Preparation of 100 nm diameter unilamellar vesicles containing zinc phthalocyanine and cholesterol for use in photodynamic therapy

An efficient (89-95% yield) and low-cost procedure to prepare unilamellar vesicles was used to incorporate zinc phthalocyanine (ZnPc), a model compound used as a phototherapeutic agent in studies aiming the use of unilamellar vesicles as delivery system for photodynamic therapy (PDT). ZnPc was incorporated in the presence or absence of cholesterol (CHOL), which improved the stability of the delivery system. The net vesicles present a mean diameter around 1000 nm, whereas in the presence of CHOL, CHOL and ZnPc, or only ZnPc, a drastic reduction in its diameter, varying between 100 and 150 nm, was observed. The incorporation of only ZnPc also results in a considerable reduction in the diameter of the liposomes suggests that ZnPc, due to its high hidrophobicity, must share the same microenvironment occupied by CHOL molecules. (C) 2004 Elsevier Ireland Ltd. All rights reserved.1331697

Photoinactivation of different human tumor cell lines and sheep red blood cells in vitro by liposome-bound Zn(II) Phthalocyanine: Effects of cholesterol

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)The in vitro photoinactivation of human tumor cell lines and sheep red blood cells (SRBC) by Zinc (II) Phthalocyanine (ZnPc) was investigated using unilamellar liposome (LUV) as delivery system, in the presence and absence of cholesterol (CHOL) in the formulation. The presence of CHOL improves the stability of the system showing to be essential for the photodynamic action of ZnPc. LUVs prepared without CHOL did not present any antiproliferative effects neither induced significant photohaemolysis. The presence of ZnPc in the culture medium caused total cell growth inhibition (TGI) only at concentrations higher than 250 mu mol dm(-3). For ZnPc in LUV/CHOL (mass ratio = 3:1), the mean TGI values for almost all studied cells were around 80 mu mol dm(-3), and 14 mu mol dm(-3) for human ovarian carcinoma (NIH: OVCAR-3) cells. The cytoplasmic components of OVCAR-3 and SRBC when irradiated in presence of ZnPc in LUV/CHOL were completely destroyed, culminating in cell swelling, lysis and death by necrosis. (C) 2010 Elsevier B.V. All rights reserved.10029299Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)Fundação de Amparo à Pesquisa do Estado de Minas Gerais (FAPEMIG)Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP

Experimental and theoretical studies of the thermal degradation of a phenolic dibenzodioxocin lignin model

International audienceA large part of biphenyl structures in lignin are etherified by alpha- and beta-carbons of another phenylpropane unit to give an eight-member ring called dibenzodioxocin. The behavior of a phenolic dibenzodioxocin lignin model, 4-(4,9-dimethoxy-2,11-n-dipropyl-6,7-dihydro-5,8-dioxa-dibenzo[a,c]cycloocten-6-yl)-2-methoxyphenol (DBDOH, 1), was studied by different mass spectrometry and thermal methods, leading to the conclusion that dibenzodioxocins are thermally unstable products. Both semi-empirical and density functional theory quantum calculations show that both C-O bonds, which connect the biphenyl part of the dibenzodioxocin molecule to the phenolic group, can be broken under increasing temperature. However, they do not play the same role since their dissociation occurs through different barrier heights. The C-O bond directly connected to the phenolic group (alpha-O-4) will dissociate first since its barrier energy for scission is lower than the other one (beta-O-4), by roughly 12 kcal mol(-1) (a parts per thousand 50 kJ mol(-1)). This conclusion is likely applicable to thermal degradation of DBDO units in lignin polymer

Effect of Nuclear Factor κB Inhibition on Serotype 9 Adeno-Associated Viral (AAV9) Minidystrophin Gene Transfer to the mdx Mouse

Gene therapy studies for Duchenne muscular dystrophy (DMD) have focused on viral vector-mediated gene transfer to provide therapeutic protein expression or treatment with drugs to limit dystrophic changes in muscle. The pathological activation of the nuclear factor (NF)-κB signaling pathway has emerged as an important cause of dystrophic muscle changes in muscular dystrophy. Furthermore, activation of NF-κB may inhibit gene transfer by promoting inflammation in response to the transgene or vector. Therefore, we hypothesized that inhibition of pathological NF-κB activation in muscle would complement the therapeutic benefits of dystrophin gene transfer in the mdx mouse model of DMD. Systemic gene transfer using serotype 9 adeno-associated viral (AAV9) vectors is promising for treatment of preclinical models of DMD because of vector tropism to cardiac and skeletal muscle. In quadriceps of C57BL/10ScSn-Dmdmdx/J (mdx) mice, the addition of octalysine (8K)–NF-κB essential modulator (NEMO)-binding domain (8K-NBD) peptide treatment to AAV9 minidystrophin gene delivery resulted in increased levels of recombinant dystrophin expression suggesting that 8K-NBD treatment promoted an environment in muscle tissue conducive to higher levels of expression. Indices of necrosis and regeneration were diminished with AAV9 gene delivery alone and to a greater degree with the addition of 8K-NBD treatment. In diaphragm muscle, high-level transgene expression was achieved with AAV9 minidystoophin gene delivery alone; therefore, improvements in histological and physiological indices were comparable in the two treatment groups. The data support benefit from 8K-NBD treatment to complement gene transfer therapy for DMD in muscle tissue that receives incomplete levels of transduction by gene transfer, which may be highly significant for clinical applications of muscle gene delivery