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Techniques for fluorescence detection of protoporphyrin IX in skin cancers associated with photodynamic therapy. 光动力疗法相关皮肤癌中原卟啉IX的荧光检测技术。
Pub Date : 2013-11-01 DOI: 10.1515/plm-2013-0030
Kishore R Rollakanti, Stephen C Kanick, Scott C Davis, Brian W Pogue, Edward V Maytin

Photodynamic therapy (PDT) is a treatment modality that uses a specific photosensitizing agent, molecular oxygen, and light of a particular wavelength to kill cells targeted by the therapy. Topically administered aminolevulinic acid (ALA) is widely used to effectively treat cancerous and precancerous skin lesions, resulting in targeted tissue damage and little to no scarring. The targeting aspect of the treatment arises from the fact that ALA is preferentially converted into protoporphyrin IX (PpIX) in neoplastic cells. To monitor the amount of PpIX in tissues, techniques have been developed to measure PpIX-specific fluorescence, which provides information useful for monitoring the abundance and location of the photosensitizer before and during the illumination phase of PDT. This review summarizes the current state of these fluorescence detection techniques. Non-invasive devices are available for point measurements, or for wide-field optical imaging, to enable monitoring of PpIX in superficial tissues. To gain access to information at greater tissue depths, multi-modal techniques are being developed which combine fluorescent measurements with ultrasound or optical coherence tomography, or with microscopic techniques such as confocal or multiphoton approaches. The tools available at present, and newer devices under development, offer the promise of better enabling clinicians to inform and guide PDT treatment planning, thereby optimizing therapeutic outcomes for patients.

光动力疗法(PDT)是一种使用特定光敏剂、分子氧和特定波长的光来杀死治疗目标细胞的治疗方式。局部给药氨基乙酰丙酸(ALA)被广泛用于有效治疗癌性和癌前皮肤病变,导致靶向组织损伤,几乎没有疤痕。这种治疗的靶向性源于ALA在肿瘤细胞中优先转化为原卟啉IX (PpIX)。为了监测组织中PpIX的量,已经开发了测量PpIX特异性荧光的技术,这为在PDT照明阶段之前和期间监测光敏剂的丰度和位置提供了有用的信息。本文综述了这些荧光检测技术的现状。非侵入性设备可用于点测量或宽视场光学成像,以监测表面组织中的PpIX。为了获得更深层的组织信息,正在开发多模态技术,将荧光测量与超声或光学相干断层扫描相结合,或与显微技术如共聚焦或多光子方法相结合。目前可用的工具和正在开发的新设备,使临床医生能够更好地告知和指导PDT治疗计划,从而优化患者的治疗结果。
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引用次数: 60
Fluorescence lifetime imaging with near-infrared dyes 近红外染料荧光寿命成像
Pub Date : 2013-02-22 DOI: 10.1515/plm-2014-0017
W. Becker, V. Shcheslavskiy
Abstract Near-infrared (NIR) dyes are used as fluorescence markers in small animal imaging and in diffuse optical tomography. In these applications it is important to know whether the dyes bind to proteins or to other tissue constituents, and whether their fluorescence lifetimes depend on the targets they bind to. Unfortunately, neither the optical beam paths nor the detectors of commonly used in confocal and multiphoton laser scanning microscopes (LSMs) directly allow for excitation and detection of NIR fluorescence. This paper presents three ways of adapting existing LSMs with time-correlated single photon counting (TCSPC) fluorescence lifetime imaging (FLIM) systems for NIR FLIM: 1) confocal systems with wideband beamsplitters and diode laser excitation, 2) confocal systems with wideband beamsplitters and one-photon excitation by titanium-sapphire lasers, and 3) two-photon systems with optical parametric oscillator (OPO) excitation and non-descanned detection. A number of NIR dyes are tested in biological tissue. All of them show clear lifetime changes depending on the tissue structures they are bound to. We therefore believe that NIR FLIM can deliver supplementary information about the tissue composition and on local biochemical parameters.
近红外(NIR)染料在小动物成像和漫射光学层析成像中被用作荧光标记。在这些应用中,重要的是要知道染料是否与蛋白质或其他组织成分结合,以及它们的荧光寿命是否取决于它们结合的目标。不幸的是,通常用于共聚焦和多光子激光扫描显微镜(lsm)的光束路径和检测器都不能直接激发和检测近红外荧光。本文提出了三种将现有lsm与时间相关单光子计数(TCSPC)荧光寿命成像(FLIM)系统相结合,用于近红外FLIM的方法:1)带宽带分束器和二极管激光激励的共聚焦系统,2)带宽带分束器和钛蓝宝石激光单光子激励的共聚焦系统,以及3)带光学参量振荡器(OPO)激励和非去扫描检测的双光子系统。许多近红外染料在生物组织中进行了测试。它们都显示出明显的寿命变化,这取决于它们所依附的组织结构。因此,我们相信近红外光谱可以提供有关组织组成和局部生化参数的补充信息。
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引用次数: 18
Low-level laser (light) therapy (LLLT) on muscle tissue: performance, fatigue and repair benefited by the power of light. 低水平激光(光)疗法(LLLT)对肌肉组织:性能,疲劳和修复受益于光的力量。
Pub Date : 2012-11-01 DOI: 10.1515/plm-2012-0032
Cleber Ferraresi, Michael R Hamblin, Nivaldo A Parizotto

The use of low level laser (light) therapy (LLLT) has recently expanded to cover areas of medicine that were not previously thought of as the usual applications such as wound healing and inflammatory orthopedic conditions. One of these novel application areas is LLLT for muscle fatigue and muscle injury. Since it is becoming agreed that mitochondria are the principal photoacceptors present inside cells, and it is known that muscle cells are exceptionally rich in mitochondria, this suggests that LLLT should be highly beneficial in muscle injuries. The ability of LLLT to stimulate stem cells and progenitor cells means that muscle satellite cells may respond well to LLLT and help muscle repair. Furthermore the ability of LLLT to reduce inflammation and lessen oxidative stress is also beneficial in cases of muscle fatigue and injury. This review covers the literature relating to LLLT and muscles in both preclinical animal experiments and human clinical studies. Athletes, people with injured muscles, and patients with Duchenne muscular dystrophy may all benefit.

低水平激光(光)治疗(LLLT)的使用最近已经扩展到覆盖以前不被认为是通常应用的医学领域,如伤口愈合和炎症性骨科疾病。其中一个新的应用领域是肌肉疲劳和肌肉损伤的LLLT。由于人们一致认为线粒体是细胞内主要的光受体,并且已知肌肉细胞中线粒体异常丰富,这表明LLLT应该对肌肉损伤非常有益。LLLT刺激干细胞和祖细胞的能力意味着肌肉卫星细胞可能对LLLT反应良好并帮助肌肉修复。此外,LLLT减轻炎症和氧化应激的能力在肌肉疲劳和损伤的情况下也是有益的。本文综述了临床前动物实验和人类临床研究中有关LLLT和肌肉的文献。运动员、肌肉受伤的人、杜氏肌营养不良症患者都可能受益。
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引用次数: 163
Far infrared radiation (FIR): its biological effects and medical applications. 远红外线辐射(FIR):其生物效应和医学应用。
Pub Date : 2012-11-01 DOI: 10.1515/plm-2012-0034
Fatma Vatansever, Michael R Hamblin

Far infrared (FIR) radiation (λ = 3-100 μm) is a subdivision of the electromagnetic spectrum that has been investigated for biological effects. The goal of this review is to cover the use of a further sub-division (3- 12 μm) of this waveband, that has been observed in both in vitro and in vivo studies, to stimulate cells and tissue, and is considered a promising treatment modality for certain medical conditions. Technological advances have provided new techniques for delivering FIR radiation to the human body. Specialty lamps and saunas, delivering pure FIR radiation (eliminating completely the near and mid infrared bands), have became safe, effective, and widely used sources to generate therapeutic effects. Fibers impregnated with FIR emitting ceramic nanoparticles and woven into fabrics, are being used as garments and wraps to generate FIR radiation, and attain health benefits from its effects.

远红外线(FIR)辐射(λ = 3-100 μm)是电磁波谱的一个分支,已被研究用于生物效应。本综述的目的是涵盖在体外和体内研究中观察到的该波段的进一步细分(3- 12 μm)的使用,以刺激细胞和组织,并被认为是某些医疗条件下有希望的治疗方式。技术进步提供了向人体输送FIR辐射的新技术。专业灯和桑拿,提供纯FIR辐射(完全消除近红外和中红外波段),已成为安全,有效,广泛使用的来源,以产生治疗效果。纤维中浸渍有FIR发射陶瓷纳米粒子并编织成织物,正被用作服装和包装,以产生FIR辐射,并从其影响中获得健康益处。
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引用次数: 226
Low intensity laser therapy accelerates muscle regeneration in aged rats. 低强度激光治疗加速老年大鼠肌肉再生。
Pub Date : 2012-10-01 DOI: 10.1515/plm-2012-0035
Fatma Vatansever, Natalia C Rodrigues, Livia L Assis, Sabrina S Peviani, Joao L Durigan, Fernando M A Moreira, Michael R Hamblin, Nivaldo A Parizotto

Background: Elderly people suffer from skeletal muscle disorders that undermine their daily activity and quality of life; some of these problems can be listed as but not limited to: sarcopenia, changes in central and peripheral nervous system, blood hypoperfusion, regenerative changes contributing to atrophy, and muscle weakness. Determination, proliferation and differentiation of satellite cells in the regenerative process are regulated by specific transcription factors, known as myogenic regulatory factors (MRFs). In the elderly, the activation of MRFs is inefficient which hampers the regenerative process. Recent studies found that low intensity laser therapy (LILT) has a stimulatory effect in the muscle regeneration process. However, the effects of this therapy when associated with aging are still unknown.

Objective: This study aimed to evaluate the effects of LILT (λ=830 nm) on the tibialis anterior (TA) muscle of aged rats.

Subjects and methods: The total of 56 male Wistar rats formed two population sets: old and young, with 28 animals in each set. Each of these sets were randomly divided into four groups of young rats (3 months of age) with n=7 per group and four groups of aged rats (10 months of age) with n=7 per group. These groups were submitted to cryoinjury + laser irradiation, cryoinjury only, laser irradiation only and the control group (no cryoinjury/no laser irradiation). The laser treatment was performed for 5 consecutive days. The first laser application was done 24 h after the injury (on day 2) and on the seventh day, the TA muscle was dissected and removed under anesthesia. After this the animals were euthanized. Histological analyses with toluidine blue as well as hematoxylin-eosin staining (for counting the blood capillaries) were performed for the lesion areas. In addition, MyoD and VEGF mRNA was assessed by quantitative polymerase chain reaction.

Results: The results showed significant elevation (p<0.05) in MyoD and VEGF genes expression levels. Moreover, capillary blood count was more prominent in elderly rats in laser irradiated groups when compared to young animals.

Conclusion: In conclusion, LILT increased the maturation of satellite cells into myoblasts and myotubes, enhancing the regenerative process of aged rats irradiated with laser.

背景:老年人患有骨骼肌疾病,影响其日常活动和生活质量;其中一些问题可以列出但不限于:肌肉减少症,中枢和周围神经系统的变化,血液灌注不足,导致萎缩的再生变化和肌肉无力。再生过程中卫星细胞的决定、增殖和分化受特定的转录因子调控,即肌源性调节因子(MRFs)。在老年人中,核磁共振成像的激活效率低下,阻碍了再生过程。近年来的研究发现,低强度激光治疗(LILT)在肌肉再生过程中具有刺激作用。然而,当与衰老相关时,这种疗法的效果仍然未知。目的:研究λ=830 nm的LILT对老年大鼠胫前肌(TA)的影响。实验对象和方法:雄性Wistar大鼠56只,分为老年和青年两组,每组28只。每组随机分为4组幼龄大鼠(3月龄),每组n=7只;4组老龄大鼠(10月龄),每组n=7只。分为低温损伤+激光照射组、单纯低温损伤组、单纯激光照射组和对照组(无低温损伤/无激光照射组)。激光治疗连续5天。第一次激光应用于损伤后24小时(第2天),第7天,在麻醉下剥离TA肌。在此之后,这些动物被安乐死。用甲苯胺蓝和苏木精-伊红染色(用于计数毛细血管)对病变区域进行组织学分析。定量聚合酶链反应检测MyoD和VEGF mRNA表达。结果:结果显示明显升高(p)。结论:LILT促进了卫星细胞向成肌细胞和肌管的成熟,促进了激光照射老年大鼠的再生过程。
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引用次数: 24
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