Fluorinated systems are defined by unique properties that offer advantages in drug delivery, material synthesis and industrial applications. In comparison to their hydrocarbon counterparts, the design of fluorinated solutes for tailored applications is limited by the inability to predict the effect of fluorination on phase behavior. This work examines and interprets the influence of fluorination on the phase behavior of fluorinated solutes and surfactants, with emphasis on their impact on vesicle bilayers. Thermodynamic partitioning of functionalized series of fluorinated and hydrocarbon nicotinate prodrugs fashioned to promote solubility in a fluorocarbon solvent (perfluorooctyl bromide; PFOB) is measured. Predictive approaches are also employed to describe partitioning of these nicotinates between immiscible phases relevant to drug delivery. The findings reveal no strong correlation of the partitioning trends with biological markers of cytotoxicity and prodrug uptake for PFOB mediated delivery. However, partitioning in model membranes (liposomes), which, increases with the hydrophobicity of the perhydrocarbon nicotinates, suggests incorporation in a cellular matrix is chain length dependent. The impact of incorporating fluorinated surfactants in catanionic vesicles, which form spontaneously in dilute aqueous solutions and serve as potential substitutes to conventional meta-stable liposome-based vesicles, is studied. Much larger isotropic vesicle regions are observed in the phase map of the partially fluorinated catanionic surfactant pair, cetylpyridinium bromide/ sodium perfluorooctanoate (CPB/SPFO) than in fully fluorinated HFDPC (1,1,2,2,-tetrahydroperfluorododecyl pyridinium chloride )/SPFO. Fluorescence probing of the vesicle bilayers suggest more fluid bilayers in CPB/SPFO than in HFDPC/SPFO due to better chain packing in the fully fluorinated bilayer. However, the vesicle region is expanded in more asymmetric fluorinated bilayers of HFDPC/SPFH (sodium perfluorohexanoate). The increased chain asymmetry in HFDPC/SPFH results in reduced packing density and more fluid bilayers than in HFDPC/SPFO. The robustness of CPB/SPFO and HFDPC/SPFO vesicles is demonstrated in the synthesis of silica hollow spheres by templating and the retention of encapsulated solutes. Higher colloidal stability of the silica spheres is achieved in HFDPC/SPFO relative to CPB/SPFO due to the barrier effect of the fluorinated bilayer. Similarly, higher solute retention in HFDPC/SPFO is observed. The modulation of phase behavior with fluorination offers opportunities in tunable applications of fluorinated bilayers.

<p>The use of catanionic drug-surfactant mixtures was proven to be an efficient novel method of obtaining prolonged drug release from gels. It was shown that various commonly used drug compounds are able to form catanionic mixtures together with oppositely charged surfactants. These mixtures exhibited interesting phase behaviour, where, among other structures, vesicles and large worm-like or branched micelles were found. The size of these aggregates makes them a potential means of prolonging the drug release from gels, as only monomer drugs in equilibrium with larger aggregates were readily able to diffuse through the gel. When the diffusion coefficient for drug release from the formulation based upon a catanionic mixture was compared to that obtained for the drug substance and gel alone, the coefficient was some 10 to 100 times smaller.</p><p>The effects of changes in the pH and ionic strength on the catanionic aggregates was also investigated, and this method of prolonging the release was found to be quite resilient to variations in both. Although the phase behaviour was somewhat affected, large micelles and vesicles were still readily found. The drug release was significantly prolonged even under physiological conditions, that is, at a pH of 7.4 and an osmolality corresponding to 0.9% NaCl.</p><p>Surfactants of low irritancy, capric and lauric acid, may successfully be used instead of the more traditional surfactants, such as sodium lauryl sulfate (SDS), and prolonged release can still be obtained with ease.</p><p>Some attempts to deduce the release mechanism from the proposed systems have also been made using transient current measurements, dielectric spectroscopy, and modelling of the release using the regular solution theory. In these studies, the previous assumptions made concerning the mechanism responsible for the release were confirmed to a large extent. Only small amounts of the drug existed in monomer form, and most seemed to form large catanionic aggregates with the oppositely charged surfactant.</p>

The use of catanionic drug-surfactant mixtures was proven to be an efficient novel method of obtaining prolonged drug release from gels. It was shown that various commonly used drug compounds are able to form catanionic mixtures together with oppositely charged surfactants. These mixtures exhibited interesting phase behaviour, where, among other structures, vesicles and large worm-like or branched micelles were found. The size of these aggregates makes them a potential means of prolonging the drug release from gels, as only monomer drugs in equilibrium with larger aggregates were readily able to diffuse through the gel. When the diffusion coefficient for drug release from the formulation based upon a catanionic mixture was compared to that obtained for the drug substance and gel alone, the coefficient was some 10 to 100 times smaller. The effects of changes in the pH and ionic strength on the catanionic aggregates was also investigated, and this method of prolonging the release was found to be quite resilient to variations in both. Although the phase behaviour was somewhat affected, large micelles and vesicles were still readily found. The drug release was significantly prolonged even under physiological conditions, that is, at a pH of 7.4 and an osmolality corresponding to 0.9% NaCl. Surfactants of low irritancy, capric and lauric acid, may successfully be used instead of the more traditional surfactants, such as sodium lauryl sulfate (SDS), and prolonged release can still be obtained with ease. Some attempts to deduce the release mechanism from the proposed systems have also been made using transient current measurements, dielectric spectroscopy, and modelling of the release using the regular solution theory. In these studies, the previous assumptions made concerning the mechanism responsible for the release were confirmed to a large extent. Only small amounts of the drug existed in monomer form, and most seemed to form large catanionic aggregates with the oppositely charged surfactant.

This thesis is focused on possibility of studying stability and fusion of catanionic vesicles with Förster resonance energy transfer. The mainly used technique was Time-Correlated single photon counting. Firstly, excitation and emission spectra of chosen probes were measured and donor-acceptor pairs were suggested: 5-hexadecanoylaminofluorescein with Octadecyl Rhodamine B, Bodipy 493/503 with rhodamine or DiI, perylene with fluorescein, DiO with DiI. Then, time-resolved measurements of suggested pairs from environment of catanionic vesicles with different content of cholesterol were made in order to track the FRET associated with fusion of vesicles. It was found out, that it is not possible to use DiO as a donor because of it’s inefficient solubilisation into vesicles. It is also not possible for Bodipy to be used as a donor, because of it‘s excimer formation. In case of using fluorescein as a donor, it was found, that there is ongoing homo-fret between fluorescein molecules. Thanks to this, fusion was tracked by addition of unstained vesicles. It was also possible to track fusion in longer period of time. Also perylene-fluorescein pair was found to be capable of tracking the fusion, but with the exception of vesicles with content of cholesterol of 43 mol. %, tracking of fusion was possible only in short period of time.

L'administration de principes actifs de manière locale et externe permet de contrôler leur biodisponibilité et de prolonger leur action thérapeutique sur le site à traiter. Cependant, la délivrance au sein des couches profondes de la peau est un challenge pour la recherche dermatologique car ce tissu représente une barrière difficilement franchissable. Il permet ainsi de protéger l'organisme mais c'est un frein pour l'administration de médicaments par voie cutanée. De nombreux systèmes ont été imaginés afin d'améliorer le passage d'actifs à travers la peau : promoteurs chimiques de pénétration, méthodes électriques, vecteurs encapsulant le principe actif. Parmi ceux-ci, les vésicules montrent un fort potentiel en particulier si elles sont capables de se déformer ou fusionner avec les lipides de la peau. Notre équipe a conçu un système de vectorisation à partir d'un tensioactif catanionique, issu de l'association de deux amphiphiles de charges opposées, capable de s'auto-associer en solution aqueuse pour former des vésicules pouvant être utilisées pour l'encapsulation de principes actifs de natures variées. Ces vecteurs ont démontré par le passé leur aptitude à interagir et fusionner avec les membranes lipidiques. Dans ces travaux, nous avons donc étudié leur capacité à améliorer la pénétration cutanée de principes actifs. Les propriétés physicochimiques des vésicules sont très bien décrites dans l'eau mais une formulation dermatologique peut impliquer la présence d'additifs tels que des promoteurs de pénétration ou des agents hydratants. Les vésicules peuvent également être incorporées dans des gels aqueux pour faciliter leur application. L'influence de ces différents additifs a donc été étudiée en caractérisant systématiquement les vésicules en termes de taille, de charge, de stabilité et en évaluant la fluidité de leur membrane. Ensuite, afin d'étudier la capacité des formulations à pénétrer dans la peau, une sonde fluorescente a été encapsulée dans les vésicules. Des expériences in vitro sur peau de cochon ont permis de prouver l'importance de l'état thermodynamique de la membrane. En effet, la rétention de la sonde dans la peau est supérieure lorsque la bicouche de la vésicule est fluide. La microscopie confocale a été utilisée pour estimer la profondeur de pénétration ainsi que les chemins préférentiellement empruntés par la sonde dans la peau. Enfin, l'application de ce système à la vectorisation d'un corticostéroïde employé dans le traitement du psoriasis a validé le potentiel des vésicules catanioniques pour des usages thérapeutiques. Ces expériences ont montré une nette augmentation de la rétention cutanée grâce à nos vecteurs, en comparaison notamment avec une solution de corticoïde additionnée d'éthanol, pourtant promoteur de pénétration reconnu. / The local and external administration of drugs allows bioavailability control and sustained therapeutic action on the treatment site. Yet, the delivery to the deep layers of the skin is a challenge for dermatological research because this tissue represents an efficient barrier. It protects the organism against aggressions but it also constrains cutaneous drug delivery. Various systems have been imagined in order to improve drug passage through the skin: chemical penetration enhancers, electrical methods, drug carriers. Among these, vesicles have a strong potential, in particular if they can deform or fuse with skin lipids. Our team has designed a drug delivery system based on a catanionic surfactant, made of two oppositely charged amphiphiles. It is able to self-associate in aqueous solutions to form vesicles used to encapsulate drugs of various nature. These carriers have shown the ability to interact and fuse with lipidic membranes. In this work, we therefore studied their capacity to improve drugs' skin penetration. The vesicles' physicochemical properties are well described in water but a dermatological formulation may contain additives such as penetration enhancers or hydrating agents. The carriers can also be incorporated into aqueous gels in order to facilitate their application. The influence of these different additives was studied systematically by characterizing the vesicles in terms of size, charge, stability and by evaluating the membrane fluidity. Then, a fluorescent probe was encapsulated into the vesicles to study the formulations' ability to penetrate the skin. In vitro experiments on pig skin proved the importance of the membrane's thermodynamic state. Indeed, the probe's skin retention is higher when the vesicles' bilayer is in a fluid state. Confocal microscopy was used to estimate the penetration depth as well as the pathways followed by the probe into the skin. Finally, this system was applied to a corticosteroid used for psoriasis treatment. It confirmed the catanionic vesicles' potential for therapeutic use. These experiments showed a clear increase in cutaneous retention thanks to our carriers, in comparison with a corticoid solution containing ethanol, which is a known penetration enhancer.

Orientador: Watson Loh / Dissertação (mestrado) - Universidade Estadual de Campinas, Instituto de Química / Made available in DSpace on 2018-08-16T00:50:27Z (GMT). No. of bitstreams: 1 Silva_AndreLuisCondeda_M.pdf: 1492752 bytes, checksum: 969a849fc8d776ed3dd74fac01294cde (MD5) Previous issue date: 2009 / Resumo: O surfatante cataniônico dodecilsulfato de dodecildimetil-n-hidroxilamônio foi preparado e caracterizado em relação às suas propriedades físico-químicas. O composto apresentou solubilidade muito baixa em água, porém com capacidade de redução da tensão superficial e interfacial entre água e xileno, em ambos os casos com lenta cinética de saturação das interfaces. A solubilidade em solventes orgânicos foi avaliada e constatamos tendência a maior solubilidade em solventes polares como clorofórmio e etanol, e no solvente aromático xileno. O estudo de comportamento termotrópico indicou a existência de fases líquido-cristalinas entre a fase sólida cristalina e a fase líquida isotrópica. A partir dos resultados deste trabalho avaliamos que as possíveis aplicações para estes surfatante estão relacionadas às situações onde a adsorção e deposição seja requerida, e em especial para a preparação de emulsões de água em óleo, com o atributo de baixa solubilidade em ambas as fases, e a possível vantagem de redução de irritabilidade dérmica para cremes e loções. O desenvolvimento desta aplicação requer avanços nos procedimentos de preparação da emulsão de forma a facilitar a adsorção do composto na interface água/óleo / Abstract: The catanionic surfactant dodecyldimehyl-n-hydroxylamonium dodecylsulphate was prepared and its physical-chemical properties were characterized. The surfactant presented very low water solubility, but with ability to reduce the surface tension and the interfacial tension between water and xylene, in both situations with slow kinetics of interface saturation. The solubility in organic solvents was evaluated and the trend for higher solubility in polar solvents like ethanol and chloroform and in the aromatic solvent xylene was noticed. The evaluation of its termotropic behavior was carried out and indicated the existence of liquid crystalline phases between the solid crystalline phase and the isotropic liquid phase. The results of this study indicate the possible applications in situations where adsorption and deposition are required, and for the preparation of water in oil emulsions, with the special feature of low solubility in both phases, with the advantage of reduced skin irritation for creams and lotions. The development of this application requires advancements in the procedures for emulsion preparation in order to allow the surfactant adsorption at the water/oil interface / Mestrado / Físico-Química / Mestre em Química

The diploma thesis deals with the preparation and characterization of catanionic complexes and their possible application to pharmaceutics and medicine. The catanionic complexes were prepared by mixing two oppositely charged surfactants with the concentration of 20 mmol·dm­3 in various volume ratios. Two systems were analyzed – CTAB + SDS and Septonex + SDS. Both systems were prepared in an aqueous and physiological environment. The turbidimetric analyses provided results of the intensity of turbidity of the prepared mixtures. The size and stability of the prepared particles were determined by DLS and ELS measurements. CTAB-rich samples showed significant viscosity changes and they were therefore characterized by rheological measurements. The results of this work show that the asymmetry of alkyl chains, surfactant selection, temperature, and ionic strength have a significant influence on the properties and self-assembly of surfactants in catanionic complexes.

O trabalho apresentado foi realizado em duas etapas independentes e baseou-se no estudo de diferentes sistemas nanométricos para viabilizar a aplicação da ftalocianina de cloro alumínio (ClAlPc) na terapia fotodinâmica (TFD) para o tratamento do câncer de pele do tipo melanoma. O fármaco fotossensibilizante (FS) utilizado apresenta propriedades físico-químicas que lhe permitem exercer sua atividade fotodinâmica com excelência, sem a interferência do cromóforo endógeno melanina existente nas células melanocíticas. Para driblar sua elevada hidrofobicidade, ClAlPc foi encapsulada em sistemas nanométricos para administração em meio fisiológico. Inicialmente nanopartículas lipídicas sólidas (NLS) foram desenvolvidas por emulsificação direta, após um estudo de elaboração do diagrama de fases. O compritol foi o lipídio sólido escolhido para compor as NLS, com diferentes concentrações de ClAlPc. Todas as formulações desenvolvidas foram devidamente caracterizadas, com tamanho médio entre 100 e 200 nm, baixa polidispersão, potencial zeta adequadamente negativo (~|30| mV), drug loading de ClAlPc entre 76-94% (com pequena redução após 24 meses) e alta eficiência de encapsulação (E.E.). A morfologia arredondada das nanopartículas foi confirmada por microscopia eletrônica de transmissão e de força atômica. A estabilidade das NLS foi de 24 meses. A avaliação da cristalinidade do lipídio revelou a integração da ClAlPc à matriz lipídica da NLS, presença de estruturas polimórficas e grau de cristalinidade adequado, sem alterações após 24 meses. Nos estudos de difusão in vitro, observou-se que ftalocianina encapsulada nas NLS acumulam-se preferencialmente na epiderme e derme do que no estrato córneo, sem traços de permeação do ativo. Foi confirmado o caráter biocompatível das NLS sobre fibroblastos NIH-3T3. A ftalocianina encapsulada nas NLS não foi tóxica na linhagem de melanoma B16-F10 na ausência de luz, porém, apresentou excelente efeito fototóxico (0,75 ?g mL-1 de ClAlPc nanoencapsulada e irradiação entre 0,5 e 2,0 J cm-2), com redução da viabilidade celular de 87%. O segundo sistema de veiculação estudado foram as vesículas cataniônicas (VesCat), que se formam espontaneamente em água com o tensoativo TriCat 12. A obtenção das vesículas contendo ClAlPc envolve uma etapa adicional, para remoção de solvente orgânico, que foi aprimorada, reduzindo o tempo de produção em 55%. As VesCat/ClAlPc obtidas mantiveram suas propriedades físico-químicas e morfologia arredondada (confirmada por microscopia eletrônica de varredura), drug loading de 47% e alta E.E. Os resultados comprovaram que a aplicação desses dois sistemas nanométricos é altamente eficiente para aplicação da TFD no tratamento do câncer de pele do tipo melanoma ou outras doenças cutâneas, apresentando características favoráveis para avanços nos estudos de fase clínica e pré-clínica. / The present work was conducted in two independent steps, which were based on the study of different nanometric systems that make feasible the application of aluminum phthalocyanine chloride (ClAlPc) in the photodynamic therapy (PDT) to the melanoma skin cancer treatment. The photosensitizer (PS) used has physical-chemical properties that allow it to perform its photodynamic activity with excellence, without the interference of the melanin, an endogenous chromophore found in melanotic cells. In order to circumvent the high PS hydrophobicity, ClAlPc was encapsulated into nanosystems to administration in physiological environment. At first, solid lipid nanoparticles (SLN) were developed by direct emulsification process after drawing up phase diagram study. The solid lipid compritol was chosen to make the SLN, produced with different ClAlPc concentrations. The developed samples were properly characterized with mean size between 100-200 nm, low polydispersity, negative zeta potential (~|30| mV), ClAlPc drug loading around 76-94% (with slight decrease after 24 months) and high encapsulation efficiency (EE). The round shape of SLN was confirmed by transmission electron microscopy and atomic force microscopy. The nanoparticles were stable for at least 24 months. The evaluation of lipid crystallinity has proved the ClAlPc integration to SLN lipid matrix, the presence of polymorphic structures and a suitable crystalline degree, without large variations after 24 months. In the in vitro diffusion studies were observed that phthalocyanine conveyed in the nanoparticles accumulates preferably in the epidermis and dermis than in the stratum corneum, without any drug permeation traits. The NLS biocompatibility was confirmed on NIH-3T3 fibroblasts. ClAlPc-loaded NLS did not exhibit toxicity on B16-F10 melanoma cell line in the dark, but it was shown their outstanding phototoxicity effect (0.75 ?g mL-1 of encapsulated ClAlPc and irradiation between 0.5 and 2.0 J cm-2) with cell viability reduction of 87%. The second drug delivery system studied were the catanionic vesicles (VesCat) that are spontaneously obtained by mixing the self-assembly surfactant TriCat 12 in water. The production of ClAlPc-loaded vesicles comprises an additional step (to remove the organic solvent) that was optimized, saving 55% of the production time. The final VesCat/ClAlPc kept their physical-chemical properties and round shape (confirmed by scanning electron microscopy), drug loading of 47% and high EE. Hence, the results have proved the great efficiency of these two nanometric systems applied in the PDT to the treatment of melanoma skin cancer and other cutaneous disease, useful features for further progress towards preclinical and clinical trials.

O trabalho apresentado foi realizado em duas etapas independentes e baseou-se no estudo de diferentes sistemas nanométricos para viabilizar a aplicação da ftalocianina de cloro alumínio (ClAlPc) na terapia fotodinâmica (TFD) para o tratamento do câncer de pele do tipo melanoma. O fármaco fotossensibilizante (FS) utilizado apresenta propriedades físico-químicas que lhe permitem exercer sua atividade fotodinâmica com excelência, sem a interferência do cromóforo endógeno melanina existente nas células melanocíticas. Para driblar sua elevada hidrofobicidade, ClAlPc foi encapsulada em sistemas nanométricos para administração em meio fisiológico. Inicialmente nanopartículas lipídicas sólidas (NLS) foram desenvolvidas por emulsificação direta, após um estudo de elaboração do diagrama de fases. O compritol foi o lipídio sólido escolhido para compor as NLS, com diferentes concentrações de ClAlPc. Todas as formulações desenvolvidas foram devidamente caracterizadas, com tamanho médio entre 100 e 200 nm, baixa polidispersão, potencial zeta adequadamente negativo (~|30| mV), drug loading de ClAlPc entre 76-94% (com pequena redução após 24 meses) e alta eficiência de encapsulação (E.E.). A morfologia arredondada das nanopartículas foi confirmada por microscopia eletrônica de transmissão e de força atômica. A estabilidade das NLS foi de 24 meses. A avaliação da cristalinidade do lipídio revelou a integração da ClAlPc à matriz lipídica da NLS, presença de estruturas polimórficas e grau de cristalinidade adequado, sem alterações após 24 meses. Nos estudos de difusão in vitro, observou-se que ftalocianina encapsulada nas NLS acumulam-se preferencialmente na epiderme e derme do que no estrato córneo, sem traços de permeação do ativo. Foi confirmado o caráter biocompatível das NLS sobre fibroblastos NIH-3T3. A ftalocianina encapsulada nas NLS não foi tóxica na linhagem de melanoma B16-F10 na ausência de luz, porém, apresentou excelente efeito fototóxico (0,75 ?g mL-1 de ClAlPc nanoencapsulada e irradiação entre 0,5 e 2,0 J cm-2), com redução da viabilidade celular de 87%. O segundo sistema de veiculação estudado foram as vesículas cataniônicas (VesCat), que se formam espontaneamente em água com o tensoativo TriCat 12. A obtenção das vesículas contendo ClAlPc envolve uma etapa adicional, para remoção de solvente orgânico, que foi aprimorada, reduzindo o tempo de produção em 55%. As VesCat/ClAlPc obtidas mantiveram suas propriedades físico-químicas e morfologia arredondada (confirmada por microscopia eletrônica de varredura), drug loading de 47% e alta E.E. Os resultados comprovaram que a aplicação desses dois sistemas nanométricos é altamente eficiente para aplicação da TFD no tratamento do câncer de pele do tipo melanoma ou outras doenças cutâneas, apresentando características favoráveis para avanços nos estudos de fase clínica e pré-clínica. / The present work was conducted in two independent steps, which were based on the study of different nanometric systems that make feasible the application of aluminum phthalocyanine chloride (ClAlPc) in the photodynamic therapy (PDT) to the melanoma skin cancer treatment. The photosensitizer (PS) used has physical-chemical properties that allow it to perform its photodynamic activity with excellence, without the interference of the melanin, an endogenous chromophore found in melanotic cells. In order to circumvent the high PS hydrophobicity, ClAlPc was encapsulated into nanosystems to administration in physiological environment. At first, solid lipid nanoparticles (SLN) were developed by direct emulsification process after drawing up phase diagram study. The solid lipid compritol was chosen to make the SLN, produced with different ClAlPc concentrations. The developed samples were properly characterized with mean size between 100-200 nm, low polydispersity, negative zeta potential (~|30| mV), ClAlPc drug loading around 76-94% (with slight decrease after 24 months) and high encapsulation efficiency (EE). The round shape of SLN was confirmed by transmission electron microscopy and atomic force microscopy. The nanoparticles were stable for at least 24 months. The evaluation of lipid crystallinity has proved the ClAlPc integration to SLN lipid matrix, the presence of polymorphic structures and a suitable crystalline degree, without large variations after 24 months. In the in vitro diffusion studies were observed that phthalocyanine conveyed in the nanoparticles accumulates preferably in the epidermis and dermis than in the stratum corneum, without any drug permeation traits. The NLS biocompatibility was confirmed on NIH-3T3 fibroblasts. ClAlPc-loaded NLS did not exhibit toxicity on B16-F10 melanoma cell line in the dark, but it was shown their outstanding phototoxicity effect (0.75 ?g mL-1 of encapsulated ClAlPc and irradiation between 0.5 and 2.0 J cm-2) with cell viability reduction of 87%. The second drug delivery system studied were the catanionic vesicles (VesCat) that are spontaneously obtained by mixing the self-assembly surfactant TriCat 12 in water. The production of ClAlPc-loaded vesicles comprises an additional step (to remove the organic solvent) that was optimized, saving 55% of the production time. The final VesCat/ClAlPc kept their physical-chemical properties and round shape (confirmed by scanning electron microscopy), drug loading of 47% and high EE. Hence, the results have proved the great efficiency of these two nanometric systems applied in the PDT to the treatment of melanoma skin cancer and other cutaneous disease, useful features for further progress towards preclinical and clinical trials.

In this master ‘s thesis, catanionic vesicles formed by the pseudo-double-chain complex CTA – DS were investigated from the point of view of microviscosity. Samplesand of cationic vesicles contained 23, 43 and 53 mol. % of cholesterol and the double-chain surfactant DODAC. Cationic vesicles were prepared for visual observation, their stability was determined by DLS and the prepared system was further investigated. Microviscosity was determined from fluorescence anisotropy. To study the outer part of the membrane, laurdan fluorescent probes were used whereas diphenylhexatriene was used for the inner part of the membrane. This method has been proven to be suitable because it reflects the conditions of the membrane. Moreover, a 1,3-bispyrenylpropane probe forming intramolecular excimers was used to study the microviscosity in the vesicle bilayer. The dicyanovinyljulolidine (DCVJ) probe was applied in the case of the molecular rotor technique. It has been shown that in the case of the DCVJ probe, the molecular rotor technique is practically unusable, due to the fact that the probe has a low quantum yield at low temperatures. Also, the excimer formation of P3P probes does not lead to the expected results. The cationic vesicles do not seem to support this formation, as they are too closely related. This type of probe can be used for the selected system with some restrictions.