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The effect of UV-Vis to near-infrared light on the biological response of human dental pulp cells

机译:UV-Vis对近红外光对人牙髓细胞生物响应的影响

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Human dental pulp cells (DPCs) were isolated and cultured in phenol-red-free α-MEM/10%-FCS at 37°C in 5% CO_2. DPCs at passages 2-4 were seeded (150μL; 25,000 cell/ml) in black 96-microwell plates with transparent bases. 24h post-seeding, cultures were irradiated using a bespoke LED array consisting of 60 LEDs (3.5mW/cm~2) of wavelengths from 400-900nm (10 wavelengths, n=6) for time intervals of up to 120s. Metabolic and mitochondrial activity was assessed via a modified MTT assay. Statistical differences were identified using multi-factorial analysis of variance and post-hoc Tukey tests (P=0.05). The biological responses were significantly dependent upon post-irradiation incubation period, wavelength and exposure time (P<0.05). At shorter wavelength irradiances (400nm), a reduction in mitochondrial activity was detected although not significant, whereas longer wavelength irradiances (at 633, 656, 781 and 799nm) significantly increased mitochondrial activity (P<0.05) in DPCs. At these wavelengths, mitochondrial activity was generally increased for exposures less than 90s with 30s exposures being most effective with 24h incubation. Increasing the post-irradiation incubation period increased the measured response and identified further significance (P<0.05). The biological responses of human DPCs were wavelength, exposure-time and incubation period dependent. The optimisation of irradiation parameters will be key to the successful application of LLLT in dentistry.
机译:将人牙髓(DPC)分离并在37℃下在5%CO_2中在37℃下以苯酚 - 红α-MEM / 10%-FC培养。通道2-4的DPC在黑色96微孔板中接种(150μl; 25,000个单元/ ml),具有透明碱基。 24小时后,使用由400-900nm(10波长,n = 6)的60个LED(3.5mW / cm〜2)组成的定制LED阵列,培养培养物,该时间间隔高达120秒。通过改性的MTT测定评估代谢和线粒体活性。使用对差异和后HOC Tukey测试的多因素分析来确定统计差异(P = 0.05)。生物反应显着取决于照射后孵育期,波长和暴露时间(P <0.05)。在较短的波长(400nm)下,检测到线粒体活性的降低虽然不显着,而较长的波长(633,656,781和799nm)显着增加了DPC的线粒体活性(P <0.05)。在这些波长下,对于少于90秒的暴露通常增加线粒体活性,30S曝光最有效,24h孵育。增加后照射后培养期增加了测量的响应并确定了进一步的意义(P <0.05)。人DPC的生物反应是波长,暴露时间和潜伏期依赖性。辐照参数的优化将是牙科LLLT成功应用的关键。

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