21 oktober 2021: Bron: 2015; 10(5): e0127219. Published online 2015 May 26

We krijgen wat vragen hoe nu precies de laserwatch precies werkt. Ik ben daarvoor op zoek gegaan naar wat meer informatie en kwam o.a. op deze studie / beschrijving. Abstract staat verderop in dit artikel.

Extracorporeal Photo-Immunotherapy for Circulating Tumor Cells

Al in 2015 publiceerden Japanse onderzoekers een manier om PDT - Foto Dynamische Therapie toe te passen op in bloed circulerende tumorcellen. Deze manier wordt Extracorporele fotoferese (ECP) genoemd. Zij gebruikten daarvoor een zoals zij dat noemen extracorporale buis, een dunne transparante medische buis en belichten het bloed dat door de buis heen stroomde met PDT met een golflengte van 660 selectieveNm. gegenereerd door een LED-array. Zoals ook de laserwatch belicht met 660 Nm..

Een blootstelling van twee minuten was voldoende om kankercelnecrose te bereiken, schrijven deze onderzoekers. Dus in 2 minuten werden tumorcellen gedood door het licht. Met de laserwatch zijn ook al enkele patiënten 'behandeld' met een soortgelijke manier waarbij deal belicht werd buiten het lichaam voordat het via een infuus het bloed in ging. In feite een min of meer zelfde manier van belichten buiten het lichaam. Maar ik zal Andrei dit artikel sturen en vragen om zijn commentaar.

Uit het studieverslag vertaald om proberen duidelijk te maken hoe deze manier van PDT - Foto Dynamische therapie dus werkt:

Hier stellen we een benadering voor met behulp van extracorporale fotodynamische therapie (PDT of foto-immunotherapie) in combinatie met antilichaamtargeting. PDT vereist drie componenten, namelijk: zuurstof, een fotosensibilisator en licht (voornamelijk in het zichtbare bereik). Deze moeten allemaal tegelijkertijd aanwezig zijn om de fotosensibilisator te activeren, reactieve zuurstof te genereren (voornamelijk singlet zuurstof O2 _) en cellen of weefsels te beschadigen. Bovendien is de toxiciteit van de reactieve zuurstofspecies gelokaliseerd in de cel in direct contact ermee, vanwege de korte (< 100 nm) diffusieafstand. [20, 21].

Deze kenmerken resulteren in een hoge doelspecificiteit met bijna nul nevenschade aan aangrenzende cellen/weefsels, waardoor PDT een effectieve en veiligere behandeling is in vergelijking met conventionele bestraling en chemotherapie. Ondanks deze voordelen kan zichtbaar licht nauwelijks door weefsel heen dringen, [22, 23] vooral in aanwezigheid van bloed (een zichtbaar lichtabsorbeerder) en water (een IR-lichtabsorbeerder) en dus is de toepassing van PDT voornamelijk beperkt tot ziekten in geopende/topische gebieden, waaronder huid, hoofd, nek, longen, tanden, enz.

In deze studie hebben we de haalbaarheid aangetoond van een nieuwe therapeutische toepassing van PDT voor hematologische pathogenen - vormen van bloedkanker, door middel van antilichaamtargeting en een extracorporaal apparaat om de limiet van de weefselpenetratiediepte van PDT te overwinnen.

We hebben een fotosensitizer (chloor E6 (Ce6)) - antilichaam (anti-CD44)-conjugaat (Ce6-CD44 Ab-conjugaat) ontwikkeld om het fotosensibiliserende middel selectief aan CTC's te leveren (in dit geval PC-3-cellen). PDT werd uitgevoerd door het bloed door een dunne transparante medische buis te laten stromen (Fig 1) en vertoonde een significant verbeterde werkzaamheid van celdoding. Een bijkomend voordeel van deze techniek is dat het antilichaam binnen enkele dagen veilig uit het lichaam kan worden verwijderd door natuurlijke antilichaamafbraakmechanismen [].

Hieronder grafisch uitgelegd hoe de Japanse onderzoekers te werk gingen.

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Schematic of the proposed device in operation.

Photosensitizer-antibody conjugate is injected prior to PDT procedure. Blood circulation was guided by medical tubing with a peristaltic pump. Extracorporeal PDT is performed as the blood flows through the tube inside a reflective chamber. The treated blood is returned to body. All procedures are accomplished in a constant flow mode.

PDT was performed using high power (maximum 100 W input) 660 nm LED array shown in Fig 2(A) (LEDwholesalers.com). Samples were placed within an aluminum foil covered styrofoam chamber shown in Fig 2(B) and Fig 2(C) and illuminated. The duration of illumination was carefully optimized for minimum exposure to PDT treatment.

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Fig 2

Extracorporeal PDT setup.

(a) 660 nm LED-array, (b) de met aluminiumfolie bedekte kamer gemaakt van piepschuimdoos met twee buizen gevuld met bloed. (c) PDT wordt uitgevoerd door de LED-array bovenop de kamer te plaatsen. Een belichting van 2 minuten werd uitgevoerd om de extracorporale bloedcirculatie te simuleren.

Naast onderstaand studieverslag zijn recentere studies gepubliceerd, ook uitgevoerd bij kankerpatiënten, die ook refereren aan bovenstaand beschreven studieverslag. Klik op de titels voor de studieverslagen. daaronder abstract plus referentielijst van bovenstaand studieverslag.

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2015; 10(5): e0127219.

Published online 2015 May 26. doi: 10.1371/journal.pone.0127219

PMCID: PMC4444246

PMID: 26011055

Extracorporeal Photo-Immunotherapy for Circulating Tumor Cells

Gwangseong Kim 1 , 2 and Angelo Gaitas 1 , 2 ,*

Hiromu Suzuki, Academic Editor

Author information Article notes Copyright and License information Disclaimer

This article has been cited by other articles in PMC.

Associated Data

Supplementary Materials

Data Availability Statement

Abstract

It is well established that metastasis through the circulatory system is primarily caused by circulating tumor cells (CTCs). In this preliminary effort, we report an approach to eliminate circulating tumor cells from the blood stream by flowing the blood though an extracorporeal tube and applying photodynamic therapy (PDT). Chlorin e6 (Ce6), a photosensitizer, was conjugated to CD44 antibody in order to target PC-3, a prostate cancer cell line. PC-3 cells were successfully stained by the Ce6-CD44 antibody conjugate. PDT was performed on whole blood spiked with stained PC-3 cells. As the blood circulated through a thin transparent medical tube, it was exposed to light of 660 nm wavelength generated by an LED array. An exposure of two minutes was sufficient to achieve selective cancer cell necrosis. In comparison, to PDT of cells growing inside a tissue culture, the PDT on thin tube exhibited significantly enhanced efficiency in cell killing, by minimizing light attenuation by blood. It suggests a new extracorporeal methodology of PDT for treating CTCs as well as other hematological pathogens.

Discussion

We have adequately demonstrated in our preliminary studies the utility of PDT in killing CTCs using a tubing and appropriate antibody binding technology. PDT is an effective alternative treatment modality that addresses several of the drawbacks of conventional treatments in cancer and in other diseases. However, the absorption of visible light by blood (especially due to the red blood cells' hemoglobins) significantly reduces the penetration of light through tissue. This is evident in our results shown in "PDT in blood" column of Fig 3, where PC-3 cells were cultured on a 12-well plate. Following a two minute illumination 2 hours later the many cells were still alive. In contrast, the cells in media were completely wiped out by the two minute illumination in the absence of blood within the first 15 minutes. These results clearly demonstarte that cellular and other components present in blood can hamper efficacy of PDT in killing your target cells.

The results, from Fig 4, where PDT was performed in a tube exhibited improved efficacy. Majority of cell death could be achieved before 1 hour following two minute illumination. We believe that the improvement came from the utilization of the narrow tube that reduced light attenuation by blood. The tube used in this study is a transparent PDMS tube with 1.02 mm inner diameter. Since the light came from all the directions surrounding the tube in the reflective chamber, the thin diameter of tube allowed for nearly the entire sample to be within the penetration depth of light. More exposure to light resulted in better outcomes with PDT. These results promise a new effective treatment modality for hematological pathogens using PDT, overcoming its penetration depth limitation.

The experimental parameters used in this investigation, such as the choice of photosensitizer, the illumination time, the antibody, the type of tube material, size of tube (length and diameter), the light source etc. need further refinement. These parameters need to be adequately addressed and optimized to obtain maximum efficacy of PDT, especially with consideration to in vivo constant flow conditions.

A similar extracorporeal approach was suggested previously by Edelson et al. [26] where the "extracorporeal photopheresis (ECP)" concept was first reported. This technology aims to destroy white blood cells in the blood. The fact that this principle is based on UV light and the photosentizer is used without prior targeting, the white blood cells (WBCs) are required to be separated by apheresis to avoid interference by red blood cells (RBCs) and thereby prevent collateral damages to other blood cells. Then the isolated WBCs are treated with UV illumination and re-injected in the body along with other blood components. This technology is used to treat cutaneous T-cell lymphoma by reducing tumor burden and graft-versus-host diseases (GVHD) in organ transplantation by suppressing immune reaction against foreign organs and is currently approved by the FDA. Our present approach has numerous advantages over photopheresis, such as 1) It does nor require time-consuming processes to separate blood cellular components, 2) Our technology achieves active targeting to specific cells of interest and finally 3) Our PDT utilizes light activation in the far-red/near infrared wavelengths that is less harmful and has deeper penetration depth.

In addition, the photosensitizer-antibody conjugates can be used as an imaging contrast to detect metastasized cancers, as well as adapted to other treatment modalities, including endoscopic photodynamic therapy and the ability to target the lymphatic system. and several other diseases. It complements the current approaches that are being used to reduce the number of CTCs in medical centers worldwide.

Acknowledgments

We would like to thank Dr. Ricky Malhotra for providing us with valuable feedback.

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Funding Statement

This research was funded by National Institutes of Health (grant # GM084520). Kytaro, Inc. provided support in the form of salaries for authors AG and GK, but did not have any additional role in the study design, data collection and analysis, decision to publish, or preparation of the manuscript. The specific roles of these authors are articulated in the ‘author contributions’ section.

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Data Availability

All relevant data are within the paper and its Supporting Information files.

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PDT, Foto Dynamische therapie, immuuntherapie, immuunstimulatie, in bloed circulerende tumorcellen, extracorporale buis, tumornecrose