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Hyaluronic acid promotes osteogenic differentiation of human amniotic mesenchymal stem cells via the TGF-β/Smad signalling pathway

This study investigated the effects of hyaluronic acid (HA), a commonly used osteogenic medium referred to as DAG, and the combined administration of HA and DAG (CG) on the osteogenic differentiation of human amniotic mesenchymal stem cells (hAMSCs), and the underlying mechanism. The phenotype of hA...

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Published in:Life sciences (1973) 2019-09, Vol.232, p.116669-116669, Article 116669
Main Authors: Zhang, Ling-Tao, Liu, Ru-Ming, Luo, Yi, Zhao, Yu-Jie, Chen, Dai-Xiong, Yu, Chang-Yin, Xiao, Jian-Hui
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cites cdi_FETCH-LOGICAL-c381t-4db277ad7b65de1bb4f03f16824431c88af056da1dc0d034dea76d9a31e09ce63
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container_title Life sciences (1973)
container_volume 232
creator Zhang, Ling-Tao
Liu, Ru-Ming
Luo, Yi
Zhao, Yu-Jie
Chen, Dai-Xiong
Yu, Chang-Yin
Xiao, Jian-Hui
description This study investigated the effects of hyaluronic acid (HA), a commonly used osteogenic medium referred to as DAG, and the combined administration of HA and DAG (CG) on the osteogenic differentiation of human amniotic mesenchymal stem cells (hAMSCs), and the underlying mechanism. The phenotype of hAMSCs was detected by flow cytometry and immunocytochemical staining. Alkaline phosphatase (ALP) and calcium deposition assays were employed for evaluating the osteogenic differentiation of hAMSCs. The expression of osteogenesis-related genes and proteins was determined by quantitative reverse transcription PCR (qRT-PCR) and Western blotting, respectively. Meanwhile, the molecular mechanism of osteogenic differentiation of hAMSCs was detected by PCR array and qRT-PCR. The results showed that treatment with CG could significantly stimulate hAMSC ALP activity and calcium deposition compared to treatment with DAG, while HA had little effect. The expression of osteogenesis-related molecules and stemness-related molecules was up-regulated at the mRNA and protein levels in all three groups, and this up-regulation was most significant in the CG group. In addition, treatment with CG significantly increased the gene expressions involved in regulation of the TGF-β/Smad signalling pathway compared to treatment with DAG. Furthermore, the pro-osteogenic differentiation effects as well as the up-regulated expression of genes observed in the CG treatment group were significantly inhibited when the cells were pre-treated with SB431542, an inhibitor of the TGF-β/Smad pathway. These results suggest that HA in combination with DAG could significantly enhance the osteogenic differentiation of hAMSCs, potentially via the TGF-β/Smad signalling pathway.
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The phenotype of hAMSCs was detected by flow cytometry and immunocytochemical staining. Alkaline phosphatase (ALP) and calcium deposition assays were employed for evaluating the osteogenic differentiation of hAMSCs. The expression of osteogenesis-related genes and proteins was determined by quantitative reverse transcription PCR (qRT-PCR) and Western blotting, respectively. Meanwhile, the molecular mechanism of osteogenic differentiation of hAMSCs was detected by PCR array and qRT-PCR. The results showed that treatment with CG could significantly stimulate hAMSC ALP activity and calcium deposition compared to treatment with DAG, while HA had little effect. The expression of osteogenesis-related molecules and stemness-related molecules was up-regulated at the mRNA and protein levels in all three groups, and this up-regulation was most significant in the CG group. In addition, treatment with CG significantly increased the gene expressions involved in regulation of the TGF-β/Smad signalling pathway compared to treatment with DAG. Furthermore, the pro-osteogenic differentiation effects as well as the up-regulated expression of genes observed in the CG treatment group were significantly inhibited when the cells were pre-treated with SB431542, an inhibitor of the TGF-β/Smad pathway. These results suggest that HA in combination with DAG could significantly enhance the osteogenic differentiation of hAMSCs, potentially via the TGF-β/Smad signalling pathway.</description><identifier>ISSN: 0024-3205</identifier><identifier>EISSN: 1879-0631</identifier><identifier>DOI: 10.1016/j.lfs.2019.116669</identifier><identifier>PMID: 31326566</identifier><language>eng</language><publisher>Netherlands: Elsevier Inc</publisher><subject>Alkaline phosphatase ; Biomedical materials ; Calcium ; Deposition ; Differentiation (biology) ; Flow cytometry ; Gene expression ; Genes ; Human amniotic mesenchymal stem cells ; Hyaluronic acid ; Hydroxyapatite ; Mesenchymal stem cells ; Mesenchyme ; Osteogenesis ; Osteogenic differentiation ; Phenotypes ; Proteins ; Reverse transcription ; Signal transduction ; Signaling ; Smad protein ; Stem cells ; TGF-β/Smad signalling pathway ; Western blotting</subject><ispartof>Life sciences (1973), 2019-09, Vol.232, p.116669-116669, Article 116669</ispartof><rights>2019 Elsevier Inc.</rights><rights>Copyright © 2019. 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These results suggest that HA in combination with DAG could significantly enhance the osteogenic differentiation of hAMSCs, potentially via the TGF-β/Smad signalling pathway.</description><subject>Alkaline phosphatase</subject><subject>Biomedical materials</subject><subject>Calcium</subject><subject>Deposition</subject><subject>Differentiation (biology)</subject><subject>Flow cytometry</subject><subject>Gene expression</subject><subject>Genes</subject><subject>Human amniotic mesenchymal stem cells</subject><subject>Hyaluronic acid</subject><subject>Hydroxyapatite</subject><subject>Mesenchymal stem cells</subject><subject>Mesenchyme</subject><subject>Osteogenesis</subject><subject>Osteogenic differentiation</subject><subject>Phenotypes</subject><subject>Proteins</subject><subject>Reverse transcription</subject><subject>Signal transduction</subject><subject>Signaling</subject><subject>Smad protein</subject><subject>Stem cells</subject><subject>TGF-β/Smad signalling pathway</subject><subject>Western blotting</subject><issn>0024-3205</issn><issn>1879-0631</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2019</creationdate><recordtype>article</recordtype><recordid>eNp9kbFuFDEQhi0EIpfAA9AgSzQ0e7HXu_auqFBEEqRIFITamrNn73xarw_bG3Q1b8SD8Ezx6gIFBZWL-f5f4_kIecPZmjMuL_frcUjrmvF-zbmUsn9GVrxTfcWk4M_JirG6qUTN2jNyntKeMda2SrwkZ4KLWrZSrsjP2yOMcwyTMxSMs_QQgw8ZEw0pY9jiMrBuGDDilB1kFyYaBrqbPUwU_ORCLoTHhJPZHT2MtOQ8NTiOiT44oHmH9P7muvr96_KrB0uT204wjm7a0gPk3Q84viIvBhgTvn56L8i360_3V7fV3Zebz1cf7yojOp6rxm5qpcCqjWwt8s2mGZgYuOzqphHcdB0MrJUWuDXMMtFYBCVtD4Ij6w1KcUHen3rLH7_PmLL2Li2LwoRhTrquJe-VVKIp6Lt_0H2YY9l7oTrVMiYULxQ_USaGlCIO-hCdh3jUnOnFkN7rYkgvhvTJUMm8fWqeNx7t38QfJQX4cAKwnOLBYdTJuHJctC6iydoG95_6RwW-o7c</recordid><startdate>20190901</startdate><enddate>20190901</enddate><creator>Zhang, Ling-Tao</creator><creator>Liu, Ru-Ming</creator><creator>Luo, Yi</creator><creator>Zhao, Yu-Jie</creator><creator>Chen, Dai-Xiong</creator><creator>Yu, Chang-Yin</creator><creator>Xiao, Jian-Hui</creator><general>Elsevier Inc</general><general>Elsevier BV</general><scope>NPM</scope><scope>AAYXX</scope><scope>CITATION</scope><scope>7QP</scope><scope>7QR</scope><scope>7TK</scope><scope>7U7</scope><scope>7U9</scope><scope>8FD</scope><scope>C1K</scope><scope>FR3</scope><scope>H94</scope><scope>P64</scope><scope>RC3</scope><scope>7X8</scope></search><sort><creationdate>20190901</creationdate><title>Hyaluronic acid promotes osteogenic differentiation of human amniotic mesenchymal stem cells via the TGF-β/Smad signalling pathway</title><author>Zhang, Ling-Tao ; 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The phenotype of hAMSCs was detected by flow cytometry and immunocytochemical staining. Alkaline phosphatase (ALP) and calcium deposition assays were employed for evaluating the osteogenic differentiation of hAMSCs. The expression of osteogenesis-related genes and proteins was determined by quantitative reverse transcription PCR (qRT-PCR) and Western blotting, respectively. Meanwhile, the molecular mechanism of osteogenic differentiation of hAMSCs was detected by PCR array and qRT-PCR. The results showed that treatment with CG could significantly stimulate hAMSC ALP activity and calcium deposition compared to treatment with DAG, while HA had little effect. The expression of osteogenesis-related molecules and stemness-related molecules was up-regulated at the mRNA and protein levels in all three groups, and this up-regulation was most significant in the CG group. In addition, treatment with CG significantly increased the gene expressions involved in regulation of the TGF-β/Smad signalling pathway compared to treatment with DAG. Furthermore, the pro-osteogenic differentiation effects as well as the up-regulated expression of genes observed in the CG treatment group were significantly inhibited when the cells were pre-treated with SB431542, an inhibitor of the TGF-β/Smad pathway. These results suggest that HA in combination with DAG could significantly enhance the osteogenic differentiation of hAMSCs, potentially via the TGF-β/Smad signalling pathway.</abstract><cop>Netherlands</cop><pub>Elsevier Inc</pub><pmid>31326566</pmid><doi>10.1016/j.lfs.2019.116669</doi><tpages>1</tpages></addata></record>
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1879-0631
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subjects Alkaline phosphatase
Biomedical materials
Calcium
Deposition
Differentiation (biology)
Flow cytometry
Gene expression
Genes
Human amniotic mesenchymal stem cells
Hyaluronic acid
Hydroxyapatite
Mesenchymal stem cells
Mesenchyme
Osteogenesis
Osteogenic differentiation
Phenotypes
Proteins
Reverse transcription
Signal transduction
Signaling
Smad protein
Stem cells
TGF-β/Smad signalling pathway
Western blotting
title Hyaluronic acid promotes osteogenic differentiation of human amniotic mesenchymal stem cells via the TGF-β/Smad signalling pathway
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