Polycaprolactone microcapsules containing citric acid and naringin for plant growth and sustainable agriculture: physico-chemical properties and release behavior.

Plant growth promoting rhizobacteria (PGPR) is an alternative to chemical fertilizers for sustainable, environment friendly agriculture. There is a need to develop strategies to potentiate the interaction between rhizobacteria and plants. Flavonoids and organic acids (components of root exudates) play specific beneficial roles as carbon sources and signal molecules in the plant - rhizobacteria interactions. The goal of this work is to encapsulate signal molecules, namely citric acid and naringin, an organic acid and a flavonoid, respectively, by a biodegradable polymer, polycaprolactone (PCL), in order to maintain the stability and activity of those signal molecules and enable their slow or controlled release over a selected period of time, according to the needs of the plants. This approach is expected to potentiate food crops, namely peanut crop, in adverse environmental conditions (water deficit), by promoting the beneficial interaction between the peanut plant (A. hypogaea) and rhizobacteria. The microcapsules (MCs) are obtained by an emulsion process combined with solvent evaporation technique and are characterized by scanning electron microscopy, thermogravimetry and Fourier transformed infrared spectroscopy. The kinetics of in vitro release of encapsulated molecules, in a period where the uptake of the compound in plants can occur, is studied. The encapsulation synthesis parameters that lead to the best encapsulation process yield and efficiency, as well as to the best final performance in terms of release, are identified. The effect of pH and molecular weight of PCL is found to mediate the release properties of the molecules for different types of soil. PCL 45000 Mw dissolved at 16% in dichloromethane leads to an encapsulation efficiency of 75% and the resulting MCs containing naringin exhibit a slow release profile for 30 days, unmodified by pH, enabling their use in soils of different characteristics. This research makes possible the manufacturing of smart materials for sustainable agriculture practices.

Biodistribution and phototherapeutic properties of Zinc (II) 2,9,16,23-tetrakis (methoxy) phthalocyanine in vivo.

Photodynamic therapy (PDT) was investigated using the phthalocyanine photosensitizer Zinc (II) 2,9,16,23-tetrakis (methoxy) phthalocyanine (ZnPc(OCH(3))(4)) on BALB/c mice. Animals bearing tumor were treated with 0.2mg/kg body weight (bw) ZnPc(OCH(3))(4) and 24h later were irradiated with 70, 140 and 210 J/cm(2) of visible light from a source delivering 39 mW/cm(2). In this study, we have tested the efficiency of ZnPc(OCH(3))(4) liposomal formulation on mice. Biodistribution studies were performed in tumor-free mice and tumor-bearing mice at various time points up to 24h after ZnPc(OCH(3))(4)-PDT treatment. The tumor sizes were evaluated over different period in parallel experiments. The maximal efficiency and selectivity of photosensitizer accumulation in tumor tissue take place at 24h after drug administration of 0.2mg/kg bw ZnPc(OCH(3))(4). In the tumor sections for biochemical studies, apoptosis was visualized by activation of caspase-3. ZnPc(OCH(3))(4)-PDT tumors showed a significant delay in growth as compared to untreated control mice. In all cases, ZnPc(OCH(3))(4)-PDT-treated tumors showed a significant regression. The results indicated a dramatic decrease of tumors size after 10 days post-irradiation with 210 J/cm(2) and no recurrence of the disease was detectable within at least 90 days. The phototherapeutic agent ZnPc(OCH(3))(4) demonstrated preferential accumulation in tumor in comparison with skin tissues, except in the case of kidney. The ratio of tumor/skin tissues ranged a value of 8. These results suggest that ZnPc(OCH(3))(4)-PDT may be of particular importance in the treatment of accessible malignancies.

Synthesis and photodynamic activity of zinc(II) phthalocyanine derivatives bearing methoxy and trifluoromethylbenzyloxy substituents in homogeneous and biological media.

Two zinc(II) phthalocyanines bearing either four methoxy (ZnPc 3) or trifluoromethylbenzyloxy (ZnPc 4) substituents have been synthesized by a two-step procedure starting from 4-nitrophthalonitrile. Absorption and fluorescence spectroscopic studies were analyzed in different media. These compounds are essentially non-aggregated in the organic solvent. Fluorescence quantum yields (phi(F)) of 0.26 for ZnPc 3 and 0.25 for ZnPc 4 were calculated in tetrahydrofuran (THF). The photodynamic activity of these compounds was compared in both THF containing photooxidizable substrates and in vitro on Hep-2 human larynx-carcinoma cell line. The production of singlet molecular oxygen, O(2)((1)Delta(g)), was determined using 9,10-dimethylanthracene yielding values of approximately 0.56 for both sensitizers. Under these conditions, the addition of beta-carotene (Car) suppresses the O(2)((1)Delta(g))-mediated photooxidation. In biological medium, no dark cytotoxicity was found for cells incubated with 0.1 microM of phthalocyanines 3 and 4 for 24 h. However, under similar conditions 0.5 microM of ZnPc 4 was toxic (70% cell survival). The uptake into Hep-2 cells was evaluated using 0.1muM of sensitizer, reaching values of approximately 0.05 nmol/10(6) cells after 3h of incubation at 37 degrees C. The cell survival after irradiation of the cultures with visible light was dependent upon both light exposure level and intracellular sensitizer concentration. A higher photocytotoxic effect was found for ZnPc 3 with respect to 4 (32%/70% cell survival after 15 min of irradiation). Also, these studies were performed treating the cells with 0.5 microM of ZnPc 3. In this case, an increase in the uptake (approximately 0.28 nmol/10(6) cells) was observed, which is accompanied by a higher photocytotoxic activity (20% cell survival). These results show that even though both sensitizer present similar photophysical properties in homogeneous medium, the photodynamic behavior in cellular media can significantly be changed.

Pharmacokinetic and tumour-photosensitizing properties of methoxyphenyl porphyrin derivative.

The photokilling activity of 5-[4-N-(2',6'-dinitro-4'-trifluoromethylphenyl) aminophenyl]-10,15,20-tris(2,4,6-trimethoxy phenyl) porphyrin (CF3) was evaluated on a Hep-2 human larynx-carcinoma cell line. An apoptotic mechanism of cell death was found at low irradiation doses. Pharmacokinetic and tumour-photosensitizing properties were studied in mice. The results show that a different mechanism of cell death can be induced depending on the irradiation doses in the photodynamic treatments with CF3, which makes this agent an interesting sensitizer for potential tumour photosensitizer.