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In vitro evaluation of the effects of low level laser therapy on cultured osteoblasts, fibroblasts and keratonicytes submitted to zoledronic acid therapy

Grant number: 09/54722-1
Support Opportunities:Regular Research Grants
Duration: April 01, 2010 - March 31, 2013
Field of knowledge:Health Sciences - Dentistry - Dental Materials
Principal Investigator:Carlos Alberto de Souza Costa
Grantee:Carlos Alberto de Souza Costa
Host Institution: Faculdade de Odontologia (FOAr). Universidade Estadual Paulista (UNESP). Campus de Araraquara. Araraquara , SP, Brazil


Oral osteonecrosis has been associated to bisphosphonates therapy. It has been demonstrated that these drugs may cause cytopathic effects to cells of bone tissue and to adjacent tissues, such as fibroblasts and keratinocytes. Some clinical studies demonstrated excellent results when the patients with osteonecrosis were submitted to low-level laser therapy. Therefore, the aims of this in vitro study are to evaluate the effects: 1) of aminobisphosphonate Zoledronate (Zometa® - Novartis, Brasil) on cultured of osteoblasts, gingival fibroblasts, and keratinocytes; 2) low-level laser therapy on these cells previously treated with Zometa® For the first experiment, the cells will be seeded (30,000 cells/cm2) in wells of 24-well plates using complete culture medium (DMEM) and incubated for 24 hours. Then, the DMEM will be replaced by a solution of Zometa® (5μM/mL) diluted in a new DMEM with no concentrations of fetal bovine serum (FBS). After 48-hour incubation, the cells will be evaluated for cell metabolism, total protein dosage, alkaline phosphatase activity and morphology will be evaluated. For the second part, the cells treated with Zometa® will be irradiated three times with LLL in specific parameters (780±3 nm, 40 mW, 3 J/cm2) every 24 hours, giving rise to the following experimental groups: G1- DMEM (control); G2- DMEM+Zometa®; G3-DMEM+Lasertherapy; G4- DMEM+Zometa®+Lasertherapy. Then, all groups will be evaluated by the same analyses described before and also by gene expression for VEGF, bFGF, Col-1, RANK, OPG, KGF e EGF (RT-PCR). The data will be analyzed by the normality, and if this is respected the chosen tests will be ANOVA, complemented by the Tukey. If normality could not be achieved, the test chosen will be Kruskal-Wallis, complemented with Mann-Whitney at significance level of 5%. (AU)

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Scientific publications (8)
(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)
BASSO, FERNANDA G.; SILVEIRA TURRIONI, ANA PAULA; HEBLING, JOSIMERI; DE SOUZA COSTA, CARLOS A.. Zoledronic Acid Inhibits Human Osteoblast Activities. Gerontology, v. 59, n. 6, p. 534-541, . (09/54722-1)
BASSO, FERNANDA G.; TURRIONI, ANA PAULA S.; HEBLING, JOSIMERI; DE SOUZA COSTA, CARLOS ALBERTO. Zoledronic acid decreases gene expression of vascular endothelial growth factor and basic fibroblast growth factor by human epithelial cells. BRITISH JOURNAL OF ORAL & MAXILLOFACIAL SURGERY, v. 51, n. 8, p. 971-973, . (09/54722-1)
BASSO, F. G.; PANSANI, T. N.; TURRIONI, A. P. S.; KURACHI, C.; BAGNATO, V. S.; HEBLING, J.; DE SOUZA COSTA, C. A.. Biostimulatory effects of low-level laser therapy on epithelial cells and gingival fibroblasts treated with zoledronic acid. Laser Physics, v. 23, n. 5, . (09/54722-1)
FERNANDA GONCALVES BASSO; TAISA NOGUEIRA PANSANI; CAMILA FAVERO DE OLIVEIRA; ANA PAULA SILVEIRA TURRIONI; DIANA GABRIELA SOARES; JOSIMERI HEBLING; CARLOS ALBERTO DE SOUZA COSTA. Cytotoxic Effects of Zoledronic Acid on Human Epithelial Cells and Gingival Fibroblasts. Brazilian Dental Journal, v. 24, n. 6, p. 551-558, . (09/54722-1)
BASSO, FERNANDA G.; OLIVEIRA, CAMILA F.; KURACHI, CRISTINA; HEBLING, JOSIMERI; DE SOUZA COSTA, CARLOS A.. Biostimulatory effect of low-level laser therapy on keratinocytes in vitro. Lasers in Medical Science, v. 28, n. 2, p. 367-374, . (09/54722-1)
BASSO, F. G.; TURRIONI, A. P. S.; HEBLING, J.; DE SOUZA COSTA, C. A.. Effects of zoledronic acid on odontoblast-like cells. ARCHIVES OF ORAL BIOLOGY, v. 58, n. 5, p. 467-473, . (09/54722-1)
FERNANDA GONÇALVES BASSO; ANA PAULA SILVEIRA TURRIONI; JOSIMERI HEBLING; CARLOS ALBERTO DE SOUZA COSTA. Inibição da atividade de osteoblastos expostos ao ácido zoledrônico. Jornal Brasileiro de Patologia e Medicina Laboratorial, v. 49, n. 5, p. 368-371, . (09/54722-1)
NOGUEIRA PANSANI, T.; GONCALVES BASSO, F.; SILVEIRA TURIRIONI, A. P.; KURACHI, C.; HEBLING, J.; DE SOUZA COSTA, C. A.. Effects of low-level laser therapy on the proliferation and apoptosis of gingival fibroblasts treated with zoledronic acid. International Journal of Oral and Maxillofacial Surgery, v. 43, n. 8, p. 1030-1034, . (09/54722-1, 10/08933-8)

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