Free 2-propen-1-amine Derivative And Inclusion Complexes With β-cyclodextrin: Scanning Electron Microscopy, Dissolution, Cytotoxicity And Antimycobacterial Activity

Inclusion complexes and physical mixtures of isomeric mixture of E/Z (50:50) of 3-(4′-bromo-[1,1′-biphenyl]-4-yl)-3-(4-bromophenyl)-N,N- dimethyl-2-propen-1-amine (BBAP) and β-cyclodextrin (β-CD) in the molar proportion of 1:1 and 1:2 were analyzed by scanning electron microscopy. The dissolution behavior of BBAP and of the inclusion complexes were also evaluated for six hours. By scanning electron microscopy (SEM), it was possible to observe an inclusion complex formed between BBAP and β-CD by co-evaporation, either in the molar proportion of 1:1 or 1:2. In the physical mixtures, no complex was observed as previously detected by physicochemical analysis. The dissolution studies showed that the inclusion complexes BBAP/β-CD 1:1 and 1:2 released respectively 49.07 ± 1.48 and 40.26 ± 3.90% of BBAP during six hours. Free BBAP was less soluble than the inclusion complex and reached 9.00 ± 0.75% of dissolution. Biological assays, such as cytotoxicity to J774 macrophages and to a permanent lung fibroblast cell line (V79), indicated that the BBAP does not exhibit any additional toxic effect with the β-CD complexes. However, the complexes were less cytotoxic to V79 cells than the free form. The BBAP/β-CD inclusion complexes were more effective (MIC) than the free compound on several mycobacteria strains. Similar behavior was observed for BBAP/β-CD complexes and rifampicin, a front-line antitubercular drug, on M. tuberculosis H37Rv growing inside J774 macrophages.155682689Bibby, D.C., Davies, N.M., Tucker, I.G., (2000) Int. J. Pharm., 197, p. 1De Souza, A.O., Sato, D.N., Aily, D.C.G., Durán, N., (1998) J. Antimicrob. Chemother., 42, p. 407Pereira, D.G., De Castro, S.L., Durán, N., (1998) Acta Tropica, 69, p. 205De Souza, A.O., Santos Júnior, R.R., Ferreira-Júlio, J.F., Rodrigues, J.A., Melo, P.S., Haun, M., Sato, D.N., Durán, N., (2001) Eur. J. Med. Chem., 36, p. 843De Souza, A.O., Hemerly, F.P., Busollo, A.C., Melo, P.S., Machado, G.M.C., Miranda, C.C., Santa-Rita, R.M., Durán, N., (2002) J. Antimicrob. Chemother., 50, p. 629De Conti, R., Gimenez, S.M.N., Haun, M., Pilli, R.A., De Castro, S.L., Durán, N., (1996) Eur. J. Med. Chem., 31, p. 915De Souza, A.O., Santos Jr., R.R., Sato, D.N., Lima, H.O.S., Andrade-Santana, M.H., Alderete, J.B., Faljoni-Alario, A., Durán, N., (2000) Abstracts of the 29 a Reunião Anual Da Sociedade Brasileira de Bioquímica, , Caxambu, BrazilHiguchi, T., Connors, K.A., (1965) Adv. Anal. Chem. Instrum., 4, p. 117Collins, L.A., Franzblau, S.G., (1997) Antimicrob. Agents Chemother., 41, p. 1004Oh, Y.K., Nix, D.E., Straubinger, R.M., (1995) Antimicrob Agents Chemother., 39, p. 2104Cingi, M.R., De Angelis, I., Fortunati, E., Reggiani, D., Bianchi, V., Tiozzo, R., Zucco, F., (1991) Toxicol. In Vitro, 5, p. 119Denizot, F., Lang, R., (1986) J. Immun. Methods, 89, p. 271Borenfreund, E., Puerner, J.A., (1984) J. Tiss. Cult. Meth., 9, p. 7Melo, P.S., Maria, S.S., Vidal, B.C., Haun, M., Durán, N., (2000) In Vitro Cell Rev. Biol. Animal, 36, p. 539Melo, P.S., Durán, N., Haun, M., (2001) Toxicology, 159, p. 135Shrivastava, R., John, G.W., Rispat, G., Chevalier, A., Massingham, R., (1991) ATLA - Alt. Lab. Anim., 19, p. 39

In Vitro Studies On The Release Of Isoniazid Incorporated In Poly(ε-caprolactone)

A polymeric micro- and nanosphere formulation using poly (ε-caprolactone) (PCL) to entrap an antituberculosis drug, isoniazid (INH), was developed and characterized. The microspheres were prepared by a solvent evaporation method using ethyl acetate, PCL and INH as the organic phase and water and Tween 40 as the aqueous phase. The nanospheres were prepared by a spontaneous emulsification solvent diffusion method using 40% ethanol in acetone (v/v), PCL and INH as the organic phase and water and Tween 40 as the aqueous phase. After freeze-drying, these systems were characterized by scanning electron microscopy (SEM), particle size analysis, determination of entrapped INH content, in vitro INH release and brine shrimp toxicity bioassay. © E.S.I.F.T. srl.185473479Fauci, A.S., Infectious diseases: Considerations for the 21st century (2004) Clin Infect Dis, 32, pp. 675-685Espinal, M.A., Kim, S.J., Suarez, P.G., Standard short-course chemotherapy for drug-resistant tuberculosis: Treatment outcomes in 6 countries (2000) JAMA, 283, pp. 2537-2545Khuller, G.K., Pandey, R., Sustained release drug delivery systems in management of tuberculosis (2003) Indian J Chest Dis Allied Sci, 45, pp. 229-230Dutt, M., Khuller, G.K., Liposomes and PLG microparticles as sustained release antitubercular drug carrier - An in vitro-in vivo study (2001) Inter J Antimicrob Agents, 18, pp. 345-1252Dutt, M., Khuller, G.K., Chemotherapy of Mycobaterium tuberculosis infections in mice with a combination of isoniazid and rifampin entrapped in poly (DL-lactide-co- glycolide) microparticles (2001) J Antimicrob Chemother, 47, pp. 829-835Ul-Ain, Q., Sharma, S., Khuller, G.K., Chemotherapeutic potential of orally administered poly (lactide-co-glycolide) microparticles containing isoniazid, rifampin, and pyrazinamide against experimental tuberculosis (2003) Antimicrob Agents Chemother, 47, pp. 3005-3007Ul-Ain, Q., Sharma, S., Khuller, G.K., Garg, S.K., Alginate-based oral drug delivery system for tuberculosis: Pharmacokinetics and therapeutic effects (2003) J Antimicrob Chemother, 51, pp. 931-938Silva, R.M.L., Evangelista, R.C., Microspheres of alginate-chitosan containing isoniazid (2003) J Microencapsul, 20, pp. 145-152Sharma, R., Saxena, D., Dwivedi, A.K., Misra, A., Inhalable microparticles containing drug combination to target alveolar macrophages for treatment of pulmonary tuberculosis (2001) Pharm Res, 18, pp. 1405-1410Zhou, H.Y., Lengsfeld, C., Claffey, D.J., Ruth, J.A., Hybertson, B., Randolph, T.W., Ng, K.Y., Manning, M.C., Hydrophobic ion pairing of isoniazid using a prodrug approach (2002) J Pharm Sci, 91, pp. 1502-1511Dutt, M., Khuller, G.K., Therapy efficacy of poly (DL-lactide-co-glycolide)-encapsulated antitubercular drugs against Mycobaterium tuberculosis infections in mice (2001) Antimicrob Agents Chemother, 45, pp. 363-366Dutt, M., Khuller, G.K., Sustained release of isoniazid from a single injectable dose of poly (DL-lactide-co-glycolide) microparticles as a therapeutic approach towards tuberculosis (2001) Intern J Antimicrob Agents, 17, pp. 115-122Durán, N., De Oliveira, A.F., De Azevedo, M.M.M., Production of micro and nanospheres of polycaprolactone in the isoniazid encapsulation as antimycobacterial agents (2002), Brazilian Patent-PIBr 2000204125. Derwent Number, 2004-571899 [56]Anisimova, Y.V., Gelperina, S.I., Peloquin, C.A., Heifets, L.B., Nanoparticles as antituberculosis drugs carrier: Effect on activated against Mycobcterium tuberculosis in human monocyte-derived macrophages (2000) J Nanoparticle Res, 2, pp. 165-171. , 2000Pandey, R., Sharma, A., Zahoor, A., Sharma, S., Khuller, G.K., Prasad, B., Poly (DL-lactide-co-glycolide) nanoparticle-based inhalable sustained drug delivery system for experimental tuberculosis (2003) J Antimicrob Chemother, 52, pp. 981-986Pandey, R., Zahoor, A., Sharma, S., Khuller, G.K., Nanoparticles encapsulated antitubercular drugs as a potential oral drug delivery system against murine tuberculosis (2003) Tuberculosis, 83, pp. 373-378Sharma, A., Sharma, S., Khuller, G.K., Lectin-functionalized poly (lactide-co-glycolide) nanoparticles as oral/aerosolized antitubercular drug carrier for treatment of tuberculosis (2004) J Antimicrob Chemother, 54, pp. 761-766Dutt, M., Khuller, G.K., Sub-cutaneous nanoparticles-based antitubercular chemotherapy in an experimental model (2004) J Antimicrob Chemother, 54, pp. 266-268Mei, N., Chen, G., Zhou, P., Chen, X., Shao, Z.-Z., Biocompatibility of poly (ε-caprolactone) scaffold modified by chitosan - The fibroblasts proliferation in vitro (2005) J Biomat Appl, 19, pp. 323-339Pitt, C.G., (1990) Biodegradable Polymers as Drug Delivery System, p. 710. , M. Chasin R. Langer (Eds.), Dekker, New YorkSinha, V.R., Bansal, K., Kaushik, R., Kumria, R., Trehan, A., Poly-ε-caprolactone microspheres and nanospheres: An overview (2004) Intern J Pharm, 278, pp. 1-23Perez, M.H., Zinutti, C., Lamprecht, A., The preparation and evaluation of poly (ε-caprolactone) microparticles containing both lipophilic and hydrophilic drug (2000) J Control Release, 65, pp. 429-438Goldstein, J., Newburg, D.E., Joy, C., (2003) Scanning Electron Microscopy and X-Rays Micro Analysis, p. 675. , Springer Sci. N.Y., USAStine, K.J., Knobler, C.M., Fluorescence Microscopy - A tool for studying the physical-chemistry of interfaces (1992) Ultramicroscopy, 47, pp. 23-34O'Donnell, P.B., McGinity, J.W., Preparation of microspheres by the solvent evaporation technique (1997) Advanced Drug Delivery Rev, 28, pp. 25-42Murakami, H., Kobayashi, M., Takeuchi, H., Kawashima, Y., Further application of a modified spontaneous emulsification solvent diffusion method to various types of PLGA and PLA polymers for preparation of nanoparticles (2000) Powder Technol, 107, pp. 137-143Mc Laughin, J.L., Colman-Saizarbitoria, T., Anderson, J.E., Three simple bioassays for natural chemical products (1995) Rev Soc Venezolana Quim, 18, pp. 13-18Sam, T.W., Toxicity testing using the brine shrimp: Artemia salina (1993) Ioactive Natural Products: Detection, Isolation ans Structural Determination, pp. 491-1454. , Ed. SM Colegate CRC Press Inc. :Ch. 18Ha, J.-H., Kim, S.-H., Han, S.-Y., Albumin release from bioerodible hydrogels based on semi-interpenetrating polymer networks composed of poly(e-caprolactone) and poly(ethylene glycol) macromer (1997) J Control Release, 49, pp. 253-262Armani, D.K., Liu, C., Microfabrication technology for polycaprolactone, a biodegradable polymer (2000) J Micromech Microeng, 10, pp. 80-84Murillo, M., Gamazo, C., Goñi, M.M., Irache, J.M., Blanco-Prieto, M.J., Development of microparticles prepared by spray-drying as a vaccine delivery system against brucellosis (2002) Inter J Pharm, 242, pp. 341-34

Characterisation And Properties Of The Inclusion Complex Of 24-epibrassinolide With β-cyclodextrin

This paper reports the first study of an inclusion complex of a brassinosteroid with β-cyclodextrin. The formation of inclusion complexes between 24-epibrassinolide and β-cyclodextrin was confirmed by their physicochemical properties and the compounds were analysed by differential scanning calorimetry, powder X-ray diffraction, nuclear magnetic resonance spectrometry and scanning electron microscopy. Theoretical calculations using the MM+ HyperChem force field showed a preference for inclusion of the side chain of the epibrassinolide molecule into the β-cyclodextrin cavity to form a 1:1 inclusion complex, although complexes involving inclusion of the steroidal nucleus also possess a favourable interaction energy. Rice lamina inclination assay, employing IAC-103 and IAC-104 cultivars, showed an improved activity for the epibrassinolide-cyclodextrin complex compared to the epibrassinolide itself. The results suggest that brassinosteroid complexation with cyclodextrins may enhance the biological activity of these plant growth regulators.373233240Ahmed, S.M., Improvement of solubility and dissolution of 19-norprogesterone via inclusion complexation (1998) J. Inclusion Phenom. Mol. Rec. Chem., 30, pp. 111-125Alberts, E., Muller, B.W., Complexation of steroid-hormones with cyclodextrin derivatives. Substituent effects of the guest molecule on solubility and stability in aqueous-solution (1992) J. Pharm. Sci., 81, pp. 756-761Braun, P., Wild, A., The influence of brassinosteroids on growth and parameters of photosynthesis of wheat and mustard plants (1984) J. Plant Physiol., 116, pp. 189-196Brutti, C., Apostolo, N.M., Ferrerotti, S.A., Llorente, B.E., Krymkiewicz, N., Micropropagation of Cynara scolymus L. employing cyclodextrins to promote rhizogenesis (2000) Sci. Hortic., 83, pp. 1-10Clouse, S.D., Sasse, J.M., Brassinosteroids: Essential regulators of plant growth and development (1998) Annual Rev. Plant Physiol. Plant Molec. Biol., 49, pp. 427-451Connors, A., The stability of cyclodextrin complexes in solution (1997) Chem. Rev., 97, pp. 1325-1357Cutler, H.G., Yokota, T., Adam, G., (1991) Brassinosteroids: Chemistry, Bioactivity and Applications, ACS Symposium Series, 474. , American Chemical Society, Washington, DCDe Azevedo, M.B.A., Alderete, J.B., Lino, A.C.S., Loh, W., Faljoni-Alario, A., Durán, N., Violacein/β-cyclodextrin inclusion complex formation studied by measurements of diffusion coefficient and circular dichroism (2000) J. Incl. Phenom. Macrocyclic Chem., 37, pp. 67-74De Azevedo, M.B.M., Alderete, J., Zullo, M.A.T., Salva, T.J.G., Duran, N., Brassinosteroids: A new class of plant hormones. The biological activity of 24-epibrassinolide and an inclusion complex of 24-epibrassinolide and β-cyclodextrin (2000) Proceed. Int'l Control. Rel. Bioact. Mater., pp. 5006-5007. , Controlled Release Society, IncDurán, N., De Azevedo, M.B.M., Zullo, M.A.T., Salva, T.J.G., Alderete, J.B., (2000), Process of cyclodextrin/brassinosteroids formulation, for agricultural application, used as plant hormones, Brazilian Patent BR9906202-ADurzan, D.J., Ventimiglia, F.F., (2000), Cyclodextrin nutrients in plant tissue cultures, US Patent US 6087176 [Chem. Abstr. 133: 88976 (2000)]Fujioka, S., Noguchi, T., Takatsuto, S., Yoshida, S., Activity of brassinosteroids in the dwarf rice lamina inclination bioassay (1998) Phytochemistry, 49, pp. 1841-1848Fujioka, S., Sakurai, A., Biosynthesis and metabolism of brassinosteroids (1997) Physiol. Plant., 100, pp. 710-715Gosset, S., Gauvrit, C., (1992), Activity enhancement of benzamide herbicides by cyclodextrins. French 9222204 A1 [Chem. Abstr. 118: 75387 (1997)]Hayashi, T., Iijima, Y., Hoshino, A., Nakamura, M., (1998), Agents and method for flowering acceleration using cinnamic acid-cyclodextrin inclusion compounds Japanese Patent. Kokai Tokkyo Koho JP 10273404 A2 [Chem. Abstr. 129: 327304 (1999)]Huet, H., Jullien, M., The β-cyclodextrins delay the germination of the somatic embryos of carrot (Daucus carota L.) (1992) Acad. Sci., Ser. III, 314, pp. 171-177Ikekawa, N., Zhao, Y., Application of 24-epibrassinolide in agriculture (1991) Brassinosteroids: Chemistry, Bioactivity and Applications. ACS Symposium Series, 474, pp. 280-305. , Cutler H.G., Yokota T. and Adam G. (eds). American Chemical Society, Washington, DCKalinch, F.N., Mandava, N.B., Todhunter, J.A., Relationship of nucleic acid metabolism to brassinolide-induced responses in bean (1985) J. Plant Physiol., 120, pp. 207-214Khripach, V.A., Zhabinskii, V.N., De Groot, A.E., (1999) Brassinosteroids, A New Class of Plant Hormones, , Academic Press, Harcourt Brace & Company, USAKoehler, G., Grabner, G., Klein, C.T.H., Marconi, G., Mayer, B., Monti, S., Structure spectroscopic properties of cyclodextrin inclusion complexes (1996) J. Inc. Phenom. Mol. Rec. Chem., 25, pp. 103-108Lipkowitz, K.B., Applications of computational chemistry to the study of cyclodextrins (1998) Chem. Rev., 98, pp. 1829-1873Mandava, N.B., Plant growth promoting brassinosteroids (1988) Ann. Rev. Plant Physiol. Plant Mol. Biol., 39, pp. 23-52Marquart, V., Adam, G., Recent advances in brassinosteroids research (1991) Chemistry of Plant Protection, Herbicide Resistance - Brassinosteroids, Gibberellins, Plant Growth Regulators, 7, pp. 104-139. , Ebing W. (ed.). Springer-Verlag, BerlinMarzona, M., Carpignano, R., Quagliotto, P., Quantitative structure-stability relationships in the inclusion complexes of steroids with cyclodextrins (1992) Annal. Di Chim., 82, pp. 517-537Okii, M., (1993), Cyclodextrins for enhanced callus tissue culture of rice Japanese Patent. Kokai Tokkyo Koho JP 05292955 A2 [Chem. Abstr. 120: 1324504 (1997)]Rajagopalan, N., Chen, S.C., Chow, W.S., A study of the inclusion complex of amphotericin-B with cyclodextrin (1986) Int. J. Pharm., 29, pp. 161-168Sairam, P.K., Effect of homobrassinolide application on plant metabolism and grain yield under irrigated and moisture stress conditions of two wheat varieties (1994) Plant Growth Regul., 14, pp. 173-181Sakurai, A., Fujioka, S., The current status of physiology and biochemistry of brassinosteroids (1993) Plant Growth Regul., 13, pp. 147-159Sasse, J.M., Recent progress in brassinosteroid research (1997) Physiol. Plant., 100, pp. 696-701Sawamoto, T., (2000), Plant growth-stimulating and disease-preventing agents containing 1-triacontanol and their manufacture Japanese Patent 2000128707 A2 [Chem. Abstr. 132:304657]Stella, V.J., Rajewski, R.A., Cyclodextrins: Their future in drug formulation and delivery (1997) Pharm. Res., 14, pp. 556-567Stella, V.J., Rao, V.M., Zannou, E.A., Zia, V., Mechanisms of drug release from cyclodextrin complexes (1999) Adv. Drugs Del. Rev., 36, pp. 3-16Szejtli, J., Szente, L., Harshegyi, J., Daroczi, I., Vorashazy, L., Torok, S., (1989), Inclusion complexes and mixtures of plant growth regulators with cyclodextrins., Hung. Patent Teljes HU 47961 A2 [Chemical Abstracts 112: 50398 (1997)]Takeno, K., Pharis, R.P., Brassinosteroid-induced bending of the leaf lamina of dwarf rice seedlings: An auxin-mediated phenomenon (1982) Plant Cell Physiol., 23, pp. 1275-1281Uden, W.V., Woerdenbag, H.J., Cyclodextrins as a useful tool for bioconversions in plant cell biotechnology (1994) Plant Cell Tissue Organ Cult., 38, pp. 103-113Vardhini, B.V., Rao, S.S.R., Effect of brassinosteroids on nodulation and nitrogenase activity in groundnut Arachis hypogaea L. (1999) Phytochemistry, 48, pp. 927-930Wada, K., Marumo, S., Abe, H., Morishita, T., Nakamura, K., Uchiyama, M., A rice lamina inclination test - A micro-quantitative bioassay for brassinosteroids (1984) Agric. Biol. Chem., 48, pp. 719-726Wada, K., Marumo, S., Ikekawa, N., Morisaki, M., Mori, K., Brassinolide and homobrassinolide promotion of lamina inclination of rice seedlings (1981) Plant Cell Physiol., 22, pp. 323-326Zhou, D., Wu, Y., Xu, Q., Yang, L., Bai, C., Tan, Z., Molecular mechanics study of the inclusion of trimethylbenzene isomers in α-cyclodextrin (2000) J. Inc. Phenom. Macrocyclic Chem., 37, pp. 273-27

Poly(ε-caprolactone)/propolis Extract: Microencapsulation And Antibacterial Activity Evaluation

Spherical and homogenous microparticles of poly(ε-caprolactone) (PCL), containing propolis were prepared by the emulsification-solvent evaporation technique. Using this method of preparation, a solid formulation of propolis, free of ethanol and suitable for manipulation and storage, was obtained from an ethanolic extract of propolis. The incorporation efficiency of propolis in the microparticles was almost 30% and around 60% of the substance was released in 48 h. In vitro propolis microparticles exhibited similar halo zones in the Petri plate test against Streptococcus mutans (GS5) with a 10-fold lower concentration than the free propolis extract showing that the encapsulated propolis in microparticles is more efficient as antibiotic.624287290Baras, B., Benoit, M.-A., Poulain-Godefroy, O., Schacht, A.-M., Capron, A., Gillard, J., Riveau, G., Vaccine properties of antigens entrapped in microparticles produced by spray-drying technique and using various polyester polymers (2000) Vaccine, 18, pp. 1495-1505Bruschi, M.L., Cardoso, M.L.C., Lucchesi, M.B., Gremião, M.P.D., Gelatin microparticles containing propolis obtained by spray-drying technique: Preparation and characterization (2003) Int J Pharm, 264, pp. 45-55Castro, S.L., Propolis: Biological and pharmacological activities. Therapeutic uses of this bee-product (2001) Ann Rev Biomed Sci, 3, pp. 49-83Gebara, E.C.E., Lima, L.A., Mayer, M.P.A., Propolis antimicrobial activity against periodontopathic bacteria (2002) Braz J Microbiol, 33, pp. 365-369Hai, Y., Zheng, W., Gao, Y., Encapsulation and properties of propolis extract powder (2002) Zhongguo Shipin Zazhish, 23, pp. 61-64Koo, H., Rosalen, P.L., Cury, J.A., Ambrosano, G.M.B., Murata, R.M., Yatsuda, R., Ikegaki, M., Park, Y.K., Effect of a new variety of Apis mellifera propolis on mutans Streptococci (2000) Current Microbiol, 41, pp. 192-196Leitao, D.P.D., Da Silva, A.A., Polizello, A.C.M., Bastos, J.K., Spadaro, A.C.C., Comparative evaluation in vitro effects of Brazilian green propolis and Baccharis dracunculifolia extracts on cariogenic factors of Streptococcus mutans (2004) Biol Pharm Bull, 27, pp. 1834-1839Maciejewicz, W., Daniewski, M., Marowski, W., GC-MS identification of flavonoids algycones isolated from propolis (2001) Chromatography, 53, pp. 343-346Melo, P.S., De Azevedo, M.M.M., Haun, M., Durán, N., Violacein and violacein-loaded poly (D,L-lactide-co-glycolide) nanoparticles induce apoptosis via a mitochondrial pathway in human leukemic cells (2005) Proc. Thirth Meeting Network of Nanobiotechnology, 3, pp. 56-57. , Research Progress São Pedro, BrazilMoreno, M.M., Isla, M.L., Samlietro, A.R., Vattuone, M.A., Comparison of the free radical-scaveging activity of propolis from several regions of Argentina (2000) J Ethnopharmacol, 71, pp. 109-114Murad, J.M., SAMVC, C., Bankova, V., Sforcin, J.M., Effects of propolis from Brazil and Bulgaria on fungicidal activity of macrophages against Paracoccidioides brasiliensis (2004) J Enthnopharmacol, 79, pp. 331-334Paraventi BH, Esposito E (2002) Atividade antimicrobiana de propolis em bacterias orais. Estudo in vitro. Anais Encontro Pesquisa Inic. Cientif., Primeiro Congresso de Odontologia da UMC, Outubro, p. 71Pepeljnjak, S., Jalsenjak, J.M., Maysinger, D., Influence of microencapsulation propolis extract on Bacillus subtilis strain IP-5832 (1981) Acta Pharm Jugoslavica, 31, pp. 27-32Sawaya, A.C.H.F., Souza, K.L., Marcucci, M.C., Cunha, I.B.S., Shimizu, M.T., Analysis of the composition of brazilian propolis extract by chromatography and evaluation of their in vitro activity against gram- positive bacteria (2004) Braz J Microbiol, 35, pp. 104-109Santoyo, S., Ga de Jalén, E., Ygatua, P., Renedo, M.J., Branco-Prieto, M.J., Optimization of topical cidofovir penetration using microparticles (2002) Int J Pharm, 242, pp. 107-113Sforcin, J.M., Fernandes Jr, A., Lopes, C.A.M., Bankova, V., Funari, S.R.C., Seasonal effect on Brazilian propolis antibacterial activity (2000) J Ethnopharmacology, 73, pp. 243-249Uzel, A., Sorkun, K., Oncag, O., Cogulu, D., Gencay, M., Salih, B., Chemical compositions and antimicrobial activities of four different Anatolian propolis samples (2005) Microbiol Res, 160, pp. 189-19

Nanocytotoxicity: Violacein And Violacein-loaded Poly (d, L-lactide-co-glycolide) Nanoparticles Acting On Human Leukemic Cells

Violacein is a compound obtained from Chromobacterium violaceum, a bacterium found in the Amazonian region. Violacein-loaded poly (D, L-lactide-co-glycolide) nanoparticles has a similar inhibitory effect evaluated by trypan blue assay on leukemic HL60 cells than the free form. However, the cytotoxic effects evaluated by phosphatase activity and MTT reduction assays were lower for the encapsulated form than for free violacein. Based on morphological changes, violacein and violacein entrapped in nanoparticles were found to induce terminal differentiation (assessed by nitro blue tetrazolium reduction) in HL60 cells. Thus, both formulations inhibit HL60 cell growth in vitro, partly by inducing cytotoxic effects and cell differentiation. Flow cytometric analysis of HL60 cells after treatment for 12 h showed that violacein-loaded PLGA induced apoptosis, with maximum cell death at a concentration of 2 μM. Violacein and violacein/PLGA induced opposite effects on the mitochondrial swelling which indicates altered mitochondrial function. The mitochondrial activity was also checked by flow cytometry studies. Labelled cells with the probe JC1 displayed a basal hypopolarized status of the mitochondria in treated cells. Based on morphological changes, alterations in phospholipid asymmetry and changes in mitochondrial polarization, violacein and nanoparticles containing violacein were found to trigger cell death by apoptosis. These methodologies are promising as diagnostic and mechanistic effects of nanoparticles in cell cultures. Copyright © 2009 American Scientific Publishers All rights reserved.52192201Durán, N., Menck, C.F.M., Chromobacterium violaceum: A review of pharmacological and industrial perspectives (2001) Crit. Rev. Microbiol., 27, p. 201Melo, P.S., Justo, G.Z., De Azevedo, M.B.M., Durán, N., Haun, M., Violacein and its complexes induce apoptosis and differentiation in HL60 cells (2003) Toxicology, 186, p. 217Dessaux, Y., Elmerich, C., Faure, D., Violacein: A molecule of biological interest originating from the soil-borne bacterium Chromobacterium violaceum (2004) La Revue de Médecine Interne, 25, p. 659Saraiva, V.S., Marshall, J.C., Cools-Lartigue, J., Burnier, M.N., Cytotoxic effects of violacein in human uveal melanoma cell lines (2004) Melanoma Res., 14, p. 421Durán, N., Justo, G.Z., Ferreira, C.V., Melo, P.S., Cordi, L., Martins, D., Violacein: Properties and biological activities (2007) Biotechnol. Appl. Biochem., 48, p. 127Shenoy, D.B., Amiji, M.M., Poly(ethylene oxide)-modified poly(ε-caprolactone) nanoparticles for targeted delivery of tamoxifen in breast cancer (2005) Int. J. Pharm., 293, p. 261Rao, G.C., Kumar, M.S., Mathivanan, N., Rao, M.E., Nanosuspensions at the most promising approach in nanoparticulate drug delivery systems (2004) Pharmazie, 59, p. 5Jin, S., Ye, K.M., Nanoparticle-mediated drug delivery and gene therapy (2007) Biotechnol. Progr., 23, p. 32Kabanov, A.V., Batrakova, E.V., Miller, D.W., Pluronic block copolymers as modulators of drug efflux transporter activity in the blood-brain barrier (2003) Advan. Drug Deliv. Rev., 55, p. 151Minko, T., Batrakova, E.V., Li, S., Li, Y.L., Pakunlu, T.I., Alakhov, V.Y., Kabanov, A.V., Pluronic block copolymers alter apoptotic signal transduction of doxorubicin in drug-resistant cancer cells (2005) J. Control. Release, 105, p. 269Rapoport, N., Marin, A.P., Timoshin, A.A., Effect of a polymeric surfactant on electron transport in HL60 cells (2000) Arch. Biochem. Biophys., 384, p. 100Kohli, E., Han, H.Y., Zeman, A.D., Vinogradov, S.V., Formulations of biodegradable Nanogel carriers with 5′- triphosphates of nucleoside analogs that display a reduced cytotoxicity and enhanced drug activity (2007) J. Control. Release, 121, p. 19Uzunoglu, S., Uslu, R., Tobu, M., Saydam, G., Terzioglu, E., Buyukkececi, F., Omay, S.B., Augmentation of methylprednisolone-induced differentiation of myeloid leukemia cells by serine/threonine protein phosphatase inhibitors (1999) Leukaemia Res., 23, p. 507Anazetti, M.C., Melo, P.S., Durán, N., Haun, M., Comparative cytotoxicity of dimethylamide-crotonin in the promyelocytic leukemia cell line (HL60) and human peripheral blood mononuclear cells (2003) Toxicology, 188, p. 261Redmond, K.M., Wilson, T.R., Johnston, P.G., Longley, D.B., Resistance mechanisms to cancer chemotherapy (2008) Frontiers Biosci., 13, p. 5138Melet, A., Song, K., Bucur, O., Jagani, Z., Grassian, A.R., Khosravi-Far, R., Apoptotic pathways in tumor progression and therapy (2008) Advan. Exper. Med. Biol., 615, p. 47Debatin, K.M., Activation of apoptosis pathways by anticancer treatment (2000) Toxicol. Lett., 112, p. 41Inayat-Hussain, S.H., Osman, A.B., Din, L.B., Taniguchi, N., Altholactone, a novel styryl-lactone induces apoptosis via oxidative stress in human HL60 leukemia cells (2002) Toxicol. Lett., 131, p. 153Kim, R., Emi, M., Tanabe, K., The role of apoptosis in câncer cell survival and therapeutic outcome (2006) Cancer Biol. Ther., 30, p. 5Anuradha, C.D., Kanno, S., Hirano, S., Oxidative damage to mitochondria is a preliminary step to caspase-3 activation in fluoride-induced apoptosis in HL60 cells (2001) Free Radic. Biol. Med., 31, p. 367Stoetzer, O.J., Progrebniak, A., Pelka-Fleischer, R., Hasmann, M., Hiddemann, W., Nuessler, V., Modulation of apoptosis by mitochondrial uncouplers: Apoptosis-delying features despite intrinsic cytotoxicity (2002) Biochem. Pharmacol., 63, p. 471Ujihara, M., Nomura, K., Yamada, O., Deura, H., Establishment of a stable HL60 subline having the potential for monocytic differentiation using granulocyte-macrophage colony-stimulating factor: Possible use for the study of monocytic differentiation and oxidative stress (1998) Atherosclerosis, 139, p. 301Sokoloski, J.A., Sartorelli, A.C., Induction of the differentiation of HL60 promyelocytic leukemia cells by nonsteroidal anti-inflammatory agents in combination with low levels of vitamin D 3 (1997) Leukemia. Res., 22, p. 153Wyllie, A.H., Kerr, J.F.K., Currie, A.R., Cell Death: The significance of apoptosis (1980) Int. Rev. Cytol., 68, p. 251Israels, L.G., Israels, E.D., Apoptosis (1999) Steam Cells, 17, p. 306Kohroki, J., Norio, M.A., Tanaka, T., Itoh, N., Inada, A., Tanaka, K., Induction of differentiation and apoptosis by dithizone in human myeloid leukemia cell lines (1998) Leukemia Res., 22, p. 405Minko, T., Kopecková, P., Kopecek, J., Preliminary evaluation of caspases-dependent apoptosis signaling pathways of free and HPMA copolymer-bound doxorubicin in human ovarian carcinoma cells (2001) J. Control. Release, 71, p. 227Minko, T., Pakunlu, R.I., Wang, Y., Khandar, J.J., Saad, M., New generation of liposomal drugs for cancer (2006) Anticancer Agents Med. Chem., 6, p. 537Kodach, L.L., Bos, C.L., Durán, N., Peppelenbosch, M.P., Ferreira, C.V., Hardwick, J.C., Violacein synegistically increases 5-fluorouracil cytotoxicity, induces apoptosis and inhibits Akt-mediated signal transduction in humancolorectal cancer cells (2006) Carcinogenesis, 27, p. 508De Carvalho, D.D., Costa, F.T.M., Durán, N., Haun, M., Cytotoxic activity of violacein in human colon cancer cells (2006) Toxicol. in Vitro, 20, p. 1514Rettori, D., Durán, N., Production, extraction and purification of violacein: An antibiotic produced by Chromobacterium violaceum (1998) World J. Microbiol. Biotechnol., 14, p. 685Ferdous, A.J., Stembridge, N.Y., Singh, M., Role of monensin PLGA polymer nanoparticles and liposomes as potentiator of ricin a immunotoxins in vitro (1998) J. Control. Release, 50, p. 71Denizot, F., Lang, R., Rapid colorimetric assay for cell growth and survival, modifications to the tetrazolium dye procedure giving improved sensitivity and reliability (1986) J. Immunol. Methods, 89, p. 271Schneider, W.C., Hogeboom, G.H., Intracellular distribution of enzymes. V. Further studies of cytochrome c in rat liver homogenates (1950) J. Biol. Chem., 183, p. 123Anazetti, M.C., Melo, P.S., Durán, N., Haun, M., Dehydrocrotonin and its derivative, dimethylamide-crotonin induce apoptosis with lipid peroxidation and activation of caspases-2, -6 and -9 in human leukemic cells HL60 (2004) Toxicology, 203, p. 123Araragi, S., Kondoh, M., Kawase, M., Saito, S., Higashimoto, M., Sato, M., Mercuric chloride induces apoptosis via a mitochondrial-dependent pathway in human leukemia cells (2003) Toxicology, 184, p. 1Fillon, S., Lang, F., Jendrossek, V., Pseudomonas aeruginosa triggered apoptosis of human epithelial cells depends on the temperature during infection (2002) Cell Physiol. Biochem., 12, p. 207Melo, P.S., Maria, S.S., Vidal, B.C., Haun, M., Durán, N., Violacein cytotoxicity and induction of apoptosis in V79 cells (2000) In Vitro Cell. Develop. Biol.-Animal, 36, p. 539Pailard, F., Finot, F., Mouche, I., Prenez, A., Vericat, J.A., Use of primary cultures of rat hepatocytes to predict toxicity in the early development of new chemical entities (1999) Toxicol. in Vitro, 13, p. 693Mosmann, T., Rapid colorimetric assay for cellular growth and survival: Application to proliferation and cytotoxicity assay (1983) J. Immunol. Methods, 65, p. 55Freire, A.C.G., Aoyama, H., Haun, M., Ferreira, C.V., Relationship between phosphatase inhibitors, okadaic acid and pervanadate, on human myeloid leukaemia cell line (2003) J. Enzyme Inhib. Med. Chem., 18, p. 425Mehar, A., Macanas-Pirard, P., Mizokami, A., Takahashi, Y., Kass, G.E.N., Coley, H.M., The effects of cyclooxygenase-2 expression in prostate cancer cells: Modulation of response to cytotoxic agents (2008) J. Pharmacol. Exper. Ther., 324, p. 1181Rossi, D., Gaidano, G., Messengers of cell death: Apoptotic signalling in health and disease (2003) Haematologica, 88, p. 212Lee, J.Y., Hwang, W.I., Lim, S.T., Antioxidant and anticancer activities of organic extracts from Platycodon grandiflorum A. de Candolle roots (2004) J. Ethnopharmacol., 93, p. 409Facompré, M., Wattez, N., Kluza, J., Lansiaux, A., Bailly, C., Relationship between cell cycle changes and variations of the mitochondrial membrane potential induced by etoposide (2000) Mol. Cell Biol. Re. Comm., 4, p. 37Greenebaum, B., Blossfield, K., Hannig, J., Carrillo, C.S., Beckett, M.A., Weichselbaum, R.R., Lee, R.C., Poloxamer 188 prevents acute necrosis of adult skeletal muscle cells following high-dose irradiation (2004) Burns, 30, p. 539Luo, J., Borgens, R., Shi, R., Polyethylene glycol immediately repairs neuronal membranes and inhibits free radical production after acute spinal cord injury (2002) J. Neurochem., 83, p. 471Marks, J.D., Pan, C., Bushell, R., Cromie, W., Lee, R.C., Amphiphilic, tri-block copolymers provide potent, membrane-targeted neuroprotection (2001) FASEB J., 15, p. 1107Maskarinec, S.A., Lee, K.Y.C., Comparative study of poloxamer insertion into lipid monolayers (1809) Langmuir, 19, p. 2003Tharmalingam, T., Ghebeth, H., Wuerz, T., Butler, M., Pluronic enhances the robustness and reduces the cell attachment of mammalian cells (2008) Mol. Biotechnol., 39, p. 16