María F Moreno García, A. Martín, Shigeko Fushimi, S. Feldman, Nina F Pastorino, Jorge N Juárez, M. V. Jammal, L. Missana
{"title":"Optimization for Bone Samples Embedded in Methyl Methacrylate","authors":"María F Moreno García, A. Martín, Shigeko Fushimi, S. Feldman, Nina F Pastorino, Jorge N Juárez, M. V. Jammal, L. Missana","doi":"10.2485/jhtb.31.181","DOIUrl":"https://doi.org/10.2485/jhtb.31.181","url":null,"abstract":"","PeriodicalId":16040,"journal":{"name":"Journal of Hard Tissue Biology","volume":"1 1","pages":""},"PeriodicalIF":0.4,"publicationDate":"2022-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"68966184","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}
K. Kawamoto, Takuma Suzuki, T. Nagano, T. Kawamoto, K. Gomi
Titanium is most widely used for implants. Almost all the histological studies of the implants have been carried out with polished specimens and the use of frozen sections are challenging. This study focused to define the formation process of bone around titanium with frozen sections made from tissue implanted with a commercially pure titanium foil (Tifoil). We inserted the Ti-foil with a 30-μm thickness into rat femurs and extracted it after 1, 3, 5, 7, 10, 15, and 30 days. After freezing, they were sliced into 3-μm thick serial frozen sections and the sections were used for hematoxylin and eosin (H&E) staining, Masson trichrome staining, Alizarin Red S staining, enzymatic staining (ALPase and TRAPase), immunostaining (osteopontin, osteocalcin, and collagen type I), and elemental analysis. At 1 day after the implantation, the area around the Ti-foil was filled with blood and there are no collagen fibers and no proliferated cells in the area. At 3 days, the new collagen fibers were observed on the marrow side of the blood clot. The proliferated cells were observed around the fibers and they were positive for immunostaining to osteopontin, osteocalcin, and collagen type I. After 5 days, calcium precipitation was observed on the newly formed collagen fibers. After 7 days, the calcification progressed and started to reach the Ti-foil surface. After 10 days, calcification progressed to the area around the Ti-foil. At 15 days, a remarkable bone resorption appears on the marrow side. At 30 days, further resorption was observed and bony tissue was seen on the Ti-foil surface only. These results clearly showed that the bone around the Ti-foil is formed after undergoing the blood clotting process around the Ti-foil, osteoblast and fibroblast proliferation, calcium precipitation on the collagen fibers, bone formation around the Ti-foil, and bone resorption by osteoclasts.
{"title":"A Study of Bone Formation around Titanium Implants Using Frozen Sections","authors":"K. Kawamoto, Takuma Suzuki, T. Nagano, T. Kawamoto, K. Gomi","doi":"10.2485/JHTB.30.165","DOIUrl":"https://doi.org/10.2485/JHTB.30.165","url":null,"abstract":"Titanium is most widely used for implants. Almost all the histological studies of the implants have been carried out with polished specimens and the use of frozen sections are challenging. This study focused to define the formation process of bone around titanium with frozen sections made from tissue implanted with a commercially pure titanium foil (Tifoil). We inserted the Ti-foil with a 30-μm thickness into rat femurs and extracted it after 1, 3, 5, 7, 10, 15, and 30 days. After freezing, they were sliced into 3-μm thick serial frozen sections and the sections were used for hematoxylin and eosin (H&E) staining, Masson trichrome staining, Alizarin Red S staining, enzymatic staining (ALPase and TRAPase), immunostaining (osteopontin, osteocalcin, and collagen type I), and elemental analysis. At 1 day after the implantation, the area around the Ti-foil was filled with blood and there are no collagen fibers and no proliferated cells in the area. At 3 days, the new collagen fibers were observed on the marrow side of the blood clot. The proliferated cells were observed around the fibers and they were positive for immunostaining to osteopontin, osteocalcin, and collagen type I. After 5 days, calcium precipitation was observed on the newly formed collagen fibers. After 7 days, the calcification progressed and started to reach the Ti-foil surface. After 10 days, calcification progressed to the area around the Ti-foil. At 15 days, a remarkable bone resorption appears on the marrow side. At 30 days, further resorption was observed and bony tissue was seen on the Ti-foil surface only. These results clearly showed that the bone around the Ti-foil is formed after undergoing the blood clotting process around the Ti-foil, osteoblast and fibroblast proliferation, calcium precipitation on the collagen fibers, bone formation around the Ti-foil, and bone resorption by osteoclasts.","PeriodicalId":16040,"journal":{"name":"Journal of Hard Tissue Biology","volume":"1 1","pages":""},"PeriodicalIF":0.4,"publicationDate":"2021-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"68963813","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}
Gaku Kusaba, S. Matsunaga, Kei Kitamura, M. Kasahara, Yoshiaki Shimoo, S. Abe, T. Nakano, Takuya Ishimoto, Atsuhiko Hikita, K. Nojima, Y. Nishii
: Our objective was to perform a quantitative evaluation of changes in the micro/nanostructural characteristics of entheses in rats with reduced masticatory muscle functional pressure, with the aim of elucidating the mechanism whereby masticatory muscle functional pressure contributes to growth and development of the mandible from a biomechanical per-spective. Male Wister rats aged 4, 11, 18 and 25 weeks were divided into a Botox group injected with a botulinum toxin serotype A formulation to reduce muscle function (BTX) and a control group (CTRL). They were euthanized 6 weeks later and bone quality at the masseter insertion at the mandibular was analyzed. In the BTX group, the number of fibrous chon drocytes at entheses was significantly lower than in the CTRL group at all ages. The diameter of collagen fiber bundles in rats in the BTX group injected with BoNT/A during their growth phase was significantly smaller than that of rats in the CTRL group. In the mandibles of rats in the CTRL group the preferential alignment was consistent with the orientations of the muscle and tendon, but in growing rats treated with BTX, it was less closely aligned with the orientations of the muscle and tendon.
{"title":"Micro/nanostructural Characteristic Changes in the Mandibles of Rats after Injection of Botulinum Neurotoxin","authors":"Gaku Kusaba, S. Matsunaga, Kei Kitamura, M. Kasahara, Yoshiaki Shimoo, S. Abe, T. Nakano, Takuya Ishimoto, Atsuhiko Hikita, K. Nojima, Y. Nishii","doi":"10.2485/JHTB.30.183","DOIUrl":"https://doi.org/10.2485/JHTB.30.183","url":null,"abstract":": Our objective was to perform a quantitative evaluation of changes in the micro/nanostructural characteristics of entheses in rats with reduced masticatory muscle functional pressure, with the aim of elucidating the mechanism whereby masticatory muscle functional pressure contributes to growth and development of the mandible from a biomechanical per-spective. Male Wister rats aged 4, 11, 18 and 25 weeks were divided into a Botox group injected with a botulinum toxin serotype A formulation to reduce muscle function (BTX) and a control group (CTRL). They were euthanized 6 weeks later and bone quality at the masseter insertion at the mandibular was analyzed. In the BTX group, the number of fibrous chon drocytes at entheses was significantly lower than in the CTRL group at all ages. The diameter of collagen fiber bundles in rats in the BTX group injected with BoNT/A during their growth phase was significantly smaller than that of rats in the CTRL group. In the mandibles of rats in the CTRL group the preferential alignment was consistent with the orientations of the muscle and tendon, but in growing rats treated with BTX, it was less closely aligned with the orientations of the muscle and tendon.","PeriodicalId":16040,"journal":{"name":"Journal of Hard Tissue Biology","volume":"1 1","pages":""},"PeriodicalIF":0.4,"publicationDate":"2021-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"68964327","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}
S. Matsuoka, H. Hasegawa, Sachie Koike, Tsutomu Koyama, T. Takeda, K. Miura, T. Eguchi, K. Hamanaka, M. Kito, J. Takahashi, Toshirou Fukushima, T. Koizumi, K. Shimizu, T. Uehara
: This study aimed to summarize the clinicopathological findings and assess the immunophenotypes for undifferen tiated pleomorphic sarcoma of soft tissue with multinucleated giant cells (UPS-ST-MGCs). We retrospectively identified five cases of UPS-ST-MGCs between 2010 and 2020, and evaluate histological and immunohistochemical findings using osteogenic markers, which were receptor activator of nuclear factor-kappa B ligand (RANKL), runt-related transcription factor 2 (RUNX2), and special AT-rich sequence-binding protein 2 (SATB2). Cases were divided into two types, based on the distribution of multinucleated giant cells (MGCs), as diffusely (MGCs diffuse type) or focally scattering (MGCs focal type). One out of five cases was classified as MGCs diffuse type and comprised relatively monotonous proliferation of atyp ical spindle cells widely expressing RANKL, RUNX2, and SATB2. This case showed aggressive clinicopathological features, such as a rapidly growing tumor with a high maximum standardized uptake value, high Ki-67 labeling index, and early postoperative recurrence, which can be called malignant giant cell tumor (GCT-ST). Conversely, the other four cases of the MGCs focal type were focally positive for RANKL, and negative for RUNX2 and STAB2, which appeared to be consistent with conventional features of undifferentiated pleomorphic sarcoma of soft tissue. Our results indicate that malignant GCT-ST can be included in UPS-ST-MGCs. Therefore, it is important to note its aggressive malignant characteristics and osteogenic differentiation. Osteogenic immunohistochemical examinations should be considered for UPS-ST-MGCs to con firm an accurate diagnosis and provide appropriate treatment.
{"title":"Undifferentiated Pleomorphic Sarcoma of Soft Tissue with Multinucleated Giant Cells with Osteogenic Phenotypes: A Mimicker of Malignant Giant Cell Tumor of Soft Tissue","authors":"S. Matsuoka, H. Hasegawa, Sachie Koike, Tsutomu Koyama, T. Takeda, K. Miura, T. Eguchi, K. Hamanaka, M. Kito, J. Takahashi, Toshirou Fukushima, T. Koizumi, K. Shimizu, T. Uehara","doi":"10.2485/jhtb.30.309","DOIUrl":"https://doi.org/10.2485/jhtb.30.309","url":null,"abstract":": This study aimed to summarize the clinicopathological findings and assess the immunophenotypes for undifferen tiated pleomorphic sarcoma of soft tissue with multinucleated giant cells (UPS-ST-MGCs). We retrospectively identified five cases of UPS-ST-MGCs between 2010 and 2020, and evaluate histological and immunohistochemical findings using osteogenic markers, which were receptor activator of nuclear factor-kappa B ligand (RANKL), runt-related transcription factor 2 (RUNX2), and special AT-rich sequence-binding protein 2 (SATB2). Cases were divided into two types, based on the distribution of multinucleated giant cells (MGCs), as diffusely (MGCs diffuse type) or focally scattering (MGCs focal type). One out of five cases was classified as MGCs diffuse type and comprised relatively monotonous proliferation of atyp ical spindle cells widely expressing RANKL, RUNX2, and SATB2. This case showed aggressive clinicopathological features, such as a rapidly growing tumor with a high maximum standardized uptake value, high Ki-67 labeling index, and early postoperative recurrence, which can be called malignant giant cell tumor (GCT-ST). Conversely, the other four cases of the MGCs focal type were focally positive for RANKL, and negative for RUNX2 and STAB2, which appeared to be consistent with conventional features of undifferentiated pleomorphic sarcoma of soft tissue. Our results indicate that malignant GCT-ST can be included in UPS-ST-MGCs. Therefore, it is important to note its aggressive malignant characteristics and osteogenic differentiation. Osteogenic immunohistochemical examinations should be considered for UPS-ST-MGCs to con firm an accurate diagnosis and provide appropriate treatment.","PeriodicalId":16040,"journal":{"name":"Journal of Hard Tissue Biology","volume":"1 1","pages":""},"PeriodicalIF":0.4,"publicationDate":"2021-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"68964827","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}
Miki Hori, Makoto Jincho, Tadasuke Hori, Hironao Sekine, A. Kato, Atsuko Ueno, T. Kawai
: Most studies of artificial intelligence in the medical field involve classification problems, but few consider recog nition of one characteristic point in images or regression analysis such as data recognition. In this research, we constructed a fundamental convolutional neural network framework for regression analysis. Images of the handwritten digit “3” from the MNIST dataset were used as training data, with the protruding middle point as an image feature point. Input images and training data (x1, y1) were connected to 6 convolutional layers and then run through 2 affine layers to produce the output data (x2, y2). The loss function was the mean radial error (MRE) between the training and output data. After machine learn ing, the error converged to 0.75 pixels on average. We expect that this algorithm can be clinically applied to points having certain characteristics in images, such as locating hard tissue lesions or recognizing measurement points in cephalograms.
{"title":"Recognition of Image One Feature Point Using Convolutional Neural Networks","authors":"Miki Hori, Makoto Jincho, Tadasuke Hori, Hironao Sekine, A. Kato, Atsuko Ueno, T. Kawai","doi":"10.2485/JHTB.30.161","DOIUrl":"https://doi.org/10.2485/JHTB.30.161","url":null,"abstract":": Most studies of artificial intelligence in the medical field involve classification problems, but few consider recog nition of one characteristic point in images or regression analysis such as data recognition. In this research, we constructed a fundamental convolutional neural network framework for regression analysis. Images of the handwritten digit “3” from the MNIST dataset were used as training data, with the protruding middle point as an image feature point. Input images and training data (x1, y1) were connected to 6 convolutional layers and then run through 2 affine layers to produce the output data (x2, y2). The loss function was the mean radial error (MRE) between the training and output data. After machine learn ing, the error converged to 0.75 pixels on average. We expect that this algorithm can be clinically applied to points having certain characteristics in images, such as locating hard tissue lesions or recognizing measurement points in cephalograms.","PeriodicalId":16040,"journal":{"name":"Journal of Hard Tissue Biology","volume":"53 1","pages":""},"PeriodicalIF":0.4,"publicationDate":"2021-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"68964272","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":"1 1","pages":""},"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}
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":"50 1","pages":""},"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}
Hiroki Nakashima, M. Yasunaga, Mizuki Yoshida, M. Yamaguchi, Saki Takahashi, H. Kajiya, Sachio Tamaoki, J. Ohno
: This study aimed to examine whether a low concentration of etoposide can accelerate osteogenesis via the activation of peptidyl-prolyl isomerase Pin1 in human MG63 cells using a genetic knockdown approach. MG63 cells treated with 0.1 μM etoposide for 24 h showed neither reduction in cell viability nor induction of cellular senescence; however, there was a significant increase in the percentage of nuclear staining with γH2AX as compared with that in untreated control cells, indicating that 0.1 μM etoposide induces weak DNA damage response (DDR) in the cells. Treatment with 0.1 μM etoposide accelerates osteogenesis in osteogenic induction medium (OIM)-cultured MG63 cells, demonstrating increased expression of Runx2 and Osterix and intense alkaline phosphatase (ALP) staining, as compared with cells treated without etoposide. In addition to those osteogenic markers, Pin1 expression was upregulated in the etoposide-treated cells, suggesting that the weak DDR may provide an interaction between Pin1 activation and osteogenic markers. The RNA interfer -ence-mediated silencing of Pin1 suppressed the expression of osteogenic markers and ALP staining in the OIM-cultured cells pretreated with 0.1 μM etoposide. Based on these findings, we suggest that a low concentration of etoposide induces mild DDR to activate Pin1, eventually promoting OIM-induced osteogenesis in the MG63 cells.
{"title":"Low Concentration of Etoposide Induces Enhanced Osteogenesis in MG63 Cells via Pin1 Activation","authors":"Hiroki Nakashima, M. Yasunaga, Mizuki Yoshida, M. Yamaguchi, Saki Takahashi, H. Kajiya, Sachio Tamaoki, J. Ohno","doi":"10.2485/JHTB.30.175","DOIUrl":"https://doi.org/10.2485/JHTB.30.175","url":null,"abstract":": This study aimed to examine whether a low concentration of etoposide can accelerate osteogenesis via the activation of peptidyl-prolyl isomerase Pin1 in human MG63 cells using a genetic knockdown approach. MG63 cells treated with 0.1 μM etoposide for 24 h showed neither reduction in cell viability nor induction of cellular senescence; however, there was a significant increase in the percentage of nuclear staining with γH2AX as compared with that in untreated control cells, indicating that 0.1 μM etoposide induces weak DNA damage response (DDR) in the cells. Treatment with 0.1 μM etoposide accelerates osteogenesis in osteogenic induction medium (OIM)-cultured MG63 cells, demonstrating increased expression of Runx2 and Osterix and intense alkaline phosphatase (ALP) staining, as compared with cells treated without etoposide. In addition to those osteogenic markers, Pin1 expression was upregulated in the etoposide-treated cells, suggesting that the weak DDR may provide an interaction between Pin1 activation and osteogenic markers. The RNA interfer -ence-mediated silencing of Pin1 suppressed the expression of osteogenic markers and ALP staining in the OIM-cultured cells pretreated with 0.1 μM etoposide. Based on these findings, we suggest that a low concentration of etoposide induces mild DDR to activate Pin1, eventually promoting OIM-induced osteogenesis in the MG63 cells.","PeriodicalId":16040,"journal":{"name":"Journal of Hard Tissue Biology","volume":"1 1","pages":""},"PeriodicalIF":0.4,"publicationDate":"2021-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"68963828","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}
: We aimed to detect the expressions of Yes-associated protein (YAP) and transcriptional co-activator with PDZ-binding motif (TAZ) in different development stages of mandibular first molar and incisor in BALB/c mice, and to un cover the action mechanisms of YAP and TAZ during tooth development. BALB/c mice were randomly divided into 5 groups, i.e. 19.5-d-old embryo, and 0-, 6-, 14- and 28-d-old mice (n=8). To detect the expression changes of YAP and TAZ during tooth development, the mandibular tissues of first molar and incisor were collected and subjected to HE staining and SABC immunohistochemical staining. YAP expression was observed in the molar gem of 19.5-d-old embryo, which increased gradually in mice on 0, 6, 14 and 28 d after birth. TAZ was expressed in granule shape in the molar germ and tooth of 19.5-d-old embryo and 0-d-old mice. Expression of TAZ was observed on 6, 14 and 28 d after birth, which was the low-est on 6 d after birth and rose on 14 and 28 d. YAP and TAZ are specifically expressed and localized during the development of mandibular first molars of mice, probably being involved in ameloblast and odontoblast differentiation and dentin calcifi -cation.
{"title":"Expressions of Yes-associated Protein and Transcriptional Co-Activator with PDZ-binding Motif during the Development of Mandibular First Molar in BALB/c Mice","authors":"Xuliang Ma, Chao Wang, Yunmeng Da, Ruiqing Cheng, Jing-ju Zhang, Xiaotong Qiao","doi":"10.2485/JHTB.30.145","DOIUrl":"https://doi.org/10.2485/JHTB.30.145","url":null,"abstract":": We aimed to detect the expressions of Yes-associated protein (YAP) and transcriptional co-activator with PDZ-binding motif (TAZ) in different development stages of mandibular first molar and incisor in BALB/c mice, and to un cover the action mechanisms of YAP and TAZ during tooth development. BALB/c mice were randomly divided into 5 groups, i.e. 19.5-d-old embryo, and 0-, 6-, 14- and 28-d-old mice (n=8). To detect the expression changes of YAP and TAZ during tooth development, the mandibular tissues of first molar and incisor were collected and subjected to HE staining and SABC immunohistochemical staining. YAP expression was observed in the molar gem of 19.5-d-old embryo, which increased gradually in mice on 0, 6, 14 and 28 d after birth. TAZ was expressed in granule shape in the molar germ and tooth of 19.5-d-old embryo and 0-d-old mice. Expression of TAZ was observed on 6, 14 and 28 d after birth, which was the low-est on 6 d after birth and rose on 14 and 28 d. YAP and TAZ are specifically expressed and localized during the development of mandibular first molars of mice, probably being involved in ameloblast and odontoblast differentiation and dentin calcifi -cation.","PeriodicalId":16040,"journal":{"name":"Journal of Hard Tissue Biology","volume":"1 1","pages":""},"PeriodicalIF":0.4,"publicationDate":"2021-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"68964216","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}
Mingmei Yang, Lin Gao, San Cai, Li Gao, Qi Zhang, Chun Gui
: MCT1 is an important regulator in glycolysis and has significant effects on inflammatory responses and osteoclast differentiation etc. This study was to study the effects and mechanism of MCT1 in chondrocytes injury and inflammatory responses. ATDC5 cells with stably transfection of MCT1shRNA were treated with 5 μg/mL of LPS. Cell viability was de termined by MTT assay. The mRNA and protein expressions were detected by qRT-PCR and western blotting, respectively. The concentrations of cytokines in culture medium were measured by ELISA. ROS generation was tested by 2,7-dichloro fluorescein diacetate (DCFH-DA). The results showed that MCT1 was increased by LPS treatment in ATDC5 cells in a dose dependent manner. MCT1 knockdown improved the survival of LPS-treated ATDC5 cells. MCT1 knockdown also de creased LPS-induced expression of pro-inflammatory cytokines (IL-1β, IL-6, IL-8 and TNFα) and oxidative stress media tors (iNOS, COX-2 and NOX-4) in ATDC5 cell. Importantly, PFKFB3 overexpression reversed the anti-inflammatory and anti-oxidative stress effects of MCT1 knockdown in LPS-induced ATDC5 cells. These results indicated that MCT1 knock out decreased the expression of inflammatory mediators and oxidative stress mediators induced by LPS through regulating PFKFB3. The study provides a potential target for the prevention or treatment of osteoarthritis (OA) and rheumatoid arthritis (RA).
{"title":"Down-Regulation of MCT1 Ameliorates LPS-Induced Cell Injury in Murine Chondrocyte-like ATDC5 Cells by Regulation of PFKFB3","authors":"Mingmei Yang, Lin Gao, San Cai, Li Gao, Qi Zhang, Chun Gui","doi":"10.2485/jhtb.30.251","DOIUrl":"https://doi.org/10.2485/jhtb.30.251","url":null,"abstract":": MCT1 is an important regulator in glycolysis and has significant effects on inflammatory responses and osteoclast differentiation etc. This study was to study the effects and mechanism of MCT1 in chondrocytes injury and inflammatory responses. ATDC5 cells with stably transfection of MCT1shRNA were treated with 5 μg/mL of LPS. Cell viability was de termined by MTT assay. The mRNA and protein expressions were detected by qRT-PCR and western blotting, respectively. The concentrations of cytokines in culture medium were measured by ELISA. ROS generation was tested by 2,7-dichloro fluorescein diacetate (DCFH-DA). The results showed that MCT1 was increased by LPS treatment in ATDC5 cells in a dose dependent manner. MCT1 knockdown improved the survival of LPS-treated ATDC5 cells. MCT1 knockdown also de creased LPS-induced expression of pro-inflammatory cytokines (IL-1β, IL-6, IL-8 and TNFα) and oxidative stress media tors (iNOS, COX-2 and NOX-4) in ATDC5 cell. Importantly, PFKFB3 overexpression reversed the anti-inflammatory and anti-oxidative stress effects of MCT1 knockdown in LPS-induced ATDC5 cells. These results indicated that MCT1 knock out decreased the expression of inflammatory mediators and oxidative stress mediators induced by LPS through regulating PFKFB3. The study provides a potential target for the prevention or treatment of osteoarthritis (OA) and rheumatoid arthritis (RA).","PeriodicalId":16040,"journal":{"name":"Journal of Hard Tissue Biology","volume":"1 1","pages":""},"PeriodicalIF":0.4,"publicationDate":"2021-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"68964300","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}
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