A new method for photodynamic therapy of melanotic melanoma -- effects of depigmentation with violet light photodynamic therapy

Melanotic melanomas have a poor response to photodynamic therapy (PDT). The reason for this is that melanin absorbs light over the entire wavelength region used for PDT (400-750 nm). Photobleaching of melanin is an approach to overcome this obstacle. In the present work, reflectance spectroscopy was applied to study depigmentation of human and murine skin with different melanin contents, and effects induced by PDT with topical application of methyl 5-aminolevulinate (MAL) on B16F10 melanotic melanomas transplanted to nude mice. Depigmentation and inhibition of tumor growth after violet light (420 nm) exposure, red light (634 nm) exposure, and combinations of both were studied. Reflectance spectroscopy was suitable for evaluation of the pigmentation of both human and murine skin. Skin depigmentation leads to increase in reflectance. PDT with violet light bleached some of the melanin in the skin above the B16F10 melanomas, and possibly also in the upper part of the melanomas. This resulted in a larger growth inhibition of tumors first given PDT with violet light and then with red light compared to treatments using the reverse order of illumination, namely, red light before violet light. It is concluded that violet light PDT can bleach melanin in melanotic tumors and therefore increase their sensitivity to red light PDT. This finding indicates a new PDT modality that can be further developed for treatment of superficial melanotic melanomas and possibly other diseases where pigmentation is a problem.     

纯氧及LED阵列光动力复合治疗设备的研制

基于变压吸附制氧原理产生纯氧,并采用微透镜阵列多光谱LED发光器件与二次透镜阵列相结合构成光动力治疗辐照器,研制一种纯氧及LED阵列光动力复合治疗设备。其辐照器输出纯氧浓度大于90%,照射光包括波长625 nm红光、465 nm蓝光和520 nm绿光,应用光排序辐照技术实现纯氧及多光谱光动力复合治疗。通过辐照器上纯氧和照射光同步输出等多种外源性给氧保持光动力治疗区域的富氧状态,解决由于乏氧影响光动力疗效的问题。辐照器光学系统解决现有技术采用LED阵列排布替代激光器作为光动力治疗光源时存在的光能利用率低、光功率密度分布不均匀、不同波长光束在目标靶面光功率密度分布曲面差异大等缺陷。     

Chlorophyll derivatives--a new photosensitizer for photodynamic therapy of cancer in mice

The in vivo photosensitizing efficacy of chlorophyll derivatives (CpD), which had been developed as a new photosensitizer, was compared with that of hematoporphyrin derivatives (HpD). A murine tumor model implanted subcutaneously with S-180 cells on the abdomen was used. The CpD or HpD was administered by intratumoral injection, and light of appropriate wavelength was irradiated on the tumor areas for 10 minutes at 1h and 24h or 24h and 48h after the injection of photosensitizer. When CpD was injected, the early irradiation group (1h and 24h) showed a 100% tumor cure rate; however, the late irradiation group (24h and 48h) showed a 60% tumor cure rate (p less than 0.01). This showed that the early irradiation with light after injection of CpD was an important factor for obtaining better results. With HpD, there was no difference in tumor cure rate between early (1h and 24h, 80%) and late irradiation (24h and 48h, 80%) groups. Thus, in early irradiation groups, the tumor cure rate using CpD (100%) was superior to that of HpD (80%) (p less than 0.05). However, in late irradiation groups, the tumor cure rate using CpD (60%) was inferior to that of HpD (80%), but this difference was not statistically significant (p greater than 0.1). Pathologic sections of these tumors were made before treatment and 48h and 3 weeks after treatment. These showed geographic necrosis at 48h after treatment and no viable tumor tissue at 3 weeks after treatment. Our results showed that CpD was as effective as HpD as a photosensitizer for in vivo photodynamic therapy.     

A new method of eliciting delayed hypersensitivity in the mouse abdominal cavity

A new method of determining delayed hypersensitivity quantitatively was investigated in mice. Mice were sensitized with 150 micrograms of ferritin and, 3 weeks later, antigen challenge was performed by implanting a sponge containing antigen in the abdominal cavity. Cells accumulated in the sponge markedly increased in number for 24-72 h after the challenge; mononuclear cells predominated by 48 h. When sensitized lymphocytes were transferred passively to a normal recipient, marked cell accumulation in the sponge was found 48 h after the challenge. Immunological specificity was confirmed in animals sensitized to antigen and receiving passive transfer of sensitized cells. Strain differences in this reaction were observed. Cortisone (20 mg/kg for 6 days before challenge) significantly decreased cell accumulation. Delayed hypersensitivity was also elicited in the ear of sensitized animals. Extracts of sponges removed from antigen-challenged mice had macrophage chemotactic activity.     

Optical dosimetry for interstitial photodynamic therapy

An approach to photodynamic treatment of tumors is the interstitial implantation of fiber optic light sources. Dosimetry is critical in identifying regions of low light intensity in the tumor which may prevent tumor cure. We describe a numerical technique for calculating light distributions within tumors, from multiple fiber optic sources. The method was tested using four translucent plastic needles, which were placed in a 0.94 X 0.94 cm grid pattern within excised Dunning R3327-AT rat prostate tumors. A cylindrical diffusing fiber tip, illuminated by 630 nm dye laser light was placed within one needle and a miniature light detector was placed within another. The average penetration depth in the tumor region between the two needles was calculated from the optical power measured by the detector, using a modified diffusion theory. Repeating the procedure for each pair of needles revealed significant variations in penetration depth within individual tumors. Average values of penetration depth, absorption coefficient, scattering coefficient, and mean scattering cosine were 0.282 cm, 0.469 cm-1, 250 cm-1 and 0.964, respectively. Calculated light distributions from four cylindrical sources in tumors gave reasonable agreement with direct light measurements using fiber optic probes.     

Design and construction of a light-delivery system for photodynamic therapy.

We have developed a device to divide the output from a dye laser into as many as eight beams of equal power with negligible total power loss. In this system, 630-nm s-plane polarized laser light was split by a series of highly polarization-sensitive plate beamsplitters. Each of the beams was coupled to a 200, 400, or 600 microm diameter optical fiber. Brewster-window-type attenuators allowed the power of each beam to be individually set. It was possible to reconfigure the device to produce four, two, or one output(s). We discuss the design requirements of the beamsplitter device and describe its construction from mostly commercially available components. An apparatus for positioning and stabilizing each optical fiber relative to the skin surface of a patient is also described. The illumination from the fiberoptic supported by such an apparatus strikes a defined surface area and is independent of patient movement. Both the beamsplitter device and the optical fiber positioner are used routinely in photodynamic therapy (PDT) of malignant tumors in the clinic and in the laboratory.     

Integrating spheres for improved skin photodynamic therapy

The prescribed radiant exposures for photodynamic therapy (PDT) of superficial skin cancers are chosen empirically to maximize the success of the treatment while minimizing adverse reactions for the majority of patients. They do not take into account the wide range of tissue optical properties for human skin, contributing to relatively low treatment success rates. Additionally, treatment times can be unnecessarily long for large treatment areas if the laser power is not sufficient. Both of these concerns can be addressed by the incorporation of an integrating sphere into the irradiation apparatus. The light fluence rate can be increased by as much as 100%, depending on the tissue optical properties. This improvement can be determined in advance of treatment by measuring the reflectance from the tissue through a side port on the integrating sphere, allowing for patient-specific treatment times. The sphere is also effective at improving beam flatness, and reducing the penumbra, creating a more uniform light field. The side port reflectance measurements are also related to the tissue transport albedo, enabling an approximation of the penetration depth, which is useful for real-time light dosimetry.     

Vascular and cellular targeting for photodynamic therapy

Photodynamic therapy (PDT) involves the combination of photosensitizers (PS) with light as a treatment, and has been an established medical practice for about 10 years. Current primary applications of PDT are age-related macular degeneration (AMD) and several types of cancer and precancer. Tumor vasculature and parenchyma cells are both potential targets of PDT damage. The preference of vascular versus cellular targeting is highly dependent upon the relative distribution of photosensitizers in each compartment, which is governed by the photosensitizer pharmacokinetic properties and can be effectively manipulated by the photosensitizer drug administration and light illumination interval (drug-light interval) during PDT treatment, or by the modification of photosensitizer molecular structure. PDT using shorter PS-light intervals mainly targets tumor vasculature by confining photosensitizer localization within blood vessels, whereas if the sensitizer has a reasonably long pharmacokinetic lifetime, then PDT at longer PS-light intervals can induce more tumor cellular damage, because the photosensitizer has then distributed into the tumor cellular compartment. This passive targeting mechanism is regulated by the innate photosensitizer physicochemical properties. In addition to the passive targeting approach, active targeting of various tumor endothelial and cellular markers has been studied extensively. The tumor cellular markers that have been explored for active photodynamic targeting are mainly tumor surface markers, including growth factor receptors, low-density lipoprotein (LDL) receptors, transferrin receptors, folic acid receptors, glucose transporters, integrin receptors, and insulin receptors. In addition to tumor surface proteins, nuclear receptors are targeted, as well. A limited number of studies have been performed to actively target tumor endothelial markers (ED-B domain of fibronectin, VEGF receptor-2, and neuropilin-1). Intracellular targeting is a challenge due to the difficulty in achieving sufficient penetration into the target cell, but significant progress has been made in this area. In this review, we summarize current studies of vascular and cellular targeting of PDT after more than 30 years of intensive efforts.     

The use of photodynamic therapy for diseases of the esophagus

This is a review of the uses, history, and current status of photodynamic therapy for diseases of the esophagus, specifically Barrett's dysplasia and early esophageal carcinoma. This paper describes the clinical experience of photodynamic therapy and compares the use of various photosensitizer drugs. Finally, important biophotonics developments are discussed, including their anticipated impact for improved endoscopic detection of dysplasia and carcinoma. In addition, methods for real-time photodynamic therapy and light dosimetry are provided in order to optimize ablation treatment outcomes while minimizing the risk of complications.     

Intrauterine light delivery for photodynamic therapy of the human endometrium

Photodynamic therapy is currently being evaluated as a minimallyinvasive procedure for endometrial ablation not requiring anaesthesia.Light penetration depths at 630, 660 and 690 nm and the optimalconfiguration of intrauterine light-diffusing fibres were determinedin 14 human uteri to assist in the design of a light intrauterinedevice. Post-menopausal ex-vivo uteri showed a significantlylower light penetration depth than pre-menopausal uteri. Witha single central diffusing fibre inserted, the fluence ratemeasured in the uterine wall at the most remote point of thecavity decreased to 1.1 ± 0.4% of that measured at closestproximity, whereas it decreased to only 40.0 ± 9.0% withthree fibres. Distension of the uterine cavity with 2 ml ofan optically clear fluid increased the fluence rate at the fundusbetween the fibres at a depth of 2 mm by a factor of 4. We concludethat in normal-sized pre-menopausal uterine cavities, threediffusing fibres will deliver an optical dose above the photodynamicthreshold level at a depth of 4 mm, even in the most remoteareas, in <30 min without causing thermal damage. For distortedand elongated cavities, either slight distension of the cavityor the insertion of a fourth diffusing fibre is required.     

A new method of sanation and drainage of the abdominal cavity in experimental generalized suppurative peritonitis

Elimination of microflora and its products from the abdomena, prevention of commissures, abscesses in postoperative peritonitis are essential for the disease outcome. The sack for this purpose made of polyvinilpirrolidone film (PVF) differs from the standard one by aceptic solution which washes the small intestine, eliminates microflora and products via the tube. This diminishes the count of microorganisms in the abdomena, arrests toxicity, activates immune system.