TY - JOUR
T1 - Macrophages enhance mesenchymal stem cell osteogenesis via down-regulation of reactive oxygen species
AU - Luo, Meng lin
AU - Jiao, Yang
AU - Gong, Wen ping
AU - Li, Yan
AU - Niu, Li na
AU - Tay, Franklin R.
AU - Chen, Ji hua
N1 - Funding Information:
This work was financially supported by the Open Project of State Key Laboratory of Military Stomatology (No. 2018KA02 ), National Natural Science Foundation of China ( 81720108011 and 81470773 ), and the program for Changjiang Scholars and Innovative Research Team in University (No. IRT13051 ).The author declared no potential conflict of interest with respect to the research, authorship, and/or publication of this work.
Funding Information:
This work was financially supported by the Open Project of State Key Laboratory of Military Stomatology (No. 2018KA02), National Natural Science Foundation of China (81720108011 and 81470773), and the program for Changjiang Scholars and Innovative Research Team in University (No. IRT13051).The author declared no potential conflict of interest with respect to the research, authorship, and/or publication of this work.
Publisher Copyright:
© 2020
PY - 2020/3
Y1 - 2020/3
N2 - Objectives: The role played by macrophages in regulating the differentiation of mesenchymal stem cells (MSCs) during wound healing and bone regeneration is increasingly being recognized. The present study compared the pro-osteogenic effects of three co-culture methods, conditioned medium generated by macrophages (CM), indirect culture (IC) or direct culture (DC) with macrophages, on bone marrow MSCs (BMMSCs). Methods: Primary BMMSCs were isolated, characterized and co-cultured with RAW264.7 mouse macrophages. Cell morphology and intracellular reactive oxygen species (ROS) levels were determined by scanning electron microscopy (SEM) and flow cytometry, respectively. Alkaline phosphatase (ALP) staining and assay, Alizarin red staining (ARS) and quantitative real-time polymerase chain reaction (qRT-PCR) were performed to evaluate osteogenic differentiation. Results: Inclusion of macrophages in any of the three co-culture methods resulted in improvement in osteogenic differentiation and mineralization of BMMSCs (DC > IC > CM), as measured by ALP staining and activity, ARS and osteoblastic gene expression (Runx2, Alp, Ocn and Bmp2). The enhanced osteogenesis was reversed with hydrogen peroxide. Macrophages reduced the increased levels of intracellular ROS generated by BMMSCs during osteogenic differentiation in a manner similar to the use of an antioxidant, N-acetyl cysteine. Conclusions: Macrophages exert an osteogenesis-enhancing effect to accelerate BMMSC osteogenesis via ROS downregulation. Clinical significance: The present findings suggest that targeting MSC-macrophage interaction is an effective strategy for regulating stem cell fate and facilitating bone regeneration.
AB - Objectives: The role played by macrophages in regulating the differentiation of mesenchymal stem cells (MSCs) during wound healing and bone regeneration is increasingly being recognized. The present study compared the pro-osteogenic effects of three co-culture methods, conditioned medium generated by macrophages (CM), indirect culture (IC) or direct culture (DC) with macrophages, on bone marrow MSCs (BMMSCs). Methods: Primary BMMSCs were isolated, characterized and co-cultured with RAW264.7 mouse macrophages. Cell morphology and intracellular reactive oxygen species (ROS) levels were determined by scanning electron microscopy (SEM) and flow cytometry, respectively. Alkaline phosphatase (ALP) staining and assay, Alizarin red staining (ARS) and quantitative real-time polymerase chain reaction (qRT-PCR) were performed to evaluate osteogenic differentiation. Results: Inclusion of macrophages in any of the three co-culture methods resulted in improvement in osteogenic differentiation and mineralization of BMMSCs (DC > IC > CM), as measured by ALP staining and activity, ARS and osteoblastic gene expression (Runx2, Alp, Ocn and Bmp2). The enhanced osteogenesis was reversed with hydrogen peroxide. Macrophages reduced the increased levels of intracellular ROS generated by BMMSCs during osteogenic differentiation in a manner similar to the use of an antioxidant, N-acetyl cysteine. Conclusions: Macrophages exert an osteogenesis-enhancing effect to accelerate BMMSC osteogenesis via ROS downregulation. Clinical significance: The present findings suggest that targeting MSC-macrophage interaction is an effective strategy for regulating stem cell fate and facilitating bone regeneration.
KW - Cell–cell interaction
KW - Macrophage
KW - Mesenchymal stem cell
KW - Osteogenic differentiation
KW - Reactive oxygen species
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U2 - 10.1016/j.jdent.2020.103297
DO - 10.1016/j.jdent.2020.103297
M3 - Article
C2 - 32057768
AN - SCOPUS:85079535780
SN - 0300-5712
VL - 94
JO - Journal of Dentistry
JF - Journal of Dentistry
M1 - 103297
ER -