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C CL2, CCL5, and IGF ‐1 participate in the immunomodulation of osteogenesis during M1/M 2 transition in vitro
Author(s) -
Córdova Luis A.,
Loi Florence,
Lin TzuHua,
Gibon Emmanuel,
Pajarinen Jukka,
Nabeshima Akira,
Lu Laura,
Yao Zhenyu,
Goodman Stuart B.
Publication year - 2017
Publication title -
journal of biomedical materials research part a
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.849
H-Index - 150
eISSN - 1552-4965
pISSN - 1549-3296
DOI - 10.1002/jbm.a.36166
Subject(s) - ccl5 , ccl2 , in vitro , alkaline phosphatase , microbiology and biotechnology , chemokine , osteoblast , chemistry , immunology , biology , inflammation , biochemistry , cytotoxic t cell , il 2 receptor , enzyme
The modulation of macrophage phenotype from pro‐inflammatory (M1) to tissue healing (M2) via exogenous addition of interleukin‐4 (IL‐4) facilitates osteogenesis; however, the molecular mediators underlying this phenomenon remain unknown. This study characterizes the IL‐4‐dependent paracrine crosstalk between macrophages and osteoprogenitors and its effect on osteogenesis in vitro . Primary murine M1 were co‐cultured with MC3T3 cells (M1‐MC3T3) in both transwell plates and direct co‐cultures. To modulate M1 to M2, M1‐MC3T3 were treated with IL‐4 (20 ng/mL) at day 3 after seeding (M1 + IL‐4‐MC3T3). Selected molecular targets were assessed at days 3 and 6 after seeding at protein and mRNA levels. Mineralization was assessed at day 21. Transwell M1 + IL‐4‐MC3T3 significantly enhanced the secretion of CCL2/MCP‐1, IGF‐1 and to a lesser degree, CCL5/RANTES at day 6. At day 3, alkaline phosphatase ( Alpl ) was upregulated in direct M1‐MC3T3. At day 6, Smurf2 and Insulin growth factor‐1 ( IGF‐1 ) were downregulated and upregulated, respectively, in direct M1 + IL‐4‐MC3T3. Finally, M1 + IL‐4‐MC3T3 increased bone matrix mineralization compared with MC3T3 cells in transwell, but this was significantly less than M1‐MC3T3. Taken together, macrophage subtypes enhanced the osteogenesis in transwell setting and the transition from M1 to M2 was associated with an increase in bone anabolic factors CCL2/MCP‐1, CCL5/RANTES and IGF‐1 in vitro . © 2017 Wiley Periodicals, Inc. J Biomed Mater Res Part A: 105A: 3069–3076, 2017.