Photo-toxic effects of 809-nm diode laser and indocyanine green on MDA-MB231 breast cancer cells.

Photodynamic therapy (PDT), due to its positive outcomes in clinical applications, easiness of practice and few side effects, is a good candidate for an efficient treatment of cancer. Indocyanine green (ICG), a water-soluble, anionic tricarbocyanine and non-toxic molecule is a promising photosensitive agent for PDT applications on tumor cells. ICG exhibits strong maximum absorption at around 805 nm which will be an advantage for its use in PDT; light at that wavelength can be used to treat deeper tumors. In this study the inhibitory growth effects of ICG-PDT on MDA-MB231 human breast cancer cells were investigated in a time course experiment. Cells were irradiated with a continuous wave diode laser (lambda=809 nm, 60 mW, 24 J cm(-2)). Cell viability was measured by MTT assay 0, 3, 6, 9, 12, 24 and 48h after light irradiation. The results showed that ICG-PDT application exerted its photo-oxidative effect on MDA-MB231 breast cancer cells immediately. Relative cell viability was determined throughout the 48h time course, and a consistent decrease was observed after ICG-PDT applications. In conclusion, ICG when used in combination with near-infrared light showed a very fast (within 3h) and persistent (up to 48h) photo-toxic effect on MDA-MB231 human breast cancer cells.

Multiple components of photodynamic therapy can phosphorylate Akt.

A growing number of clinically relevant molecular and cellular responses are observed following photodynamic therapy (PDT). PDT-mediated oxidative stress and PDT-induced tissue hypoxia can elicit the transcriptional and/or translational expression of genes associated with cellular stress, inflammation, angiogenesis, immuno-modulation, apoptosis and signal transduction. One of the signaling molecules activated by oxidative stress is Akt/protein kinase B. Phosphorylation of Akt/protein kinase B activates this signaling molecule and induces a survival response in effected cells and tissue. We hypothesized that PDT using Photofrin (PH) as the photosensitizer could also induce increased levels of Akt phosphorylation. Results from our initial set of experiments demonstrated that in vitro and in vivo PDT treatments induced Akt phosphorylation. Interestingly, incubation of mouse and human breast cancer cells with the porphyrin-based photosensitizer, PH, increased the expression of Akt phosphorylation in the absence of light. Exposure of the corresponding mouse and human-derived breast cancer tumors growing in mice to 630 nm light in the absence of PH administration also induced Akt phosphorylation. These results demonstrate that individual components of the PDT process, photosensitizer alone and light alone, as well as the complete PDT procedure can activate the Akt signaling pathway.

Closure of skin incisions by 980-nm diode laser welding.

A 980-nm diode laser is proposed to be an alternative welding laser in dermatology due to its optimal penetration in tissue. An in vivo predosimetry study was done to estimate the optimal laser energy delivery conditions (6 W, 400 ms). Next, in vivo experiments were comparatively performed to examine healing of wounds closed either with suture or laser welding. One-centimeter-long, full-thickness incisions were done on the dorsal side of Wistar rats. Wounds were surgically removed at 1, 4, 7, 14, and 21 days postoperatively. Macroscopic examinations showed that welding had minimal scarring and a fine quality healing. According to histological (hematoxylin and eosin staining) results, change of epidermal thickness and granulation tissue formation through 21 days of healing period showed similarities in both methods. Epidermal thickness of welded wounds decreased from 62.46+/-6.87 microm (first day) to 36.49+/-0.92 microm (21st day) and that of sutured wounds decreased from 62.94+/-13.53 microm (first day) to 37.88+/-7.41 microm (21st day). At day 14, epidermal thickness of sutured wounds (61.20+/-6.60 microm) were higher than welded wounds (49.69+/-6.31 microm) (p<0.05). Besides, granulation values were greater for the sutured wounds but the difference was statistically significant (p<0.05) only for the seventh day (197,190.29+/-.89,554.96 microm(2) for sutured wounds, 138,433.1+/-51,077.17 microm(2) for welded wounds). Those differences indicate a faster recovery with laser welding. It is concluded that tissue welding with a 980-nm diode laser can be a good candidate for tissue welding applications with accelerated and improved healing, but further investigations are in progress for clinical use.

The biological effects of 632.8-nm low energy He-Ne laser on peripheral blood mononuclear cells in vitro.

The aim of this study was to examine the proliferation of peripheral blood mononuclear cells due to the low energy 632.8-nm He-Ne laser application. The results of previous studies supported the hypothesis that low level laser therapy (LLLT) might have an increasing effect on the proliferation of lymphocytes and production of cytokines. The effect of laser irradiation was investigated by comparing the proliferation of peripheral blood mononuclear cells (PBMC) with a mitogenic stimulator, PHA (phytohemaglutinin) and laser irradiation. PBMCs of control samples, only laser irradiated samples, PHA included samples and both PHA included and laser irradiated samples were quantified and compared. Results of [3H] Thymidine test, 20 s laser irradiated and 40 s laser irradiated samples' proliferation were found statistically higher than control samples. There was no significant difference between control and 60 s laser irradiated samples. PHA also showed its ability to proliferate cells. PHA included samples and both PHA included and laser irradiated samples' proliferation was higher than both control and only laser irradiated samples. Our results showed that He-Ne laser application enhanced the proliferation significantly. Moreover, laser dose was noted as a significant parameter. On the other hand, LLLT by itself was found less effective than PHA.

Development of a surgical diode-laser system: controlling the mode of operation.

OBJECTIVE: The aim of this study was to develop a microcontroller based surgical diode laser system and to test it at two different modes (continuous [CW] and modulated) in vitro on lamb liver tissue. BACKGROUND DATA: In laser surgery, depending on the properties of laser source (wavelength, power, application time, and mode of operation), the effects observed on the tissue may change from carbonization to hyperthermia. The aim is to remove the target tissue without giving any thermal damage to the surrounding tissue. Carbonization should be avoided, thus controlling the mode of operation is very crucial. METHODS: The system consisted of a microcontroller based control unit, 980-nm high-power diode laser source, and fiber delivery unit. This system has the capability of delivering different modes of laser energy to the target tissue ranging from CW to 20-Hz modulated beams. The surgical diode laser system was tested on liver tissue in vitro. Efficiency of laser-tissue interaction was quantified in terms of thermal alteration per unit energy and corresponding carbonization level. RESULTS: Modulated mode resulted in larger coagulated area with minimum carbonizations. Carbonized area/thermally altered area (CarbA/TAA) ratio for CW mode of operation at 16 J is 0.35; however, this ratio was found to be 0.05 at modulated mode, when even 10 times higher energy (160 J) was delivered to the target tissue. CONCLUSION: Results emphasized the significance of mode of operation as well as the other laser parameters. Modulated mode was found to be a promising regime for safer laser surgery.

The 980-nm diode laser for brain surgery: histopathology and recovery period.

The 980-nm diode laser has been under investigation for neurosurgery because of a local peak in the absorption spectra of water around this wavelength. This work was carried out to examine the extent of thermal changes and the recovery process of laser-induced brain lesions. In order to study the quality of the lesions, a conventional monopolar electrocoagulation technique was applied comparatively. An in vivo stereotaxic neurosurgical procedure was performed on Wistar rats. Bilateral brain lesions of the same size (2-3 mm diameter) were created with a diode laser and via electrocoagulation. Subjects were sacrificed 0, 2 and 7 days after surgery in order to observe the healing process of the necrotic tissue. The surgical after-effects of both types of lesions were identified through immunohistochemical staining with CD68 macrophage marker and haematoxylin eosin (H&E). CD68 was found to be more efficient than H&E in determining the thermally altered areas. Histological examinations showed that the 980-nm diode laser system has a remarkable ablating ability with minimal thermal damage of nearby tissue.