著者

Satoko Iwasa Hirotaka Ota Kensuke Nishio Mariko Ohtsu Masafumi Kusunoki Takahiro Gojoubori Tetsuo Shirakawa Masatake Asano

出版者

日本大学歯学部

雑誌

Journal of Oral Science (ISSN:13434934)

巻号頁・発行日

vol.58, no.3, pp.317-323, 2016 (Released:2016-09-25)

参考文献数

25

被引用文献数

4

---

Toll-like receptors (TLR) recognize microbe-associated molecular patterns and induce the innate immune response. Among them, TLR5 recognizes the Gram-negative bacterial component flagellin. The aim of this study was to examine the expression of TLR5 in mouse salivary gland (SG). The SG was excised from 8- to 10-week-old female C57BL/6 mice. Salivary gland epithelial cells (SGECs) were purified and subjected to reverse transcription polymerase chain reaction (RT-PCR). Western blotting was performed to detect TLR5 expression at the protein level in several organs. The localization of TLR5 in SG was examined using immunohistochemical staining. The responsiveness of SGECs to flagellin was further examined by evaluating the induction of CXCL1 by real-time PCR and immunoprecipitation followed by Western blotting. TLR5 expression in SG was confirmed at the gene and protein levels. Immunohistochemical staining detected TLR5 in both acinic and ductal cells of the sublingual gland, but not in serous acinic cells of the submandibular gland. Although TLR5 was detected throughout the cytoplasm in ductal cells, positive staining was observed on the basal side of the mucous acinic cells. The purified SGECs responded to flagellin and induced the production of CXCL1. These findings suggest that TLR5 is functionally expressed in the SG and responds to its cognate ligand flagellin. (J Oral Sci 58, 317-323, 2016)

著者

Kensuke Nishio Keiko Motozawa Daisuke Omagari Takahiro Gojoubori Takayuki Ikeda Masatake Asano Nobuhito Gionhaku

出版者

日本大学歯学部

雑誌

Journal of Oral Science (ISSN:13434934)

巻号頁・発行日

vol.58, no.1, pp.59-65, 2016 (Released:2016-03-26)

参考文献数

39

被引用文献数

25

---

Matrix metalloproteinases (MMPs) and tumor-associated macrophages (TAMs) play important roles in tumor growth. The present study investigated the expression levels of MMP2 and MMP9 in relation to the distribution of TAMs in the primary and metastatic regions of oral squamous cell carcinoma. Twenty-nine cases of oral squamous cell carcinoma (OSCC) with regional lymph node metastasis were selected from available documents in the archives of the Department of Pathology, Nihon University School of Dentistry. Four-micrometer-thick sections were prepared from the primary and metastatic regions. Each section was subjected to immunohistochemical staining using anti-MMP2, anti-MMP9, and anti-CD68 antibodies. The distribution and localization of MMPs and TAMs were compared between primary and metastatic regions. The expression levels of both MMPs were higher in the metastatic regions of lingual and gingival cancers. Statistically significant differences were observed in both T1 and T2 cases. In contrast to the higher expression of MMPs in metastatic regions, a higher number of TAMs were distributed in the primary regions. From these results, MMP expression levels and the numbers of TAMs were expected to have an inverse relationship between the primary and metastatic regions of OSCC. (J Oral Sci 58, 59-65, 2016)

著者

Eri Sata Leo Takada Ryo Kaetsu Mai Fukasawa Mariko Ohtsu Mitsuru Motoyoshi Masatake Asano

出版者

Nihon University School of Dentistry

雑誌

Journal of Oral Science (ISSN:13434934)

巻号頁・発行日

pp.19-0477, (Released:2020-06-04)

参考文献数

14

被引用文献数

3

---

Interleukin-1α (IL-1α) is produced inside cells in its precursor form (pIL-1α). Enzymatic cleavage yields mature (mIL-1α) and the propiece of IL-1α (ppIL-1α), which are thought to be localized in the nucleus, because of the presence of nuclear localizing signals. Studies of ppIL-1α function have been hampered by the lack of a ppIL-1α-specific antibody (Ab). In the present study, the authors generated anti-ppIL-1α Ab by using recombinant histidine-tagged ppIL-1α (His-ppIL-1α) as an immunogen. Rabbits were immunized with His-ppIL-1α, and affinity-purified Ab was obtained. Ab reactivity and specificity were examined by Western blotting. The antibody successfully recognized transfectant-derived green fluorescence protein (GFP)-tagged ppIL-1α but not GFP. A sandwich enzyme-linked immunosorbent assay (ELISA) system established by biotinylating the anti-ppIL-1α Ab successfully detected GFP-ppIL-1α. The Ab and ELISA system allows functional analysis of ppIL-1α and improves understanding of ppIL-1α.

著者

Atsushi Tateno Masatake Asano Daisuke Akita Taku Toriumi Niina Tsurumachi-Iwasaki Tomohiko Kazama Yoshinori Arai Taro Matsumoto Koichiro Kano Masaki Honda

出版者

Nihon University School of Dentistry

雑誌

Journal of Oral Science (ISSN:13434934)

巻号頁・発行日

pp.18-0458, (Released:2019-10-21)

参考文献数

45

被引用文献数

13

---

Tissue engineering is a promising approach to supplement existing treatment strategies for craniofacial bone regeneration. In this study, a type I collagen scaffold made from a recombinant peptide (RCP) with an Arg-Gly-Asp motif was developed, and its effect on regeneration in critical-size mandibular bone defects was evaluated. Additionally, the combined effect of the scaffold and lipid-free dedifferentiated fat (DFAT) cells was assessed. Briefly, DFAT cells were separated from mature adipocytes by using a ceiling culture technique based on buoyancy. A 3 cm × 4 cm critical-size bone defect was created in the rat mandible, and regeneration was evaluated by using RCP with DFAT cells. Then, cultured DFAT cells and adipose-derived stem cells (ASCs) were seeded onto RCP scaffolds (DFAT/RCP and ASC/RCP) and implanted into the bone defects. Micro-computed tomography imaging at 8 weeks after implantation showed significantly greater bone regeneration in the DFAT/RCP group than in the ASC/RCP and RCP-alone groups. Similarly, histological analysis showed significantly greater bone width in the DFAT/RCP group than in the ASC/RCP and RCP-alone groups. These findings suggest that DFAT/RCP is effective for bone formation in critical-size bone defects and that DFAT cells are a promising source for bone regeneration.

著者

Satsuki Saito Mariko Ohtsu Masatake Asano Tomohiko Ishigami

出版者

Nihon University School of Dentistry

雑誌

Journal of Oral Science (ISSN:13434934)

巻号頁・発行日

pp.18-0411, (Released:2019-09-20)

参考文献数

24

被引用文献数

4

---

Accurate evaluation of the anti-cancer effects of ouabain, a cardiac glycoside, requires an understanding of its signaling pathway. This study examined the effects of ouabain stimulation on spontaneous interleukin (IL)-8 and IL-1α secretion in the HSC3 oral squamous cell carcinoma cell line. IL-8 secretion was reduced and IL-1α secretion was increased in the cells. Real-time polymerase chain reaction confirmed that these changes were regulated at the transcriptional level. Further analysis revealed that ouabain stimulation induced phosphorylation of activator protein (AP)-1 components (c-Jun and c-Fos) but not nuclear factor kappa B (NF-κB) components (p65 and p50). A luciferase assay demonstrated that the NF-κB-binding site located at 1 kb upstream of the TATA box in the IL-8 gene contributed to the reduction in IL-8 secretion. Pre-incubation of the cells with BAPTA-AM and L-glutathione increased IL-8 secretion, which indicates that Ca2+ ions and reactive oxygen species are associated with the ouabain-mediated reduction in IL-8 levels. The inhibitory effect of ouabain was attributed to reduced nuclear translocation of the NF-κB p65 subunit. Taken together, these findings indicate that ouabain exerts opposing effects on transcription factors NF-κB and AP-1.

著者

Hisanori Ito Takashi Kifune Misa Ishiyama Satoko Iwasa Hiroki Takei Tomokazu Hasegawa Masatake Asano Tetsuo Shirakawa

出版者

Nihon University School of Dentistry

雑誌

Journal of Oral Science (ISSN:13434934)

巻号頁・発行日

vol.60, no.4, pp.544-551, 2018 (Released:2018-12-27)

参考文献数

30

被引用文献数

5

---

Hypoxia after traumatic injuries to a tooth is one of the causes of subsequent root resorption. Inflammatory cytokines produced under hypoxic conditions are associated with root resorption, but the mechanism has not been fully understood. In this study, the role of hypoxia-inducible factor-1 (HIF-1) signaling in the regulation of CCAAT (cytosine-cytosine-adenosine-adenosine-thymidine)/enhancer-binding protein-β (C/EBPβ) and the receptor activator of nuclear factor kappa-B ligand (RANKL) expressions in immortalized human periodontal ligament (PDL) cells was investigated. PDL cells cultured under a hypoxic condition showed an increase in the expression of C/EBPβ and RANKL messenger RNAs (mRNAs), whereas the expression of osteoprotegerin and HIF-1α mRNAs was unaffected. Hypoxia had no effects on the secretion of interleukin (IL)-1β, IL-6, IL-8, IL-17A, tumor necrosis factor-alpha, macrophage migration inhibitory factor, monocyte chemoattractant protein-1, and macrophage colony-stimulating factor in the culture media. Treatment of the cells with dimethyloxaloylglycine, a competitive HIF prolyl hydroxylase inhibitor, significantly increased the expression of C/EBPβ and RANKL mRNAs. This suggested that the hypoxia-induced elevation of C/EBPβ and RANKL mRNAs was dependent on the HIF-1 activity. PDL cells transfected with a specific small interfering RNA designed to target the C/EBPβ gene showed a significant suppression of the RANKL mRNA. These findings indicated that C/EBPβ may play an important role in tooth root resorption via RANKL activation in hypoxia-exposed PDL cells.

著者

Takashi Kifune Hisanori Ito Misa Ishiyama Satoko Iwasa Hiroki Takei Tomokazu Hasegawa Masatake Asano Tetsuo Shirakawa

出版者

Nihon University School of Dentistry

雑誌

Journal of Oral Science (ISSN:13434934)

巻号頁・発行日

vol.60, no.4, pp.519-525, 2018 (Released:2018-12-27)

参考文献数

32

被引用文献数

11

---

Hypoxia induces complex cellular responses that are mediated by a key transcription factor, hypoxia-inducible factor-1 (HIF-1). HIF-1 promotes production of cytokines and angiogenic factors and contributes to recovery of injured tissues. In the present study, expressions of angiogenin (ANG) and vascular endothelial growth factor (VEGF), which are potent angiogenic factors in mammalian tissues, were examined in immortalized fibroblasts exposed to hypoxia. After 24 h of exposure to hypoxia, ANG and VEGF mRNAs expressions were significantly elevated in periodontal ligament (PDL) fibroblasts but not in embryonic fibroblasts. Hypoxia also increased productions of ANG and VEGF proteins in PDL fibroblasts. HIF-1α mRNA expression was not affected by hypoxia in either fibroblast, although HIF-1α protein expression was enhanced after exposure to hypoxia. Treatment of PDL fibroblasts with dimethyloxaloylglycine, a prolyl hydroxylase inhibitor that stabilizes the HIF-1α protein, significantly increased expressions of ANG and VEGF mRNAs under normoxia. This suggests that stabilization of HIF-1α is crucial for upregulation of ANG and VEGF in PDL fibroblasts. These results indicate that, under hypoxic conditions, HIF-1α upregulates synthesis of ANG and VEGF in PDL fibroblasts and promotes angiogenesis.

著者

Miho Obayashi-Ishii Satsuki Saito Daisuke Omagari Masatake Asano Tomohiko Ishigami

出版者

Nihon University School of Dentistry

雑誌

Journal of Oral Science (ISSN:13434934)

巻号頁・発行日

pp.17-0380, (Released:2018-08-25)

参考文献数

29

被引用文献数

2

---

Magnetic attachments are commonly used for overdentures. The deleterious effects of exposure to magnetic flux on human health have not been substantiated so far; nevertheless, there is a need to understand the extent of magnetic field exposure in the oral area resulting from the use of magnetic attachments. The purpose of this study was to investigate the influence of a magnetic field on oral squamous cell carcinoma. Tumor cells cultured on a magnetic plate were compared with those not cultured on a magnetic plate (controls). The cells were seeded at a density of 1 × 105 cells/well and cultured for 6 days. The influence of the magnetic field on cytokine production was examined by cytokine array analysis. Secretion of platelet-derived growth factor-AA (PDGF-AA) was measured by enzyme-linked immunosorbent assay and Western blotting. The expression of PDGF-AA messenger RNA was examined by real-time polymerase chain reaction, whereas nuclear factor-kappa B activity was measured by luciferase assay. The results indicated that the magnetic field inhibited the secretion of PDGF-AA, thereby inhibiting PDGF-AA-induced expression, thus reducing the risk of cancer recurrence.

著者

Mariko Ohtsu Muneaki Tamura Hideto Sasaki Shuichi Sato Masatake Asano

出版者

日本大学歯学部

雑誌

Journal of Oral Science (ISSN:13434934)

巻号頁・発行日

vol.59, no.3, pp.453-456, 2017 (Released:2017-09-14)

参考文献数

16

被引用文献数

1

---

The effects of bittern water (BW), obtained from the ocean floor, on cariogenic bacteria and saliva secretion were examined. Streptococcus mutans was mixed with BW for 1, 3, 5, 10, and 20 min to explore the bactericidal effects of BW against cariogenic bacteria. Bacterial viability was calculated by counting the number of colony-forming units on Brain Heart Infusion agar plates. The results indicated a bacterial viability of more than 35% even after 20 min of incubation. Subsequently, the effects of BW on saliva secretion and the salivary concentration of secretory IgA (sIgA) were examined. Gargling with BW significantly augmented saliva secretion. Although the sIgA concentration was reduced, the total sIgA secreted into saliva was increased significantly. Our findings indicate that the use of BW may be a new strategy for the treatment of various oral diseases, including dry mouth.

著者

Tomoyasu Kiyokawa Mitsuru Motoyoshi Mizuki Inaba Remi Sano Akari Saiki Go Torigoe Masatake Asano Noriyoshi Shimizu

出版者

日本大学歯学部

雑誌

Journal of Oral Science (ISSN:13434934)

巻号頁・発行日

vol.59, no.3, pp.447-451, 2017 (Released:2017-09-14)

参考文献数

22

被引用文献数

2

---

The purpose of this experiment was to investigate whether low-intensity pulsed ultrasound (LIPUS) irradiation can inhibit dentoalveolar ankylosis in transplanted rat teeth. LIPUS irradiation (the pulsed ultrasound signal had a frequency of 3.0 MHz, a spatial average intensity of 30 mW/cm2, and a pulse ratio of 1:4) was performed on the face over the re-planted teeth of rats for 4 weeks. After the rats were euthanized, we measured mobility (Periotest value [PTV]) of the transplanted and control teeth using a Periotest. Finally, we performed histological evaluation to detect ankylosis. PTVs tended to be significantly lower for re-planted teeth than for control teeth. Histological evaluation revealed that the roots of all re-planted teeth were coalescent with alveolar bone. Furthermore, no ankylosis was observed in three-fifths of the re-planted teeth following LIPUS irradiation. These results indicate the potential efficacy of LIPUS to inhibit dentoalveolar ankylosis.

著者

Masao Araki Shunsuke Namaki Toshihiko Amemiya Kunihito Matsumoto Kazuya Honda Yoshiyuki Yonehara Naoyuki Matsumoto Masatake Asano

出版者

日本大学歯学部

雑誌

Journal of Oral Science (ISSN:13434934)

巻号頁・発行日

vol.58, no.4, pp.533-537, 2016 (Released:2016-12-23)

参考文献数

32

被引用文献数

3

---

The occurrence of ameloblastic fibro-odontoma (AFO) in the oral region is unusual and accounts for 1-3% of all odontogenic tumors. AFO presents mixed radiopaque patterns within the lesion with diverse findings; therefore, it is important to compare this tumor with other odontogenic tumors that radiographically present with calcified bodies. Herein, we observed the calcification patterns within the lesion in seven AFO cases (five males, two females; mean age, 8.3 years; age range, 4-13 years). Periapical and panoramic radiographs were obtained from all seven cases. Five cases underwent conventional computed tomography (CT) scanning, and one underwent cone beam CT. Classification of the calcifications primarily involved the following two characteristics on the X-rays: appearance and location of the lesions. All seven cases were located in the molar regions of the mandible in association with impacted teeth. The calcification patterns of these AFOs were mixed or inhomogeneous within the lesion with various findings, including complex odontoma-like calcifications. However, the patterns differed between panoramic radiography and CT in some cases. The radiolucent lesions in AFO demonstrated varying calcification patterns and were associated with impacted teeth on the CT images.(J Oral Sci 58, 533-537, 2016)

著者

Taiichi Kitano Yoshikazu Mikami Takashi Iwase Masatake Asano Kazuo Komiyama

出版者

日本大学歯学部

雑誌

Journal of Oral Science (ISSN:13434934)

巻号頁・発行日

vol.58, no.2, pp.163-169, 2016 (Released:2016-06-25)

参考文献数

24

被引用文献数

10

---

Porphyromonas gingivalis is important in the development of marginal periodontitis. However, the precise role and localization of P. gingivalis in chronic periapical periodontitis remain unclear. Thus, methods that can detect P. gingivalis in formalin-fixed and paraffin-embedded (FFPE) tissue samples are needed. We assessed a technique combining loop-mediated isothermal amplification (LAMP) with PCR (PCR-LAMP) for detection of P. gingivalis, using 110 FFPE tissue samples of chronic apical periodontitis. PCR-LAMP specifically detected P. gingivalis with high sensitivity in FFPE tissue samples, and the sensitivity of the technique was higher than that of PCR or LAMP alone. The results of immunohistochemistry (IHC) confirmed the specificity of PCR-LAMP. IHC showed that P. gingivalis was localized in a granular layer of chronic apical periodontitis, a region that correlated with the localization of macrophages. This is the first study to describe the localization of P. gingivalis in human periapical periodontitis. In conclusion, PCR-LAMP was an effective tool for detecting P. gingivalis in periapical periodontitis. In addition, IHC results improve our understanding of the role of P. gingivalis in the progression of periapical periodontitis. (J Oral Sci 58, 163-169, 2016)