{"title":"Inhibitory Effects of Resveratrol on Inflammatory Response in Rat Dental Pulp","authors":"L. Hu, Hao Shen, Shiliang Guo","doi":"10.2485/jhtb.31.135","DOIUrl":"https://doi.org/10.2485/jhtb.31.135","url":null,"abstract":"","PeriodicalId":16040,"journal":{"name":"Journal of Hard Tissue Biology","volume":null,"pages":null},"PeriodicalIF":0.4,"publicationDate":"2022-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"68965997","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}
Yusuke Oomura, S. Matsunaga, M. Okamura, Taiki Suzuki, N. Kasahara, S. Abe, Takeshi Nomura
: The objective of this study was to determine the effect of zoledronic acid on mandibular bone quality in osteo porotic model mice. Zoledronic acid was administered to ovariectomized mice, and mandibular bone was harvested. Pol ished specimens were prepared, and the biological apatite (BAp) crystal alignment and changes in collagen fiber bundles in the alveolar and basal regions of the mandible were analyzed. It was found that ovariectomy increased BAp crystal align -ment. The administration of zoledronic acid post-ovariectomy normalized BAp crystal alignment in the basal region of the mandible. However, BAp crystal alignment in the alveolar region decreased significantly. Ovariectomy decreased the diam eters of collagen fiber bundles in both the alveolar and basal regions and significantly increased their lengths. The adminis tration of zoledronic acid post-ovariectomy decreased both the diameters and lengths of collagen fiber bundles. These re sults showed that the microarchitecture of the mandibular bone changes to compensate for osteoporosis-induced loss of bone mass and adapts to the load environment resulting from mastication. Interestingly, they suggested that zoledronic acid severely reduces the bone quality of osteoporotic alveolar bone in a site-specific manner.
{"title":"Effect of Zoledronic Acid on Bone Structure of the Mandible in Ovariectomized Mice","authors":"Yusuke Oomura, S. Matsunaga, M. Okamura, Taiki Suzuki, N. Kasahara, S. Abe, Takeshi Nomura","doi":"10.2485/jhtb.31.207","DOIUrl":"https://doi.org/10.2485/jhtb.31.207","url":null,"abstract":": The objective of this study was to determine the effect of zoledronic acid on mandibular bone quality in osteo porotic model mice. Zoledronic acid was administered to ovariectomized mice, and mandibular bone was harvested. Pol ished specimens were prepared, and the biological apatite (BAp) crystal alignment and changes in collagen fiber bundles in the alveolar and basal regions of the mandible were analyzed. It was found that ovariectomy increased BAp crystal align -ment. The administration of zoledronic acid post-ovariectomy normalized BAp crystal alignment in the basal region of the mandible. However, BAp crystal alignment in the alveolar region decreased significantly. Ovariectomy decreased the diam eters of collagen fiber bundles in both the alveolar and basal regions and significantly increased their lengths. The adminis tration of zoledronic acid post-ovariectomy decreased both the diameters and lengths of collagen fiber bundles. These re sults showed that the microarchitecture of the mandibular bone changes to compensate for osteoporosis-induced loss of bone mass and adapts to the load environment resulting from mastication. Interestingly, they suggested that zoledronic acid severely reduces the bone quality of osteoporotic alveolar bone in a site-specific manner.","PeriodicalId":16040,"journal":{"name":"Journal of Hard Tissue Biology","volume":null,"pages":null},"PeriodicalIF":0.4,"publicationDate":"2022-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"68966275","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}
Yue Cai, Xue-xin Tan, Li Zhao, Ran Zhang, Tong Zhu, Yang Du, Xukai Wang
{"title":"Withdrawal: Synthesis of a Novel bFGF/nHAP/COL Bone Tissue Engineering Scaffold for Mandibular Defect Regeneration in a Rabbit Model [Journal of Hard Tissue Biology 27(1) Vol.27, 2018, pp 85-94]","authors":"Yue Cai, Xue-xin Tan, Li Zhao, Ran Zhang, Tong Zhu, Yang Du, Xukai Wang","doi":"10.2485/jhtb.31.61","DOIUrl":"https://doi.org/10.2485/jhtb.31.61","url":null,"abstract":"","PeriodicalId":16040,"journal":{"name":"Journal of Hard Tissue Biology","volume":null,"pages":null},"PeriodicalIF":0.4,"publicationDate":"2022-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"68966479","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}
Fumitaka Yoshino, Rina Sasaki, Yuka Asada, K. Shiozaki, S. Shimoda, Takatsugu Yamamoto
In this study, we immersed amorphous calcium phosphate (ACP) powder in biochemical buffer solutions and performed analysis of its solubility and phase transformation of the precipitate. After preparing ACP powder that contains no impurities, we used 4-(2-hydroxyethyl)-1-piperazinetethanesulfonic acid (HEPES) buffer, one of the good buffers, as a buffer solution and measured the amount of calcium ions eluted from ACP and other calcium phosphate crystals. ACP was immersed in the buffer solution at 5°C, 20°C, and 37°C, and the amount of eluted calcium ions was measured from 15 min to 24 h thereafter. The precipitated solid phase was analyzed using X-ray diffraction and its morphology was observed using transmission electron microscopy. The precipitation of hydroxyapatite (HAp) was observed after 15 min in HEPES buffer solution. Furthermore, in this experimental group, the precipitates of the sample incubated in HEPES buffer solution at 37°C for 24 h produced the largest HAp crystals. From these results we concluded that ACP immersed in HEPES buffer solution easily releases calcium ions and phosphate ions, and a rapid phase transformation to HAp occurs. Moreover, we assume that, in addition to the thermodynamic effect, the crystal growth of HAp is enhanced by the buffer solution.
{"title":"Studies on Change in Solubility over Time of the Bioactive Material Amorphous Calcium Phosphate and Precipitation of Hydroxyapatite","authors":"Fumitaka Yoshino, Rina Sasaki, Yuka Asada, K. Shiozaki, S. Shimoda, Takatsugu Yamamoto","doi":"10.2485/jhtb.31.1","DOIUrl":"https://doi.org/10.2485/jhtb.31.1","url":null,"abstract":"In this study, we immersed amorphous calcium phosphate (ACP) powder in biochemical buffer solutions and performed analysis of its solubility and phase transformation of the precipitate. After preparing ACP powder that contains no impurities, we used 4-(2-hydroxyethyl)-1-piperazinetethanesulfonic acid (HEPES) buffer, one of the good buffers, as a buffer solution and measured the amount of calcium ions eluted from ACP and other calcium phosphate crystals. ACP was immersed in the buffer solution at 5°C, 20°C, and 37°C, and the amount of eluted calcium ions was measured from 15 min to 24 h thereafter. The precipitated solid phase was analyzed using X-ray diffraction and its morphology was observed using transmission electron microscopy. The precipitation of hydroxyapatite (HAp) was observed after 15 min in HEPES buffer solution. Furthermore, in this experimental group, the precipitates of the sample incubated in HEPES buffer solution at 37°C for 24 h produced the largest HAp crystals. From these results we concluded that ACP immersed in HEPES buffer solution easily releases calcium ions and phosphate ions, and a rapid phase transformation to HAp occurs. Moreover, we assume that, in addition to the thermodynamic effect, the crystal growth of HAp is enhanced by the buffer solution.","PeriodicalId":16040,"journal":{"name":"Journal of Hard Tissue Biology","volume":null,"pages":null},"PeriodicalIF":0.4,"publicationDate":"2022-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"68965873","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}
Le Wang, Xingqiang Wang, Rui Liang, Shusen Wang, Jinglin Cao
{"title":"A Comparison of Mesenchymal Stem Cells from Human Adipose Tissues by Resection and by Liposuction","authors":"Le Wang, Xingqiang Wang, Rui Liang, Shusen Wang, Jinglin Cao","doi":"10.2485/jhtb.31.15","DOIUrl":"https://doi.org/10.2485/jhtb.31.15","url":null,"abstract":"","PeriodicalId":16040,"journal":{"name":"Journal of Hard Tissue Biology","volume":null,"pages":null},"PeriodicalIF":0.4,"publicationDate":"2022-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"68966064","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 aim of this study was to identify genes that are prominently expressed in rat bone marrow cell-derived osteoblasts during the initial phase of low-intensity pulsed ultrasound (LIPUS) exposure. Bone marrow cells were obtained from three Sprague-Dawley rats (8-week-old, male), and cell cultures were prepared by suspension in osteogenic medium. After cultures were established, test cultures were exposed to LIPUS from the bottom of the cell culture plate for 15 min/d on days 1–4 (LIPUS group). LIPUS signals were transmitted at a frequency of 3 MHz and a spatial average intensity of 40 mW/cm 2 . The control group was not exposed to LIPUS. On day 14, alizarin red S staining was performed to detect calcifi -cation. On day 4, total RNA was extracted from both cultures, hybridized to microarray slides, and the resulting data set was analyzed. Genes exhibiting a fold-change ≥2 and a p-value <0.05 (LIPUS vs. control) were identified as differentially expressed genes. Pathway analysis was performed on genes whose expression increased in the LIPUS group. The cellular areas stained with alizarin red S were significantly larger in the LIPUS group than in the control group on day 14. LIPUS exposure increased the expression of genes related to type II interferon signaling, and endochondral ossification was ob served after 4 d of culture. The results demonstrated that LIPUS exposure activated the immune response and promoted osteoblast differentiation.
本研究的目的是鉴定在低强度脉冲超声(LIPUS)暴露初始阶段在大鼠骨髓细胞来源的成骨细胞中显著表达的基因。取3只8周龄雄性Sprague-Dawley大鼠骨髓细胞，在成骨培养基中悬浮培养细胞。培养完成后，在第1-4天(LIPUS组)将实验培养物从细胞培养板底部暴露于LIPUS中15 min/d。LIPUS信号以3 MHz的频率和40 mW/ cm2的空间平均强度传输。对照组不使用LIPUS。第14天，茜素红S染色检测钙化。第4天，从两个培养物中提取总RNA，将其杂交到微阵列载玻片上，并分析结果数据集。fold-change≥2且p值<0.05 (LIPUS vs. control)的基因被鉴定为差异表达基因。对LIPUS组中表达增高的基因进行通路分析。第14天，LIPUS组茜素红S染色的细胞面积明显大于对照组。LIPUS暴露增加了II型干扰素信号相关基因的表达，培养4 d后观察到软骨内成骨。结果表明，LIPUS暴露激活了免疫反应，促进了成骨细胞的分化。
{"title":"Gene Expression in Early Stages of Low-Intensity Pulsed Ultrasound Exposure on Bone Marrow Cells","authors":"Daisuke Yamaguchi, Kazuo Takeuchi, Atsuko Ueno, Masataka Yamaguchi, H. Murakami, Suguru Kimoto","doi":"10.2485/jhtb.31.23","DOIUrl":"https://doi.org/10.2485/jhtb.31.23","url":null,"abstract":": The aim of this study was to identify genes that are prominently expressed in rat bone marrow cell-derived osteoblasts during the initial phase of low-intensity pulsed ultrasound (LIPUS) exposure. Bone marrow cells were obtained from three Sprague-Dawley rats (8-week-old, male), and cell cultures were prepared by suspension in osteogenic medium. After cultures were established, test cultures were exposed to LIPUS from the bottom of the cell culture plate for 15 min/d on days 1–4 (LIPUS group). LIPUS signals were transmitted at a frequency of 3 MHz and a spatial average intensity of 40 mW/cm 2 . The control group was not exposed to LIPUS. On day 14, alizarin red S staining was performed to detect calcifi -cation. On day 4, total RNA was extracted from both cultures, hybridized to microarray slides, and the resulting data set was analyzed. Genes exhibiting a fold-change ≥2 and a p-value <0.05 (LIPUS vs. control) were identified as differentially expressed genes. Pathway analysis was performed on genes whose expression increased in the LIPUS group. The cellular areas stained with alizarin red S were significantly larger in the LIPUS group than in the control group on day 14. LIPUS exposure increased the expression of genes related to type II interferon signaling, and endochondral ossification was ob served after 4 d of culture. The results demonstrated that LIPUS exposure activated the immune response and promoted osteoblast differentiation.","PeriodicalId":16040,"journal":{"name":"Journal of Hard Tissue Biology","volume":null,"pages":null},"PeriodicalIF":0.4,"publicationDate":"2022-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"68966392","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}
E. Kobayashi, Haruka Takahashi, Kensuke Igarashi, Hiroki Katagiri, M. Haga-Tsujimura, I. Ogura, K. Nakahara, A. Tanaka
: In 2014, the American Association of Oral and Maxillofacial Surgery (AAOMS) recommended surgical treatment for medication-related osteonecrosis of the jaw (MRONJ) patients classified as Stage 3. In 2016, the Japanese position pa per recommended surgical treatment classified as Stage 2. Some systematic review reported effectiveness of surgical treat ment for MRONJ. However there is no concerted consciousness on the treatment methods of MRONJ patients in the pres-ent. This study aimed to retrospectively elucidate clinical outcomes of the surgical treatment of MRONJ patients under the same criteria. This study included 86 patients in 40 osteoporosis patients (5 men and 35 women, average age: 78.8 years) and 46 cancer patients (18 men and 28 women, average age: 67.7 years). The outcome was classified into two categories: Healing or No healing in each stage. Among 86 patients, MRONJ was found in 91 jaws, of which 69 jaws (75.8%) were surgically operated. Total healing rate of surgical treatment was 85.5%, and total healing rate of conservative treatment was 4.5%. It was suggested that surgical treatment for MRONJ had a high clinical response rate in all stages.
{"title":"Prognosis of Medication-Related Osteonecrosis of the Jaw with Surgical Treatment","authors":"E. Kobayashi, Haruka Takahashi, Kensuke Igarashi, Hiroki Katagiri, M. Haga-Tsujimura, I. Ogura, K. Nakahara, A. Tanaka","doi":"10.2485/jhtb.31.39","DOIUrl":"https://doi.org/10.2485/jhtb.31.39","url":null,"abstract":": In 2014, the American Association of Oral and Maxillofacial Surgery (AAOMS) recommended surgical treatment for medication-related osteonecrosis of the jaw (MRONJ) patients classified as Stage 3. In 2016, the Japanese position pa per recommended surgical treatment classified as Stage 2. Some systematic review reported effectiveness of surgical treat ment for MRONJ. However there is no concerted consciousness on the treatment methods of MRONJ patients in the pres-ent. This study aimed to retrospectively elucidate clinical outcomes of the surgical treatment of MRONJ patients under the same criteria. This study included 86 patients in 40 osteoporosis patients (5 men and 35 women, average age: 78.8 years) and 46 cancer patients (18 men and 28 women, average age: 67.7 years). The outcome was classified into two categories: Healing or No healing in each stage. Among 86 patients, MRONJ was found in 91 jaws, of which 69 jaws (75.8%) were surgically operated. Total healing rate of surgical treatment was 85.5%, and total healing rate of conservative treatment was 4.5%. It was suggested that surgical treatment for MRONJ had a high clinical response rate in all stages.","PeriodicalId":16040,"journal":{"name":"Journal of Hard Tissue Biology","volume":null,"pages":null},"PeriodicalIF":0.4,"publicationDate":"2022-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"68966464","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. Miyazawa, Takeo Sekiya, Misuzu Kawaguchi, Yuji Kojima, M. Tabuchi, Takuma Sato, T. Kawai, S. Goto
: Bone morphogenetic protein (BMP) is the only cytokine that induces heterotopic new bone. BMP has been classi-fied as a subfamily of the TGF superfamily. The most important and essential aspect for the clinical application of BMP is the replenishment and reinforcement of bone defects and fragile areas with new bone. However, it is difficult to ascertain long term clinical results. In this study, we examined the osteogenic potential in mouse femoral fascia of crude BMP ex-tracted from bovine bone that had been stored at room temperature for 25 years. The results showed vigorous osteogenesis. This study demonstrates the long-term stability of crude BMP. study, we examined the osteogenic potential of crude BMP (cBMP) ex-tracted from and the material exhibited vigorous This study reports the long-term stability of the cBMP osteogenic potential.
{"title":"Vigorous Osteoinductivity Observed in Crude Bone Morphogenetic Protein Stored for 25 Years after Extraction at Room Temperature","authors":"K. Miyazawa, Takeo Sekiya, Misuzu Kawaguchi, Yuji Kojima, M. Tabuchi, Takuma Sato, T. Kawai, S. Goto","doi":"10.2485/jhtb.31.253","DOIUrl":"https://doi.org/10.2485/jhtb.31.253","url":null,"abstract":": Bone morphogenetic protein (BMP) is the only cytokine that induces heterotopic new bone. BMP has been classi-fied as a subfamily of the TGF superfamily. The most important and essential aspect for the clinical application of BMP is the replenishment and reinforcement of bone defects and fragile areas with new bone. However, it is difficult to ascertain long term clinical results. In this study, we examined the osteogenic potential in mouse femoral fascia of crude BMP ex-tracted from bovine bone that had been stored at room temperature for 25 years. The results showed vigorous osteogenesis. This study demonstrates the long-term stability of crude BMP. study, we examined the osteogenic potential of crude BMP (cBMP) ex-tracted from and the material exhibited vigorous This study reports the long-term stability of the cBMP osteogenic potential.","PeriodicalId":16040,"journal":{"name":"Journal of Hard Tissue Biology","volume":null,"pages":null},"PeriodicalIF":0.4,"publicationDate":"2022-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"68966419","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}
Qingchao Wu, I. Yamawaki, Y. Taguchi, Kei Shiomi, Daisuke Kimura, Tsurayuki Takahashi, M. Umeda
Patients with diabetes mellitus (DM) are at an increased risk of tooth loss compared to healthy individuals. Although studies human subjects suggest that diabetes control is affected by periodontitis, there is scarce mechanistic evidence supporting its biological plausibility. Therefore, using type 2 DM rat bone marrow mesenchymal stromal cells (BMMSCs) were incubated with four concentrations of glucose (5.5, 8.0, 12.0 or 24.0 mM), the effects of different glucose concentrations on BMMSCs stemness and osteogenesis were evaluated. High glucose concentrations decreased the fluorescence intensity of β-actin, STRO-1, CD73 and CD90. Moreover, cell proliferation decreased at high glucose concentrations. Alkaline phosphatase activity was decreased at 12.0 mM and 24.0 mM. In contrast, osteocalcin production and calcium deposition were considerably increased at 24.0 mM. Differences in the calcium/phosphate ratio associated with various glucose concentrations were similar to calcium deposition. The mRNA expression of Runx2 and inflammatory cytokines increased with increasing glucose concentration. The RANKL/OPG ratio decreased at high glucose concentrations. A high glucose concentration increased hard tissue formation, but the quality and stemness of the mineralized tissue decreased. Thus, hard tissue had a high risk of bone resorption in the case of uncontrolled diabetes even if periodontal treatment stabilized state of periodontitis for a moment.
{"title":"Glucose Affects the Quality and Properties of Hard Tissue in Diabetes Mellitus Model","authors":"Qingchao Wu, I. Yamawaki, Y. Taguchi, Kei Shiomi, Daisuke Kimura, Tsurayuki Takahashi, M. Umeda","doi":"10.2485/jhtb.31.29","DOIUrl":"https://doi.org/10.2485/jhtb.31.29","url":null,"abstract":"Patients with diabetes mellitus (DM) are at an increased risk of tooth loss compared to healthy individuals. Although studies human subjects suggest that diabetes control is affected by periodontitis, there is scarce mechanistic evidence supporting its biological plausibility. Therefore, using type 2 DM rat bone marrow mesenchymal stromal cells (BMMSCs) were incubated with four concentrations of glucose (5.5, 8.0, 12.0 or 24.0 mM), the effects of different glucose concentrations on BMMSCs stemness and osteogenesis were evaluated. High glucose concentrations decreased the fluorescence intensity of β-actin, STRO-1, CD73 and CD90. Moreover, cell proliferation decreased at high glucose concentrations. Alkaline phosphatase activity was decreased at 12.0 mM and 24.0 mM. In contrast, osteocalcin production and calcium deposition were considerably increased at 24.0 mM. Differences in the calcium/phosphate ratio associated with various glucose concentrations were similar to calcium deposition. The mRNA expression of Runx2 and inflammatory cytokines increased with increasing glucose concentration. The RANKL/OPG ratio decreased at high glucose concentrations. A high glucose concentration increased hard tissue formation, but the quality and stemness of the mineralized tissue decreased. Thus, hard tissue had a high risk of bone resorption in the case of uncontrolled diabetes even if periodontal treatment stabilized state of periodontitis for a moment.","PeriodicalId":16040,"journal":{"name":"Journal of Hard Tissue Biology","volume":null,"pages":null},"PeriodicalIF":0.4,"publicationDate":"2022-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"68966460","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}
Huai-chun Chang, Tingting Jiang, Liang Kou, Duo Li, Xinchen Yu, Youqin Li, Lei Zhang
: We aimed to evaluate the regulatory effects of miR-148a-3p on stem cell osteogenic differentiation and enamel development by targeting runt-related transcription factor 2 (RUNX2) and E-cadherin, respectively. TargetScan software was utilized to predict the binding sites between miR-148a-3p and osteogenic marker gene RUNX2 or E-cadherin. The changes in miR-148a-3p expression during osteogenic differentiation of epidermal stem cells were detected. After transfec tion with miR-148a-3p mimics or miR-148a-3p inhibitor, epidermal stem cells were induced towards osteogenic differentia tion, and the changes in RUNX2 expression were measured. The changes in miR-148a-3p expression during enamel devel opment regulated by epidermal stem cells were determined. After liposome-mediated transfection with miR-148a-3p mimics or miR-148a-3p inhibitor, epidermal stem cells were induced towards ameloblast development. Cell proliferation and apoptosis abilities were tested using methyl thiazolyl tetrazolium assay and flow cytometry, respectively. The expres sion of miR-148a-3p was detected by RT-PCR. The protein expressions of Wnt1, β-catenin, RUNX2 and E-cadherin were measured by Western blotting. Epidermal stem cells differentiated into osteoblasts through osteogenic induction culture. On 5, 12, 15 and 30 d, epidermal stem cells gradually differentiated into osteoblasts through epithelial aggregation and depres sion, mesenchymal aggregation, and dentin and enamel secretion. After transfection, compared with negative control (NC) group, the cell viability of miR-148-3p group significantly decreased (P<0.05), and the apoptosis rate increased (P<0.01). The viability of miR-148-3p inhibitor group significantly increased (P<0.05), while the apoptosis rate reduced (P<0.01). The dual-luciferase reporter assay showed that miR-148a-3p targeted Wnt1. Compared with NC group, the expression of miR-148a-3p significantly rose in miR-148a-3p group (P<0.001), and the protein expression levels of Wnt1, β-catenin, RUNX2 and E-cadherin significantly decreased. Compared with NC group, the expression of miR-148a-3p in miR-148a-3p inhibitor group significantly decreased (P<0.05), and the protein expression levels of Wnt1, β-catenin, RUNX2 and E-cad herin significantly increased. MiR-148a-3p is highly expressed in and regulates osteogenic differentiation and enamel de velopment through targeting RUNX2 and E-cadherin respectively via the Wnt1/β-catenin pathway, which plays an impor tant role in cell differentiation, stem cell proliferation and enamel development.
{"title":"MiR-148a-3p Regulates Stem Cell Osteogenic Differentiation and Enamel Development by Targeting Runt-Related Transcription Factor 2 and E-cadherin via the Wnt1/β-catenin Signaling Pathway","authors":"Huai-chun Chang, Tingting Jiang, Liang Kou, Duo Li, Xinchen Yu, Youqin Li, Lei Zhang","doi":"10.2485/jhtb.31.141","DOIUrl":"https://doi.org/10.2485/jhtb.31.141","url":null,"abstract":": We aimed to evaluate the regulatory effects of miR-148a-3p on stem cell osteogenic differentiation and enamel development by targeting runt-related transcription factor 2 (RUNX2) and E-cadherin, respectively. TargetScan software was utilized to predict the binding sites between miR-148a-3p and osteogenic marker gene RUNX2 or E-cadherin. The changes in miR-148a-3p expression during osteogenic differentiation of epidermal stem cells were detected. After transfec tion with miR-148a-3p mimics or miR-148a-3p inhibitor, epidermal stem cells were induced towards osteogenic differentia tion, and the changes in RUNX2 expression were measured. The changes in miR-148a-3p expression during enamel devel opment regulated by epidermal stem cells were determined. After liposome-mediated transfection with miR-148a-3p mimics or miR-148a-3p inhibitor, epidermal stem cells were induced towards ameloblast development. Cell proliferation and apoptosis abilities were tested using methyl thiazolyl tetrazolium assay and flow cytometry, respectively. The expres sion of miR-148a-3p was detected by RT-PCR. The protein expressions of Wnt1, β-catenin, RUNX2 and E-cadherin were measured by Western blotting. Epidermal stem cells differentiated into osteoblasts through osteogenic induction culture. On 5, 12, 15 and 30 d, epidermal stem cells gradually differentiated into osteoblasts through epithelial aggregation and depres sion, mesenchymal aggregation, and dentin and enamel secretion. After transfection, compared with negative control (NC) group, the cell viability of miR-148-3p group significantly decreased (P<0.05), and the apoptosis rate increased (P<0.01). The viability of miR-148-3p inhibitor group significantly increased (P<0.05), while the apoptosis rate reduced (P<0.01). The dual-luciferase reporter assay showed that miR-148a-3p targeted Wnt1. Compared with NC group, the expression of miR-148a-3p significantly rose in miR-148a-3p group (P<0.001), and the protein expression levels of Wnt1, β-catenin, RUNX2 and E-cadherin significantly decreased. Compared with NC group, the expression of miR-148a-3p in miR-148a-3p inhibitor group significantly decreased (P<0.05), and the protein expression levels of Wnt1, β-catenin, RUNX2 and E-cad herin significantly increased. MiR-148a-3p is highly expressed in and regulates osteogenic differentiation and enamel de velopment through targeting RUNX2 and E-cadherin respectively via the Wnt1/β-catenin pathway, which plays an impor tant role in cell differentiation, stem cell proliferation and enamel development.","PeriodicalId":16040,"journal":{"name":"Journal of Hard Tissue Biology","volume":null,"pages":null},"PeriodicalIF":0.4,"publicationDate":"2022-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"68966038","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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