Department of Pediatric Dentistry, Orthodontics and Collective Health, Bauru School of Dentistry, University of São Paulo, Alameda Dr. Octávio Pinheiro Brisolla, 9-75, Bauru, São Paulo, 17012-901, Brazil.
Sacred Heart University, Bauru, São Paulo, Brazil.
Lasers Med Sci. 2021 Oct;36(8):1751-1758. doi: 10.1007/s10103-021-03291-4. Epub 2021 Apr 1.
This study aimed to evaluate the effect of different photobiomodulation (PBM) radiant exposures on the viability, proliferation, and gene expression of pulp fibroblasts from human primary teeth (HPF) involved in the pulp tissue repair. HPF were irradiated with Laser InGaAlP (Twin Flex Evolution, MMOptics®) at 660-nm wavelength (red); single time, continuous mode, 0.04-cm laser tip area, and 0.225-cm laser tip diameter, keeping the distance of 1 mm between the laser beam and the cell culture. The doses used were between 1.2 and 6.2 J/cm and were evaluated at the 6 h, 12 h, and 24 h after PBM. MTT and crystal violet assays evaluated the cell viability and proliferation. RT-PCR verified VEGF and FGF-2 mRNA expression. A blinded examiner analyzed the data through two-way ANOVA followed by Tukey test (p < 0.05). The groups with higher powers (10 mW, 15 mW, 20 mW, and 25 mW), shortest application periods (10 s), and radiant exposures between 2.5 and 6.2 J/cm exhibited statistically higher viability than that of the groups with small power (5 mW), longer application period (50 s), and radiant exposure of 6.2 J/cm (p < 0.05). VEGF and FGF-2 mRNA expression were observed at the three evaluated periods (6 h, 12 h, and 24 h) and the highest expression was in the shortest period (p < 0.05). All radiant exposures maintained HPF viable. The period of 6 h after irradiation showed statistically greater gene expression for both growth factors than other periods. VEGF mRNA had no differences among the dosimetries studied. The best radiant exposures for FGF-2 gene expression were 2.5 J/cm and 3.7 J/cm.
本研究旨在评估不同光生物调节(PBM)辐射剂量对参与牙髓组织修复的人原代牙髓细胞(HPF)活力、增殖和基因表达的影响。HPF 用波长为 660nm(红色)的激光 InGaAlP(Twin Flex Evolution,MMOptics®)照射;单次、连续模式,激光尖端面积 0.04cm²,激光尖端直径 0.225cm,激光束与细胞培养物之间的距离保持 1mm。使用的剂量在 1.2 至 6.2J/cm 之间,并在 PBM 后 6 小时、12 小时和 24 小时进行评估。MTT 和结晶紫测定法评估细胞活力和增殖。RT-PCR 验证 VEGF 和 FGF-2 mRNA 表达。通过双因素方差分析和 Tukey 检验(p<0.05),由一名盲法检查者分析数据。高功率(10mW、15mW、20mW 和 25mW)、最短应用时间(10s)和辐射剂量在 2.5 至 6.2J/cm 之间的组,与低功率(5mW)、较长应用时间(50s)和辐射剂量为 6.2J/cm 的组相比,细胞活力显著更高(p<0.05)。在三个评估时间段(6 小时、12 小时和 24 小时)观察到 VEGF 和 FGF-2 mRNA 表达,最短时间表达最高(p<0.05)。所有辐射剂量均使 HPF 保持活力。照射后 6 小时的时间段显示两种生长因子的基因表达均显著高于其他时间段。在研究的剂量中,VEGF mRNA 没有差异。FGF-2 基因表达的最佳辐射剂量为 2.5J/cm 和 3.7J/cm。