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Kang Xu, Tingwen Zhou, Yuming Huang, Qingjia Chi, Jiawei Shi, Peng Zhu, Nianguo Dong
Exploring effective therapies for delaying calcific heart valve disease (CHVD) is the focus of current cardiovascular clinical and scientific research. In this study, human aortic valve interstitial cells (hVICs) were isolated from patients with CHVD. After expansion, cultured hVICs were induced with the tumor necrosis factor-alpha (TNF-α) with or without anthraquinone emodin (EMD) treatments. Cytotoxicity and flow cytometric analysis were used to assess cell growth, while Alizarin Red S staining was used to detect hVICs calcification...
2018: Frontiers in Pharmacology
Lei Wang, Jian Cheng, Zhen Hua, Mingming Liu, Haiyan Xu, Yong Ma, Guicheng Huang, Guoqing Mao
α-melanocyte stimulating hormone (α-MSH) is a member of the melanocortin family, that has displayed important biological functions in diverse cells and tissues. The purpose of this study is to test the effect of α-MSH on the differentiation and mineralization of osteoblast cells. The expression of the α-MSH membrane receptor MC1R but not MC2R, MC3R, or MC4R increased distinctively during the osteogenic differentiation from 3, 7 to 14 d. Treatment with α-MSH promoted the differentiation and mineralization of MC3T3-E1 cells by increasing the activity of ALP, enhancing Alizarin Red S staining, and stimulating the expression of osteogenic genes, including ALP1, osteocalcin (Bglap2), and osterix (Sp7)...
November 26, 2018: European Journal of Pharmacology
Yadong Liu, Tao Yang, Ting Chen, Jun Hao, Yu Gai, Weiguo Zhang
Lonicera japonica has been used in traditional Chinese medicine as an important medicinal plant, with the ability to inhibit osteoclast development and bone loss. However, it is not clear which active ingredient exerts these effects. (R)‑dehydroxyabscisic alcohol β‑D‑apiofuranosyl‑(1ˮ→6')‑β‑D‑glucopyranoside (DAG) is an active constituent isolated from Lonicera japonica. In the present study, the ST2 bone marrow stromal cell line was treated by DAG at different concentrations and the osteoblastic differentiation was explored by ELISA assay, Von Kossa staining, Alizarin Red S staining, reverse transcription‑quantitative polymerase chain reaction and western blot analysis...
November 23, 2018: Molecular Medicine Reports
Tangzhao Liang, Wenling Gao, Lei Zhu, Jianhua Ren, Hui Yao, Kun Wang, Dehai Shi
This study aimed to evaluate the effect of tissue inhibitor of metalloproteinase-1 (TIMP-1) on the proliferation and osteogenic differentiation potential of human bone marrow-derived MSCs (hBMSCs). hBMSCs with stable TIMP-1 overexpression or TIMP-1 knockdown were generated. Osteogenic differentiation was assessed by Alizarin Red S staining, alkaline phosphatase (ALP) activity and expression of specific markers. Compared to the vehicle controls, TIMP-1 knockdown significantly promoted the growth of hBMSCs. TIMP-1 knockdown up-regulated β-catenin and cyclin D1 proteins...
November 25, 2018: Bioscience Reports
Karen Chang, Rung-Shu Chen, Fu-Hsiung Chang, Min-Huey Chen
PURPOSE: The purposes of this study are to explore the roles of microRNA-218 (miR-218) delivered by a newly designed magnetic nanocarrier: GCC-Fe3 O4 (GCC-Fe) in dentinogenesis potentials of human dental pulp stem cells (DPSCs). METHODS: Human DPSCs were obtained from impacted wisdom teeth of healthy donors under the permission of National Taiwan University Hospital institutional review board (NTUH IRB). Meanwhile, the transfection efficiency of GCC-Fe was evaluated...
November 21, 2018: Journal of the Formosan Medical Association, Taiwan Yi Zhi
Jin-Sun Lee, Hong Jae Lee, Jae Won Lee, Sang Cheon Lee, Jung Sun Heo
BACKGROUND/AIMS: This study investigated the effect of inducible nitric oxide synthase-loaded mineralized nanoparticles (iNOS-MNPs) on the osteogenic differentiation of mouse embryonic stem cells (ESCs). METHODS: We prepared iNOS-MNPs using an anionic block copolymer template-mediated calcium carbonate (CaCO3) mineralization process in the presence of iNOS. iNOS-MNPs were spherical and had a narrow size distribution. iNOS was stably loaded within MNPs without denaturation...
2018: Cellular Physiology and Biochemistry
Chenyue Liu, Ruyuan Zhu, Haixia Liu, Lin Li, Beibei Chen, Qiangqiang Jia, Lili Wang, Rufeng Ma, Simin Tian, Min Wang, Min Fu, Jianzhao Niu, Alexander N Orekhov, Sihua Gao, Dongwei Zhang, Baosheng Zhao
Purpose: The present study is aimed to explore whether the aqueous extract of Mori Folium (MF) exhibits bone protective effect by regulating calcium and redox homeostasis in diabetic rats, and to identify the signaling pathways involved in this process. Methods: Diabetic rats were established using high-sugar and high-fat diet and streptozotocin (STZ) (30 mg/kg for 3 consecutive days). The serum levels of osteocalcin (OC), insulin-like growth factor-1 (IGF-1), tartrate-resistant acid phosphatase (TRAP), phosphorus (P), calcium (Ca), 1,25-dihydroxyvitamin D3 [1,25(OH)2 D3 ], parathormone (PTH), advanced glycation end products (AGEs), superoxide dismutase (SOD), and malondialdehyde (MDA), total antioxidant capacity (TAC), 8-hydroxy-2'-deoxyguanosine (8-OH-dG), and interleukin 6 (IL-6) were determined by ELISA or biochemical assays...
2018: Frontiers in Pharmacology
Liying Wang, Fan Wu, Chen Liu, Yang Song, Jiawen Guo, Yanwei Yang, Yinong Qiu
The aim of this in vitro study was to evaluate the effects of low-level laser therapy (LLLT) at different energy intensities on proliferation and osteogenic differentiation of bone marrow mesenchymal stem cells (BMSCs) under healthy and inflammatory microenvironments. Human BMSCs and BMSCs from inflammatory conditions (i-BMSCs, BMSCs treated with tumor necrosis factor α; TNF-α) were subject to LLLT (Nd:YAG;1064 nm) at different intensities. We designed one control group (without irradiation) and four testing groups (irradiation at 2, 4, 8, and 16 J/cm2 ) for both BMSCs and i-BMSCs...
November 19, 2018: Lasers in Medical Science
Kyeong-Min Kim, Wan-Jin Jeon, Eun-Jung Kim, Won-Gu Jang
AIMS: CREB (cAMP response element-binding protein)-regulated transcription coactivator (CRTC2) has been reported to act as a coactivator of CREB during gluconeogenesis. The role of CRTC2 in osteoblastic differentiation has not yet been elucidated. The aim of this study is to identify the mechanism of CRTC2 in osteoblast differentiation. MAIN METHODS: The mRNA expression was determined by RT-PCR and qPCR. Protein levels were measured using Western blot assay. Alkaline phosphatase (ALP) staining was performed to evaluate ALP activity...
December 1, 2018: Life Sciences
Giulia Ghiacci, Simone Lumetti, Edoardo Manfredi, Daniele Mori, Guido Maria Macaluso, Roberto Sala
Stanozolol (ST) is a synthetic androgen with high anabolic potential. Although it is known that androgens play a positive role in bone metabolism, ST action on bone cells has not been sufficiently tested to support its clinical use for bone augmentation procedures. OBJECTIVE: This study aimed to assess the effects of ST on osteogenic activity and gene expression in SaOS-2 cells. MATERIAL AND METHODS: SaOS-2 deposition of mineralizing matrix in response to increasing doses of ST (0-1000 nM) was evaluated through Alizarin Red S and Calcein Green staining techniques at 6, 12 and 24 days...
November 8, 2018: Journal of Applied Oral Science: Revista FOB
Hamada Elsayed, Giulia Brunello, Chiara Gardin, Letizia Ferroni, Denis Badocco, Paolo Pastore, Stefano Sivolella, Barbara Zavan, Lisa Biasetto
Titanium implant surface modifications have been widely investigated to favor the process of osseointegration. The present work aimed to evaluate the effect of sphene (CaTiSiO₅) biocoating, on titanium substrates, on the in vitro osteogenic differentiation of Human Adipose-Derived Stem Cells (hADSCs). Sphene bioceramic coatings were prepared using preceramic polymers and nano-sized active fillers and deposited by spray coating. Scanning Electron Microscopy (SEM) analysis, surface roughness measurements and X-ray diffraction analysis were performed...
November 9, 2018: Materials
S-L Tang, Q-H Huang, L-G Wu, C Liu, A-L Cai
OBJECTIVE: Ankylosing spondylitis (AS) is a spastic and spinal joint disease with the characteristic of pathological ossification. Bioinformatics analysis demonstrated that there is a complementary binding site between microRNA-124 (miR-124) and the 3'-UTR of glycogen synthase kinase-3β (GSK-3β) mRNA. We aimed to investigate the role of miR-124 in regulating GSK-3β expression, Wnt/β-catenin pathway activity, and osteoblast differentiation of spinal ligament fibroblasts. PATIENTS AND METHODS: The ligament tissues of AS and the femoral neck fracture patients were collected...
October 2018: European Review for Medical and Pharmacological Sciences
Cheng-Cong Wu, Fang Wang, Shu Rong, Jing Ren, Jian-Shan Wan, Li-Xiang Shi, Zhen Wu, Tao Liu, Qiang Li
Bone marrow mesenchymal stem cells (BMSCs) are considered the most important seed cells in bone tissue engineering. The present study aimed to investigate the potential of rabbit BMSCs in osteogenesis after the transfection of human BMP-2 and EGFP recombinant adenovirus. Rabbit BMSCs were isolated and the surface stem cell makers, including CD29, CD44 and CD45 were detected by flow cytometry. The expression of BMP-2 mRNA and protein in BMSCs were detected by reverse transcription-quantitative polymerase chain reaction and western blot analysis, respectively...
November 2018: Experimental and Therapeutic Medicine
Valentina Palmieri, Marta Barba, Lorena Di Pietro, Claudio Conti, Marco De Spirito, Wanda Lattanzi, Massimiliano Papi
Graphene and graphene oxide can promote the adhesion, growth and differentiation of mesenchymal stem cells. Further, graphene surface coatings accelerate the differentiation of human mesenchymal stem cells acting as osteogenic inducers. Quantification of the osteogenic induction is conventionally performed with Alizarin Red S (ARS), an anthraquinone derivative used to identify calcium deposits in tissue sections and cell cultures. The ARS staining is quite versatile because the dye forms an Alizarin Red S⁻calcium complex that can be extracted from the stained monolayer of cells and readily assayed by absorbance measurements...
October 26, 2018: International Journal of Molecular Sciences
Bin Ren, Volker M Betz, Christian Thirion, Michael Salomon, Volkmar Jansson, Peter E Müller, Oliver B Betz
Bone can be engineered in vivo by implantation of gene-activated muscle tissue fragments. This expedited approach may be further improved by use of muscle tissue with attached fascia. The aim of this in vitro study was to provide an in depth comparison of the osteogenic differentiation capacity of muscle alone and muscle with fascia after BMP-2 transduction. Skeletal muscle tissue from rats was cut into pieces with and without a fascia layer on the surface. Adenoviral BMP-2 or GFP vectors were used for transduction...
October 27, 2018: Gene Therapy
Nicola Keay, Gavin Francis, Karen Hind
Objectives: To evaluate the efficacy of a sport-specific energy availability (EA) questionnaire, combined with clinical interview, for identifying male athletes at risk of developing bone health, endocrine and performance consequences of relative energy deficiency in sports (RED-S). Methods: Fifty competitive male road cyclists, recruited through links of participants in a pilot study, were assessed by a newly developed sport-specific questionnaire and clinical interview (SEAQ-I) and received dual energy X-ray absorptiometry (DXA) bone mineral density (BMD) and body composition scans and blood tests for endocrine markers...
2018: BMJ Open Sport & Exercise Medicine
Bao-Liang Wang, Zhi Wang, Xi Nan, Qing-Cai Zhang, Wei Liu
MicroRNAs (miRNAs) play critical roles in various biological processes including cell differentiation. Some researchers suggested that the p38 mitogen-activated protein kinases (MAPK) signaling pathway had an effect on regulating the odontoblastic differentiation of human dental pulp stem cells (hDPSCs). This study focuses on the effects of miR-143-5p on hDPSCs by regulating the p38 MAPK signaling pathway. The targeting relationship of MAPK14 and miR-143-5p targets were verified by TargetScan and dual-luciferase reporter gene assay...
October 26, 2018: Journal of Cellular Physiology
Dongxia Chen, Jingjin Zhao, Liangliang Zhang, Rongjun Liu, Yong Huang, Chuanqing Lan, Shulin Zhao
Calcium ion (Ca2+ ) plays crucial roles in the signal transduction pathways associated with various physiological and pathological events. Monitoring intracellular Ca2+ is of great significance for cell biology research. Here, we report the use of biomass quantum dots (BQDs) as a fluorescent reporter for imaging of intracellular Ca2+ , based on the inner-filter-mediated luminescence which was assisted by a Ca2+ chelator alizarin red S (ARS). BQDs were prepared by hydrothermal heating of capsicum. The absorption of ARS overlaps with the excitation of the BQDs, the fluorescence of BQDs being quenched through the inner-filter effect...
November 6, 2018: Analytical Chemistry
Jung-Sun Moon, Seong-Duk Kim, Hyun-Mi Ko, Young-Jun Kim, Sun-Hun Kim, Min-Seok Kim
The transcription factor Twist1 is known to be closely associated with the formation of bone by mesenchymal stem cells and osteoblasts; however, the role of Twist1 in cementogenesis has not yet been determined. This study was undertaken to elucidate the roles of Twist1 in cementoblast differentiation by means of the gain- or loss-of-function method. We used alkaline phosphatase (ALP) and alizarin red S staining and quantitative real-time reverse transcriptase polymerase chain reaction (qRT-PCR) to determine whether the forced transient expression or knock-down of Twist1 in a mouse cementoblast cell line, OCCM-30, could affect cementogenic differentiation...
October 17, 2018: Dentistry journal
Eirini Athanasiadou, Maria Paschalidou, Anna Theocharidou, Nikolaos Kontoudakis, Konstantinos Arapostathis, Athina Bakopoulou
OBJECTIVE: To investigate the biological interactions of a calcium silicate based cement (Biodentine™) with Stem Cells from Human Exfoliated Deciduous teeth (SHED), focusing on viability/proliferation, odontogenic differentiation, biomineralization and elemental release/exchange. METHODS: Biodentine™ specimens were used directly or for eluate preparation at serial dilutions (1:1-1:64). SHED cultures were established from deciduous teeth of healthy children. Viability/proliferation and morphological characteristics were evaluated by live/dead fluorescent staining, MTT assay and Scanning Electron Microscopy...
October 10, 2018: Dental Materials: Official Publication of the Academy of Dental Materials
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