Взаимосвязь спектроскопических и структурных свойств j-агрегатов индоцианина зеленого

Indocyanine green (ICG), when in free form in a liquid, can form stable nanoparticle structures or colloidal solution, while changing its spectroscopic properties. In the work, the aggregation degree and the average size of nanoparticles depending on the concentration of a colloidal solution of indocyanine green (ICG NPs) in the form of J-aggregates were investigated by various methods based on light scattering. The size of nanoparticles is an important parameter from the point of view of clinical application, because the technique of intravenous administration of drugs, in order to avoid microvascular thrombosis and embolism, provides dosage forms with inclusions of individual molecules or their clusters, not exceeding 500 nm diameter. In turn, small nanoparticles less than 30 nm lead to prolonged circulation of the drug in the body with an increased possibility of permeation into cells of healthy tissue. In the course of studies, it was found that an increase in the concentration of ICG NPs in the solution leads to an increase in the average size of spontaneously formed J-aggregates, which, in turn, leads to a decrease in the absorption coefficient in the aggregates. Presumably, this phenomenon, i.e. the established nonlinear dependence of the J-aggregate absorption on its size, can be explained by the formation of absorption centers on the J-aggregate surface in the form of mobile surface molecules. The threshold range of ICG molecule concentration was determined, at which there is a transition from aggregation with an increase in size with a slow addition of ICG J-aggregate molecules in height to a rapid addition in width.Индоцианин зеленый (ICG), находясь в растворе, способен образовывать стабильные структуры наночастиц или коллоидный раствор, изменяя при этом свои спектроскопические свойства. В работе различными методами, основанными на светорассеянии, были исследованы степень агрегации и средний размер наночастиц в зависимости от концентрации коллоидного раствора наночастиц индоцианина зеленого (ICG NPs) в форме J-агрегатов. Размер наночастиц представляет собой важный параметр с точки зрения клинического применения, так как техника внутривенного введения препаратов, с целью избежания тромбозов микрососудов и эмболии, предусматривает лекарственные формы с включениями, в виде отдельных молекул или их кластеров, не превышающими в диаметре 500 нм. С другой стороны, наночастицы размером менее 30 нм длительно циркулируют в организме и могут проникать в клетки здоровой ткани. В ходе исследований, было установлено, что увеличение концентрации ICG NPs в растворе ведет к увеличению среднего размера спонтанно формируемых J-агрегатов, что в свою очередь ведет к уменьшению коэффициента поглощения в агрегатах. Предположительно, нелинейная зависимость поглощения J-агрегата от его размера, может быть объяснен формированием центров поглощения на поверхности J-агрегата в виде подвижных поверхностных молекул. Был определен пороговый диапазон концентрации молекул ICG, при котором происходит переход от агрегации с увеличением размера с медленным прибавлением молекул J-агрегата ICG в высоту, но с быстрым прибавлением в ширину

Quantum dots in axillary lymph node mapping: Biodistribution study in healthy mice

<p>Abstract</p> <p>Background</p> <p>Breast cancer is the first cause of cancer death among women and its incidence doubled in the last two decades. Several approaches for the treatment of these cancers have been developed. The axillary lymph node dissection (ALND) leads to numerous morbidity complications and is now advantageously replaced by the dissection and the biopsy of the sentinel lymph node. Although this approach has strong advantages, it has its own limitations which are manipulation of radioactive products and possible anaphylactic reactions to the dye. As recently proposed, these limitations could in principle be by-passed if semiconductor nanoparticles (quantum dots or QDs) were used as fluorescent contrast agents for the <it>in vivo </it>imaging of SLN. QDs are fluorescent nanoparticles with unique optical properties like strong resistance to photobleaching, size dependent emission wavelength, large molar extinction coefficient, and good quantum yield.</p> <p>Methods</p> <p>CdSe/ZnS core/shell QDs emitting around 655 nm were used in our studies. 20 μL of 1 μM (20 pmol) QDs solution were injected subcutaneously in the anterior paw of healthy nude mice and the axillary lymph node (ALN) was identified visually after injection of a blue dye. <it>In vivo </it>fluorescence spectroscopy was performed on ALN before the mice were sacrificed at 5, 15, 30, 60 min and 24 h after QDs injection. ALN and all other organs were removed, cryosectioned and observed in fluorescence microscopy. The organs were then chemically made soluble to extract QDs. Plasmatic, urinary and fecal fluorescence levels were measured.</p> <p>Results</p> <p>QDs were detected in ALN as soon as 5 min and up to 24 h after the injection. The maximum amount of QDs in the ALN was detected 60 min after the injection and corresponds to 2.42% of the injected dose. Most of the injected QDs remained at the injection site. No QDs were detected in other tissues, plasma, urine and feces.</p> <p>Conclusion</p> <p>Effective and rapid (few minutes) detection of sentinel lymph node using fluorescent imaging of quantum dots was demonstrated. This work was done using very low doses of injected QDs and the detection was done using a minimally invasive method.</p

Foscan® (mTHPC) photosensitized macrophage activation: enhancement of phagocytosis, nitric oxide release and tumour necrosis factor-α-mediated cytolytic activity

Photodynamic activation of macrophage-like cells contributes to an effective outcome of photodynamic therapy (PDT) treatment. The possibility for an enhancement of macrophage activity by photosensitization with meta-tetra(hydroxyphenyl)chlorin (mTHPC) (1 μg ml−1) was studied in U937, monocyte cell line differentiated into macrophages (U937Φ cells). Phagocytic activity of U937Φ cells was evaluated by flow-cytometry monitoring of ingestion of fluorescein-labelled Escherichia coli particles. Increase in irradiation fluence up to 3.45 mJ cm−2 (corresponding irradiation time 15 s) resulted in significant increase in fluorescence signal (145%), while at higher light fluences the signal lowered down to the untreated control values. A light energy-dependent production of tumour necrosis factor-alpha (TNF-α) by photosensitized macrophages was further demonstrated using the L929 assay. The maximum TNF-α mediated cytolysis was observed at 28 mJ cm−2 and was 1.7-fold greater than that in control. In addition, we demonstrated a fluence-dependent increase in nitric oxide (NO) production by mTHPC-photosensitized macrophages. NO release increased gradually and reached a plateau after irradiation fluence of 6.9 mJ cm−2. Cytotoxicity measurements indicated that the observed manifestations of mTHPC-photosensitized macrophage activation took place under the sublethal light doses. The relevance of the present findings to clinical mTHPC-PDT is discussed. © 1999 Cancer Research Campaig

Relationship between subcellular localisation of Foscan® and caspase activation in photosensitised MCF-7 cells

The present study investigates the relationship between the subcellular localisation of Foscan® and intrinsic apoptotic pathway post Foscan®-based photodynamic therapy (PDT). With this purpose, mammary carcinoma MCF-7 cells were incubated with Foscan® for 3 or 24 h and then subjected to equitoxic light doses. Fluorescence microscopy revealed very good Foscan® co-localization to endoplasmic reticulum (ER) and Golgi apparatus after 3 h incubation with MCF-7 cells. Progressive increase in incubation time shows leakage of Foscan® from Golgi apparatus. Twenty-four hours incubation yielded a fluence-dependent enhanced induction of the ER-resident glucose-regulated protein 78 (Bip/GRP78), along with a weak mitochondrial damage, thus underscoring the ER as the main site of photodamage after prolonged incubation. Analysis of events implicated in apoptotic pathway after 24 h incubation demonstrated photodamage to Bcl-2 protein in total cellular extract, but not in the mitochondrial fraction. We further determined an increase in caspases-7 and -6 activation, which was strongly related to the expression of GRP78. The above findings demonstrate that Foscan® localisation in ER improves the photoactivation of the caspase-7 apoptotic pathway, which is poorly related to mitochondrial damage

Parameters affecting photodynamic activity of Foscan® or meta-tetra(hydroxyphenyl)chlorin (mTHPC

Abstract. In vitro photodynamic activity of Foscan or meta-tetra(hydroxyphenyl)chlorin (mTHPC) was approximately five times lower in the presence of protein in comparison with serum-free medium. Photocytotoxicity does not depend on the incubation time of the dye with cells. The in vitro results are discussed from the point of view of photodynamically active aggregated species. The photodynamic activity of mTHPC was elevated in vivo by macroscopic measurement of the necrosis depth after tumoral resection using an in vivo staining procedure with Evans blue dye. Only tumours from treated animals presented measurable necrosis areas, mostly localised in the surface around the irradiated site with a mean depth of 3.0 0.3 mm. The photodynamic activity was found to be significantly higher when using low irradiance (32 mW/cm 2 ) than when using a higher one (160 mW/cm 2 ). These results were not related to intratumoral mTHPC photodestruction analysed by in vivo fluorescence spectral analysis

Cyclodextrin-based photoactive liposomal nanoparticles for tumor targeting

The present study is aimed at the development of drug-in-cyclodextrin-in-liposome (DCL) nanoconstruct by coupling two independent delivery systems: cyclodextrin/mTHPC inclusion complexes and liposomal vesicles to improve the transport of mTHPC to the target tissue and to strengthen its intra-tissue accumulation in the tumor. Liposomes offer an excellent opportunity to achieve selective drug, targeting what is expected to prevent local irritation and reduce drug toxicity. Сyclodextrins (CDs) have been utilized as independent carriers for improvement of pharmaceutical properties such as solubility, stability, and bioavailability of various drug molecules, including mTHPC. Therefore, we assumed that encapsulation of CD-complexed drug into liposomes might increase drug loading capacity, entrapment efficiency, may restrain the dissociation of drug-CD complexes and prolong its systemic circulation. DCL nanoparticles have been prepared with various compositions to optimize the structure aiming to alter more favorably the distribution of temoporfin in tumor tissue. To enhance the encapsulation efficiency, double loaded DCLs, which include mTHPC in lipid bilayer along with (CD-mTHPC) inclusion complexes in the inner aqueous lumen, were prepared. It was demonstrated that DCLs possessed higher serum stability compared with conventional mTHPC liposomes (Foslip®). In fine, we showed that the presence of serum in the medium less affected cellular uptake of mTHPC delivered by double loaded MDCL compared with Foslip®

Optimization of temoporfin biodistribution by cyclodextrins-based nanostructures

Молекулярная биофизикаThis work was supported by Belarusian Republican Foundation for Fundamental Research (BRFFR) (grant numbers M17MC-028, Б17-106)

Drug delivery to solid tumors: the predictive value of the multicellular tumor spheroid model for nanomedicine screening

Marie Millard,1,2 Ilya Yakavets,1&ndash;3 Vladimir Zorin,3,4 Aigul Kulmukhamedova,1,2,5 Sophie Marchal,1,2 Lina Bezdetnaya1,2 1Centre de Recherche en Automatique de Nancy, Centre National de la Recherche Scientifique UMR 7039, Universit&eacute; de Lorraine, 2Research Department, Institut de Canc&eacute;rologie de Lorraine, Vand&oelig;uvre-l&egrave;s-Nancy, France; 3Laboratory of Biophysics and Biotechnology, 4International Sakharov Environmental Institute, Belarusian State University, Minsk, Belarus; 5Department of Radiology, Medical Company Sunkar, Almaty, Kazakhstan Abstract: The increasing number of publications on the subject shows that nanomedicine is an attractive field for investigations aiming to considerably improve anticancer chemotherapy. Based on selective tumor targeting while sparing healthy tissue, carrier-mediated drug delivery has been expected to provide significant benefits to patients. However, despite reduced systemic toxicity, most nanodrugs approved for clinical use have been less effective than previously anticipated. The gap between experimental results and clinical outcomes demonstrates the necessity to perform comprehensive drug screening by using powerful preclinical models. In this context, in vitro three-dimensional models can provide key information on drug behavior inside the tumor tissue. The multicellular tumor spheroid (MCTS) model closely mimics a small avascular tumor with the presence of proliferative cells surrounding quiescent cells and a necrotic core. Oxygen, pH and nutrient gradients are similar to those of solid tumor. Furthermore, extracellular matrix (ECM) components and stromal cells can be embedded in the most sophisticated spheroid design. All these elements together with the physicochemical properties of nanoparticles (NPs) play a key role in drug transport, and therefore, the MCTS model is appropriate to assess the ability of NP to penetrate the tumor tissue. This review presents recent developments in MCTS models for a better comprehension of the interactions between NPs and tumor components that affect tumor drug delivery. MCTS is particularly suitable for the high-throughput screening of new nanodrugs. Keywords: nanodrug, tridimensional model, distribution, accumulation, cytotoxicit