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Hyperglycemia reduces integrin subunits alpha v and alpha 5 on the surface of dermal fibroblasts contributing to deficient migration

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Abstract

Deficient wound healing is a common multifactorial complication in diabetic patients, but the cellular and molecular mechanisms involved are poorly defined. In the present study, we analyzed the effects of hyperglycemia on integrins expression in rat dermal fibroblasts and addressed its role in cell adhesion and migration. Diabetes Mellitus was induced in rats by streptozotocin injection and maintained for 30 days. Primary cultures of dermal fibroblasts from control and diabetic rats were maintained under low glucose (5 mM d-glucose) or high glucose (30 mM d-glucose) for 7 days. Cell adhesion and migration were studied by kymography, transwell, and time-lapse assays, and the expressions of integrin subunits αv and α5 were studied by immunocytochemistry and western blotting. Fibroblasts derived from diabetic rats confirmed a reduced migration speed and delayed spreading compared to fibroblasts derived from control rats. The membrane fraction of diabetic-derived fibroblasts showed a decrease of integrin subunits α5 and αv, which was confirmed by immunocytochemistry assays. A reduction in the pericellular fibronectin matrix was also observed. The exposure of diabetic-derived cells to a higher concentration of exogenous fibronectin improved migration velocity and the expression of αv but did not completely restore their migration capacity. In conclusion, the mechanisms involved in the deleterious effects of Diabetes Mellitus on wound healing include the ability of fibroblasts to secrete and to adhere to fibronectin.

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Acknowledgments

The authors thank the excellent technical assistance from Gabriella Malheiros. We also thank the technical support of Mario Cruz, from CEFAP-USP for confocal microscopy and Henrique Rofatto from Butantan Institute, São Paulo, Brazil.

Funding

FAPESP (2012/15963-6; 2012/03990-9), CNPq (448052/2014-8).

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Authors and Affiliations

  1. Department of Cell and Developmental Biology, Biomedical Sciences Institute, University of São Paulo, Av. Prof. Lineu Prestes 1524, Cidade Universitária, São Paulo, SP, CEP 05508-000, Brazil

    Maira Estanislau S. Almeida, Kelly S. Monteiro & Marinilce F. Santos

  2. Laboratory of Pathophysiology, Butantan Institute, São Paulo, Brazil

    Ellen E. Kato & Sandra C. Sampaio

  3. Department of Immunology, Biomedical Sciences Institute, University of São Paulo, São Paulo, Brazil

    Tarcio T. Braga & Niels O. S. Câmara

  4. Department of Morphological Sciences, Institute of Basic Health Sciences, Federal University of Rio Grande do Sul, Porto Alegre, Brazil

    Marcelo L. Lamers

Authors
  1. Maira Estanislau S. Almeida
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  2. Kelly S. Monteiro
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  3. Ellen E. Kato
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  7. Marcelo L. Lamers
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  8. Marinilce F. Santos
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Correspondence to Marinilce F. Santos.

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Almeida, M.E.S., Monteiro, K.S., Kato, E.E. et al. Hyperglycemia reduces integrin subunits alpha v and alpha 5 on the surface of dermal fibroblasts contributing to deficient migration. Mol Cell Biochem 421, 19–28 (2016). https://doi.org/10.1007/s11010-016-2780-4

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  • DOI: https://doi.org/10.1007/s11010-016-2780-4

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