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CCL2 promotes osteogenesis by facilitating macrophage migration during acute inflammation

Novel minimally invasive strategies are needed to obtain robust bone healing in complex fractures and bone defects in the elderly population. Local cell therapy is one potential option for future treatment. Mesenchymal stromal cells (MSCs) are not only involved in osteogenesis but also help direct t...

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Autores principales: Toya, Masakazu, Zhang, Ning, Tsubosaka, Masanori, Kushioka, Junichi, Gao, Qi, Li, Xueping, Chow, Simon Kwoon-Ho, Goodman, Stuart B.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Frontiers Media S.A. 2023
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10348816/
https://www.ncbi.nlm.nih.gov/pubmed/37457301
http://dx.doi.org/10.3389/fcell.2023.1213641
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author Toya, Masakazu
Zhang, Ning
Tsubosaka, Masanori
Kushioka, Junichi
Gao, Qi
Li, Xueping
Chow, Simon Kwoon-Ho
Goodman, Stuart B.
author_facet Toya, Masakazu
Zhang, Ning
Tsubosaka, Masanori
Kushioka, Junichi
Gao, Qi
Li, Xueping
Chow, Simon Kwoon-Ho
Goodman, Stuart B.
author_sort Toya, Masakazu
collection PubMed
description Novel minimally invasive strategies are needed to obtain robust bone healing in complex fractures and bone defects in the elderly population. Local cell therapy is one potential option for future treatment. Mesenchymal stromal cells (MSCs) are not only involved in osteogenesis but also help direct the recruitment of macrophages during bone regeneration via MSC-macrophage crosstalk. The C-C motif chemokine ligand 2 (CCL2) is an inflammatory chemokine that is associated with the migration of macrophages and MSCs during inflammation. This study investigated the use of CCL2 as a therapeutic target for local cell therapy. MSCs and macrophages were isolated from 10 to 12 week-old BALB/c male mice. Genetically modified CCL2 over-expressing MSCs were produced using murine CCL2-secreting pCDH-CMV-mCCL2-copGFP expressing lentivirus vector. Osteogenic differentiation assays were performed using MSCs with or without macrophages in co-culture. Cell migration assays were also performed. MSCs transfected with murine CCL2-secreting pCDH-CMV-mCCL2-copGFP expressing lentivirus vector showed higher levels of CCL2 secretion compared to unaltered MSCs (p < 0.05). Genetic manipulation did not affect cell proliferation. CCL2 did not affect the osteogenic ability of MSCs alone. However, acute (1 day) but not sustained (7 days) stimulation with CCL2 increased the alizarin red-positive area when MSCs were co-cultured with macrophages (p < 0.001). Both recombinant CCL2 (p < 0.05) and CCL2 released from MSCs (p < 0.05) facilitated macrophage migration. We demonstrated that acute CCL2 stimulation promoted subsequent osteogenesis in co-culture of MSCs and macrophages. Acute CCL2 stimulation potentially facilitates osteogenesis during the acute inflammatory phase of bone healing by directing local macrophage migration, fostering macrophage-MSC crosstalk, and subsequently, by activating or licensing of MSCs by macrophage pro-inflammatory cytokines. The combination of CCL2, MSCs, and macrophages could be a potential strategy for local cell therapy in compromised bone healing.
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spelling pubmed-103488162023-07-15 CCL2 promotes osteogenesis by facilitating macrophage migration during acute inflammation Toya, Masakazu Zhang, Ning Tsubosaka, Masanori Kushioka, Junichi Gao, Qi Li, Xueping Chow, Simon Kwoon-Ho Goodman, Stuart B. Front Cell Dev Biol Cell and Developmental Biology Novel minimally invasive strategies are needed to obtain robust bone healing in complex fractures and bone defects in the elderly population. Local cell therapy is one potential option for future treatment. Mesenchymal stromal cells (MSCs) are not only involved in osteogenesis but also help direct the recruitment of macrophages during bone regeneration via MSC-macrophage crosstalk. The C-C motif chemokine ligand 2 (CCL2) is an inflammatory chemokine that is associated with the migration of macrophages and MSCs during inflammation. This study investigated the use of CCL2 as a therapeutic target for local cell therapy. MSCs and macrophages were isolated from 10 to 12 week-old BALB/c male mice. Genetically modified CCL2 over-expressing MSCs were produced using murine CCL2-secreting pCDH-CMV-mCCL2-copGFP expressing lentivirus vector. Osteogenic differentiation assays were performed using MSCs with or without macrophages in co-culture. Cell migration assays were also performed. MSCs transfected with murine CCL2-secreting pCDH-CMV-mCCL2-copGFP expressing lentivirus vector showed higher levels of CCL2 secretion compared to unaltered MSCs (p < 0.05). Genetic manipulation did not affect cell proliferation. CCL2 did not affect the osteogenic ability of MSCs alone. However, acute (1 day) but not sustained (7 days) stimulation with CCL2 increased the alizarin red-positive area when MSCs were co-cultured with macrophages (p < 0.001). Both recombinant CCL2 (p < 0.05) and CCL2 released from MSCs (p < 0.05) facilitated macrophage migration. We demonstrated that acute CCL2 stimulation promoted subsequent osteogenesis in co-culture of MSCs and macrophages. Acute CCL2 stimulation potentially facilitates osteogenesis during the acute inflammatory phase of bone healing by directing local macrophage migration, fostering macrophage-MSC crosstalk, and subsequently, by activating or licensing of MSCs by macrophage pro-inflammatory cytokines. The combination of CCL2, MSCs, and macrophages could be a potential strategy for local cell therapy in compromised bone healing. Frontiers Media S.A. 2023-06-30 /pmc/articles/PMC10348816/ /pubmed/37457301 http://dx.doi.org/10.3389/fcell.2023.1213641 Text en Copyright © 2023 Toya, Zhang, Tsubosaka, Kushioka, Gao, Li, Chow and Goodman. https://creativecommons.org/licenses/by/4.0/This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner(s) are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.
spellingShingle Cell and Developmental Biology
Toya, Masakazu
Zhang, Ning
Tsubosaka, Masanori
Kushioka, Junichi
Gao, Qi
Li, Xueping
Chow, Simon Kwoon-Ho
Goodman, Stuart B.
CCL2 promotes osteogenesis by facilitating macrophage migration during acute inflammation
title CCL2 promotes osteogenesis by facilitating macrophage migration during acute inflammation
title_full CCL2 promotes osteogenesis by facilitating macrophage migration during acute inflammation
title_fullStr CCL2 promotes osteogenesis by facilitating macrophage migration during acute inflammation
title_full_unstemmed CCL2 promotes osteogenesis by facilitating macrophage migration during acute inflammation
title_short CCL2 promotes osteogenesis by facilitating macrophage migration during acute inflammation
title_sort ccl2 promotes osteogenesis by facilitating macrophage migration during acute inflammation
topic Cell and Developmental Biology
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10348816/
https://www.ncbi.nlm.nih.gov/pubmed/37457301
http://dx.doi.org/10.3389/fcell.2023.1213641
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