatic diagnosis of periodontal bone loss and staging of periodontitis.

AIMS: Oral health problems such as periodontal disease, dental caries, and tooth loss have been suggested to have associations with cardiovascular disease. This study aimed to evaluate whether oral hygiene behaviour can alleviate cardiovascular risk associated with oral health status using a nationwide population-based cohort. METHODS AND RESULTS: The data of 247 696 healthy adults aged 40 years or older who underwent an oral health screening programme and had no history of major cardiovascular events were extracted from the National Health Insurance System-National Health Screening Cohort. After a median follow-up of 9.5 years, 14 893 major cardiovascular events occurred including cardiac death, myocardial infarction, stroke, and heart failure. The risk of cardiovascular events was higher when a subject had periodontal disease, a higher number of dental caries, or more tooth loss. Performing one more tooth brushing a day was associated with a 9% significantly lower risk of cardiovascular events after multivariable adjustment. Regular dental visits (once a year or more) for professional cleaning were also shown to reduce cardiovascular risk by 14%. Improved oral hygiene behaviours were shown to attenuate the cardiovascular risk originating from periodontal disease, dental caries, and tooth loss. CONCLUSION: Oral hygiene care such as frequent tooth brushing and regular dental visits for professional cleaning reduced the risk of future cardiovascular events in healthy adults. This study also suggests that improved oral hygiene behaviour may modify the association between oral health and cardiovascular diseases.

The platelet-rich fibrin (PRF) is known as a rich source of autologous cytokines and growth factors and universally used for tissue regeneration in current clinical medicine. However, the microstructure of PRF has not been fully investigated nor have been studied the key molecules that differ PRF from platelet-rich plasma. We fabricated PRF under Choukroun's protocol and produced its extract (PRFe) by freezing at -80°C. The conventional histological, immunohistological staining, and scanning electron microscopy images showed the microstructure of PRF, appearing as two zones, the zone of platelets and the zone of fibrin, which resembled a mesh containing blood cells. The PRFe increased proliferation, migration, and promoted differentiation of the human alveolar bone marrow stem cells (hABMSCs) at 0.5% concentration in vitro. From the results of proteome array, matrix metalloproteinase 9 (MMP9) and Serpin E1 were detected especially in PRFe but not in concentrated platelet-rich plasma. Simultaneous elevation of MMP9, CD44, and transforming growth factor β-1 receptor was shown at 0.5% PRFe treatment to the hABMSC in immunoblot. Mineralization assay showed that MMP9 directly regulated mineralization differentiation of hABMSC. Transplantation of the fresh PRF into the mouse calvarias enhanced regeneration of the critical-sized defect. Our results strongly support the new characteristics of PRF as a bioscaffold and reservoir of growth factors for tissue regeneration.

OBJECTIVE: To determine the success rate and the factors related to the success rate of orthodontic miniscrew implants (OMI) placed at the attached gingiva of the posterior buccal region. MATERIALS AND METHODS: Four hundred eighty OMI placed in 209 orthodontic patients were examined retroactively. The sample was divided into young patients (range 10-18 years, N = 108) and adult patients (range 19-64 years, N = 109). The placement site was divided into three interdental areas from the first premolar to the second molar in the maxilla and mandible. According to soft tissue management, the samples were divided into incision and nonincision groups. Chi-square tests and multiple logistic regression analyses were used. RESULTS: The overall success rate was 83.8%. Dislodgement of the OMI occurred most frequently in the first 1-2 months, and more than 90% of the failures occurred within the first 4 months. Sex, age, jaw, soft tissue management, and placement side did not show any difference in the success rate. Placement site, however, showed a significant difference in the mandible of adult patients. There was no difference in the success rate in the maxilla. CONCLUSIONS: Placement site is one of the important factors for success rate of OMI.

BACKGROUND: Although mice have long served as an animal model for periodontitis, information on the composition of their indigenous oral microbiota is limited. The aim of the current study was to characterize mouse oral bacterial flora by applying extensive parallel pyrosequencing using the latest model pyrosequencer, a Roche/454 Genome Sequencer FLX Titanium. In addition, the effect of Toll-like receptor (TLR) 2 deficiency on oral microbiota was evaluated. RESULTS: Eight oral bacterial communities of wild-type (n = 4) and TLR2 knock-out (n = 4) C57BL/6 mice were characterized by analyzing 80,046 reads of 16S rRNA genes obtained by pyrosequencing. Excluding the PCR primers, the average length of each sequencing product was 443 bp. The average species richness of the murine oral bacterial communities was estimated to be about 200, but the communities were dominated by only two main phyla and several species. Therefore, the bacterial communities were relatively simple. The bacterial composition of the murine oral microbiota was significantly different from that of humans, and the lack of TLR2 had a negligible effect on the murine oral microbiota. CONCLUSION: Pyrosequencing using the Roche/454 FLX Titanium successfully characterized mouse oral bacterial communities. The relatively simple oral bacterial communities of mice were not affected by TLR2 deficiency. These findings will provide a basis for future studies on the role of periodontal pathogens in the murine model of periodontitis.

OBJECTIVE: To compare the accuracy and computational efficiency of two of the latest deep-learning algorithms for automatic identification of cephalometric landmarks. MATERIALS AND METHODS: A total of 1028 cephalometric radiographic images were selected as learning data that trained You-Only-Look-Once version 3 (YOLOv3) and Single Shot Multibox Detector (SSD) methods. The number of target labeling was 80 landmarks. After the deep-learning process, the algorithms were tested using a new test data set composed of 283 images. Accuracy was determined by measuring the point-to-point error and success detection rate and was visualized by drawing scattergrams. The computational time of both algorithms was also recorded. RESULTS: The YOLOv3 algorithm outperformed SSD in accuracy for 38 of 80 landmarks. The other 42 of 80 landmarks did not show a statistically significant difference between YOLOv3 and SSD. Error plots of YOLOv3 showed not only a smaller error range but also a more isotropic tendency. The mean computational time spent per image was 0.05 seconds and 2.89 seconds for YOLOv3 and SSD, respectively. YOLOv3 showed approximately 5% higher accuracy compared with the top benchmarks in the literature. CONCLUSIONS: Between the two latest deep-learning methods applied, YOLOv3 seemed to be more promising as a fully automated cephalometric landmark identification system for use in clinical practice.

PURPOSE: This study aimed to identify the effects of the sintering conditions of dental zirconia on the grain size and translucency. MATERIALS AND METHODS: Ten specimens of each of two commercial brands of zirconia (Lava and KaVo) were made and sintered under five different conditions. Microwave sintering (MS) and conventional sintering (CS) methods were used to fabricate zirconia specimens. The dwelling time was 20 minutes for MS and 20 minutes, 2, 10, and 40 hours for CS. The density and the grain size of the sintered zirconia blocks were measured. Total transmission measurements were taken using a spectrophotometer. Two-way analysis of variance model was used for the analysis and performed at a type-one error rate of 0.05. RESULTS: There was no significant difference in density between brands and sintering conditions. The mean grain size increased according to sintering conditions as follows: MS-20 min, CS-20 min, CS-2 hr, CS-10 hr, and CS-40 hr for both brands. The mean grain size ranged from 347-1,512 nm for Lava and 373-1,481 nm for KaVo. The mean light transmittance values of Lava and KaVo were 28.39-34.48% and 28.09-30.50%, respectively. CONCLUSION: Different sintering conditions resulted in differences in grain size and light transmittance. To obtain more translucent dental zirconia restorations, shorter sintering times should be considered.

CONTEXT: Despite the growing use of virtual patients (VPs) in medical education, few studies have explored the features and effectiveness of VP-based medical communication skills training. We undertook a systematic review to summarise the design and evaluation of VP-based medical communication skills training systems in order to identify features of successful cases. METHODS: Following PRISMA guidelines, we searched four databases for studies published between 2006 and 2018. Using a refined classification scheme, we extracted data on instructional design (scenario and instructional intervention), technological design (modality and interaction), and evaluation (user experience, learning effectiveness and evaluator). We assessed the quality of studies using the Medical Education Research Study Quality Instrument (MERSQI) and the QualSyst standard assessment criteria. RESULTS: A total of 14 studies were included for review. Of these, 85.7% (n = 12) were quantitative and 71.4% (n = 10) involved undergraduate students. The most common VP training scenario was history taking followed by the delivery of bad news. Diverse instructional interventions, including tutorials, learning activities, and feedback, were embedded in the VPs. The first-person perspective animated within-screen size VP was a popular technological feature. Most evaluations concerned the reality of simulation (for user experience) and skill in expressing empathy (as a learning outcome). Of the eight comparative studies, half reported significant attitude or skill improvements in the VP group. The distinct features of VPs shown to be effective were well-designed instructional interventions (eg, a pre-activity with a protocol-informed tutorial), and post-activity (eg, debrief or reflection), scaffolding and human feedback, but not system feedback. CONCLUSIONS: Evidence-based VP training can enable students to gain communication skills in a safe and affordable learning environment. Elaborate technology alone cannot guarantee effective learning, but evidence-based instructional interventions can facilitate its optimal use and bring about better learning outcomes.

OBJECTIVE: To identify the influence of prosthetic features through a comprehensive analysis with other known risk factors. MATERIALS AND METHODS: A total of 169 patients (n = implants: 349) was retrospectively included in the present study. Peri-implantitis was diagnosed based on peri-implant bone loss and probing depth. Using radiographs taken 1 and 5 years following prosthesis insertion, the following features were determined: peri-implant marginal bone loss (MBL), emergence angle (EA), emergence profile (EP) and crown/implant ratio (CIR). The splinted position of prosthesis was also recorded. Multivariable generalized estimating equation was used to analyse the influence of each feature on the prevalence of peri-implantitis. The final prediction model was constructed by Cox proportional hazard regression analysis. RESULTS: The EA showed a significant correlation with MBL. A statistically greater prevalence of peri-implantitis was observed if EA ≥ 30 degrees, when EP is convex and in middle implant splinted with both mesial and distal adjacent implants in bone-level implant. A similar correlation was not observed in tissue-level implants. CIR had no significant effect on the prevalence of peri-implantitis. CONCLUSION: Over-contoured implant prosthesis is a critical local confounder for peri-implantitis. The implant splinted to both mesial and distal adjacent implant has a higher risk of peri-implantitis.

BACKGROUND: The present study was undertaken to evaluate the potential of periodontal ligament stem cells (PDLSCs) and bone marrow SCs (BMSCs) on alveolar bone regeneration in a canine peri-implant defect model. METHODS: Four adult, male beagle dogs were used in this study. Autologous BMSCs from the iliac crests and PDLSCs from extracted teeth were cultured. Three months after extraction, BMSC- and PDLSC-loaded hydroxyapatite/beta-tricalcium phosphate (HA/TCP) (test groups) and cell-free HA/TCP (control group) were implanted in three rectangular, saddle-like peri-implant defects, respectively. The left side of the mandible was initially prepared, and after 8 weeks, the right side was also prepared. The animals were sacrificed after an 8-week healing period. Undecalcified ground sections were prepared. New bone formation and bone-to-implant contact (BIC) were measured histomorphometrically. BMSCs and PDLSCs were fluorescently labeled and traced. RESULTS: Alveolar bone regeneration in surgically created peri-implant saddle-like defects was more effective in test groups than the control group. The BMSC group had the highest new bone formation (34.99% and 40.17% at healing times of 8 and 16 weeks, respectively) followed by the PDLSC group (31.90% and 36.51%) and control group (23.13% and 28.36%), respectively. Test groups exhibited a significantly higher new bone formation than the control group at 8 weeks, but the same was true for only the BMSC group at 16 weeks (P <0.05). Fluorescently labeled cells were identified adjacent to HA/TCP carriers and, partly, near connective tissues and osteoids. CONCLUSION: This study demonstrated the feasibility of using stem cell-mediated bone regeneration to treat peri-implant defects.

Periodontal disease is a major public health issue, and various periodontal therapies have been performed to regenerate periodontal tissues. The periodontium is a complex structure composed of specialized tissues that support the teeth, and most periodontal surgeries are invasive procedures, including a resection of the gingiva or the alveolar bone. The periodontal wound healing process is slightly different from cutaneous wound healing and is similar to fetal healing, being almost scar-free. The aim of this review article is to provide an overview of periodontal wound healing and discuss various surgical and pharmaceutical approaches to achieve stable wound healing and improve the treatment outcomes. In addition, detrimental and limiting factors that induce a compromised prognosis are discussed, along with the perspective and future direction for successful periodontal tissue regeneration.

Abstract Objectives The aim of this study was to prospectively evaluate the clinical efficacy and histological outcome of the autogenous tooth graft material (Auto BT ) compared to that of anorganic bovine bone (Bio‐Oss ® , Geistlich, Switzerland) in post‐extraction alveolar bone augmentation. Patients and Methods A total of 33 graft sites in 24 patients were included in this study. Auto BT was used in 21 sites of 15 patients and Bio‐Oss ® was used in 12 sites of 9 patients for alveolar bone augmentation 2–4 weeks after dental extraction. Vertical dimension of grafted bone was measured both at the time of graft placement and at implant fixture placement after 6 months. Trephine cores were harvested for the histomorphometric evaluations during implant placement when feasible. The primary stability of implant fixture was also measured. Results Infection of graft material or graft bed was not observed and graft sites healed without any notable complications in both groups. The vertical dimensions of alveolar bone increased by 5.38 ± 2.65 mm in Auto BT group and 6.56 ± 3.54 mm in Bio‐Oss ® group at 6 months post‐extraction. Histomorphometrically, new bone formation of Auto BT ‐grafted site was 31.24 ± 13.87% while that of Bio‐Oss ® was 35.00 ± 19.33%. The implant stability quotient ( ISQ ) of implants placed in Auto BT ‐grafted sites measured 72.80 ± 10.81 while those placed in Bio‐Oss ® ‐grafted sites measured 70.0 ± 12.86. There were no statistically significant differences between measurements of the two groups. Conclusion Autogenous demineralized dentin matrix from extracted tooth grafted to extraction sockets for the augmentation of vertical dimension was as effective as augmentation using anorganic bovine bone. Both groups showed favorable wound healing, similar amount of implant stability, and histologically confirmed new bone formation. Thus, the results of this study suggest that autogenous tooth graft material is a viable option for alveolar bone augmentation following dental extraction.

Complement-C1q TNF-related protein 1 (CTRP1), a member of the CTRP superfamily, is expressed at high levels in adipose tissues of obese Zucker diabetic fatty (fa/fa) rats, and CTRP1 expression is induced by proinflammatory cytokines, including TNF-alpha and IL-1beta. In the present study, we investigated stimulation of aldosterone production by CTRP1, since it was observed that CTRP1 was specifically expressed in the zona glomerulosa of the adrenal cortex, where aldosterone is produced. Increased aldosterone production by CTRP1 in cells of the human adrenal cortical cell line H295R was dose-dependent. Expression levels of aldosterone synthase CYP11B2 were examined to investigate the molecular mechanisms by which CTRP1 enhances the production of aldosterone. The expression of CYP11B2 was greatly increased by treatment with CTRP1, as was the expression of the transcription factors NGFIB and NURR1, which play critical roles in stimulation of CYP11B2 gene expression. It was also revealed that angiotensin II-induced aldosterone production is, at least in part, mediated by the stimulation of CTRP1 secretion, not by the increase of CTRP1 mRNA transcription. In addition, the levels of CTRP1 were significantly up-regulated in hypertensive patients' serum. As CTRP1 was highly expressed in obese subjects as well as up-regulated in hypertensive patients, CTRP1 may be a newly identified molecular link between obesity and hypertension.