Ling-Tao Zhang1, Ru-Ming Liu1, Yi Luo1, Yu-Jie Zhao2, Dai-Xiong Chen1, Chang-Yin Yu3, Jian-Hui Xiao4. 1. Zunyi Municipal Key Laboratory of Medicinal Biotechnology, Center for Translational Medicine, Affiliated Hospital of Zunyi Medical University, 149 Dalian Road, Zunyi 563003, PR China. 2. Department of Laboratory Medicine, Affiliated Hospital of Zunyi Medical University, 149 Dalian Road, Zunyi 563003, PR China. 3. Zunyi Municipal Key Laboratory of Medicinal Biotechnology, Center for Translational Medicine, Affiliated Hospital of Zunyi Medical University, 149 Dalian Road, Zunyi 563003, PR China; Department of Neurology, Affiliated Hospital of Zunyi Medical University, 149 Dalian Road, Zunyi 563003, PR China. Electronic address: yuchangyin68@163.com. 4. Zunyi Municipal Key Laboratory of Medicinal Biotechnology, Center for Translational Medicine, Affiliated Hospital of Zunyi Medical University, 149 Dalian Road, Zunyi 563003, PR China. Electronic address: jhxiao@zmc.edu.cn.
Abstract
AIMS: 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. MAIN METHODS: 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. KEY FINDINGS: 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. SIGNIFICANCE: 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.
AIMS: 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. MAIN METHODS: 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. KEY FINDINGS: 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. SIGNIFICANCE: 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.
Authors: Xifeng Liu; Matthew N George; Linli Li; Darian Gamble; A Lee Miller Ii; Bipin Gaihre; Brian E Waletzki; Lichun Lu Journal: ACS Biomater Sci Eng Date: 2020-07-09
Authors: Aidan M Kirkham; Adrian J M Bailey; Alvin Tieu; Harinad B Maganti; Joshua Montroy; Risa Shorr; T Mark Campbell; Dean A Fergusson; Manoj M Lalu; Heidi Elmoazzen; David S Allan Journal: Stem Cell Rev Rep Date: 2021-07-27 Impact factor: 5.739