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Beneficial Role of Low-Intensity Laser Irradiation on Neural β-tubulin III Protein Expression in Human Bone Marrow Multipotent Mesenchymal Stromal Cells

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Abstract

The purpose of the present study was to evaluate the neural protein expression pattern of human multipotent mesenchymal stromal cells (hMSCs) treated with forskolin (free-form/FF). The study investigated forskolin’s capacity to enhance intracellular levels of cyclic adenosine monophosphate (cAMP) by activating adenylate cyclase and probably by inducing neuron-like cells in vitro. In addition, because nanotechnology is a growing field of tissue engineering, we also assessed the action of a new system called the nanostructured-forskolin (NF) to examine the improvement of drug delivery. Afterwards, the cells were submitted to low-level laser irradiation to evaluate possible photobiostimulatory effects. Investigations using the immunofluorescence by confocal microscopy and Western blot methods revealed the expression of the neuronal marker β-tubulin III. Fluorescence intensity quantification analysis using INCell Analyzer System for β-tubulin III was used to examine significant differences. The results showed that after low-level laser irradiation exposure, there was a tendency to increase the β-tubulin III expression in all groups, as expected in the photobiostimulation process. Notably, this process induced for irradiation was more pronounced in irradiated nanoforskolin cells (INF) compared to non-irradiated free-forskolin control cells (NFFC). However, there was also an increase in β-tubulin III protein expression in the groups: irradiated nanocontrol cells (INC) compared to non-irradiated free-forskolin control cells (NFF) and after treatment with non-irradiated free-forskolin (NFF) and non-irradiated nanoforskolin (NNFC). We concluded that the methods using low-level laser irradiation and/or nanoparticles showed an up-regulation of neural-protein expression in hMSCs that could be used to facilitate cellular therapy protocols in the near future.

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Acknowledgements

The authors are grateful to Hospital Amaral de Carvalho (Jau, São Paulo) and are especially grateful to Drs Antônio Cesarino Mota and Vergílio Rensi Colturato for providing bone marrow samples from donors as well as Nayara Rezende for help with the nanoforskolin and Sandra Navarro Brescian for artwork. I want to thank Fernanda Udinal for language advice and Priscilla Carnavale Gomes Ferreira for reviewing the manuscript. Valéria Ferreira-Silva is grateful for a postdoctoral fellowship from Coordenacão de Aperfeicoamento de Pessoal de Nível Superior (CAPES). This work was supported by Financiadora de Estudos e Projetos (FINEP 01.10.0758.01), CEFAP-USP (São Paulo), Instituto Nacional de Ciência e Tecnologia, Célula-Tronco e Terapia Celular (INCTC), Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq), and Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP# 2013/50181-1), Brazil.

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Authors and Affiliations

  1. Department of Chemistry, Centre of Nanotechnology and Tissue Engineering-Photobiology and Photomedicine Research Group, Faculty of Philosophy, Sciences and Letters of Ribeirão Preto, University of São Paulo, Ribeirão Preto, 14040-901, Brazil

    Valéria Ferreira-Silva, Andrielle Castilho-Fernandes & Antonio C. Tedesco

  2. Centre for Cell Therapy and Regional Blood Centre, National Institute of Science and Technology in Stem Cell and Cell Therapy, Medical School, University of São Paulo, Ribeirão Preto, 14051-140, Brazil

    Valéria Ferreira-Silva, Danielle A.R. Magalhães, Maristela D. Orellana & Dimas T. Covas

  3. Department of Bioprocess and Biotechnology, School of Pharmaceutical Sciences, São Paulo State University (UNESP), Araraquara, 14800-903, SP, Brazil

    Fernando L. Primo

  4. Department of Cell and Molecular Biology and Pathogenic Bioagents, Medical School, University of São Paulo, Ribeirão Preto, 14049-900, Brazil

    Munira M.A. Baqui

  5. Institute of Biomedical Science, Department of Immunology, University of São Paulo, São Paulo, 05508-900, Brazil

    Mario C. Cruz & Niehls O. S. Câmara

  6. Department of Clinical Medicine, Medical School, University of São Paulo, Ribeirão Preto, 14048-900, Brazil

    Dimas T. Covas

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  1. Valéria Ferreira-Silva
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Ferreira-Silva, V., Primo, F.L., Baqui, M.M. et al. Beneficial Role of Low-Intensity Laser Irradiation on Neural β-tubulin III Protein Expression in Human Bone Marrow Multipotent Mesenchymal Stromal Cells. Stem Cell Rev and Rep 14, 585–598 (2018). https://doi.org/10.1007/s12015-017-9796-3

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