RESUMO
Systemic therapy is generally required for breast cancer. However, treatment toxicity and side effects are a concern, especially for triple-negative breast cancer (TNBC), a subtype that usually develops resistance to chemotherapy. To overcome this issue, new nanoformulations capable of targeting cancer cells have been developed and alternative biomarkers have been explored as target molecules for TNBC management. In this study, we performed anin vivoassay in a murine orthotopic TNBC model to evaluate the targeting ability of anti-carcinoembryonic antigen (anti-CEA) loaded nanoparticles (labelled MFCEA), which had been previously synthetized by our research group. 4T1 cells were injected in the mammary gland of balb-c mice, and tumors were evaluated for CEA expression by immunohistochemistry. Tumor-bearing mice received targeted (MFCEA) and non-targeted (MF) nanoparticles intraperitoneally. Tumors were removed 1, 4, 15 and 24 h after treatment, and Prussian blue iron staining was performed. Our results showed, as far as we know for the first time, that 4T1 induced tumors are CEA positive, and this opens up new prospects for treating TNBC. Furthermore, MFCEA nanoparticles were able to target malignant tissue and were retained in the tumor for longer than MF nanoparticles. The retention property of MFCEA, together with the absence of toxicity observed in the MTT assay, make these nanoparticles a promising device for management of CEA positive tumors and perhaps for TNBC. Nevertheless, further studies must be carried out to improve their performance and ensure safety for clinical studies.
Assuntos
Nanopartículas , Neoplasias de Mama Triplo Negativas , Animais , Antígeno Carcinoembrionário/uso terapêutico , Linhagem Celular Tumoral , Humanos , Ferro , Camundongos , Nanopartículas/química , Neoplasias de Mama Triplo Negativas/tratamento farmacológico , Neoplasias de Mama Triplo Negativas/metabolismo , Neoplasias de Mama Triplo Negativas/patologiaRESUMO
BACKGROUND: Magnetic fluids containing superparamagnetic iron oxide nanoparticles represent an attractive platform as nanocarriers in chemotherapy. Recently, we developed a formulation of maghemite nanoparticles coated with rhodium (II) citrate, which resulted in in vitro cytotoxicity enhanced up to 4.6 times when compared to free rhodium (II) citrate formulation on breast carcinoma cells. In this work, we evaluate the antitumor activity and toxicity induced by these formulations in Balb/c mice bearing orthotopic 4T1 breast carcinoma. METHODS: Mice were evaluated with regard to the treatments' toxicity through analyses of hemogram, serum levels of alanine aminotransferase, iron, and creatinine; DNA fragmentation and cell cycle of bone marrow cells; and liver, kidney and lung histology. In addition, the antitumor activity of rhodium (II) citrate and maghemite nanoparticles coated with rhodium (II) citrate was verified by tumor volume reduction, histology and immunohistochemistry. RESULTS: Regarding the treatments' toxicity, no experimental groups had alterations in levels of serum ALT or creatinine, and this suggestion was corroborated by the histopathologic examination of liver and kidney of mice. Moreover, DNA fragmentation frequency of bone marrow cells was lower than 15% in all experimental groups. On the other hand, the complexes rhodium (II) citrate-functionalized maghemite and free rhodium (II) citrate led to a marked growth inhibition of tumor and decrease in CD31 and Ki-67 staining. CONCLUSIONS: In summary, we demonstrated that both rhodium (II) citrate and maghemite nanoparticles coated with rhodium (II) citrate formulations exhibited antitumor effects against 4T1 metastatic breast cancer cell line following intratumoral administration. This antitumor effect was followed by inhibition of both cell proliferation and microvascularization and by tumor tissue injury characterized as necrosis and fibrosis. Remarkably, this is the first published report demonstrating the therapeutic efficacy of maghemite nanoparticles coated with rhodium (II) citrate. This treatment prolonged the survival period of treated mice without inducing apparent systemic toxicity, which strengthens its use for future breast cancer therapeutic applications.
Assuntos
Antineoplásicos/farmacologia , Compostos Férricos/química , Nanopartículas de Magnetita/química , Ródio/farmacologia , Alanina Transaminase/sangue , Animais , Neoplasias da Mama/tratamento farmacológico , Neoplasias da Mama/patologia , Ciclo Celular/efeitos dos fármacos , Linhagem Celular Tumoral , Proliferação de Células/efeitos dos fármacos , Ácido Cítrico/química , Ácido Cítrico/farmacologia , Creatinina/sangue , Fragmentação do DNA/efeitos dos fármacos , Feminino , Compostos Férricos/análise , Humanos , Imuno-Histoquímica , Ferro/sangue , Antígeno Ki-67/análise , Rim/efeitos dos fármacos , Rim/metabolismo , Fígado/efeitos dos fármacos , Fígado/metabolismo , Camundongos , Camundongos Endogâmicos BALB C , Molécula-1 de Adesão Celular Endotelial a Plaquetas/análise , Ródio/química , Raios UltravioletaRESUMO
Nanosized maghemite particles were synthesized, precoated (with dimercaptosuccinic acid) and surface-functionalized with anticarcinoembryonic antigen (anti-CEA) and successfully used to target cell lines expressing the CEA, characteristic of colorectal cancer (CRC) cells. The as-developed nanosized material device, consisting of surface decorated maghemite nanoparticles suspended as a biocompatible magnetic fluid (MF) sample, labeled MF-anti-CEA, was characterized and tested against two cell lines: a high-CEA expressing cell line (LS174T) and a low-CEA expressing cell line (HCT116). Whereas X-ray diffraction was used to assess the average core size of the as-synthesized maghemite particles (average 8.3 nm in diameter), dynamic light scattering and electrophoretic mobility measurements were used to obtain the average hydrodynamic diameter (550 nm) and the zeta-potential (-38 mV) of the as-prepared and maghemite-based nanosized device, respectively. Additionally, surface-enhanced Raman spectroscopy (SERS) was used to track the surface decoration of the nanosized maghemite particles from the very first precoating up to the attachment of the anti-CEA moiety. The Raman peak at 1655 cm(-1), absent in the free anti-CEA spectrum, is the signature of the anti-CEA binding onto the precoated magnetic nanoparticles. Whereas MTT assay was used to confirm the low cell toxicity of the MF-anti-CEA device, ELISA and Prussian blue iron staining tests performed with both cell lines (LS174T and HCT116) confirm that the as-prepared MF-anti- CEA is highly specific for CEA-expressing cells. Finally, transmission electron microscopy analyses show that the association with anti-CEA seems to increase the number of LS174T cells with internalized maghemite nanoparticles, whereas no such increase seems to occur in the HCT116 cell line. In conclusion, the MF-anti-CEA sample is a biocompatible device that can specifically target CEA, suggesting its potential use as a theragnostic tool for CEA-expressing tumors, micrometastasis, and cancer-circulating cells.
Assuntos
Anticorpos Monoclonais/uso terapêutico , Antígeno Carcinoembrionário/imunologia , Neoplasias Colorretais/tratamento farmacológico , Neoplasias Colorretais/patologia , Nanopartículas de Magnetita/química , Nanopartículas de Magnetita/uso terapêutico , Anticorpos Monoclonais/imunologia , Linhagem Celular Tumoral , Compostos Férricos/química , Humanos , Nanocápsulas/uso terapêuticoRESUMO
Antitumor activities have been described in selol, a hydrophobic mixture of molecules containing selenium in their structure, and also in maghemite magnetic nanoparticles (MNPs). Both selol and MNPs were co-encapsulated within poly(lactic-co-glycolic acid) (PLGA) nanocapsules for therapeutic purposes. The PLGA-nanocapsules loaded with MNPs and selol were labeled MSE-NC and characterized by transmission and scanning electron microscopy, electrophoretic mobility, photon correlation spectroscopy, presenting a monodisperse profile, and positive charge. The antitumor effect of MSE-NC was evaluated using normal (MCF-10A) and neoplastic (4T1 and MCF-7) breast cell lines. Nanocapsules containing only MNPs or selol were used as control. MTT assay showed that the cytotoxicity induced by MSE-NC was dose and time dependent. Normal cells were less affected than tumor cells. Cell death occurred mainly by apoptosis. Further exposure of MSE-NC treated neoplastic breast cells to an alternating magnetic field increased the antitumor effect of MSE-NC. It was concluded that selol-loaded magnetic PLGA-nanocapsules (MSE-NC) represent an effective magnetic material platform to promote magnetohyperthermia and thus a potential system for antitumor therapy.
Assuntos
Neoplasias da Mama/terapia , Hipertermia Induzida/métodos , Nanopartículas de Magnetita/química , Nanopartículas de Magnetita/uso terapêutico , Nanocápsulas/química , Nanocápsulas/uso terapêutico , Compostos de Selênio/administração & dosagem , Animais , Antineoplásicos/uso terapêutico , Neoplasias da Mama/patologia , Linhagem Celular Tumoral , Sobrevivência Celular/efeitos dos fármacos , Terapia Combinada , Feminino , Nanopartículas de Magnetita/ultraestrutura , Camundongos , Nanocápsulas/ultraestrutura , Resultado do TratamentoRESUMO
Breast tumors represent the most common malignant tumors. Current treatments for humans and pets rely on tumor excision and adjuvant chemotherapy, which may affect both cancer cells and normal cells. Photodynamic therapy (PDT) is an approved treatment modality for a variety of cancers and was recently recommended as a first-line treatment for non-melanoma skin cancers for humans. The main purpose of the present study was to determine the efficacy of PDT using aluminum-chloride-phthalocyanine that is encapsulated in liposomes and LED as a light source to kill naturally occurring female dog breast cancer in vitro. The cytotoxicity behavior of the encapsulated photosensitizer in the dark and under irradiation using the 670 nm laser were investigated using classical trypan blue and MTT cell viability tests, acridine orange and ethidium bromide staining to label organelles, and cell morphology. Cell morphology was evaluated using light and electron microscopy. Our results demonstrate a reduced cell viability that is associated with morphologic alterations. The neoplasic cell destruction was predominantly mediated via a necrotic process, which was assayed using acridine orange and ethidium bromide staining. These findings were confirmed using light and electronic microscopy. The photosensitizer or laser irradiation alone did not induce cytotoxicity or morphological alterations, indicating the safety and efficacy of PDT with chloro-aluminum-phthalocyanine that was encapsulated in liposomes for the treatment of breast cancer cells in vitro.
Assuntos
Indóis/farmacologia , Lipossomos/farmacologia , Neoplasias Mamárias Animais/tratamento farmacológico , Compostos Organometálicos/farmacologia , Fotoquimioterapia/métodos , Animais , Antineoplásicos/química , Antineoplásicos/farmacologia , Apoptose/efeitos dos fármacos , Sobrevivência Celular/efeitos dos fármacos , Cães , Feminino , Indóis/química , Lipossomos/química , Neoplasias Mamárias Animais/patologia , Camundongos , Microscopia Eletrônica de Transmissão , Microscopia de Contraste de Fase , Células NIH 3T3 , Necrose , Compostos Organometálicos/química , Fármacos Fotossensibilizantes/química , Fármacos Fotossensibilizantes/farmacologia , Células Tumorais CultivadasRESUMO
Magnetic resonance is used to investigate biodistribution aspects of dextran-coated magnetite nanoparticles (9.4 nm core diameter) in both liver and spleen from 5 minutes up to 6 months after intravenous administration of a magnetic fluid sample in female Swiss mice. Using magnetic resonance data important parameters such as the absorption half-life (t 1/2 = 12 +/- 2 min in the liver and t 1/2 = 11 +/- 2 min in the spleen), the peak time (1.7 +/- 0.2 h in the liver and 1.9 +/- 0.2 h in the spleen), and the disposition half-life of the dextran-coated magnetite nanoparticles in mice organs (t 1/2 = 70 +/- 10 h in the liver and t 1/2 = 32 +/- 7 h in the spleen) were assessed. In addition, light and electron microscopy showed several aspects that may be related to the iron metabolism. Microscopic analysis also revealed that although magnetite nanoparticles or iron released from them are retained in the organism for a long period of time, no morphologic alteration is induced by the intravenous administration of the magnetic fluid sample, evidencing its biocompatibility. The used tests may represent an adequate methodology for nanotoxicology evaluation.
Assuntos
Dextranos/farmacocinética , Portadores de Fármacos/farmacocinética , Nanopartículas de Magnetita/química , Animais , Dextranos/química , Portadores de Fármacos/química , Feminino , Histocitoquímica , Injeções Intravenosas , Fígado/metabolismo , Espectroscopia de Ressonância Magnética , Camundongos , Microscopia Eletrônica , Modelos Animais , Tamanho da Partícula , Fotomicrografia , Baço/metabolismo , Distribuição TecidualRESUMO
BACKGROUND: The magnetic albumin nanosphere (MAN), encapsulating maghemite nanoparticles, was designed as a magnetic drug delivery system (MDDS) able to perform a variety of biomedical applications. It is noteworthy that MAN was efficient in treating Ehrlich's tumors by the magnetohyperthermia procedure. METHODS AND MATERIALS: In this study, several nanotoxicity tests were systematically carried out in mice from 30 minutes until 30 days after MAN injection to investigate their biocompatibility status. Cytometry analysis, viability tests, micronucleus assay, and histological analysis were performed. RESULTS: Cytometry analysis and viability tests revealed MAN promotes only slight and temporary alterations in the frequency of both leukocyte populations and viable peritoneal cells, respectively. Micronucleus assay showed absolutely no genotoxicity or cytotoxicity effects and histological analysis showed no alterations or even nanoparticle clusters in several investigated organs but, interestingly, revealed the presence of MAN clusters in the central nervous system (CNS). CONCLUSION: The results showed that MAN has desirable in vivo biocompatibility, presenting potential for use as a MDDS, especially in CNS disease therapy.
Assuntos
Materiais Biocompatíveis/toxicidade , Sistemas de Liberação de Medicamentos/métodos , Nanopartículas de Magnetita/toxicidade , Soroalbumina Bovina/toxicidade , Análise de Variância , Animais , Química Encefálica/efeitos dos fármacos , Sobrevivência Celular/efeitos dos fármacos , Feminino , Citometria de Fluxo , Histocitoquímica , Contagem de Leucócitos , Leucócitos/efeitos dos fármacos , Teste de Materiais , Camundongos , Testes para Micronúcleos , Nanosferas/toxicidade , Testes de ToxicidadeRESUMO
Strong evidence indicates that reactive oxygen species (ROS) play an important role in the initiation as well as the promotion phase of carcinogenesis. Studies support the role of ROS in cancer, in part, by showing that dietary antioxidants act as cancer-preventive agents. Although results are promising, the research on this topic is still controversial. Thus, the aim of this study was to investigate whether vitamins C, E and pequi oil can, individually, provide prevention and/or be used afterward as an adjuvant in cancer therapy. Ehrlich solid tumor-bearing mice received antioxidant as follows: before tumor inoculation, before and after tumor inoculation (continuous administration), and after tumor inoculation; morphometric analyses of tumor, genotoxicity and hematology were then carried out. Antioxidant administrations before tumor inoculation effectively inhibited its growth in the three experimental protocols, but administrations after the tumor's appearance accelerated tumor growth and favored metastases. Continuous administration of pequi oil inhibited the tumor's growth, while the same protocol with vitamins E and C accelerated it, favoring metastasis and increasing oxidative stress on erythrocytes. Except for continuous administration with vitamin E, the development of ascites tumor metastases was linked with increased inflammation. Results suggest that the efficiency and applicability of antioxidants in the medical clinic can depend not only on the nature of the antioxidant, the type and stage of cancer being treated and the prevailing oxygen partial pressure in the tissues, but also on the type of antioxidant therapy chosen.
Assuntos
Antioxidantes/uso terapêutico , Ácido Ascórbico/uso terapêutico , Carotenoides/uso terapêutico , Óleos de Plantas/uso terapêutico , Vitamina E/uso terapêutico , Animais , Contagem de Células Sanguíneas , Carcinoma de Ehrlich/patologia , Carcinoma de Ehrlich/secundário , Ensaio Cometa , Ericales/química , Feminino , Camundongos , Neoplasias/prevenção & controle , Espécies Reativas de Oxigênio/farmacologia , Vitamina E/farmacologiaRESUMO
OBJECTIVES: The present study reports on the preparation and testing of a desoxycholate amphotericin B (D-AMB) sustained delivery system based on poly(lactic-co-glycolic acid) (PLGA) and dimercaptosuccinic acid (DMSA) polymeric blends (Nano-D-AMB) aimed at reducing the number of AMB administrations required to treat mycosis. METHODS: BALB/c mice were infected with the yeast Paracoccidioides brasiliensis intravenously to mimic the chronic form of paracoccidioidomycosis. At 30 days post-infection, the animals were treated with Nano-D-AMB [6 mg/kg of encapsulated D-AMB, intraperitoneally (ip), interval of 72 h] or D-AMB (2 mg/kg, ip, interval of 24 h). Drug efficacy was investigated by the fungal burden recovery from tissues. Toxicity was assessed by renal and hepatic biochemical parameters, physical appearance of the animals and haematological investigation. The control groups used were non-infected and the infected mice mock treated with PBS. RESULTS: Nano-D-AMB presented results comparable to free D-AMB, with a marked antifungal efficacy. The Nano-D-AMB-treated group presented lower loss of body weight and absence of stress sign (piloerection and hypotrichosis) observed after D-AMB treatment. No renal [blood urea nitrogen (BUN), creatinine] or hepatic (pyruvic and oxalacetic glutamic transaminases) biochemical abnormalities were found. The micronucleus assay showed no significant differences in both the micronucleus frequency and percentage of polychromatic erythrocytes for Nano-D-AMB, indicating the absence of genotoxicity and cytotoxic effects. CONCLUSIONS: The D-AMB-coated PLGA-DMSA nanoparticle showed antifungal efficacy, fewer undesirable effects and a favourable extended dosing interval. Nano-D-AMB comprises an AMB formulation able to lessen the number of drug administrations. Further studies would elucidate whether Nano-D-AMB would be useful to treat systemic fungal infections such as paracoccidioidomycosis, candidiasis, aspergillosis and cryptococcosis.
Assuntos
Anfotericina B/uso terapêutico , Ácido Desoxicólico/uso terapêutico , Ácido Láctico/uso terapêutico , Nanopartículas/uso terapêutico , Paracoccidioides/efeitos dos fármacos , Paracoccidioidomicose/tratamento farmacológico , Ácido Poliglicólico/uso terapêutico , Succímero/uso terapêutico , Anfotericina B/administração & dosagem , Anfotericina B/efeitos adversos , Animais , Peso Corporal , Medula Óssea/efeitos dos fármacos , Medula Óssea/patologia , Medula Óssea/fisiologia , Contagem de Colônia Microbiana , Ácido Desoxicólico/administração & dosagem , Ácido Desoxicólico/efeitos adversos , Combinação de Medicamentos , Feminino , Rim/efeitos dos fármacos , Rim/fisiologia , Ácido Láctico/administração & dosagem , Ácido Láctico/efeitos adversos , Fígado/efeitos dos fármacos , Fígado/microbiologia , Fígado/fisiologia , Pulmão/microbiologia , Pulmão/patologia , Camundongos , Camundongos Endogâmicos BALB C , Nanopartículas/administração & dosagem , Nanopartículas/efeitos adversos , Ácido Poliglicólico/administração & dosagem , Ácido Poliglicólico/efeitos adversos , Copolímero de Ácido Poliláctico e Ácido Poliglicólico , Succímero/administração & dosagem , Succímero/efeitos adversos , Resultado do TratamentoRESUMO
In this study a magnetic nanoemulsion (MNE) was developed from a mixture of two components, namely biodegradable surfactants and biocompatible citrate-coated cobalt ferrite-based magnetic fluid, for entrapment of Zn(II)-Phthalocyanine (ZnPc), the latter a classical photosensitizer (PS) species used in photodynamic therapy (PDT) procedures. The sample's stability was evaluated as a function of time using photocorrelation spectroscopy (PCS) for determination of the average hydrodynamic diameter, diameter dispersion and zeta potential. The ZnPc-loaded magneto nanoemulstion (ZnPc/MNE) formulation was evaluated in vitro assays to access the phototoxicity and the effect of application of AC magnetic fields (magnetohyperthermia damage) after incubation with J774-A1 macrophages cells. Darkness toxicity, phototoxicity and AC magnetic field exposures revealed an enhancement response for combined photodynamic and magnetohyperthermia (MHT) processes, indicating the presence of the synergic effect.