Trials of a Fluorescent Endoscopic Video System for Diagnosis and Treatment of the Head and Neck Cancer

This article presents the results of intraoperative fluorescent diagnostics via the endoscopic system for assessing the quality of photodynamic therapy (PDT) of head and neck cancer. The diagnosis and PDT procedures were performed on the five patients with malignant neoplasms of the vocal cords, lateral surface of the tongue, and trachea and cancer of the left parotid salivary gland. Molecular form of chlorin E6 (Ce6) was intravenously administered with a 1.0–1.1 mg/kg concentration for PDT. Fluorescent diagnostics (FD) was conducted before PDT and after PDT procedures. Control of PDT efficiency was carried out by evaluating the photobleaching of the drug (photosensitizer). The method of intraoperative fluorescent imaging allows determining the exact location of the tumor and its boundaries. The assessment of photosensitizer photobleaching in real time regime allows making quick decisions during PDT procedure, which helps improving the quality of patients’ treatment. The results showed the convenience of endoscopic fluorescent video system in various nosologies of head and neck cancer. Therefore, this diagnostic approach will improve the effectiveness of cancer treatment

Analysis of Fluorescence Decay Kinetics of Indocyanine Green Monomers and Aggregates in Brain Tumor Model In Vivo

Spectroscopic approach with fluorescence time resolution allows one to determine the state of a brain tumor and its microenvironment via changes in the fluorescent dye’s fluorescence lifetime. Indocyanine green (ICG) is an acknowledged infra-red fluorescent dye that self-assembles into stable aggregate forms (ICG NPs). ICG NPs aggregates have a tendency to accumulate in the tumor with a maximum accumulation at 24 h after systemic administration, enabling extended intraoperative diagnostic. Fluorescence lifetime analysis of ICG and ICG NPs demonstrates different values for ICG monomers and H-aggregates, indicating promising suitability for fluorescent diagnostics of brain tumors due to their affinity to tumor cells and stability in biological tissue

Comparitive accumulation study of chlorin group photosensitizers on monolayer and multicellular tumor spheroids of cell culture

International audienceThe comparitive analysis of the new photosensitizer for photodynamic therapy were conducted for increasing of oncological diseases efficient treatment

Phototherapy of Brain Tumours Using a Fibre Optic Neurosystem

In this work, a new approach was tested to assess the cellular composition of tissues by time-resolved methods of fluorescence analysis of exogenous and endogenous fluorophores. First of all, the differences in fluorescence kinetics of endogenous fluorophores (coenzymes NADH and FAD) in tumour and immunocompetent cells were determined. After that, differences in fluorescence kinetics of photosensitizer 5 ALA-induced protoporphyrin IX were established due to its different metabolism in cells of different phenotypes. Kinetics of photoluminescence of NADH and FAD coenzymes as well as photosensitizer were studied by means of two different methods: time-resolved spectroscopy based on a streak-camera and fibre optic neuroscopy, which served to perform process monitoring and regular fluorescence diagnosis of the probed region. Time-resolved fluorescence microscopy (FLIM) was used as a control technique. Time-resolved spectroscopic fluorescence lifetime analysis was performed on sexually mature female rats induced with glioma C6 brain tumour under in vivo conditions; thus, under conditions where the immune system actively intervenes in the process of oncogenesis. In this regard, the aim of the study was to recognize the cellular composition of the brain tumour tissue, namely the ratio of cancer and immunocompetent cells and their mutual localization. Understanding the role of the immune system thus provides new ways and approaches for further diagnosis and therapy, making tumour-associated immune cells a prime target for modern therapies

Phototherapy of Brain Tumours Using a Fibre Optic Neurosystem

In this work, a new approach was tested to assess the cellular composition of tissues by time-resolved methods of fluorescence analysis of exogenous and endogenous fluorophores. First of all, the differences in fluorescence kinetics of endogenous fluorophores (coenzymes NADH and FAD) in tumour and immunocompetent cells were determined. After that, differences in fluorescence kinetics of photosensitizer 5 ALA-induced protoporphyrin IX were established due to its different metabolism in cells of different phenotypes. Kinetics of photoluminescence of NADH and FAD coenzymes as well as photosensitizer were studied by means of two different methods: time-resolved spectroscopy based on a streak-camera and fibre optic neuroscopy, which served to perform process monitoring and regular fluorescence diagnosis of the probed region. Time-resolved fluorescence microscopy (FLIM) was used as a control technique. Time-resolved spectroscopic fluorescence lifetime analysis was performed on sexually mature female rats induced with glioma C6 brain tumour under in vivo conditions; thus, under conditions where the immune system actively intervenes in the process of oncogenesis. In this regard, the aim of the study was to recognize the cellular composition of the brain tumour tissue, namely the ratio of cancer and immunocompetent cells and their mutual localization. Understanding the role of the immune system thus provides new ways and approaches for further diagnosis and therapy, making tumour-associated immune cells a prime target for modern therapies

Changes in Spectral Fluorescence Properties of a Near-Infrared Photosensitizer in a Nanoform as a Coating of an Optical Fiber Neuroport

In this work, we tested a new approach to assess the presence of inflammatory process in the implant area using spectral methods and the technique of fiber fluorescence analysis of photosensitizers in nanoform. First of all, the spectral characteristics of the photosensitizer when interacting with the porous surface of the implant, based on hydroxyapatite under in vitro and in vivo conditions, were determined. Thus, it was shown that spectral characteristics of photosensitizers can be used for judgement on the process of inflammation in the implant area and thus on the local presence of the immunocompetent cells. The analysis was performed at a sufficient depth in the biotissue by using the near-infrared spectral region, as well as two different methods: fiber-based laser spectroscopy and fiber-optic neuroscopy, which served to monitor the process and regular fluorescence diagnosis of the studied area. Fluorescence spectroscopic analysis was performed on experimental animals in vivo, i.e., under conditions of active immune system intervention, as well as on cell cultures in vitro in order to judge the role of the immune system in the interaction with the implant in comparison. Thus, the aim of the study was to determine the relationship between the fluorescence signal of nanophotosensitizers in the near infrared spectral region and its parameters with the level of inflammation and the type of surface with which the photosensitizer interacts in the implant area. Thus, fiber-optic control opens up new approaches for further diagnosis and therapy in the implant area, making immune cells a prime target for advanced therapies

The research of chlorine e6 distribution and accumulation in multicellular tumor spheroid model

International audienceSquamous cell carcinoma is the most common type of oral and oropharyngeal cancers. Incomplete surgical removal of the tumor is often the cause of local recurrence of the disease and appearance of metastases. We have developed the novel endoscope fluorescence video system for visualization of micrometer size objects in real-time mode. The novel system was tested on three-dimensional models of cancer cells. Chlorine e6 (Ce6) was used as a photosensitizer. The accumulation of Ce6 in cancer cells was assessed by its fluorescence excited at 635 nm. It was shown that the 500 µm multicellular tumor spheroids could be easily detected with a good resolution and sensitivity. We suppose that the new endoscope video system could be useful for developing the intraoperative fluorescence navigation method using Ce6-mediated techniques that is able to visualize the small cancer cell clusters

Comparison of the Capabilities of Spectroscopic and Quantitative Video Analysis of Fluorescence for the Diagnosis and Photodynamic Therapy Control of Cholangiocellular Cancer

Cholangiocellular cancer (CCC) is a malignant neoplasm of the hepatobiliary system that is difficult to diagnose and treat. Currently, the most effective treatment of CCC is demonstrated under the control by fluorescent diagnosis. Photodynamic therapy (PDT) has also shown good results in the treatment of this disease, and fluorescence analysis of the photosensitizer is a good approach to control PDT. This article presents the results of a comparison of spectroscopic and quantitative video-fluorescent analysis of chlorin e6 photosensitizer fluorescence in vivo during cholangiocellular cancer surgery. Spectroscopic analysis provides accurate information about the concentration of the photosensitizer in the tumor, while the video-fluorescence method is convenient for visualizing tumor margins. A direct correlation is shown between these two methods when comparing the fluorescence signals before and after PDT. The applied paired Student’s t-test shows a significant difference between fluorescence signal before and after PDT in both diagnostic methods. In this regard, video-fluorescence navigation is not inferior in accuracy, sensitivity, or efficiency to spectroscopic methods

3D spheroid cultures for evaluation of nanophotosensitizers accumulation

Published in Journal of Physics: Conference Series, 1439:012032, IOPScience, 2020International audienceCurrent paper presents the results of the usage of indocyanine green and pheophorbide nanoform on 2D and 3D models of FaDu cells culture. The 2D model or monolayer was used for investigation of nanoparticles distribution within individual cells and their organelles. The 3D model or multicellular tumor spheroids were used for estimation of cells' metabolic processes by the investigation of the nanophotosensitizers fluorescence distribution within spheroid's layers. It was shown that pheophorbide nanoparticles are accumulated in the external cell layers of spheroids, indocyanine green nanoparticles distribution demonstrates completely opposite status – in the central part of the spheroid

Investigation of Ce6 accumulation and distribution in cell cultures of head and neck cancers

Article issued from an Internet Report. Published in Proceedings of SPIE, Volume 11065, Saratov Fall Meeting 2018: Optical and Nano-Technologies for Biology and MedicineInternational audienceCurrent paper presents the results of the chlorine e6 (Ce6) study on 2D and 3D models of FaDu cells culture. The 2D model or monolayer was used for investigation of Ce6 distribution within individual cells and their organelles. The 3D model or multicellular tumor spheroids were used for estimation of cells’ metabolic processes by the investigation of the Ce6 fluorescence distribution within spheroid's layers and Ce6 fluorescence lifetime. It was shown that 3D cell cultures and Сe6 allows estimating the cells’ metabolic processes better than in 2D monolayer cell cultures. Also, this model allows estimating the photodynamic effect depending on the proximity to the surface of different areas inside the heterogeneous 3D structure