RESUMO
This paper evaluates how effectively chloroaluminum phthalocyanine (ClAlPc) entrapped in colloidal nanocarriers, such as nanocapsule (NC) and nanoemulsion (NE), induces photodamage in human prostate cancer cells (LNCaP) during photodynamic therapy (PDT). The MTT cell viability assay showed that both ClAlPc-NC and ClAlPc-NE induced phototoxicity and efficiently killed LNCaP cells at low ClAlPc-NC and ClAlPc-NE concentrations (0.3µgmL-1) as well as under low light doses of 4Jcm-2 and 7Jcm-2, respectively, upon PDT with a 670-nm diode laser line. Confocal imaging studies indicated that ClAlPc-NC and ClAlPc-NE were preferentially localized in the perinuclear region of LNCaP cells both in the dark and upon irradiation with laser light. After PDT treatment, ClAlPc-NC-treated LNCaP cells exhibited a higher green fluorescence signal, possibly due to the larger shrinkage of the actin cytoskeleton, compared to ClAlPc-NE-treated LNCaP cells. Additionally, ClAlPc-NC or ClAlPc-NE and mitochondria showed a relatively high co-localization level. The cellular morphology did not change in the dark, but confocal micrographs recorded after PDT revealed that LNCaP cells treated with ClAlPc-NC or ClAlPc-NE underwent morphological alterations. Our preliminary in vitro studies reinforced the hypothesis that biocompatible theranostic ClAlPc-loaded nanocarriers could act as an attractive photosensitizer system in PDT and could serve as an interesting molecular probe for the early diagnosis of prostate cancer and other carcinomas.
Assuntos
Portadores de Fármacos , Células Epiteliais/efeitos dos fármacos , Indóis/farmacologia , Mitocôndrias/efeitos dos fármacos , Nanocápsulas/química , Compostos Organometálicos/farmacologia , Fármacos Fotossensibilizantes/farmacologia , Citoesqueleto de Actina/efeitos dos fármacos , Linhagem Celular Tumoral , Sobrevivência Celular/efeitos dos fármacos , Emulsões , Células Epiteliais/patologia , Células Epiteliais/ultraestrutura , Humanos , Luz , Masculino , Microscopia Confocal , Mitocôndrias/patologia , Mitocôndrias/ultraestrutura , Nanocápsulas/administração & dosagem , Fotoquimioterapia/métodos , Próstata/efeitos dos fármacos , Próstata/patologia , Próstata/ultraestrutura , Nanomedicina Teranóstica/métodosRESUMO
A sensitive and reproducible method by microextraction packed sorbent and liquid chromatography with UV detection (MEPS/LC-UV) is described for the determination of new generation antidepressants (sertraline, mirtazapine, fluoxetine, citalopram and paroxetine) in human plasma samples. The MEPS variables, such as sample volume, pH, number of extraction cycles (draw-eject), and desorption conditions (solvent and solvent volume of elution) influenced the MEPS/LC efficiency significantly. Important factors in the optimization of MEPS efficiency, as well as washing steps and carryover effect are discussed. The analyses were carried out using small sample volumes (400 microL), and in a short time period (3 min for the entire sample preparation step). The MEPS/LC-UV method was shown to be linear at concentrations ranging from the limit of quantification (LOQ) to 1000 ng mL(-1). The LOQ values ranged from 10 to 25 ng mL(-1). The inter-day precision of the method presented coefficient of the variation ranging from 1.3% to 8.7%. On the basis of analytical validation, it is shown that the MEPS/LC-UV methodology is adequate for antidepressant analysis, from therapeutic to toxic levels. In order to evaluate the proposed method for clinical use, the MEPS/LC-UV method was applied to analysis of plasma samples from elderly depressed patients.