Evaluation of the influence of the irradiation with laser in low intensity in the proliferative and metabolic behavior of human osteoblasts in vitro

• Grant number: 07/08367-0
• Support type: Regular Research Grants
• Duration: July 01, 2008 - December 31, 2010
• Field of knowledge: Health Sciences - Dentistry
• Principal Investigator: Camila Peres Buzalaf
• Grantee: Camila Peres Buzalaf
• Home Institution: Faculdade de Odontologia de Bauru (FOB). Universidade de São Paulo (USP). Bauru , SP, Brazil

Abstract

The therapy with laser in low intensity has been used for acceleration of the healing of wounds, however of its mechanisms of action they are very not clarified, what it reflects the divergence of results in literature. The positive influence of the laser in osteoblasts would be of great utility in the field of bone regeneration. The discovery on which proteins would have its modified profiles and expression modified under the action of the laser, could contribute for the agreement of its mechanisms of action in the remodeling bone. Ahead of these facts, the objective of the present project will be to evaluate the behavior of human osteoblasts submitted the different intensities of irradiation with laser. Cells will be pellets 103 (cellular viability) 5x103 (enzymes and blots) hFOB 1,19 in plates of 96 wells, which will be radiated with red laser (660nm) and infra-red ray (780nm) in the 5 times of 3 and J/cm². The conditional and osteoblasts ways will be collected and evaluated how much to the cellular viability (counting of cells, reduction of the MTT and Neutral Red) and activity of enzymes (total acid phosphatase (FAT), lisossomal phosphatase (FAL) and phosphotyrosine phosphatase (PTP)). The synthesis of some transcripts and proteins (metalloproteinases of matrix (MMPs 2 and 9), activator of the receiver of the factor nuclear kappa B ligand (RANKL) and osteoprotegerina (OPG)) will be evaluated by means of zymografia, RT-PCR and proteomic. After the preparation of proteins of the human osteoblasts, the same ones will be submitted to electrophoresis 2D and zymografia. The gels of the analyzed groups have controlled and treat they will be scanned using the ImageScanner and with software ImageMaster Platinum. Spots of interest will be excises and analyzed for spectrometry of mass (MALDI-TOF) for identification of proteins. The RT-PCR will be carried through for quantification of transcripts of MMP-2 and 9, RANKL and OPG. The statistical analyses will be made through the tests ANOVA, Kruskal-Wallis, and have tested "t", in accordance with its indication (p<0.05). (AU)

Scientific publications

(References retrieved automatically from Web of Science and SciELO through information on FAPESP grants and their corresponding numbers as mentioned in the publications by the authors)

DE OLIVEIRA, FLAVIA AMADEU; MATOS, ADRIANA ARRUDA; MATSUDA, SANDRA SATIKO; RABELO BUZALAF, MARILIA AFONSO; BAGNATO, VANDERLEY SALVADOR; DE ANDRADE MOREIRA MACHADO, MARIA APARECIDA; DAMANTE, CARLA ANDREOTTI; DE OLIVEIRA, RODRIGO CARDOSO; PERES-BUZALAF, CAMILA. Low level laser therapy modulates viability, alkaline phosphatase and matrix metalloproteinase-2 activities of osteoblasts. JOURNAL OF PHOTOCHEMISTRY AND PHOTOBIOLOGY B-BIOLOGY, v. 169, p. 35-40, APR 2017. Web of Science Citations: 6.

OLIVEIRA, FLAVIA A.; MATOS, ADRIANA A.; SANTESSO, MARIANA R.; TOKUHARA, CINTIA K.; LEITE, ALINE L.; BAGNATO, VANDERLEY S.; MACHADO, MARIA A. A. M.; PERES-BUZALAF, CAMILA; OLIVEIRA, RODRIGO C. Low intensity lasers differently induce primary human osteoblast proliferation and differentiation. JOURNAL OF PHOTOCHEMISTRY AND PHOTOBIOLOGY B-BIOLOGY, v. 163, p. 14-21, OCT 2016. Web of Science Citations: 16.

RICCI VOLPATO, LUIZ EVARISTO; DE OLIVEIRA, RODRIGO CARDOSO; ESPINOSA, MARIANO MARTINEZ; BAGNATO, VANDERLEY SALVADOR; MACHADO, MARIA A. A. M. Viability of fibroblasts cultured under nutritional stress irradiated with red laser, infrared laser, and red light-emitting diode. Journal of Biomedical Optics, v. 16, n. 7 JUL 2011. Web of Science Citations: 26.