骨－歯根膜線維の複合組織形成による三次元的な歯周組織再生技術の開発

Periodontal tissue is a distinctive tissue structure composed three-dimensionally of cementum, periodontal ligament (PDL) and alveolar bone. Severe periodontal diseases cause fundamental problems for oral function and general health, and conventional dental treatments are insufficient for healing to healthy periodontal tissue. Cell sheet technology has been used in many tissue regenerations, including periodontal tissue, to transplant appropriate stem/progenitor cells for tissue regeneration of a target site as a uniform tissue. However, it is still difficult to construct a three-dimensional structure of complex tissue composed of multiple types of cells, and the transplantation of a single cell sheet cannot sufficiently regenerate a large-scale tissue injury. Here, we fabricated a three-dimensional complex cell sheet composed of a bone-ligament structure by layering PDL cells and osteoblast-like cells on a temperature responsive culture dish. Following ectopic and orthotopic transplantation, only the complex cell sheet group was demonstrated to anatomically regenerate the bone-ligament structure along with the functional connection of PDL-like fibers to the tooth root and alveolar bone. This study represents successful three-dimensional tissue regeneration of a large-scale tissue injury using a bioengineered tissue designed to simulate the anatomical structure

CGRPは三叉神経節衛星グリア細胞からのサイトカイン遊離と口腔顔面侵害性疼痛を誘発する

Neuron-glia interactions contribute to pain initiation and sustainment. Intra-ganglionic (IG) secretion of calcitonin gene-related peptide (CGRP) in the trigeminal ganglion (TG) modulates pain transmission through neuron-glia signaling, contributing to various orofacial pain conditions. The present study aimed to investigate the role of satellite glial cells (SGC) in TG in causing cytokine-related orofacial nociception in response to IG administration of CGRP. For that purpose, CGRP alone (10 μL of 10-5 M), Minocycline (5 μL containing 10 μg) followed by CGRP with one hour gap (Min + CGRP) were administered directly inside the TG in independent experiments. Rats were evaluated for thermal hyperalgesia at 6 and 24 h post-injection using an operant orofacial pain assessment device (OPAD) at three temperatures (37, 45 and 10 ℃). Quantitative real-time PCR was performed to evaluate the mRNA expression of IL-1β, IL-6, TNF-α, IL-1 receptor antagonist (IL-1RA), sodium channel 1.7 (NaV 1.7, for assessment of neuronal activation) and glial fibrillary acidic protein (GFAP, a marker of glial activation). The cytokines released in culture media from purified glial cells were evaluated using antibody cytokine array. IG CGRP caused heat hyperalgesia between 6–24 h (paired-t test, p < 0.05). Between 1 to 6 h the mRNA and protein expressions of GFAP was increased in parallel with an increase in the mRNA expression of pro-inflammatory cytokines IL-1β and anti-inflammatory cytokine IL-1RA and NaV1.7 (one-way ANOVA followed by Dunnett’s post hoc test, p < 0.05). To investigate whether glial inhibition is useful to prevent nociception symptoms, Minocycline (glial inhibitor) was administered IG 1 h before CGRP injection. Minocycline reversed CGRP-induced thermal nociception, glial activity, and down-regulated IL-1β and IL-6 cytokines significantly at 6 h (t-test, p < 0.05). Purified glial cells in culture showed an increase in release of 20 cytokines after stimulation with CGRP. Our findings demonstrate that SGCs in the sensory ganglia contribute to the occurrence of pain via cytokine expression and that glial inhibition can effectively control the development of nociception

神経障害性疼痛における三叉神経筋内のIL-10とCXCL2

Many trigeminal neuropathic pain patients suffer severe chronic pain. The neuropathic pain might be related with cross-excitation of the neighboring neurons and satellite glial cell (SGCs) in the sensory ganglia and increasing the pain signals from the peripheral tissue to the central nervous system. We induced trigeminal neuropathic pain by infraorbital nerve constriction injury (IONC) in Sprague-Dawley rats. We tested cytokine (CXCL2 and IL-10) levels in trigeminal ganglia (TGs) after trigeminal neuropathic pain induction, and the effect of direct injection of the anti-CXCL2 and recombinant IL-10 into TG. We found that IONC induced pain behavior. Additionally, IONC induced satellite glial cell activation in TG and cytokine levels of TGs were changed after IONC. CXCL2 levels increased on day 1 of neuropathic pain induction and decreased gradually, with IL-10 levels showing the opposite trend. Recombinant IL-10 or anti-CXCL2 injection into TG decreased pain behavior. Our results show that IL-10 or anti-CXCL2 are therapy options for neuropathic pain

Dentin-pulp regeneration by 3D layered cell sheet

The dentin-pulp complex is a unique structure in teeth that contains both hard and soft tissues. Generally, deep caries and trauma cause damage to the dentin-pulp complex, and if left untreated, this damage will progress to irreversible pulpitis. The aim of this study was to fabricate a layered cell sheet composed of rat dental pulp (DP) cells and odontogenic differentiation of pulp (OD) cells and to investigate the ability to regenerate the dentin-pulp complex in a scaffold tooth. We fabricated two single cell sheets composed of DP cells (DP cell sheet) or OD cells (OD cell sheet) and a layered cell sheet made by layering both cells. The characteristics of the fabricated cell sheets were analyzed using light microscopy, scanning electron microscopy (SEM), hematoxylin-eosin (HE) staining, and immunohistochemistry (IHC). Furthermore, the cell sheets were transplanted into the subrenal capsule of immunocompromised mice for 8 weeks. Following this, the regenerative capacity to form dentin-like tissue was evaluated using micro-computed tomography (Micro-CT), HE staining, and IHC. The findings of SEM and IHC confirmed that layered cell sheets fabricated by stacking OD cells and DP cells maintained their cytological characteristics. Micro-CT of layered cell sheet transplants revealed a mineralized capping of the access cavity in the crown area, similar to that of natural dentin. In contrast, the OD cell sheet group demonstrated the formation of irregular fragments of mineralized tissue in the pulp cavity, and the DP cell sheet did not develop any hard tissue. Moreover, bone volume/tissue volume (BV/TV) showed a significant increase in hard tissue formation in the layered cell sheet group compared to that in the single cell sheet group (p<0.05). HE staining also showed a combination of soft and hard tissue formation in the layered cell sheet group. Furthermore, IHC confirmed that the dentin-like tissue generated from the layered cell sheet expressed characteristic markers of dentin but not bone equivalent to that of a natural tooth. In conclusion, this study demonstrates the feasibility of regenerating dentin-pulp complex using a bioengineered tissue designed to simulate the anatomical structure

Relationship between second language speaking and writing skills and modality preference of university EFL students

This study compared the performance of second language learners between the written and spoken modalities in terms of three lexical aspects (fluency, lexical complexity, and grammatical complexity) for the same picture description task. It also examined their modality preference and whether it corresponded with their actual performance. The participants were 26 Japanese university EFL students. They described a six-panel cartoon orally and in writing, answered a questionnaire about their modality preference and educational experiences, and had a follow-up interview. Analysis of their spoken and written descriptions using four linguistic measures (text length, MTLD textual lexical diversity, content word frequency, and average sentence length) revealed that they used a wider variety of vocabulary and shorter sentences in writing than in speaking. About 70% of them preferred the written modality, but half of the students performed equally well in both modalities in terms of the four measures. Further analysis suggested that the variety of words available to learners in each modality may have affected their modality preference and that a study-abroad experience might have affected it as well

スチームコンベクションオーブン調理と真空調理による水溶性ビタミンの調理損失の比較：かぼちゃおよび大根の煮物について

The purpose of this study is to compare and study water-soluble vitamin losses before and after cooking pumpkin and Japanese radish (daikon) with a steam convection oven (SC cooking) and a vacuum-packed pouch (V-P P cooking). Pumpkin was cut into 2.5 cm cubes and daikon into 2.0 cm cubes. For SC cooking, pumpkin was heated for 15 minutes and daikon for 60 minutes with liquid seasoning. For V-P P cooking, pumpkin and daikon were vacuum-packed with liquid seasoning and heated under the same conditions as SC cooking. For pumpkin, percentages of niacin, folic acid and pantothenic acid in uncooked samples, as well as those cooked by SC cooking and V-P P cooking were compared. For daikon, percentages of vitamins B1, B2, B6 and C, niacin, folic acid and pantothenic acid in uncooked samples, as well as those cooked by SC cooking and V-P P cooking were compared. Setting the percentages of vitamins remained in uncooked samples as 100 %, those in SC and V-P P cooked samples of pumpkin were calculated as follows: niacin 93.1 %, 85.4 %; folic acid 84.0 %, 74.4 %; and pantothenic acid 97.9 %, 89.1 %, respectively. Percentages of vitamins in SCand V-P P cooked samples of daikon were as follows: vitamin B1 97.4 %, 94.0 %; vitamin B2 not detected, not detected; vitamin B6 66.6 %, 65.2 %; vitamin C 43.3 %, 52.2 %; niacin 80.0 %, 66.7 %; folic acid 37.7 %, 37.7 %; and pantothenic acid 72.0 %, 65.4 %, respectively.These results indicated that vitamin losses measured in this study were attributable to the cooking process, suggesting that the degree of vitamin losses due to the cooking process varied according to the characteristics of vitamins and cooking methods.論

Stature estimation from anatomical landmarks in femur using postmortem CT

Objective: The present study aims to assess the correlation between stature and partial femur measurements in Japanese population, using three-dimensional (3D) computed tomographic (CT) images. Method: A total of 195 cadavers (105 males, 90 females) underwent postmortem CT scanning and subsequent forensic autopsy between April 2013 and June 2014. Five femur length measurements were taken from 3D CT volume rendering (VR) images according to three anatomical landmarks: the greater trochanter, the lesser trochanter, and the adductor tubercle. The correlations between stature and each of the femur length measurements were assessed with Spearman's rank correlation test, and showed significant correlations between stature and femur length measurements. Results: Using a mixed-sex regression formula, the R2 was from 0.644 to 0.766 and actual difference was less than 4.6 cm. Conclusion: Present study showed that femur length measurements, according to the anatomical landmarks obtained from 3DCT VR images, were useful for estimation of the stature, particularly in cases when a total femur bone was not available