Effect of photobiomodulation on the viability of osteoblasts and fibroblasts submitted to alendronate sodium or zoledronic acid

M. Brozoski, Natalia Caroline Aguiar Tartaroti, Andreia Aparecida Trainá, M. Deboni, M. Marques, Maria da Graça Naclério Homem
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Abstract

The goal of this study is to evaluate the effect of photobiomodulation therapy (PBMT) on the viability of osteoblasts and cultured fibroblasts in different concentrations of alendronate or zoledronic acid. Two cell lines: osteoblast-like mouse cells (OSTEO 1) and human buccal mucosa fibroblast (FMM1) were used. Cells were submitted to different concentrations of bisphosphonates (1 μM, 10 μM, and 100 μM sodium alendronate and 3 μM, 5 μM and 10 μM zoledronic acid) for 24 hours. Next, the cultures received PBMT. The irradiations were applied with a diode laser (InGaAIP, 660 nm, 30 mW, spot 0.028 cm2) in continuous, punctual and contact mode at two energy densities: 5 J/cm2 (4.5 s) or 10 Jcm2 (9s) with 6 hours-intervals. Cell viability was determined by mitochondrial activity assay (MTT) 24 h after the last irradiation. The data were compared by the one way- ANOVA, complemented by the Tukey’s test (p < 0.05). Sodium alendronate at concentrations of 100 μM and 10 μM and zoledronic acid at 10 μM concentration showed higher long-term toxicity. The cellular viability of the PBMT treated group was significantly higher than that of the negative control group. The same occurred with the osteoblasts treated with the highest concentrations of the drug (5 and 10 μM), despite not reaching the cell viability of the positive control group, it presented greater viability than the negative control where the cells were not irradiated. In the groups submitted to zoledronic acid, positive controls presented greater cell viability. We concluded that under the parameters applied in this study, PBMT at an energy density of 5 J/cm2 was able to revert the toxicity of sodium alendronate applied at the higher concentrations in both cell types, whereas zoledronic acid toxicity, regardless of its concentrations, was not influenced by PBMT.
光生物调节对阿仑膦酸钠或唑来膦酸对成骨细胞和成纤维细胞活力的影响
本研究的目的是评估光生物调节疗法(PBMT)对不同浓度阿仑膦酸或唑来膦酸对成骨细胞和培养成纤维细胞活力的影响。采用小鼠成骨样细胞(osteo1)和人颊粘膜成纤维细胞(FMM1)两种细胞系。将细胞置于不同浓度的双膦酸盐(1 μM、10 μM和100 μM阿仑膦酸钠和3 μM、5 μM和10 μM唑来膦酸)中24小时。接下来,对培养物进行PBMT。用二极管激光器(InGaAIP, 660 nm, 30 mW,光斑0.028 cm2)以连续,准时和接触模式照射,能量密度为5 J/cm2 (4.5 s)或10 Jcm2 (9s),间隔6小时。末次照射后24 h,采用线粒体活性测定法(MTT)测定细胞活力。资料比较采用单因素方差分析,并辅以Tukey检验(p < 0.05)。100 μM和10 μM浓度的阿仑膦酸钠和10 μM浓度的唑来膦酸表现出较高的长期毒性。PBMT处理组细胞活力显著高于阴性对照组。用最高浓度的药物(5 μM和10 μM)处理的成骨细胞也出现了同样的情况,尽管没有达到阳性对照组的细胞活力,但其细胞活力高于未照射的阴性对照组。在给予唑来膦酸的组中,阳性对照呈现更高的细胞活力。我们得出结论,在本研究中应用的参数下,能量密度为5 J/cm2的PBMT能够恢复两种细胞类型中较高浓度的阿仑膦酸钠的毒性,而唑来膦酸的毒性,无论其浓度如何,都不受PBMT的影响。
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