Lee Chia-Fen, Lee Chi-Jui, Chen Chin-Tin, Huang Ching-Tsan
Institute of Microbiology and Biochemistry, National Taiwan University, Taipei 106, Taiwan, ROC.
J Photochem Photobiol B. 2004 Jul 19;75(1-2):21-5. doi: 10.1016/j.jphotobiol.2004.04.003.
To demonstrate photodynamic antimicrobial chemotherapy (PACT) against planktonic and biofilm cultures of Pseudomonas aeruginosa, using photoporphyrin IX which could be endogenously synthesized by administrating delta-aminolaevulinic acid (delta-ALA), and a light emitted diode (LED) array to photoactivate the photosensitizer. P. aeruginosa suspended cells or biofilms, grown on a rotating disk reactor, were treated by different concentrations of delta-ALA in the dark for 1 h, followed by LED irradiation for various time. Regrowth experiments were conducted by placed PACT-treated disks back to a sterile reactor. Viable cells were determined by serial dilution and plate counts. Both P. aeruginosa planktonic and biofilm cells were inhibited by PACT with light doses or photosensitizer concentrations increasing. Treatments of planktonic cells with 10 mM delta-ALA and incident dose 240 J cm(-2) or 7.5 mM ALA and incident dose 360 J cm(-2) led to completely photoinactivation. No viable biofilm cells were found after treatment of 20 mM delta-ALA and incident dose 240 J cm(-2). However, regrowth was observed once PACT-treated biofilms were put back to a sterile reactor. Regrowth could be prevented only if biofilm samples were treated PACT twice. delta-ALA-mediated PACT on P. aeruginosa planktonic and biofilm cells was effective, though the detailed mechanism still required further investigation.
为了证明光动力抗菌化疗(PACT)对铜绿假单胞菌浮游菌和生物膜培养物的作用,使用可通过给予δ-氨基乙酰丙酸(δ-ALA)内源性合成的光卟啉IX,以及发光二极管(LED)阵列来光激活光敏剂。在旋转盘式反应器上生长的铜绿假单胞菌悬浮细胞或生物膜,在黑暗中用不同浓度的δ-ALA处理1小时,然后进行不同时间的LED照射。通过将经PACT处理的圆盘放回无菌反应器中进行再生长实验。通过系列稀释和平板计数来测定活细胞。随着光剂量或光敏剂浓度的增加,PACT对铜绿假单胞菌浮游菌和生物膜细胞均有抑制作用。用10 mM δ-ALA和入射剂量240 J cm(-2)或7.5 mM ALA和入射剂量360 J cm(-2)处理浮游细胞导致完全光灭活。用20 mM δ-ALA和入射剂量240 J cm(-2)处理后未发现活的生物膜细胞。然而,一旦将经PACT处理的生物膜放回无菌反应器中,就会观察到再生长。只有对生物膜样品进行两次PACT处理才能防止再生长。δ-ALA介导的PACT对铜绿假单胞菌浮游菌和生物膜细胞有效,尽管详细机制仍需进一步研究。