Evaluation of low doses of gamma irradiation in the formation of mineralization nodules in osteoblasts culture

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dc.contributor.authorTARGINO, BARBARA
dc.contributor.authorPINTO, THAIS L.
dc.contributor.authorSILVA, EVILY F.
dc.contributor.authorSOMESSARI, E.S.R.
dc.contributor.authorBELLINI, MARIA H.
dc.contributor.authorAFFONSO, REGINA
dc.coverageNacionalpt_BR
dc.creator.eventoANAIS DA SOCIEDADE BRASILEIRA DE BIOCIENCIAS NUCLEARESpt_BR
dc.date.accessioned2018-04-11T18:36:23Z
dc.date.available2018-04-11T18:36:23Z
dc.date.evento09-11 de outubro, 2017pt_BR
dc.description.abstractIntroduction: Osteoblasts are specialized fibroblasts that secrete and mineralize the bone matrix. The mineralized extracellular matrix is mainly composed of type I collagen, osteocalcin, and the inorganic mineral hydroxylapatite1. The use of radiation as therapy in some cancers causes great bone loss. However, low dose radiation may have the opposite effect. Low dose X-irradiation on osteoblastic culture had effects on proliferation and differentiation with increase of mineralization nodules2. However, there is little information on the potential therapeutic efficacy of low-dose gamma-irradiation in the formation of mineralization nodules. Objective: To evaluate the effects of irradiation with 60Co γ-rays in low doses in the formation of mineralization nodules in culture of osteoblasts. Methods: MC3T3-E1 cells were bought by the Banco de Células do Rio de Janeiro, Brazil (MC3T3-E1 Subclone 14). The cells were cultured in α-MEM medium consisting of 10% FBS and without β-glycerophosphate and L-ascorbic acid (GIBCO, Custom Product, Catalog No. A1049001) (Zhao Y, Guan H, Liu S et al. Biol. Pharm. Bull. 2005, 28(8):1371-1376). Plating efficiency assays: cells were plated at a density of 100 cell/plate into 60 mm Petri dishes. After 14 days the places were stained with violet crystal and the colonies were counted. -glycerophosphate and 50 mg/ml ascorbic acid, and analyzed on days 7, 14 and 21. Osteoblast culture irradiation assay: cells were plated at a density of 1x 105 cells/plate on 60 mm dishes and the next day were irradiated by 60Co source with 0 (as the control), 0.5, 1.0, 1.5 and 2.0 Gy using the GammaCell 220 – Irradiation Unit of Canadian-Atomic Energy Commission Ltd. (CTR-IPEN). On day 21 of culture, undifferentiated (without ascorbic acid and β-glycerophosphate), differentiating cells (0 Gy) and irradiated cells at different doses, the medium was removed, cells were washed with phosphate buffer saline, fixed with 70% ethyl alcohol and stained with Alizarin red S (Sigma). All in three biological replicates (a total of 54 samples) and multiple comparisons were assessed by One-way ANOVA followed by Bonferroni´s tests with GraphPad Prism version 6.0 software. P< 0.05 was considered statistically significant. Results: Plating efficiency (PF) analysis is generally considered to be the gold standard of assays for testing the sensitivity of cell lines to ionizing radiation or other cytotoxic agents in vitro. The results obtained were a PF of 30% for non-irradiated culture, however, the irradiated culture obtained 40% in relation to the no-irradiated one, already with 0.5 Gy, and this percentage was maintained in the other larger doses. Regarding the evaluation of the formation of mineralization nodules, significant difference in 0.5 Gy group was observed compared with the control group (0 Gy), 64.7±1.8 and 53.0±0.9, respectively. The groups of 1.0, 1.5 and 2.0 Gy obtained a decrease in the mineralization nodules. The data obtained with increasing irradiation produced an increase of mineralization nodules up to 0.5 Gy and in the higher doses had a decrease. Applying the data in a non-linear function it is observed that the line has a decreasing tendency with the negative angular coefficient. This analysis is in agreement with the hormesis model, in which low doses induce a stimulatory effect while high doses cause inhibition4. Conclusions: This study is one among the first that investigating the biophysics of low-dose gamma-irradiation on MC3T3-E1 culture, focusing on the potential applications in bone replacement therapy.pt_BR
dc.event.siglaSBBNpt_BR
dc.format.extent116-116pt_BR
dc.identifier.citationTARGINO, BARBARA; PINTO, THAIS L.; SILVA, EVILY F.; SOMESSARI, E.S.R.; BELLINI, MARIA H.; AFFONSO, REGINA. Evaluation of low doses of gamma irradiation in the formation of mineralization nodules in osteoblasts culture. In: ANAIS DA SOCIEDADE BRASILEIRA DE BIOCIENCIAS NUCLEARES, 09-11 de outubro, 2017, São Paulo, SP. <b>Abstract...</b> Rio de Janeiro, RJ: Sociedade Brasileira de Biociências Nucleares, 2017. p. 116-116. Disponível em: http://repositorio.ipen.br/handle/123456789/28823.
dc.identifier.orcidhttps://orcid.org/0000-0002-9264-4262
dc.identifier.orcidhttps://orcid.org/0000-0003-2852-6189
dc.identifier.urihttp://repositorio.ipen.br/handle/123456789/28823
dc.localRio de Janeiro, RJpt_BR
dc.local.eventoSão Paulo, SPpt_BR
dc.publisherSociedade Brasileira de Biociências Nuclearespt_BR
dc.rightsopenAccesspt_BR
dc.titleEvaluation of low doses of gamma irradiation in the formation of mineralization nodules in osteoblasts culturept_BR
dc.typeResumo de eventos científicospt_BR
dspace.entity.typePublication
ipen.autorBARBARA TARGINO
ipen.autorEVILY FERNANDES DA SILVA
ipen.autorREGINA AFFONSO
ipen.autorMARIA HELENA BELLINI MARUMO
ipen.autorELIZABETH SEBASTIANA RIBEIRO SOMESSARI
ipen.autorTHAIS HELENA LAZARI PINTO
ipen.codigoautor14575
ipen.codigoautor14376
ipen.codigoautor1547
ipen.codigoautor1242
ipen.codigoautor1544
ipen.codigoautor14372
ipen.contributor.ipenauthorBARBARA TARGINO
ipen.contributor.ipenauthorEVILY FERNANDES DA SILVA
ipen.contributor.ipenauthorREGINA AFFONSO
ipen.contributor.ipenauthorMARIA HELENA BELLINI MARUMO
ipen.contributor.ipenauthorELIZABETH SEBASTIANA RIBEIRO SOMESSARI
ipen.contributor.ipenauthorTHAIS HELENA LAZARI PINTO
ipen.date.recebimento18-04pt_BR
ipen.event.datapadronizada2017pt_BR
ipen.identifier.ipendoc24637pt_BR
ipen.notas.internasAbstractpt_BR
ipen.type.genreResumo
relation.isAuthorOfPublication421f198f-1405-4d37-93a3-42c10413ff66
relation.isAuthorOfPublication90f484f8-13c9-41d9-9717-621452f80b4b
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relation.isAuthorOfPublication6a450cbf-91b1-4386-b4a8-7304afac99cc
relation.isAuthorOfPublication155e1a85-0635-407b-920c-00af43266aca
relation.isAuthorOfPublication27226bc3-b38c-4a38-bc93-e7a7ddfc80aa
relation.isAuthorOfPublication.latestForDiscovery421f198f-1405-4d37-93a3-42c10413ff66
sigepi.autor.atividadeTARGINO, BARBARA:14575:820:Spt_BR
sigepi.autor.atividadeSILVA, EVILY F.:14376:820:Npt_BR
sigepi.autor.atividadeSOMESSARI, E.S.R.:1544:220:Npt_BR
sigepi.autor.atividadeBELLINI, MARIA H.:1242:820:Npt_BR
sigepi.autor.atividadeAFFONSO, REGINA:1547:820:Npt_BR
sigepi.autor.atividadePINTO, THAIS L.:14372:-1:N
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