Let-7c-5p associate with inhibition of phenobarbital-induced cell proliferation in human palate cells.

Cleft palate (CP) is one of the most common congenital diseases, and is accompanied by a complicated etiology. Medical exposure in women is among one of the reasons leading to CP. Recently, it has been reported that microRNA (miRNA) plays a crucial role in palate formation and the disruption of miRNA that influence the development of CP. Although association with pharmaceuticals and miRNAs were suggested, it has remained largely unknow. The aim of the current investigation is to elucidate upon the miRNA associated with the inhibition of phenobarbital (PB)-induced cell proliferation in human embryonic palatal mesenchymal (HEPM) cells. We showed that PB inhibited HEPM cell viability in a dose-dependent manner. We demonstrated that PB treatment suppressed cyclin-D1 expression in HEPM cells. Furthermore, PB upregulated let-7c-5p expression and downregulated the expression of two downstream genes (BACH1 and PAX3). Finally, we demonstrated that the let-7c-5p inhibitor alleviated PB-induced inhibition of cell proliferation and altered BACH1 and PAX3 expression levels. These results suggest that PB suppresses cell viability by modulating let-7c-5p expression.

Involvement of microRNA-4680-3p against phenytoin-induced cell proliferation inhibition in human palate cells.

Cleft palate (CP) is one of the most common birth defects and is caused by a combination of genetic and/or environmental factors. Environmental factors such as pharmaceutical exposure in pregnant women are known to induce CP. Recently, microRNA (miRNA) was found to be affected by environmental factors. The aim of the present study was to investigate the involvement of miRNA against phenytoin (PHE)-induced inhibition of proliferation in human embryonic palatal mesenchymal (HEPM) cells. We demonstrated that PHE inhibited HEPM cell proliferation in a dose-dependent manner. We found that treatment with PHE downregulated cyclin-D1 and cyclin-E expressions in HEPM cells. Furthermore, PHE increased miR-4680-3p expression and decreased two downstream genes (ERBB2 and JADE1). Importantly, an miR-4680-3p-specific inhibitor restored HEPM cell proliferation and altered expression of ERBB2 and JADE1 in cells treated with PHE. These results suggest that PHE suppresses cell proliferation via modulation of miR-4680-3p expression.

Copper-induced diurnal hepatic toxicity is associated with Cry2 and Per1 in mice.

BACKGROUND: This study aimed to investigate diurnal variations in copper-induced hepatic toxicity and the molecular mechanisms underlying this chronotoxicity. METHODS: Male C57BL/6J mice were intraperitoneally injected with copper chloride (CuCl2) at zeitgeber time 2 (ZT2) or 14 (ZT14), twice per week for 5 or 8 weeks. Seventy-two hours after the final CuCl2 injection, the mice were euthanized, and plasma samples were collected. The livers and kidneys were collected and weighed. In vitro experiments were performed to assess cell viability and fluctuations in clock gene expression levels in Hepa1-6 cells after CuCl2 treatment. We examined copper homeostasis- and apoptosis-related genes under clock genes overexpression. RESULTS: Repeated CuCl2 administration for 8 weeks resulted in more severe toxicity at ZT14 compared to ZT2. CuCl2 administration at ZT14 elevated plasma aspartate aminotransferase, hepatic tumor necrosis factor-α, and interleukin-6 for 5 weeks, whereas the toxic effects of CuCl2 administration at ZT2 were weaker. Moreover, CuCl2 treatment inhibited Hepa1-6 cell viability in a dose-dependent manner. We observed increased expression of three clock genes (Ciart, Cry2, and Per1) after CuCl2 treatment. Among them, overexpression of Cry2 and Per1 accelerated CuCl2-induced inhibition of Hepa1-6 cell viability. Moreover, we found that the overexpression of Cry2 and Per1 regulates cleaved caspase-3 by modulating the copper transporter genes ATP7B and CTR1. CONCLUSION: These results suggest that CuCl2-induced diurnal toxicity is associated with Cry2 and Per1 expression through the regulation of copper transporter genes in mice.

Ridge preservation with the socket-shield technique and immediate provisionalization for delayed implant placement.

AIM: The present case report with medium-term follow-up aims to report on the successful outcomes of ridge preservation using anorganic bovine bone mineral (ABBM) combined with the socket-shield technique (SST) in conjunction with immediate ovate pontic provisionalization of the extraction site, followed by delayed flapless implant placement and immediate implant provisionalization in the rehabilitation of the maxillary anterior esthetic region. Clinical protocol: Three consecutive patients with failing maxillary anterior teeth were treated with this protocol. The alveolar ridge dimension was preserved utilizing ABBM, combined with the retention of a partial tooth root segment in the buccal portion of the extraction socket, to minimize ridge resorption following tooth extraction. An immediate fixed provisional restoration with an ovate base in direct contact with the extraction site was fabricated and delivered immediately to maintain the soft tissue contour. At 4 months postextraction, flapless implant placement was performed via a computer-generated surgical guide, along with immediate implant provisionalization. Definitive restorations were delivered 4 months following implant placement. RESULTS: At the 2-year follow-up after loading, prosthetic and implant success was demonstrated, with a favorable esthetic outcome and a high level of patient satisfaction. CONCLUSIONS: The immediate ovate pontic provisionalization in conjunction with ridge preservation combining ABBM and the SST, followed by flapless computer-guided implant placement with an immediate implant provisionalization protocol, as proposed in this article, offers a viable method to minimize alveolar ridge resorption as well as optimize esthetic outcomes.

A Novel 3D Volumetric Method of Analyzing Gingival Positional and Dimensional Alterations During Orthodontic Therapy: A Pilot Study.

The aim of the present study was to develop a 3D digital image-analysis method to quantitatively assess gingival changes after clear-aligner orthodontic therapy. Using teeth as fixed reference points, 3D image analysis tools have been used to quantify mucosal level changes after specific therapies. This technology has not been applied to orthodontic therapy, primarily because orthodontic tooth movement precludes using teeth as fixed reference points. Rather than superimposing the pre- and posttherapy volumes for the entire dentition, the methodology presented herein superimposed the pre- and post-therapy volumes for individual teeth. The lingual tooth surfaces, which remained unaltered, were used as fixed references. Intraoral scans taken before and after clear-aligner orthodontic therapy were imported for comparison. Volumes were created for each 3D image and were superimposed in a 3D image-analysis software that allowed quantitative measurements. The results demonstrated this technique's ability to measure very small changes in the apicocoronal position of the gingival zenith, as well as alterations of gingival margin thickness, following clear-aligner orthodontic therapy. The present 3D image-analysis method offers a useful tool for investigating the periodontal dimensional and positional changes that accompany orthodontic therapy.

Involvement of Bmal1 and Clock in Bromobenzene Metabolite-Induced Diurnal Renal Toxicity.

Circadian rhythms are endogenous oscillators that regulate 24 h behavioral and physiological processes. Our previous investigation demonstrated that bromobenzene metabolite (4-bromocatechol: 4-BrCA) exhibited chronotoxicity (i.e., the nephrotoxicity induced by 4-BrCA was observed during the dark phase, while not observed at light phase in mice). However, the molecular mechanism is still unknown. The aim of the present study is to investigate the cellular molecule(s) involved in the 4-BrCA-induced nephrotoxicity using mouse renal cortex tubular cell lines (MuRTE61 cells). We found that 4-BrCA showed dose dependent (0.01-1 mM) cell proliferation defect in MuRTE61 cells. By treating with 0.03 mM 4-BrCA, we demonstrated that major clock genes (Bmal1, Clock, Cry1, Cry2, Per1, and Per2) were significantly downregulated. Interestingly, the expression levels of two genes, Bmal1 and Clock, continued to decrease after 3 h of treatment with 4-BrCA, while Cry1, Per1, and Per2 were unchanged until 24 h of treatment. Moreover, BMAL1 and CLOCK levels are higher at light phase. We speculated that BMAL1 and CLOCK might function defensively against 4-BrCA-induced nephrotoxicity since the expression levels of Bmal1 and Clock were rapidly decreased. Finally, overexpression of Bmal1 and Clock restored 4-BrCA-induced cell proliferation defect in MuRTE61 cells. Taken together, our results suggest that Bmal1 and Clock have protective roles against 4-BrCA-induced nephrotoxicity.

Sasa veitchii extracts protect phenytoin-induced cell proliferation inhibition in human lip mesenchymal cells through modulation of miR-27b-5p.

A cleft lip, with or without a cleft palate, is a common birth defect caused by environmental factors or genetic mutations. Environmental factors, such as pharmaceutical exposure in pregnant women, are known to induce cleft lip, with or without cleft palate in the child. This study aimed to investigate the protective effect of Sasa veitchii extract (SE) on phenytoin-induced inhibition of cell proliferation in human lip mesenchymal cells (KD cells) and human embryonic palatal mesenchymal cells (HEPM cells). We demonstrated that cell proliferation was inhibited by phenytoin in a dose-dependent manner in both KD and HEPM cells. Co-treatment with SE restored phenytoin-induced toxicity in KD cells but did not protect HEPM cells against phenytoin-induced toxicity. Several microRNAs (miR-27b, miR-133b, miR-205, miR-497-5p, and miR-655-3p) is reported to associate with cell proliferation in KD cells. We measured the seven kinds of microRNAs (miR27b-3p, miR-27b-5p, miR-133b, miR-205-3p, miR-205-5p, miR-497-5p, and miR-655-3p) and found that SE suppressed miR-27b-5p induced by phenytoin in KD cells. Furthermore, co-treatment with SE enhanced the expression of miR-27b-5p downstream genes (PAX9, RARA, and SUMO1). These results suggest that SE protects phenytoin-induced cell proliferation inhibition by modulating miR-27b-5p.

Clinical efficacy of soft-tissue augmentation on tissue preservation at immediate implant sites: A randomized controlled trial.

AIM: This randomized controlled trial aimed to investigate the efficacy of soft-tissue augmentation (STA) with a subepithelial connective tissue graft (SCTG) or an acellular dermal matrix (ADM) on reducing tissue alterations at an immediate implant site. MATERIALS AND METHODS: This trial had three groups: (i) immediate implant with SCTG (ICT group); (ii) immediate implant with ADM (IAD group); (iii) immediate implant without STA (control group). Forty-six patients were randomly assigned to each group. Implants were placed at the maxillary anterior or premolar areas and restored after the 6-month visit. Clinical outcomes, including buccal soft-tissue contour, peri-implant mucosal level, soft-tissue thickness and keratinized tissue width, were measured at baseline and at 3-, 6- and 12-month follow-up visits. Radiographic bone levels were measured at baseline and at 6- and 12-month follow-up visits. Patient-reported outcomes were also collected. RESULTS: STA procedures increased peri-implant mucosal thickness and maintained buccal soft-tissue contours. Compared to the control group, STA groups did not prevent peri-implant mucosal recession or interproximal bone resorption. Generally, no significant differences in clinical outcomes were detected between the ICT and IAD groups. Most patients were highly satisfied with the immediate implant procedure and outcomes without significant differences between groups. CONCLUSIONS: STA at immediate implant sites enhanced soft-tissue thickness and maintained soft-tissue contours but did not prevent peri-implant mucosal recession or interproximal bone resorption. Long-term follow-up should be performed since these results were reported for only up to 1 year.

A three-dimensional cephalometric analysis of Japanese adults and its usefulness in orthognathic surgery: A retrospective study.

This study aimed to establish a three-dimensional (3D) cephalometric analysis of craniofacial morphology and discuss its theoretical usefulness in orthognathic patients. Cone-beam computed tomography (CBCT) images of Japanese subjects with skeletal Class I malocclusion before treatment were selected from among 1000 patients so that samples matched a historic 2D cephalometric cohort with normal occlusion using propensity score matching. In each CBCT image, 67 3D measurements were calculated based on manually identified landmarks. The mean and standard deviation of the measurements were calculated and used as the normative range for each sex. To confirm the usefulness of the 3D measurements, pre- and post-treatment CT data of nine jaw deformity patients who underwent orthognathic surgery with two-dimensional planning (2DP) in the past were used. Pre- and post-treatment CT values were evaluated with a paired t-test as well as a Z-score, which was calculated using the aforementioned normative range, and then categorized into five groups ("deteriorated", "no improvement", "over-treatment", "no change", "improvement") with -1 < Z-score < 1 considered normal. Fifty-six patients were matched to normal skeletal 1 subjects. The normative range of 67 items indicating 3D craniofacial morphology of the Japanese was calculated. Postoperatively, the horizontal position of the pogonion to the mid-sagittal plane significantly decreased (p = 0.043) and "improved"; however, the ramus axis on the right side significantly increased (p = 0.005) and "deteriorated". Maxillary yaw and the horizontal position of the gonion also tended to "deteriorated". The normative range for the 3D cephalometric analysis in Japanese has been established. Given findings of deteriorated maxillomandibular yawing after surgery when using conventional 2DP, 3D cephalometric measurements should be used when planning jaw positions after surgery for orthognathic patients.

Surface-based 3-dimensional cephalometry: An objective analysis of cranio-mandibular morphology.

INTRODUCTION: The purpose of this study was to apply a previously reported homologous model for quantifying and visualizing the three-dimensional (3D) cranio-mandibular morphology. METHODS: Twenty-eight 3D cone-beam computed tomography (CBCT) images of Japanese patients (7 to 13 years of age) showing skeletal Class 1 malocclusions were used in this study. Wire mesh fitting was conducted for each patient based on the assignment of landmarks to each 3D surface (homologous modeling). Cranial and mandibular images were generated using surface rendering of the CBCT images. The mean and standard deviation of each point on the wire mesh were calculated as the normative mean. Two operators conducted the process twice for seven patients, with a one-week interval between fittings. The intra- and inter-examiner reliability were determined using the 95% confidence interval minimal detectable change (MDC95) for random error and intra-class correlation coefficients (ICCs). RESULTS: The intra- and inter-examiner reliability of the mesh fitting method were almost perfect. Normative ranges of the cranial and mandibular surfaces of Japanese were determined. CONCLUSIONS: A clinical method that enables practitioners to quantify and visualize the hard tissues of a patient's face in three dimensions with almost perfect reliability is presented. This method allows practitioners to evaluate how patients' skeletal characteristics differ from normative means in three dimensions.

Long-term Outcomes of Autotransplantation of Teeth: A Case Series.

The aim of autotransplantation of teeth (ATT) is to replace a lost tooth with a functional tooth within the same patient. Although it has recently become more of a recognized and viable treatment approach in dentistry, the long-term outcomes are still not well-documented. The principal author (M.T.) has performed more than 1000 ATTs for reasons such as treating missing teeth, deep caries, poor endodontic results, and periodontitis over the past 30 years in private practice. During the course of private practice, 2 separate analyses were performed on a total of 319 cases with follow-up ranging from 2-26 years. The results showed a tendency toward higher success rates in younger patients; the success rate was highest in ATTs performed on immature teeth (about 95%), about 90% in patients younger than 30 years of age and approximately 80% in patients older than 30. The failures were most often caused by replacement resorption (ie, ankylosis-related resorption). The purpose of this case series was to show successful long-term outcomes of ATT as well as to provide clinical insights and describe tendencies noted over the course of 30 years of performing ATTs.

Long-term outcomes of autotransplantation of teeth: A case series.

The aim of autotransplantation of teeth (ATT) is to replace a lost tooth with a functional tooth within the same patient. Although it has recently become more of a recognized and viable treatment approach in dentistry, the long-term outcomes are still not well-documented. The principal author (MT) has performed more than 1000 ATTs for reasons such as treating missing teeth, deep caries, poor endodontic results, and periodontitis over the past 30 years in private practice. During the course of private practice, 2 separate analyses were performed on a total of 319 cases with follow-up ranging from 2 to 26 years. The results showed a tendency toward higher success rates in younger patients; the success rate was highest in ATTs performed on immature teeth (about 95%), about 90% in patients younger than 30 years of age and approximately 80% in patients older than 30. The failures were most often caused by replacement resorption (ie, ankylosis-related resorption). The purpose of this case series was to show successful long-term outcomes of ATT as well as to provide clinical insights and describe tendencies noted over the course of 30 years of performing ATTs.

Amino acid composition and amino acid-metabolic network in supragingival plaque.

Dental plaque metabolizes both carbohydrates and amino acids. The former can be degraded to acids mainly, while the latter can be degraded to various metabolites, including ammonia, acids and amines, and associated with acid-neutralization, oral malodor and tissue inflammation. However, amino acid metabolism in dental plaque is still unclear. This study aimed to elucidate what kinds of amino acids are available as metabolic substrates and how the amino acids are metabolized in supragingival plaque, by a metabolome analysis. Amino acids and the related metabolites in supragingival plaque were extracted and quantified comprehensively by CE-TOFMS. Plaque samples were also incubated with amino acids, and the amounts of ammonia and amino acid-related metabolites were measured. The concentration of glutamate was the highest in supragingival plaque, while the ammonia-production was the highest from glutamine. The obtained metabolome profile revealed that amino acids are degraded through various metabolic pathways, including deamination, decarboxylation and transamination and that these metabolic systems may link each other, as well as with carbohydrate metabolic pathways in dental plaque ecosystem. Moreover, glutamine and glutamate might be the main source of ammonia production, as well as arginine, and contribute to pH-homeostasis and counteraction to acid-induced demineralization in supragingival plaque.