The effects of immunomodulation by macrophage subsets on osteogenesis in vitro

Stem Cell Research & Therapy
Florence LoiZhenyu Yao

Abstract

Bone formation and remodeling are influenced by the inflammatory state of the local microenvironment. In this regard, macrophages are postulated to play a crucial role in modulating osteogenesis. However, the differential effects of macrophage subsets and their plasticity on bone formation are currently unknown. Polarized primary murine macrophages and preosteoblastic MC3T3 cells were co-cultured to investigate the effect of non-activated M0, pro-inflammatory M1, and tissue-regenerative M2 macrophages on the osteogenic ability of MC3T3-E1 cells in vitro. Furthermore, to model the physiological transition from inflammation to tissue regeneration, M1-MC3T3 co-cultures were treated with interleukin-4 (IL-4) at different time points to modulate the M1 phenotype towards M2. Macrophage phenotypic markers were assessed by flow cytometry and enzyme-linked immunosorbent assay. A time course study of osteogenic markers at different time points was conducted: alkaline phosphatase (ALP) mRNA levels were evaluated at week 1, ALP activity and osteocalcin and osteopontin mRNA levels at week 2, and matrix mineralization and osteocalcin and osteopontin protein concentrations at week 3. Supernatant collected 72 hours after seeding or IL-4 treatm...Continue Reading

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Citations

Sep 27, 2016·Stem Cells International·Angela P Bastidas-CoralJenneke Klein-Nulend
Dec 16, 2016·Journal of Biomedical Materials Research. Part a·Jukka PajarinenStuart B Goodman
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Jul 12, 2019·Frontiers in Immunology·Lan XiaoYin Xiao
Aug 21, 2019·Frontiers in Endocrinology·Lan Xiao, Yin Xiao
Jul 22, 2019·Journal of Biological Inorganic Chemistry : JBIC : a Publication of the Society of Biological Inorganic Chemistry·Chenglong LiYandong Mu
Jul 17, 2020·Frontiers in Bioengineering and Biotechnology·Stuart B Goodman, Tzuhua Lin
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Oct 15, 2019·Frontiers in Bioengineering and Biotechnology·Stuart B GoodmanTzuhua Lin

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Methods Mentioned

BETA
flow cytometry
PCR
Assay
enzyme-linked immunosorbent assay
ELISA

Software Mentioned

FlowJo
GraphPad Prism

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