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Fibroblast contributes for osteoblastic phenotype in a MAPK-ERK and sonic hedgehog signaling-independent manner

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Abstract

We hypothesized that a crosstalk between osteoblast and fibroblast (FB) exists, which contributes to bone as a dynamic tissue. Cell-free supernatants were harvested from fibroblast cultures and later subject pre-osteoblasts to investigate there capacity to modulate cell viability and differentiation mechanisms, reporting the possible involvement of Shh signaling as a paracrine mechanism. By exploring immunoblotting technology, we have shown that FB-released factors interfere with osteoblast metabolism by up-regulating the phosphorylation of FAK and Rac-1 proteins at the early stage and later contribute to osteoblast differentiation by up-modulating alkaline phosphatase (ALP) and in vitro mineralization. We also found that Shh signaling was not required during osteoblastic differentiation promoted by the FB-released factors as well as MAPK-ERK phosphorylation, while pre-osteoblast cultures subjected to osteogenic medium (O.M.) require downstream transducers of Shh, such as Patched and Gli-1, and MAPK-ERK. Altogether, our results indicate for the first time a possible mechanism involved in the crosstalk between fibroblasts and osteoblasts, as it was possible to observe trophic factors released by fibroblasts interfering decisively in osteoblast metabolism in a Shh-independent manner. This study collaborates the body of work that indicates paracrine signaling molecules participate in the crosstalk among bone-resident cells and explains, at least partially, the biological mechanisms responsible for bone tissue dynamism, opening new avenues to understand etiologies of bone diseases.

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Acknowledgements

The authors are grateful to FAPESP (#2015/00581-9; #2014/22689-3) and CNPq (#477452/2012-4) for the financial support. W.F.Z. is a PQ-2 fellow from CNPq (#301966/2015-0).

Funding

This study was funded by the Fundação de Amparo à Pesquisa do Estado de São Paulo - FAPESP (grants #2015/00581-9; #2014/22689-3) and the Conselho Nacional de Desenvolvimento Científico e Tecnológico - CNPq (grant #477452/2012-4).

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Authors and Affiliations

  1. Bioassays and Cell Dynamics Lab, Department of Chemistry and Biochemistry, Bioscience Institute, UNESP, Botucatu, SP, Brazil

    Celio J. da Costa Fernandes, Augusto Santana do Nascimento, Rodrigo A. da Silva & Willian F. Zambuzzi

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  1. Celio J. da Costa Fernandes
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  2. Augusto Santana do Nascimento
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  3. Rodrigo A. da Silva
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  4. Willian F. Zambuzzi
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Correspondence to Willian F. Zambuzzi.

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All the authors declare to have no conflict of interest with the materials used in the present study.

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da Costa Fernandes, C.J., do Nascimento, A.S., da Silva, R.A. et al. Fibroblast contributes for osteoblastic phenotype in a MAPK-ERK and sonic hedgehog signaling-independent manner. Mol Cell Biochem 436, 111–117 (2017). https://doi.org/10.1007/s11010-017-3083-0

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  • DOI: https://doi.org/10.1007/s11010-017-3083-0

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