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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2018-08-01
Autores
Ferreira-Silva, Valéria
Primo, Fernando L. [UNESP]
Baqui, Munira M.A.
Magalhães, Danielle A.R.
Orellana, Maristela D.
Castilho-Fernandes, Andrielle
Cruz, Mario C.
Câmara, Niehls O. S.
Covas, Dimas T.
Tedesco, Antonio C.
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Resumo
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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Palavras-chave
Forskolin, Low-intensity laser irradiation, Multipotent mesenchymal stromal cells, Nano-drug delivery, Nanotechnology, β-Tubulin III
Como citar
Stem Cell Reviews and Reports, v. 14, n. 4, p. 585-598, 2018.