M. Okamura, Taiki Suzuki, Yusuke Oomura, S. Matsunaga, Takeshi Nomura
Bisphosphonate (BP) formulations are drugs that improve bone strength by suppressing osteoclast activation, preventing fractures of the vertebrae and the femoral head, but their side effects include osteonecrosis of the jaw (ONJ). In this case it is known as medication-related osteonecrosis of the jaw (MRONJ), and pathological and microbiological investigations have suggested that infection is one major causative factor. However, many points regarding the etiology of ONJ and its causative factors remain unclear. In this study, we administered BP to model mice and exposed their jaws to bacterial infection to produce a mouse model of BRONJ, and analyzed their bone structure, including an analysis of the quality of bone surrounding extraction cavities. We found that mice not exposed to bacterial infection did not develop ONJ, and that those mice exposed to bacterial infection that did develop ONJ exhibited abnormal collagen fiber arrangement and poor bioapatite crystal alignment. An analysis of areas of bone surrounding poorly healed extraction cavities also revealed that its quality was poor. These results showed that although BP use increases bone mineral density, it reduces the alignment of collagen fibers and decreases bone quality. Zoledronate (Zol) alone resulted in epithelial healing, but reduced bone quality. In addition, it was suggested that bacterial infection could develop into a condition similar to BRONJ.
{"title":"Effect of Bacterial Infection on Bone Quality and Structure in Osteonecrosis of the Jaw by Bisphosphonate (BP) Administration","authors":"M. Okamura, Taiki Suzuki, Yusuke Oomura, S. Matsunaga, Takeshi Nomura","doi":"10.2485/jhtb.30.323","DOIUrl":"https://doi.org/10.2485/jhtb.30.323","url":null,"abstract":"Bisphosphonate (BP) formulations are drugs that improve bone strength by suppressing osteoclast activation, preventing fractures of the vertebrae and the femoral head, but their side effects include osteonecrosis of the jaw (ONJ). In this case it is known as medication-related osteonecrosis of the jaw (MRONJ), and pathological and microbiological investigations have suggested that infection is one major causative factor. However, many points regarding the etiology of ONJ and its causative factors remain unclear. In this study, we administered BP to model mice and exposed their jaws to bacterial infection to produce a mouse model of BRONJ, and analyzed their bone structure, including an analysis of the quality of bone surrounding extraction cavities. We found that mice not exposed to bacterial infection did not develop ONJ, and that those mice exposed to bacterial infection that did develop ONJ exhibited abnormal collagen fiber arrangement and poor bioapatite crystal alignment. An analysis of areas of bone surrounding poorly healed extraction cavities also revealed that its quality was poor. These results showed that although BP use increases bone mineral density, it reduces the alignment of collagen fibers and decreases bone quality. Zoledronate (Zol) alone resulted in epithelial healing, but reduced bone quality. In addition, it was suggested that bacterial infection could develop into a condition similar to BRONJ.","PeriodicalId":16040,"journal":{"name":"Journal of Hard Tissue Biology","volume":null,"pages":null},"PeriodicalIF":0.4,"publicationDate":"2021-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"68964921","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
M. Murakami, K. Nagano, K. Hamaoka, Daisuke Kato, T. Kawai, H. Murakami, Y. Hasegawa
: Ozone water has long been known as a bactericidal disinfectant. However, the bactericidal effect of ozone water on bacteria associated with oral diseases has not been thoroughly examined. Further, although oral bacteria reside in biofilms, few studies have explored the effects of ozone water on biofilms. In this study, we aimed to investigate the bactericid al effect of ozone water on bacteria and bacterial biofilms associated with oral diseases. We examined the bactericidal and cleaning effects of ozone water on pathogenic bacteria associated with oral diseases ( Staphylococcus aureus , Pseudomonas aeruginosa , Streptococcus mutans , and Porphyromonas gingivalis ) under planktonic and biofilm growth conditions. When planktonic bacteria were exposed to 5-ppm ozone water, a remarkable antibacterial activity was observed against all of the tested bacterial species. Contrarily, biofilms showed high resistance to ozone water; the bacterial load only slightly de creased even after repeated exposure to ozone water. However, when ozone water was continuously applied at a low flow rate to the biofilms on polystyrene disks, the number of bacteria on the disks was significantly decreased. Our results have shown that the continuous application of ozone water can eliminate oral disease-related bacteria even in biofilms. washed between treatments, bacterial numbers were also affected by the washing effect of water and ozone water treatments. There were no signs of a reduction in viable counts of S. aureus , P. aeruginosa , and S. mutans biofilms even after three treatments with ozone water, as well as with water. P . gingivalis strains showed a slight decrease in CFUs after water and ozone water treatments.
{"title":"Ozone Water Bactericidal and Cleaning Effects on Oral Diseases-related Planktonic and Bacterial Biofilms","authors":"M. Murakami, K. Nagano, K. Hamaoka, Daisuke Kato, T. Kawai, H. Murakami, Y. Hasegawa","doi":"10.2485/JHTB.30.27","DOIUrl":"https://doi.org/10.2485/JHTB.30.27","url":null,"abstract":": Ozone water has long been known as a bactericidal disinfectant. However, the bactericidal effect of ozone water on bacteria associated with oral diseases has not been thoroughly examined. Further, although oral bacteria reside in biofilms, few studies have explored the effects of ozone water on biofilms. In this study, we aimed to investigate the bactericid al effect of ozone water on bacteria and bacterial biofilms associated with oral diseases. We examined the bactericidal and cleaning effects of ozone water on pathogenic bacteria associated with oral diseases ( Staphylococcus aureus , Pseudomonas aeruginosa , Streptococcus mutans , and Porphyromonas gingivalis ) under planktonic and biofilm growth conditions. When planktonic bacteria were exposed to 5-ppm ozone water, a remarkable antibacterial activity was observed against all of the tested bacterial species. Contrarily, biofilms showed high resistance to ozone water; the bacterial load only slightly de creased even after repeated exposure to ozone water. However, when ozone water was continuously applied at a low flow rate to the biofilms on polystyrene disks, the number of bacteria on the disks was significantly decreased. Our results have shown that the continuous application of ozone water can eliminate oral disease-related bacteria even in biofilms. washed between treatments, bacterial numbers were also affected by the washing effect of water and ozone water treatments. There were no signs of a reduction in viable counts of S. aureus , P. aeruginosa , and S. mutans biofilms even after three treatments with ozone water, as well as with water. P . gingivalis strains showed a slight decrease in CFUs after water and ozone water treatments.","PeriodicalId":16040,"journal":{"name":"Journal of Hard Tissue Biology","volume":null,"pages":null},"PeriodicalIF":0.4,"publicationDate":"2021-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"68965061","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Naofumi Yamada, T. Yamasaki, Kana Yamawaki, Minami Nakagiri, Hideyuki Ito, Yoshimasa Nakamura, T. Nakanishi
Rosemary is used as an herb in cooking and aromatic oils, and has long been used as a medicinal herb because of its functional components. According to folklore, monks recommended a remedy of rosemary in alcohol (“Hungarian water”) to a Hungarian queen suffering from limb numbness, resulting in instant recovery and rejuvenation. This is why the queen married a 20-year-old king of Holland while in her seventies. In order to clarify the “rejuvenation” mechanisms of traditional Hungarian water, we examined the anti-oxidant activity, anti-glycation activity, inhibition of melanin production and suppression of tumor growth of ethanol extracts of rosemary in this study. It was found that the ethanol extract had high anti-oxidant activity, high anti-glycation activity, high inhibition of melanin production and high suppression of tumor cell growth.
{"title":"Functional Evaluation of the Ethanol Extracts from Rosmarinus officinalis L. (Rosemary)","authors":"Naofumi Yamada, T. Yamasaki, Kana Yamawaki, Minami Nakagiri, Hideyuki Ito, Yoshimasa Nakamura, T. Nakanishi","doi":"10.2485/jhtb.30.291","DOIUrl":"https://doi.org/10.2485/jhtb.30.291","url":null,"abstract":"Rosemary is used as an herb in cooking and aromatic oils, and has long been used as a medicinal herb because of its functional components. According to folklore, monks recommended a remedy of rosemary in alcohol (“Hungarian water”) to a Hungarian queen suffering from limb numbness, resulting in instant recovery and rejuvenation. This is why the queen married a 20-year-old king of Holland while in her seventies. In order to clarify the “rejuvenation” mechanisms of traditional Hungarian water, we examined the anti-oxidant activity, anti-glycation activity, inhibition of melanin production and suppression of tumor growth of ethanol extracts of rosemary in this study. It was found that the ethanol extract had high anti-oxidant activity, high anti-glycation activity, high inhibition of melanin production and high suppression of tumor cell growth.","PeriodicalId":16040,"journal":{"name":"Journal of Hard Tissue Biology","volume":null,"pages":null},"PeriodicalIF":0.4,"publicationDate":"2021-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"68965174","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
: To detect the expression of miR-590 in oral lichen planus (OLP) tissues and oral squamous cell carcinoma (OSCC) tissues, and to analyze the correlations with clinicopathological characteristics and prognosis of OSCC patients. The oral mucosa tissues were selected from 180 patients with OLP or OSCC. They were divided into OLP group (n=92) and OSCC group (n=88), and 40 healthy volunteers with normal oral mucosa tissues were set as control group. Human tongue squamous cell carcinoma cell line SCC9 and human oral keratinocyte HOK were used. The expressions of miR-590 in tissues and cells were detected using qPCR. SCC9 cells were transfected with small interfering (si)-miR-590 and si-NC plasmids. MTT assay, flow cytometry and Transwell assay were used to detect cell proliferation, apoptosis, migration and invasion abilities, respectively. The expression levels of apoptosis-, migration- and invasion-related proteins were examined using Western blotting. Control, OLP and OSCC groups displayed successively increased expression of miR-590, suggesting that the expression was related to the TNM stage and lymph node metastasis of OSCC patients (P<0.05). Following transfection with si-miR-590, the proliferation, migration and invasion abilities of SCC9 cells were weakened significantly, while the apoptosis rate rose (P<0.05). The expression levels of Bcl-2, N-cadherin and vimentin dropped significantly, whereas those of Bax and E-cadherin increased (P<0.05). MiR-590 is highly expressed in OSCC and SCC9 cells. Silencing miR-590 can suppress the proliferation, migration and invasion and promote the apoptosis of SCC9 cells.
{"title":"Expressions of miR-590 in Oral Lichen Planus and Oral Squamous Cell Carcinoma Tissues and Clinical Values","authors":"Wanlu Chen, Yong Zhou, Zhongxiong Ma, Yunde Xie","doi":"10.2485/jhtb.30.363","DOIUrl":"https://doi.org/10.2485/jhtb.30.363","url":null,"abstract":": To detect the expression of miR-590 in oral lichen planus (OLP) tissues and oral squamous cell carcinoma (OSCC) tissues, and to analyze the correlations with clinicopathological characteristics and prognosis of OSCC patients. The oral mucosa tissues were selected from 180 patients with OLP or OSCC. They were divided into OLP group (n=92) and OSCC group (n=88), and 40 healthy volunteers with normal oral mucosa tissues were set as control group. Human tongue squamous cell carcinoma cell line SCC9 and human oral keratinocyte HOK were used. The expressions of miR-590 in tissues and cells were detected using qPCR. SCC9 cells were transfected with small interfering (si)-miR-590 and si-NC plasmids. MTT assay, flow cytometry and Transwell assay were used to detect cell proliferation, apoptosis, migration and invasion abilities, respectively. The expression levels of apoptosis-, migration- and invasion-related proteins were examined using Western blotting. Control, OLP and OSCC groups displayed successively increased expression of miR-590, suggesting that the expression was related to the TNM stage and lymph node metastasis of OSCC patients (P<0.05). Following transfection with si-miR-590, the proliferation, migration and invasion abilities of SCC9 cells were weakened significantly, while the apoptosis rate rose (P<0.05). The expression levels of Bcl-2, N-cadherin and vimentin dropped significantly, whereas those of Bax and E-cadherin increased (P<0.05). MiR-590 is highly expressed in OSCC and SCC9 cells. Silencing miR-590 can suppress the proliferation, migration and invasion and promote the apoptosis of SCC9 cells.","PeriodicalId":16040,"journal":{"name":"Journal of Hard Tissue Biology","volume":null,"pages":null},"PeriodicalIF":0.4,"publicationDate":"2021-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"68965208","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Maki Nagasaki, Kumiko Nakai, Hideki Tanaka, Manami Ozaki, Kengo Kato, R. Koshi, M. Maeno, S. Nishikubo, T. Kawato, M. Tonogi
: Preexisting diseases, such as diabetes and chronic inflammation in periodontal tissue, are risk factors associated with bisphosphonate-related osteonecrosis of the jaw. Osteoblasts produce prostaglandin (PG)E 2 via cyclooxygenases (COX), and the autocrine action of PGE 2 impacts the function of osteoblasts, including receptor activator of NF-kappa B li gand (RANKL) and osteoprotegerin (OPG) production. This study assessed the effects of the stimulation of zoledronate in the presence of lipopolysaccharide (LPS) and high concentrations of glucose on the expression of COX-2, RANKL, and OPG, in addition to PGE 2 production in osteoblasts. MG-63 cells were cultured in medium containing 1 µg/ml LPS, 25 mM glucose (high glucose), and/or zoledronate (1×10 -8 , 1×10 -7 , 1×10 -6 , 5×10 -6 , or 1×10 -5 M). The mRNA expression of COX-2, RANKL, and OPG genes was determined by real-time polymerase chain reaction. The concentrations of RANKL and OPG protein and PGE 2 in the culture supernatant were examined by enzyme-linked immunosorbent assay. Zoledronate at a con centration of 5×10 -6 M overwhelmingly increased COX-2 mRNA expression. The expression levels of RANKL and OPG as well as PGE 2 production was significantly increased in cells stimulated with 5×10 -6 M zoledronate in the presence of LPS and high glucose than in the unstimulated cells (control). NS398, a specific inhibitor of COX-2, blocked the stimulatory ef fects of zoledronate (in the presence of LPS and high glucose) on PGE 2 production and the protein expression levels of RANKL and OPG. The ratio of RANKL/OPG was also increased following zolendronate stimulation. In addition, a signifi cant difference was observed not in the stimulation with zoledronate alone, but by the stimulation of zoledronate in the pres ence of LPS and high glucose as compared that in controls. These results suggest that LPS and high concentrations of glu cose enhances zoledronate-induced increase in RANKL/OPG ratio via the autocrine action of NS398-blocked PGE 2 in osteoblasts. assessed the effects of three types of bisphosphonate, zoledronate, alen dronate, and clodronate, on mRNA expression involved in the osteoblast differentiation and function in MG-63 cells and human primary osteo -blasts. In their experiments, treatment with zoledronate at concentrations ranging from 1×10 -5 to 1×10 -9 M decreased the expression of osteoblast differentiation-related transcription factors, collagenous, and non-colla genous protein, bone morphogenic protein, and ALPase, whereas the expression levels of transforming growth factor β and vascular endothe lial growth factor were increased. They also revealed that the effects of zoledronate on these mRNA expression levels was found to be generally dose-dependent. In the present study, the amount of total RNA eluted from cells was less obvious in the stimulation of 1×10 -5 M zoledronate than in the case of a stimulation under or equal to 5×10 -6 M (data not shown), and COX-2 expression le
{"title":"Lipopolysaccharide and High Concentrations of Glucose Enhances Zoledronate-induced Increase in RANKL/OPG Ratio by Upregulating PGE2 Production in Osteoblasts","authors":"Maki Nagasaki, Kumiko Nakai, Hideki Tanaka, Manami Ozaki, Kengo Kato, R. Koshi, M. Maeno, S. Nishikubo, T. Kawato, M. Tonogi","doi":"10.2485/JHTB.30.37","DOIUrl":"https://doi.org/10.2485/JHTB.30.37","url":null,"abstract":": Preexisting diseases, such as diabetes and chronic inflammation in periodontal tissue, are risk factors associated with bisphosphonate-related osteonecrosis of the jaw. Osteoblasts produce prostaglandin (PG)E 2 via cyclooxygenases (COX), and the autocrine action of PGE 2 impacts the function of osteoblasts, including receptor activator of NF-kappa B li gand (RANKL) and osteoprotegerin (OPG) production. This study assessed the effects of the stimulation of zoledronate in the presence of lipopolysaccharide (LPS) and high concentrations of glucose on the expression of COX-2, RANKL, and OPG, in addition to PGE 2 production in osteoblasts. MG-63 cells were cultured in medium containing 1 µg/ml LPS, 25 mM glucose (high glucose), and/or zoledronate (1×10 -8 , 1×10 -7 , 1×10 -6 , 5×10 -6 , or 1×10 -5 M). The mRNA expression of COX-2, RANKL, and OPG genes was determined by real-time polymerase chain reaction. The concentrations of RANKL and OPG protein and PGE 2 in the culture supernatant were examined by enzyme-linked immunosorbent assay. Zoledronate at a con centration of 5×10 -6 M overwhelmingly increased COX-2 mRNA expression. The expression levels of RANKL and OPG as well as PGE 2 production was significantly increased in cells stimulated with 5×10 -6 M zoledronate in the presence of LPS and high glucose than in the unstimulated cells (control). NS398, a specific inhibitor of COX-2, blocked the stimulatory ef fects of zoledronate (in the presence of LPS and high glucose) on PGE 2 production and the protein expression levels of RANKL and OPG. The ratio of RANKL/OPG was also increased following zolendronate stimulation. In addition, a signifi cant difference was observed not in the stimulation with zoledronate alone, but by the stimulation of zoledronate in the pres ence of LPS and high glucose as compared that in controls. These results suggest that LPS and high concentrations of glu cose enhances zoledronate-induced increase in RANKL/OPG ratio via the autocrine action of NS398-blocked PGE 2 in osteoblasts. assessed the effects of three types of bisphosphonate, zoledronate, alen dronate, and clodronate, on mRNA expression involved in the osteoblast differentiation and function in MG-63 cells and human primary osteo -blasts. In their experiments, treatment with zoledronate at concentrations ranging from 1×10 -5 to 1×10 -9 M decreased the expression of osteoblast differentiation-related transcription factors, collagenous, and non-colla genous protein, bone morphogenic protein, and ALPase, whereas the expression levels of transforming growth factor β and vascular endothe lial growth factor were increased. They also revealed that the effects of zoledronate on these mRNA expression levels was found to be generally dose-dependent. In the present study, the amount of total RNA eluted from cells was less obvious in the stimulation of 1×10 -5 M zoledronate than in the case of a stimulation under or equal to 5×10 -6 M (data not shown), and COX-2 expression le","PeriodicalId":16040,"journal":{"name":"Journal of Hard Tissue Biology","volume":null,"pages":null},"PeriodicalIF":0.4,"publicationDate":"2021-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"68965252","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Z. Li, Ming Dong, Wen Li, Ling Li, Xinxin Yu, Y. Kong, H. Kong
: The sinonasal inverted papilloma is one of the more common benign tumors of the nasal cavity and sinus. It origi-nates from the schneiderian membrane. It has the characteristics of easy recurrence and malignant transformation. This experiment found that Cytokeratin 8 expression increased in the sinonasal inverted papilloma. To better detect the role of Cytokeratin 8 in the proliferation and deterioration of the sinonasal inverted papilloma, we divided the sinonasal inverted papilloma into NIPN, NIPAP, and NIPAH by histological. The expression of Cytokeratin 8 in the sinonasal inverted papilloma was detected by immunohistochemistry. We found that the increased expression of Cytokeratin 8 was consistent with the invasion of sinonasal squamous cell carcinoma. In vitro , it was found that after Cytokeratin 8 interference, the proliferative and invasion of head and neck squamous cells were decreased. Cytokeratin 8 played an important role of sinonasal inverted papilloma malignant transformation to sinonasal squamous cell carcinoma. Inhibition of Cytokeratin 8 could diminish the proliferation and block the invasion of head and neck squamous cells.
{"title":"Cytokeratin 8 Promoted Sinonasal Inverted Papilloma Malignant Transformation to SNSCC","authors":"Z. Li, Ming Dong, Wen Li, Ling Li, Xinxin Yu, Y. Kong, H. Kong","doi":"10.2485/JHTB.30.45","DOIUrl":"https://doi.org/10.2485/JHTB.30.45","url":null,"abstract":": The sinonasal inverted papilloma is one of the more common benign tumors of the nasal cavity and sinus. It origi-nates from the schneiderian membrane. It has the characteristics of easy recurrence and malignant transformation. This experiment found that Cytokeratin 8 expression increased in the sinonasal inverted papilloma. To better detect the role of Cytokeratin 8 in the proliferation and deterioration of the sinonasal inverted papilloma, we divided the sinonasal inverted papilloma into NIPN, NIPAP, and NIPAH by histological. The expression of Cytokeratin 8 in the sinonasal inverted papilloma was detected by immunohistochemistry. We found that the increased expression of Cytokeratin 8 was consistent with the invasion of sinonasal squamous cell carcinoma. In vitro , it was found that after Cytokeratin 8 interference, the proliferative and invasion of head and neck squamous cells were decreased. Cytokeratin 8 played an important role of sinonasal inverted papilloma malignant transformation to sinonasal squamous cell carcinoma. Inhibition of Cytokeratin 8 could diminish the proliferation and block the invasion of head and neck squamous cells.","PeriodicalId":16040,"journal":{"name":"Journal of Hard Tissue Biology","volume":null,"pages":null},"PeriodicalIF":0.4,"publicationDate":"2021-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"68965469","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
The present study investigated the changes in Indian hedgehog (Ihh), periarticular cell-derived parathyroid hormone-related protein (PTHrP), and runt-related transcription factor 2 (Runx2) in the temporomandibular joint (TMJ) cartilage with posttraumatic osteoarthritis (PTOA). The miniature pigs were randomly divided into two groups: the experimental group (EG) (n=8) and the control group (CG) (n=4). The left side of EG had type B intracapsular fractures with anterior disc displacement (ICF+DD), while the right side only had type B intracapsular fractures (ICF), and the CG was a blank control. The production of Ihh, PTHrP, and Runx2 was detected by immunohistochemistry staining and real-time polymerase chain reaction (PCR) at weeks 4 and 12 post-surgery. The expression of Ihh, PTHLH /PTHrP, and Runx2 in the EG was significantly lower than that in the CG at 4 and 12 weeks after the operation (P<0.05). Moreover, significant differences were detected between ICF+DD and ICF (P<0.05). Ihh, PTHHL/PTHrP, and Runx2 proteins affect the endochondral osteogenesis of TMJ and play a significant role in PTOA. Our findings suggested that the interaction mechanism among Ihh, PTHLH/PTHrP, and Runx2 is activated when posttraumatic osteoarthritis (PTOA) occurs, but how they regulate each other remains to be investigated.
{"title":"Posttraumatic Osteoarthritis of Temporomandibular Joint in Miniature Pigs","authors":"N. Sun, D. He, Chi Yang, Qing Zhou","doi":"10.2485/JHTB.30.79","DOIUrl":"https://doi.org/10.2485/JHTB.30.79","url":null,"abstract":"The present study investigated the changes in Indian hedgehog (Ihh), periarticular cell-derived parathyroid hormone-related protein (PTHrP), and runt-related transcription factor 2 (Runx2) in the temporomandibular joint (TMJ) cartilage with posttraumatic osteoarthritis (PTOA). The miniature pigs were randomly divided into two groups: the experimental group (EG) (n=8) and the control group (CG) (n=4). The left side of EG had type B intracapsular fractures with anterior disc displacement (ICF+DD), while the right side only had type B intracapsular fractures (ICF), and the CG was a blank control. The production of Ihh, PTHrP, and Runx2 was detected by immunohistochemistry staining and real-time polymerase chain reaction (PCR) at weeks 4 and 12 post-surgery. The expression of Ihh, PTHLH /PTHrP, and Runx2 in the EG was significantly lower than that in the CG at 4 and 12 weeks after the operation (P<0.05). Moreover, significant differences were detected between ICF+DD and ICF (P<0.05). Ihh, PTHHL/PTHrP, and Runx2 proteins affect the endochondral osteogenesis of TMJ and play a significant role in PTOA. Our findings suggested that the interaction mechanism among Ihh, PTHLH/PTHrP, and Runx2 is activated when posttraumatic osteoarthritis (PTOA) occurs, but how they regulate each other remains to be investigated.","PeriodicalId":16040,"journal":{"name":"Journal of Hard Tissue Biology","volume":null,"pages":null},"PeriodicalIF":0.4,"publicationDate":"2021-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"68965775","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Daisuke Yamaguchi, K. Takeuchi, Atsuko Ueno, Daisuke Kato, Shin Miyamae, H. Murakami
: Drug repositioning (DR) is a strategy to explore new medicinal effects from existing approved drugs whose safety and pharmacokinetics have already been established. We focused on geranylgeranylacetone (GGA), which is known as a heat shock proteins (HSPs) inducing agent. GGA is mainly used as a gastric mucosal protective agent; however, its effects on bone tissues have not been studied. Therefore, we hypothesized that “GGA induces HSPs in osteoblasts thereby promotes cell differentiation”, and administered GGA to MC3T3E-1 cells to examine cell responses. Methods: MC3T3E-1 were cultured in osteogenic medium. After the cultures were established, test cultures were exposed to GGA (GGA group). Cell proliferation, collage synthesis and ALP activity were measured on days 7 and 14 of culture. Alizarin Red S staining was performed on days 21 of culture. Results: On days 14 of culture, cell proliferation and collage synthesis were signifi cantly higher in the GGA group than in the control group (P<0.05). On days 7 and 14 of culture, ALP activity was signifi cantly higher in the GGA group than in the Control group (P<0.05). On days 28 of culture, the Alizarin Red S stained areas were significantly higher in the GGA group than in the control group (P<0.05). Conclusion: GGA promoted the differentia tion of MC3T3E-1 in an in vitro cell culture model. We hypothesized that GGA upregulates HSPs and promotes osteoblast differentiation and tested our hypothesis using a series of in vitro experiments. We investigated the responses and phenotypic changes occurring in MC3T3E-1 mouse osteoblast-like cells exposed to GGA. A 10 -3 M GGA concentration was applied to all cell cultures as preliminary assays demonstrated that it was the minimum concentration necessary to produce different calcification levels (unpub lished data).
药物重新定位(Drug repositioning, DR)是一种从已经获得批准且安全性和药代动力学已经确定的药物中探索新疗效的策略。我们重点研究了香叶乙酸酮(GGA)，它被称为热休克蛋白(HSPs)诱导剂。GGA主要用作胃粘膜保护剂;然而，其对骨组织的影响尚未被研究。因此，我们假设“GGA在成骨细胞中诱导热休克蛋白，从而促进细胞分化”，并将GGA注入MC3T3E-1细胞，观察细胞反应。方法:在成骨培养基中培养MC3T3E-1。培养建立后，将实验培养物暴露于GGA (GGA组)。在培养第7天和第14天测定细胞增殖、拼贴合成和碱性磷酸酶活性。第21天进行茜素红S染色。结果:培养第14天，GGA组细胞增殖和拼贴合成显著高于对照组(P<0.05)。培养第7天和第14天，GGA组ALP活性显著高于对照组(P<0.05)。培养第28天，GGA组茜素红S染色面积显著高于对照组(P<0.05)。结论:GGA促进MC3T3E-1在体外细胞培养模型中的分化。我们假设GGA上调热休克蛋白并促进成骨细胞分化，并通过一系列体外实验验证了我们的假设。我们研究了暴露于GGA的MC3T3E-1小鼠成骨细胞样细胞的反应和表型变化。10 -3 M GGA浓度应用于所有细胞培养，初步分析表明，这是产生不同钙化水平所需的最低浓度(未发表的数据)。
{"title":"Experimental Repositioning of Geranylgeranylacetone to Enhance Bone Remodeling","authors":"Daisuke Yamaguchi, K. Takeuchi, Atsuko Ueno, Daisuke Kato, Shin Miyamae, H. Murakami","doi":"10.2485/JHTB.30.1","DOIUrl":"https://doi.org/10.2485/JHTB.30.1","url":null,"abstract":": Drug repositioning (DR) is a strategy to explore new medicinal effects from existing approved drugs whose safety and pharmacokinetics have already been established. We focused on geranylgeranylacetone (GGA), which is known as a heat shock proteins (HSPs) inducing agent. GGA is mainly used as a gastric mucosal protective agent; however, its effects on bone tissues have not been studied. Therefore, we hypothesized that “GGA induces HSPs in osteoblasts thereby promotes cell differentiation”, and administered GGA to MC3T3E-1 cells to examine cell responses. Methods: MC3T3E-1 were cultured in osteogenic medium. After the cultures were established, test cultures were exposed to GGA (GGA group). Cell proliferation, collage synthesis and ALP activity were measured on days 7 and 14 of culture. Alizarin Red S staining was performed on days 21 of culture. Results: On days 14 of culture, cell proliferation and collage synthesis were signifi cantly higher in the GGA group than in the control group (P<0.05). On days 7 and 14 of culture, ALP activity was signifi cantly higher in the GGA group than in the Control group (P<0.05). On days 28 of culture, the Alizarin Red S stained areas were significantly higher in the GGA group than in the control group (P<0.05). Conclusion: GGA promoted the differentia tion of MC3T3E-1 in an in vitro cell culture model. We hypothesized that GGA upregulates HSPs and promotes osteoblast differentiation and tested our hypothesis using a series of in vitro experiments. We investigated the responses and phenotypic changes occurring in MC3T3E-1 mouse osteoblast-like cells exposed to GGA. A 10 -3 M GGA concentration was applied to all cell cultures as preliminary assays demonstrated that it was the minimum concentration necessary to produce different calcification levels (unpub lished data).","PeriodicalId":16040,"journal":{"name":"Journal of Hard Tissue Biology","volume":null,"pages":null},"PeriodicalIF":0.4,"publicationDate":"2021-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"68963601","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
H. Ishii, Mizuki Yoshida, H. Kajiya, Satoru Matsuo, Masako Toda-Nakamura, Nana Mori-Yamamoto, Seiichi Fujisaki, K. Oka, M. Ozaki, J. Ohno
: In this study, we determined whether cisplatin can induce epithelial-mesenchymal transition (EMT) via the activation of Sonic hedgehog (Shh) or glioma-associated antigen-1 (Gli1) signaling pathway in mouse Hertwig’s epithelial root sheath (HERS) cells using a genetic knockdown approach. HERS cells treated with a low concentration of cisplatin (0.5 µM) for 24 h showed no reduction in the cell viability; however, there was a significant increase in the percentages of nu clear staining with γH2AX as compared to that with untreated control cells, indicating that 0.5 µM cisplatin induces DNA damage. Further, 0.5-µM cisplatin-treated cells provided an induction of EMT, showing decreased and increased expression of epithelial and mesenchymal markers, respectively. Enhancement in the EMT activity in cisplatin-treated HERS cells was correlated with increased expression of Shh and accelerated translocation and accumulation of Gli1 expression into the nucleus. The RNA interference-mediated silencing of Gli1 suppressed the acceleration of EMT in cisplatin-treated HERS cells; this was confirmed by no down-regulation or up-regelation in the expression of E-cadherin and vimentin, respectively, along with no increased expression of Snail expression. These findings suggest that the activation of Shh/Gli1 signaling pathway may be required for the enhancement of EMT in cisplatin-treated HERS cells. concentration the activation Shh/Gli1 signaling pathway by of DNA damage. by the following observations: (a) low concentration of cisplatin (0.5 µM) induces DNA damage; however, this concentration does not reduce the cell viability; (b) cisplatin-induced EMT is examined on the basis of downregulated and up-regulated expression of epithelial and mesenchymal markers, respectively; (c) the cispla-tin-treated HERS cells increased Shh and Gli1 expressions; (d) Transfection of Gli1 siRNA into the HERS cells reveals a significant suppression of cisplatin-induced EMT.
{"title":"Cisplatin-Induced Sonic Hedgehog Signaling Mediates Epithelial-Mesenchymal Transition in Hertwig’s Epithelial Root Sheath Cells","authors":"H. Ishii, Mizuki Yoshida, H. Kajiya, Satoru Matsuo, Masako Toda-Nakamura, Nana Mori-Yamamoto, Seiichi Fujisaki, K. Oka, M. Ozaki, J. Ohno","doi":"10.2485/JHTB.30.115","DOIUrl":"https://doi.org/10.2485/JHTB.30.115","url":null,"abstract":": In this study, we determined whether cisplatin can induce epithelial-mesenchymal transition (EMT) via the activation of Sonic hedgehog (Shh) or glioma-associated antigen-1 (Gli1) signaling pathway in mouse Hertwig’s epithelial root sheath (HERS) cells using a genetic knockdown approach. HERS cells treated with a low concentration of cisplatin (0.5 µM) for 24 h showed no reduction in the cell viability; however, there was a significant increase in the percentages of nu clear staining with γH2AX as compared to that with untreated control cells, indicating that 0.5 µM cisplatin induces DNA damage. Further, 0.5-µM cisplatin-treated cells provided an induction of EMT, showing decreased and increased expression of epithelial and mesenchymal markers, respectively. Enhancement in the EMT activity in cisplatin-treated HERS cells was correlated with increased expression of Shh and accelerated translocation and accumulation of Gli1 expression into the nucleus. The RNA interference-mediated silencing of Gli1 suppressed the acceleration of EMT in cisplatin-treated HERS cells; this was confirmed by no down-regulation or up-regelation in the expression of E-cadherin and vimentin, respectively, along with no increased expression of Snail expression. These findings suggest that the activation of Shh/Gli1 signaling pathway may be required for the enhancement of EMT in cisplatin-treated HERS cells. concentration the activation Shh/Gli1 signaling pathway by of DNA damage. by the following observations: (a) low concentration of cisplatin (0.5 µM) induces DNA damage; however, this concentration does not reduce the cell viability; (b) cisplatin-induced EMT is examined on the basis of downregulated and up-regulated expression of epithelial and mesenchymal markers, respectively; (c) the cispla-tin-treated HERS cells increased Shh and Gli1 expressions; (d) Transfection of Gli1 siRNA into the HERS cells reveals a significant suppression of cisplatin-induced EMT.","PeriodicalId":16040,"journal":{"name":"Journal of Hard Tissue Biology","volume":null,"pages":null},"PeriodicalIF":0.4,"publicationDate":"2021-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"68963979","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Ryoichi Sato, Yasuhiro Namura, N. Tanabe, M. Sakai, A. Utsu, Keiko Tomita, N. Suzuki, M. Motoyoshi
: In recent years, atmospheric pressure plasma jet (APPJ) has been widely developed for various medical applica-tions, such as medical equipment sterilization, gene transfection, cell proliferation, and wound healing. In particular, non-thermal APPJ enables direct treatment of the biological system without any thermal-associated damage. The effect of cells on APPJ depends upon the gas species used in the treatment. However, the mechanisms underlying osteoblast differen tiation mediated by APPJ with nitrogen are yet to be studied. This study investigated the effects of nitrogen-APPJ on osteo blast differentiation by assessing the transcription factors, extracellular matrix proteins (ECMPs), alkaline phosphatase (ALP) activity, and the mRNA and protein expressions of ALP, inducible nitric oxide synthase (iNOS), and cyclooxygen-ase-2 (COX-2), in an osteoblast mouse cell line. We found that nitrogen-APPJ induced osteoblast differentiation-related transcription factors (runt-related transcription factor 2 [Runx2] and osterix), osteocalcin (OCN), and ALP activity, as well as reduced the mRNA and protein expressions of iNOS and COX-2. Thus, we concluded that APPJ affects differentiation of the osteoblast cells.
{"title":"Atmospheric Pressure Plasma Treatment with Nitrogen Induces Osteoblast Differentiation and Reduces iNOS and COX-2 Expressions","authors":"Ryoichi Sato, Yasuhiro Namura, N. Tanabe, M. Sakai, A. Utsu, Keiko Tomita, N. Suzuki, M. Motoyoshi","doi":"10.2485/JHTB.30.131","DOIUrl":"https://doi.org/10.2485/JHTB.30.131","url":null,"abstract":": In recent years, atmospheric pressure plasma jet (APPJ) has been widely developed for various medical applica-tions, such as medical equipment sterilization, gene transfection, cell proliferation, and wound healing. In particular, non-thermal APPJ enables direct treatment of the biological system without any thermal-associated damage. The effect of cells on APPJ depends upon the gas species used in the treatment. However, the mechanisms underlying osteoblast differen tiation mediated by APPJ with nitrogen are yet to be studied. This study investigated the effects of nitrogen-APPJ on osteo blast differentiation by assessing the transcription factors, extracellular matrix proteins (ECMPs), alkaline phosphatase (ALP) activity, and the mRNA and protein expressions of ALP, inducible nitric oxide synthase (iNOS), and cyclooxygen-ase-2 (COX-2), in an osteoblast mouse cell line. We found that nitrogen-APPJ induced osteoblast differentiation-related transcription factors (runt-related transcription factor 2 [Runx2] and osterix), osteocalcin (OCN), and ALP activity, as well as reduced the mRNA and protein expressions of iNOS and COX-2. Thus, we concluded that APPJ affects differentiation of the osteoblast cells.","PeriodicalId":16040,"journal":{"name":"Journal of Hard Tissue Biology","volume":null,"pages":null},"PeriodicalIF":0.4,"publicationDate":"2021-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"68964145","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
京公网安备 11010802042870号
京ICP备2023020795号-1
扫码关注我们
求助内容：
应助结果提醒方式：