ZNF862 induces cytostasis and apoptosis via the p21-RB1 and Bcl-xL-Caspase 3 signaling pathways in human gingival fibroblasts.

OBJECTIVE: This study investigates the effects of ZNF862 on the proliferation and apoptosis of human gingival fibroblasts and their related mechanisms. BACKGROUND: As a major transcription factor family, zinc finger proteins (ZFPs) regulate cell differentiation, growth, and apoptosis through their conserved zinc finger motifs, which allow high flexibility and specificity in gene regulation. In our previous study, ZNF862 mutation was associated with hereditary gingival fibromatosis. Nevertheless, little is known about the biological function of ZNF862. Therefore, this study was aimed to reveal intracellular localization of ZNF862, the influence of ZNF862 on the growth and apoptosis of human gingival fibroblasts (HGFs) and its potential related mechanisms. METHODS: Immunohistochemistry, immunofluorescence staining, and western blotting were performed to determine the intracellular localization of ZNF862 in HGFs. HGFs were divided into three groups: ZNF862 overexpression group, ZNF862 interference group, and the empty vector control group. Then, the effects of ZNF862 on cell proliferation, migration, cell cycle, and apoptosis were evaluated. qRT-PCR and western blotting were performed to further explore the mechanism related to the proliferation and apoptosis of HGFs. RESULTS: ZNF862 was found to be localized in the cytoplasm of HGFs. In vitro experiments revealed that ZNF862 overexpression inhibited HGFs proliferation and migration, induced cell cycle arrest at the G0/G1-phase and apoptosis. Whereas, ZNF862 knockdown promoted HGFs proliferation and migration, accelerated the transition from the G0/G1 phase into the S and G2/M phase and inhibited cell apoptosis. Mechanistically, the effects of ZNF862 on HGFs proliferation and apoptosis were noted to be dependent on inhibiting the cyclin-dependent kinase inhibitor 1A (p21)-retinoblastoma 1 (RB1) signaling pathway and enhancing the B-cell lymphoma-extra-large (Bcl-xL)-Caspase 3 signaling pathway. CONCLUSION: Our results for the first time reveal that ZNF862 is localized in the cytoplasm of HGFs. ZNF862 can inhibit the proliferation of HGFs by inhibiting the p21-RB1 signaling pathway, and it also promotes the apoptosis of HGFs by enhancing the Bcl-xL-Caspase 3 signaling pathway.

[An analysis of the wall width before and after root canal therapy in mandibular second molars with C-shaped canals].

PURPOSE: To measure the wall width in mandibular second molars with C-shaped canals before and after root canal therapy using cone-beam CT(CBCT). METHODS: A total of 55 mandibular second molars from 38 patients which met the criteria for inclusion at Nanjing Stomatological Hospital, Affiliated Stomatological Hospital of Medical School of Nanjing University from January 2020 to July 2021 were analyzed．From this sample, ten teeth had been treated, while another 45 of them not. CT images of the teeth were reestablished by Mimics software 20.0. Then we made a section every 1mm perpendicular to the long axis of the teeth from apex to pulp floor. The first slice from apex to pulp chamber was named the slice 1. Along the slice 1 to crown 1 mm was called slice 1, and so forth. The wall thickness at different locations of all the slices was measured. The data was entered into SPSS 20.0 software package for analysis. RESULTS: Regardless of whether the teeth were treated or not, both the mesial and distal canal walls' average width were thicker than 1mm in all slices. At the same time, the mesial and distal canal walls' width were thicker than the width of buccal and lingual canal walls in all the slices from C-shaped root canal, except slices which were near pulp chamber(P＜0.05). As for the C-shaped root canals without root canal therapy, the width of lingual wall in the slice 1 to 4, as well as apex third root, was thinner than 1 mm. The width of buccal canal wall was thicker than the width of lingual canal wall in all slices except slice 11 and 12. As for the C-shaped root canals with root canal therapy, the width of buccal canal wall in slice 1 to 5, equivalent of apex half root, and the width of lingual wall in the slice 1 to 7, amount to apex two-thirds of root, was thinner than 1 mm. The width of buccal canal wall was thicker than the width of lingual wall in all slices except slice 1 and 9. There was no significant difference between the distal canal walls' width of C-shaped canals with and without root canal therapy(P＞0.05) . There was significant difference between the buccal canal walls' width of C-shaped canals with and without root canal therapy, as same as the mesial canal walls' width and the width of lingual canal wall (P＜0.05). CONCLUSIONS: The lingual canal walls' width in apex third root of C-shaped root canal were thin before canal preparation. The buccal walls' width in apex half root and the lingual canal walls' in apex two-thirds of root of C-shaped root canal were thin after canal preparation.

New-designed 3D printed surgical guide promotes the accuracy of endodontic microsurgery: a study of 14 upper anterior teeth.

We aimed to design a novel three-dimensional (3D) printed surgical guide and evaluate its accuracy in assisting endodontic microsurgeries. A new 3D printed surgical guide was designed by computer-aided design and computer-aided manufacturing (CAD/CAM) technology and applied to 7 patients who underwent endodontic microsurgeries of upper anterior teeth from 2020.01 to 2020.12 as the experimental group. 7 patients who suffered from endodontic microsurgeries operated by the same surgeon without using the surgical guide from 2019.01 to 2019.12 were selected as the control group. Cone beam computed tomography (CBCT) was performed more than 12 months after operation, and the accuracy of apical resection was compared between the two groups. The accuracy of the microsurgery focused on the length and angle of the root apical resection. In the study, CBCT data and oral digital scanning data were used to reconstruct 3D models of periapical lesions with soft and hard tissue information, based on which we designed the new 3D printed surgical guides. The guides were successfully applied to the apectomy in endodontic microsurgeries. The deviation of the apical resection length of the experimental group (0.467 ± 0.146 mm) was better than that of the control group (1.743 ± 0.122 mm) (P < 0.0001), and the deviation of the apical resection angle of the experimental group (9.711 ± 3.593°) was significantly less than that of the control group (22.400 ± 3.362°) (P < 0.0001). The 3D-printed surgical guide could effectively guide endodontic microsurgery and improve its accuracy by fixing both the position and the angle of apectomy. The new type of surgical guide could accurately localize the root apex and guide the apical resection.

TAS2R supports odontoblastic differentiation of human dental pulp stem cells in the inflammatory microenvironment.

BACKGROUND: Inflammatory microenvironment promotes odontoblastic differentiation in human dental pulp stem cells (hDPSCs), but the regulatory mechanisms remain unclear. In this study, we aimed to explore the role of TAS2R in odontoblastic differentiation of hDPSCs in the inflammatory microenvironment. METHODS: Microarray analysis was performed to explore the differential mRNA profiles in inflammatory and healthy pulp tissues from the patients. hDPSCs isolated from the healthy pulp tissues were stimulated by LPS, TNFα and IL-6, respectively, to verify the effect of TAS2R. The expression markers related to odontoblastic differentiation of hDPSCs were observed by qPCR and chemical staining methods. TAS2R10 was overexpressed or silenced to observe the effect on odontoblastic differentiation of hDPSCs under LPS stimulation. The G protein and intracellular Ca2+ were detected, respectively, by qPCR and Fluo-4AM Ca2+ fluorescent probe. RESULTS: The expression of TAS2R was significantly upregulated in the inflammatory pulp tissues. In vitro, 5 subtypes of TAS2R mRNA expressions including TAS2R10, TAS2R14, TAS2R19, TAS2R30 and TAS2R31 in hDPSCs increased under the stimulation of LPS, TNFα or IL-6. In odontoblastic differentiation medium, we found LPS, TNFα or IL-6 stimulation promoted odontoblastic differentiation of hDPSCs. TAS2R10 overexpression in hDPSCs significantly increased the expression markers related to odontoblastic differentiation, whereas TAS2R10 silencing revealed the opposite effect. Furthermore, G protein was activated, and at the same time, intracellular Ca2+ enhanced when TAS2R10 was overexpressed, but decreased when TAS2R10 was silenced. CONCLUSIONS: This study demonstrated that TAS2R was found to be expressed in hDPSCs, and TAS2R promoted odontoblastic differentiation of hDPSCs by mediating the increase in intracellular Ca2+ via the G protein-coupled receptors (GPCR) conventional signaling pathway in inflammatory microenvironment, which may be a potential target for the development of effective conservative treatments for dental pulp repair.

Corrigendum: Experimental Periodontitis Deteriorated Atherosclerosis Associated With Trimethylamine N-Oxide Metabolism in Mice.

[This corrects the article DOI: 10.3389/fcimb.2021.820535.].

Low-level controllable blue LEDs irradiation enhances human dental pulp stem cells osteogenic differentiation via transient receptor potential vanilloid 1.

Human dental pulp stem cells (hDPSCs) have attracted tremendous attention in tissue regeneration engineering due to their excellent multidirectional differentiation potential. Photobiomodulation (PBM) using low-level light-emitting diodes (LEDs) or lasers has been proved to promote the osteogenesis of mesenchymal stem cells. However, the effect of LEDs on osteogenic differentiation of hDPSCs has little published data. In this work, the effect of blue LEDs with different energy densities of 2, 4, 6, 8, 10 J/cm2 on osteogenic differentiation of hDPSCs was examined by using in vitro ALP staining, ALP activity, mineralization, and real-time PCR. The results showed that compared with the control group, osteogenic differentiation was significantly enhanced in blue LEDs treated groups. As the energy density increased, the level of osteogenesis initially increased and then decreased reaching the highest level at 6 J/cm2. Transient receptor potential vanilloid 1 (TRPV1), a Ca2+ ion channel, was believed to be a potential player in osteogenesis by photobiomodulation. By immunofluorescence assay, calcium influx assay, PCR, and ALP staining, it was shown that blue LEDs irradiation can increase the activity of TRPV1 and intracellular calcium levels similarly to the agonist of TRPV1 capsaicin. Additionally, pretreatment with capsazepine, a selective TRPV1 inhibitor, was able to abrogate the osteogenic effect of blue LEDs. In conclusion, these findings proposed that blue LEDs can promote the osteogenesis of hDPSCs within the appropriate range (4-8 J/cm2) during culture of osteogenic medium, and TRPV1/Ca2+ may be an essential signaling pathway involved in blue LEDs-induced osteogenesis, providing new insights for the use of hDPSCs in tissue regeneration engineering.

Skin-Attachable Sensors for Biomedical Applications.

With the growing concern about human health issues, especially during the outbreak of the COVID-19 pandemic, the demand for personalized healthcare regarding disease prevention and recovery is increasing. However, tremendous challenges lie in both limited public medical resources and costly medical diagnosis approaches. Recently, skin-attachable sensors have emerged as promising health monitoring platforms to overcome such difficulties. Owing to the advantages of good comfort and high signal-to-noise ratio, skin-attachable sensors enable household, real-time, and long-term detection of weak physiological signals to efficiently and accurately monitor human motion, heart rate, blood oxygen saturation, respiratory rate, lung and heart sound, glucose, and biomarkers in biomedical applications. To further improve the integration level of biomedical skin-attachable sensors, efforts have been made in combining multiple sensing techniques with elaborate structural designs. This review summarizes the recent advances in different functional skin-attachable sensors, which monitor physical and chemical indicators of the human body. The advantages, shortcomings, and integration strategies of different mechanisms are presented. Specially, we highlight sensors monitoring pulmonary function such as respiratory rate and blood oxygen saturation for their potential usage in the COVID-19 pandemic. Finally, the future development of skin-attachable sensors is envisioned.

Experimental Periodontitis Deteriorated Atherosclerosis Associated With Trimethylamine N-Oxide Metabolism in Mice.

BACKGROUND: Periodontitis is considered a risk factor for atherosclerosis, but the mechanism is not clear. It was reported that oral administration of Porphyromonas gingivalis altered the gut microbiota in mice. Gut dysbiosis and the intestinal metabolite trimethylamine N-oxide (TMAO) were verified to be associated with atherosclerosis. Therefore, the possible TMAO-related mechanism between periodontitis and atherosclerosis needs to be explored. METHODS: Experimental periodontitis was established by oral administration of P. gingivalis for 2 months in ApoE-/- mice. Mouse hemi-mandibles were scanned using Micro-CT. Quantification of TMAO was performed using liquid chromatography-tandem mass spectrometry. Mouse feces were collected and the bacterial DNA was extracted, then the gut microbiota was analyzed using 16S rRNA genes. Atherosclerotic lesion areas were quantified. Livers, small intestines, and large intestines were analyzed for gene expression. RESULTS: Aggravated atherosclerosis plaques were found in experimental periodontitis mice. Plasma TMAO, a pathogenic factor of atherosclerosis, was initially found to be increased in periodontitis mice. Changes in the composition and abundance of the intestinal microflora of periodontitis mice were found. Flavin monooxygenase 3 (FMO3), the catalyzing enzyme of TMAO in the liver, was significantly increased, accompanied by an increase of IL-6 in liver, the abnormal intestinal integrity and enhanced plasma LPS. The IL-6 and LPS were verified to be able to increase FMO3 in HepG2 cells. CONCLUSION: Our research discovered that experimental periodontitis in ApoE-/- mice induced gut dysbiosis and an increase in TMAO. These results suggest a possible mechanism by which periodontitis may accelerate atherosclerosis by influencing the intestinal microbes and the metabolism, which were triggered by inflammation of the liver and intestine.

[Guided endodontic access of calcified root canal by laser melting templates].

OBJECTIVE: The aim of this study is to treat calcified root canal and assess the accuracy of guided endodontics using laser melting templates. METHODS: Two cases with calcified anterior teeth were treated with laser melting templates. Cone beam computed tomography (CBCT) was used to scan the maxillary teeth of patients before surgery to establish the root canal system model. The maxillary impression of the patient was made and it was scanned by a 3D scanner. The CBCT scans were matched with surface scans of plaster model. Mimics 19.0 and 3-matic 11.0 software were used to design the virtual planning to access cavities. The templates were produced by laser melting 3D printing. Access cavity was prepared under the guidance of laser melting template. Then the deviations of planned and prepared cavities in three dimensions and angle were measured. RESULTS: The two teeth obtained satisfactory results. The first case had a angle deviation of 1.77°, a drilling base deviation of 0.403-0.497 mm, and a tip of 0.433-0.537 mm. The second case had a angle deviation of 3.26°, a drill base deviation of 0.18-0.347 mm, and a tip of 0.310-0.463 mm. CONCLUSIONS: Laser melting template-guided endodontics is an effective technique for the treatment of calcified root canal and can be used as a new strategy for the treatment of calcified canal.

Evaluation of the Effect of Implants Placed in Preserved Sockets Versus Fresh Sockets on Tissue Preservation and Esthetics: A Meta-analysis and Systematic Review.

OBJECTIVES: To compare the success rates, tissue preservation, and esthetics of implants placed in fresh and preserved sockets and to evaluate the factors influencing the outcomes. METHODS: Medline, Embase, CENTRAL, Wanfang, and China National Knowledge Infrastructure (CNKI) databases were searched electronically, and a manual search was conducted as well. Studies that compared the implant success rate, tissue preservation, and patient-related outcomes such as complications and esthetic outcomes of immediate implant placement (IIP) and alveolar ridge preservation (ARP) were included. A subgroup analysis according to the follow-up period, socket conditions, and regenerative strategies was performed to investigate how these factors influence the prognosis. RESULTS: A total of 12 studies with 588 implants, where 298 are implants after ARP and the remaining 290 are after IIP. The IIP was performed in 58.4% of 250 implants inserted in the sockets with an intact buccal wall, whereas the percentage declined to 41.9% when the buccal wall was defective. The implant success rate was similar between ARP and IIP for an intact buccal wall but different for a defective buccal wall (ARP 98.6% vs IIP 89.6%). Moreover, hard-tissue preservation and the Pink Esthetic Score (PES) of the ARP group were significantly better than those of the IIP group in the molar region (P < .05). Also short-term complications showed no significant differences in the ARP group (P = .06). In the anterior region, there appeared to be no significant difference in hard- and soft-tissue preservation PES and patient-related outcomes between the 2 protocols. CONCLUSION: An alveolar bone defect might reduce the success rate of IIP. Further studies on the tissue preservation and esthetics of implants placed by IIP and ARP are still needed.

Application of OSCE for Stage Assessment in Standardized Training for Oral Residents.

The aim of this study was to establish and implement an objective structured clinical examination (OSCE) for stage assessment in standardized training for oral residents, evaluating its validity and suitability. An OSCE was established for stage assessment of 158 residents in a program in Jiangsu Province, China. Its validity and suitability were evaluated using indices such as reliability and discrimination coefficients of assessment results. The established OSCE had eight stations: interpretation of auxiliary examination results, basic knowledge, doctor-patient communication and clinical examination, clinical case analysis, medical record and prescription writing, public skills, first aid skills, and specialist's operation skills. The mean overall score and consistency coefficient (Cronbach's alpha) for the test subjects were 83.64±3.69 points and 0.732, respectively. The developed OSCE was reasonably established, with credible assessment results, and indices such as mean and discrimination coefficient of test scores from each station were appropriate. Therefore, the proposed protocol was found to be reliable and suitable.

Correction: Luteolin selectively kills STAT3 highly activated gastric cancer cells through enhancing the binding of STAT3 to SHP-1.

The PDF and HTML versions of the article have been updated to include the Creative Commons Attribution 4.0 International License information.

[An exploration on computerized examination system of dental objective structured clinical examination].

PURPOSE：In view of the present problems of clinical skill examination, in order to evaluate the candidate's clinical ability comprehensively and objectively, we constructed and implemented dental objective structured clinical examination system mainly based on computer. METHODS：This system was a multi-station examination which had 3 parts and 8 sites. The stations were composed of a variety of testing methods, including choice questions, auxiliary examinations, case analysis and basic operation skill. RESULTS: By using the computerized dental objective structured clinical examination system, we could smoothly test the ability of stomatological students in a relatively short time. The clinical ability, learning ability and professional quality could be evaluated more objectively in many ways. CONCLUSIONS：This system help students adapt to early examination of medical practitioners, improve students' clinical skills, and is worthy of wide application.

Luteolin selectively kills STAT3 highly activated gastric cancer cells through enhancing the binding of STAT3 to SHP-1.

The antitumor effect of luteolin, a plant flavonoid, in gastric cancer (GC) cells has not been fully understood. Here we show that luteolin selectively kills STAT3 overactivated GC cells that are often drug resistant. The treatment of luteolin in these GC cells significantly inhibited STAT3 phosphorylation and reduced the expression of STAT3 targeting gene Mcl-1, Survivin and Bcl-xl. Silencing of SHP-1, a protein tyrosine phosphatase, abolished the inhibitory effect of luteolin on STAT3 and cell apoptosis, suggesting that SHP-1 is crucial in luteolin-mediated cellular function. Moreover, this luteolin effect of STAT3 dephosphorylation by SHP-1 involved in HSP-90, which protected STAT3 phosphorylation by forming HSP-90/STAT3 complex. Thus, luteolin inhibited STAT3 activation through disrupting the binding of HSP-90 to STAT3, which promoted its interaction to SHP-1, resulted in the dephosphorylation of STAT3. The GC cell xenograft mouse model confirmed the effectiveness of luteolin induced inhibition of tumor growth in vivo.

Heme oxygenase-1 induction attenuates imiquimod-induced psoriasiform inflammation by negative regulation of Stat3 signaling.

Heme oxygenase-1 (HO-1), a stress-inducible protein with a potential anti-inflammatory effect, plays an important role in skin injury and wound healing. However, the function of HO-1 in cutaneous inflammatory diseases, such as psoriasis, remains unknown. The abnormal activation of Stat3, a known transcription factor that induces inflammation and regulates cell differentiation, is directly involved in the pathogenesis and development of psoriasis. Hence, targeting Stat3 is potentially beneficial in the treatment of psoriasis. In this study, HO-1 activation significantly alleviated the disease-related pathogenesis abnormality. To determine the mechanism by which HO-1 exerts immune protection on Th17-related cytokines, IL6/IL22-induced Stat3 activation was significantly suppressed, accompanied by decreased cell proliferation and reversed abnormal cell proliferation. Importantly, HO-1-induced Stat3 suppression was mediated through the activation of protein tyrosine phosphatase SHP-1. Overall, our study provides direct evidence indicating that HO-1 might be a useful therapeutic target for psoriasis. SHP-1-mediated suppression of Stat3 activation after HO-1 activation is a unique molecular mechanism for the regulation of Stat3 activation.

Anti-inflammatory effects of luteolin on experimental autoimmune thyroiditis in mice.

Hashimoto's thyroiditis (HT) is the most common organ-specific autoimmune disease and is believed to be a predominately T cell-mediated autoimmunity. Signal transducer and activator of transcription (STAT)3 is a crucial transcription factor of T cell-mediated immunity, with key roles in the proliferation and migration of T helper (Th) cells, differentiation of Th cells into Th17 cells, and the balance between Treg cells and Th17 cells. Flavonoid luteolin has been shown to markedly inhibit Tyr705 activation/phosphorylation of STAT3 and exert anti-inflammatory effects in multiple sclerosis. In the present study, the effect of luteolin on experimental autoimmune thyroiditis (EAT) was analyzed in C57BL/6 mice. Hematoxylin and eosin examination showed that luteolin attenuated lymphocytic infiltration and follicle destruction in thyroid glands. Immunohistochemistry results demonstrated that luteolin significantly reduced the phosphorylation of STAT3 within the thyroid. An in vitro study was carried out in a RAW264.7 macrophage cell line. Western blot findings demonstrated that luteolin significantly inhibited interferon-γ-induced increases in cyclooxygenase 2, phosphorylated STAT1 and phosphorylated STAT3 expression levels and the secretion of the proinflammatory cytokine tumor necrosis factor-α in supernatants. The present findings indicated that luteolin may exert potent anti-inflammatory effects on murine EAT, which may provide a novel therapeutic medication strategy for the early intervention of HT.

Analysis of Mtwo rotary instrument separation during endodontic therapy: a retrospective clinical study.

To analyze the incidence of instrument separation (IS) and the factors influencing it, when associated with Mtwo rotary system (VDW, Munich, Germany) during endodontic therapy. A retrospective study involving a total of 24,108 root canals (11,036 endodontic treated teeth) was conducted at Nanjing Stomatology Hospital between January 2011 and March 2013. The information included were tooth type, root canal curvature, number of fractured instruments, length of the separated fragments, and the distance from broken tip to apex. The incidence of IS was observed to be 2.2 % according to the number of teeth and 1.0 % according to the number of root canals. Many of the separated fragments were 2-4 mm in length and the mean length was 3.07 ± 1.46 mm, and 78.4 % of fractures occurred in the apex. The mean length of separated fragments in severely curved canals was maximum, while ultra-severe curved canals was observed to be minimum. Mtwo instruments demonstrated an extremely low fracture rate during endodontic therapy. Molar teeth (especially lower molars) and the degree of canal curvature had a significant effect on the incidence of IS.

[The influence of age upon circadian rhythm of human pulp sensibility].

OBJECTIVE: To explore the influence of age on the circadian rhythm of pulp sensibility, guide the diagnosis and treatment of dental and endodontic diseases. METHODS: The first lower molars of young, middle-aged, and aged volunteers were inspected for the threshold of pulp sensitivity. Each inspection was implemented every 4 hours earlier, totally 7 times during 24 hours. All values of pulp sensibility threshold from each volunteer were analyzed by Halberg methods for cosinor-rhythmometry. The chronobiology characteristics of pulp sensibility were compared among young, middle-aged, and aged. RESULTS: The pulp sensibility threshold values of the young, middle-aged, and aged indicated to have the circadian rhythm alternation in period of 24 hours, with fitting well to a cosine curve. The trend of rhythm curve was similar to all three age groups. The acrophase and bathyphase appeared at 0:00 and 12:00 separately. The values and amplitudes of pulp threshold sensibility showed to be: young>aged> middle-aged. CONCLUSION: The circadian rhythm of pulp sensibility changes according with age. The pulp sensibility threshold value is lower in aged people than in the young, and the lowest sensibility threshold is in middle-aged people. Besides, the extent of rhythm fluctuation is the least in middle-aged people.