Astuty Sri Dewi, Baktir Afaf, Astuti Suryani Dyah
Doctoral Program of Mathematics and Natural Science, Faculty of Science and Technology, Universitas Airlangga, Surabaya, Indonesia.
Department of Physics of Hasanuddin University, Makassar, Indonesia.
J Lasers Med Sci. 2019 Summer;10(3):215-224. doi: 10.15171/jlms.2019.35. Epub 2019 Jul 6.
Photodynamic inactivation has been developed to kill pathogenic microbes. In addition, some techniques have been introduced to minimize the biofilm resistance to antifungal properties in inhibiting cell growth. The principle of photodynamic inactivation different to antifungal drugs therapy which is resistant to biofilms. The presence of reactive oxygen species (ROS) that generating in photodynamic inactivation mechanisms can be damaging of biofilm cells and the principle of light transmission that could be penetrating in matrix layers of extracellular polymeric substance (EPS) until reaching the target cells at the base layers of biofilm. The present work aims to explore the potential of chlorophyll extract of papaya leaf as an exogenous photosensitizer to kill the biofilms after being activated by the laser. The potential of chlorophyll photosensitizer was evaluated based on the efficacy of inactivation biofilm cell through a cell viability test and an organic compound test. The treatment of photoinactivation was administered to 12 groups of biofilm for four days using the 445 nm laser and the 650 nm laser. The 445 nm and 650 nm lasers activated the chlorophyll extract of the papaya leaf (0.5 mg/L) at the same energy density. The energy density variation was determined as 5, 10, 20, 30 and 40 J/cm with the duration of exposure of each laser adjusted to the absorbance percentage of chlorophyll extract of the papaya leaf. The absorbance percentage of chlorophyll extracts of the papaya leaf on wavelengths of 650 nm and 445 nm respectively were 22.26% and 60.29%, respectively. The most effective treated group was a group of the laser with the addition of chlorophyll, done by the 650 nm lasers with inactivation about 32% (=0.001), while the 445 nm lasers only 25% (=0.061). The maximum malondialdehyde levels by treatment of the laser 650 nm were (0.046±0.004) nmol/mg. The use of chlorophyll extract of the papaya leaf as a photosensitizer, resulted in the maximum spectrum of absorption at 414 nm and 668 nm, which produced a maximum reduction effect after photoinactivation up to 32% (with chlorophyll) and 25% (without chlorophyll). The utilization of chlorophyll extract of the papaya leaf would increase the antifungal effects with the activation by the diode laser in the biofilm of .
光动力灭活技术已被用于杀灭致病微生物。此外,还引入了一些技术来最小化生物膜对抑制细胞生长的抗真菌特性的抗性。光动力灭活的原理与抗真菌药物疗法不同,后者对生物膜具有抗性。光动力灭活机制中产生的活性氧(ROS)的存在会损害生物膜细胞,并且光传输原理可以穿透细胞外聚合物(EPS)的基质层,直至到达生物膜基层的靶细胞。本研究旨在探索木瓜叶叶绿素提取物作为外源性光敏剂在激光激活后杀灭生物膜的潜力。基于通过细胞活力测试和有机化合物测试对生物膜细胞进行灭活的效果,评估叶绿素光敏剂的潜力。使用445nm激光和650nm激光对12组生物膜进行为期四天的光灭活处理。445nm和650nm激光以相同的能量密度激活木瓜叶的叶绿素提取物(0.5mg/L)。能量密度变化确定为5、10、20、30和40J/cm²,每种激光的照射时间根据木瓜叶叶绿素提取物的吸光度百分比进行调整。木瓜叶叶绿素提取物在650nm和445nm波长处的吸光度百分比分别为22.26%和60.29%。最有效的处理组是添加叶绿素的激光组,由650nm激光完成,灭活率约为32%(=0.001),而445nm激光仅为25%(=0.061)。650nm激光处理后的最大丙二醛水平为(0.046±0.004)nmol/mg。使用木瓜叶叶绿素提取物作为光敏剂,在414nm和668nm处产生最大吸收光谱,光灭活后产生的最大还原效果高达32%(有叶绿素)和25%(无叶绿素)。利用木瓜叶叶绿素提取物并通过二极管激光激活将增加对生物膜的抗真菌效果。