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Available evidence suggest that exposure to PM2.5 during pregnancy is associated with reduced cognitive function in offspring. This study aimed to investigate the effects of maternal exposure to PM2.5 on offspring cognitive function and to elucidate the underlying mechanisms. In this work, pregnant C57BL/6 female mice were exposed to concentrated ambient PM2.5 or filtered air from day 0.5 (=vaginal plug) to day 15.5 in the Shanghai Meteorological and Environmental Animal Exposure System, and offspring cerebellar tissues were collected on embryonic day 15.5, as well as postnatal days 0, 10 and 42. The mean PM2.5 concentrations exposed to the pregnant mice were 73.06 ± 4.90 µg/m3 and 11.15 ± 2.71 µg/m3 in the concentrated ambient PM2.5 and filtered air chambers, respectively. Maternal concentrated PM2.5 exposure was negatively correlated with offspring spatial memory significantly as assessed by the Morris water maze. Compared with the filtered air group, PM2.5-exposed offspring mice had reduced cerebellar microglia. Both RNA and protein levels of IL-8 and TNF-α were elevated in the concentrated ambient PM2.5 group. PM2.5 exposure increased the level of 8-OHG in miRNA of microglia and Purkinje cells in 6-week-old offspring. The level of prostaglandin F2α (8-iso-PGF2Aα) in the cerebellum was increased at different growing stages of offspring after gestational exposure of PM2.5. These results suggested that maternal air pollution exposure might cause inflammatory damage and oxidative stress to the cerebellum, contributing to reduced cognitive performance in mice offspring.
Assuntos
Poluentes Atmosféricos , Disfunção Cognitiva , Humanos , Gravidez , Feminino , Camundongos , Animais , Exposição Materna , Material Particulado , Doenças Neuroinflamatórias , Camundongos Endogâmicos C57BL , China , Estresse Oxidativo , CerebeloRESUMO
Haemorrhagic stroke represents a significant public health burden, yet our knowledge and ability to treat this type of stroke are lacking. Previously we showed that we can target ischaemic-stroke lesions by selective translocation of lipid nanoparticles through the site of blood-brain barrier (BBB) disruption. The data we presented in this study provide compelling evidence that haemorrhagic stroke in mice induces BBB injury that mimics key features of the human pathology and, more importantly, provides a gate for entry of lipid nanoparticles-based therapeutics selectively to the bleeding site. Methods: Haemorrhagic stroke was induced in mice by intra-striatal collagenase injection. lipid nanoparticles were injected intravenously at 3 h, 24 h & 48 h post-haemorrhagic stroke and accumulation in the brain studied using in-vivo optical imaging and histology. BBB integrity, brain water content and iron accumulation were characterised using dynamic contrast-enhanced MRI, quantitative T1 mapping, and gradient echo MRI. Results: Using in-vivo SPECT/CT imaging and optical imaging revealed biphasic lipid nanoparticles entry into the bleeding site, with an early phase of increased uptake at 3-24 h post-haemorrhagic stroke, followed by a second phase at 48-72 h. Lipid nanoparticles entry into the brain post-haemorrhage showed an identical entry pattern to the trans-BBB leakage rate (Ktrans [min-1]) of Gd-DOTA, a biomarker for BBB disruption, measured using dynamic contrast-enhanced MRI. Discussion: Our findings suggest that selective accumulation of liposomes into the lesion site is linked to a biphasic pattern of BBB hyper-permeability. This approach provides a unique opportunity to selectively and efficiently deliver therapeutic molecules across the BBB, an approach that has not been utilised for haemorrhagic stroke therapy and is not achievable using free small drug molecules.
Assuntos
Acidente Vascular Cerebral Hemorrágico , Acidente Vascular Cerebral , Animais , Barreira Hematoencefálica/patologia , Encéfalo/diagnóstico por imagem , Encéfalo/patologia , Humanos , Lipossomos , Imageamento por Ressonância Magnética/métodos , Camundongos , Nanopartículas , Acidente Vascular Cerebral/diagnóstico por imagem , Acidente Vascular Cerebral/patologiaRESUMO
Modulation of peripheral immune cells in the spleen plays a key role in many life-threatening conditions such as stroke. Immune cell changes can lead to the excessive release of pro-inflammatory cytokines into the circulation and preferential loss of innate immune cells which can further exacerbate tissue damage and predispose patients to infectious complications. Reversing these processes represents an attractive treatment strategy and has shown to have beneficial effects in animal models of ischemic stroke, sepsis, traumatic brain injury (TBI) as well as myocardial infraction (MI). However, systemic interventions are often challenging to deliver due to the non-selective broad range of action of many treatments. More selective targeted treatment approaches are therefore desirable. The spleen is considered a natural filtration site for many nanomaterials due to the spontaneous tendency of this organ to filter blood-borne molecules. This selective targeting of nanomaterials to the spleen therefore offers considerable potential in the management of many conditions affected by peripheral inflammation. In this review, we will explore the key nanomaterials-related parameters that mediate splenic targeting and how these could influence the actual localization and function of nanomaterials once in the spleen. We aim to emphasize the potential of utilising nanomaterials as selective tools for peripheral immunomodulation to accelerate clinical translation.
Assuntos
Baço , Acidente Vascular Cerebral , Animais , Citocinas , Imunomodulação , Nanomedicina , Acidente Vascular Cerebral/terapiaRESUMO
Despite considerable efforts, the properties that drive the cytotoxicity of engineered nanomaterials (ENMs) remain poorly understood. Here, the authors inverstigate a panel of 31 ENMs with different core chemistries and a variety of surface modifications using conventional in vitro assays coupled with omics-based approaches. Cytotoxicity screening and multiplex-based cytokine profiling reveals a good concordance between primary human monocyte-derived macrophages and the human monocyte-like cell line THP-1. Proteomics analysis following a low-dose exposure of cells suggests a nonspecific stress response to ENMs, while microarray-based profiling reveals significant changes in gene expression as a function of both surface modification and core chemistry. Pathway analysis highlights that the ENMs with cationic surfaces that are shown to elicit cytotoxicity downregulated DNA replication and cell cycle responses, while inflammatory responses are upregulated. These findings are validated using cell-based assays. Notably, certain small, PEGylated ENMs are found to be noncytotoxic yet they induce transcriptional responses reminiscent of viruses. In sum, using a multiparametric approach, it is shown that surface chemistry is a key determinant of cellular responses to ENMs. The data also reveal that cytotoxicity, determined by conventional in vitro assays, does not necessarily correlate with transcriptional effects of ENMs.
RESUMO
The development of effective therapies for stroke continues to face repeated translational failures. Brain endothelial cells form paracellular and transcellular barriers to many blood-borne therapies, and the development of efficient delivery strategies is highly warranted. Here, in a mouse model of stroke, we show selective recruitment of clinically used liposomes into the ischemic brain that correlates with biphasic blood brain barrier (BBB) breakdown. Intravenous administration of liposomes into mice exposed to transient middle cerebral artery occlusion took place at early (0.5 and 4 h) and delayed (24 and 48 h) time points, covering different phases of BBB disruption after stroke. Using a combination of in vivo real-time imaging and histological analysis we show that selective liposomal brain accumulation coincides with biphasic enhancement in transcellular transport followed by a delayed impairment to the paracellular barrier. This process precedes neurological damage in the acute phase and maintains long-term liposomal colocalization within the neurovascular unit, which could have great potential for neuroprotection. Levels of liposomal uptake by glial cells are similarly selectively enhanced in the ischemic region late after experimental stroke (2-3 days), highlighting their potential for blocking delayed inflammatory responses or shifting the polarization of microglia/macrophages toward brain repair. These findings demonstrate the capability of liposomes to maximize selective translocation into the brain after stroke and identify two windows for therapeutic manipulation. This emphasizes the benefits of selective drug delivery for efficient tailoring of stroke treatments.
Assuntos
Barreira Hematoencefálica/metabolismo , Lipossomos , Acidente Vascular Cerebral/metabolismo , Animais , Modelos Animais de Doenças , Sistemas de Liberação de Medicamentos , Infarto da Artéria Cerebral Média/metabolismo , Lipossomos/química , Lipossomos/farmacocinética , Lipossomos/farmacologia , Masculino , Camundongos , Camundongos Endogâmicos C57BL , Transcitose/efeitos dos fármacosRESUMO
Hybrids composed of liposomes (L) and metallic nanoparticles (NPs) hold great potential for imaging and drug delivery purposes. However, the efficient incorporation of metallic NPs into liposomes using conventional methodologies has so far proved to be challenging. In this study, we report the fabrication of hybrids of liposomes and hydrophobic gold NPs of size 2-4 nm (Au) using a microfluidic-assisted self-assembly process. The incorporation of increasing amounts of AuNPs into liposomes was examined using microfluidics and compared to L-AuNP hybrids prepared by the reverse-phase evaporation method. Our microfluidics strategy produced L-AuNP hybrids with a homogeneous size distribution, a smaller polydispersity index, and a threefold increase in loading efficiency when compared to those hybrids prepared using the reverse-phase method of production. Quantification of the loading efficiency was determined by ultraviolet spectroscopy, inductively coupled plasma mass spectroscopy, and centrifugal field flow fractionation, and qualitative validation was confirmed by transmission electron microscopy. The higher loading of gold NPs into the liposomes achieved using microfluidics produced a slightly thicker and more rigid bilayer as determined with small-angle neutron scattering. These observations were confirmed using fluorescent anisotropy and atomic force microscopy. Structural characterization of the liposomal-NP hybrids with cryo-electron microscopy revealed the coexistence of membrane-embedded and interdigitated NP-rich domains, suggesting AuNP incorporation through hydrophobic interactions. The microfluidic technique that we describe in this study allows for the automated production of monodisperse liposomal-NP hybrids with high loading capacity, highlighting the utility of microfluidics to improve the payload of metallic NPs within liposomes, thereby enhancing their application for imaging and drug delivery.
Assuntos
Ouro/química , Dispositivos Lab-On-A-Chip , Lipossomos/química , Nanopartículas Metálicas/química , Técnicas Analíticas MicrofluídicasRESUMO
The self-assembled layered adsorption of proteins onto nanoparticle (NP) surfaces, once in contact with biological fluids, is termed the "protein corona" and it is gradually seen as a determinant factor for the overall biological behavior of NPs. Here, the previously unreported in vivo protein corona formed in human systemic circulation is described. The human-derived protein corona formed onto PEGylated doxorubicin-encapsulated liposomes (Caelyx) is thoroughly characterized following the recovery of liposomes from the blood circulation of ovarian carcinoma patients. In agreement with previous investigations in mice, the in vivo corona is found to be molecularly richer in comparison to its counterpart ex vivo corona. The intravenously infused liposomes are able to scavenge the blood pool and surface-capture low-molecular-weight, low-abundance plasma proteins that cannot be detected by conventional plasma proteomic analysis. This study describes the previously elusive or postulated formation of protein corona around nanoparticles in vivo in humans and illustrates that it can potentially be used as a novel tool to analyze the blood circulation proteome.
Assuntos
Lipossomos/química , Polietilenoglicóis/química , Coroa de Proteína/química , Adsorção , Doxorrubicina/química , Humanos , Nanopartículas/químicaRESUMO
The prominent discrepancy between the significant investment towards plasma biomarker discovery and the very low number of biomarkers currently in clinical use stresses the need for discovery technologies. The discovery of protein biomarkers present in human blood by proteomics is tremendously challenging, owing to the large dynamic concentration range of blood proteins. Here, we describe the use of blood-circulating lipid-based nanoparticles (NPs) as a scavenging tool to comprehensively analyse the blood proteome. We aimed to exploit the spontaneous interaction of NPs with plasma proteins once injected in the bloodstream, known as 'protein corona', in order to facilitate the capture of tumor-specific molecules. We employed two different tumor models, a subcutaneous melanoma model (B16-F10) and human lung carcinoma xenograft model (A549) and comprehensively compared by mass spectrometry the in vivo protein coronas formed onto clinically used liposomes, intravenously administered in healthy and tumor-bearing mice. The results obtained demonstrated that blood-circulating liposomes surface-capture and amplify a wide range of different proteins including low molecular weight (MW) and low abundant tumor specific proteins (intracellular products of tissue leakage) that could not be detected by plasma analysis, performed in comparison. Most strikingly, the NP (liposomal) corona formed in the xenograft model was found to consist of murine host response proteins, as well as human proteins released from the inoculated and growing human cancer cells. This study offers direct evidence that the in vivo NP protein corona could be deemed as a valuable tool to enrich the blood proteomic analysis and to allow the discovery of potential biomarkers in experimental disease models.
Assuntos
Biomarcadores Tumorais/sangue , Proteínas Sanguíneas/análise , Lipossomos/metabolismo , Neoplasias Pulmonares/sangue , Melanoma Experimental/sangue , Coroa de Proteína/análise , Células A549 , Animais , Biomarcadores Tumorais/metabolismo , Proteínas Sanguíneas/metabolismo , Feminino , Humanos , Lipossomos/sangue , Neoplasias Pulmonares/metabolismo , Melanoma Experimental/metabolismo , Camundongos Endogâmicos C57BL , Nanopartículas/metabolismo , Coroa de Proteína/metabolismoRESUMO
With the extensive use of silver nanoparticles (AgNPs) in various consumer products their potential toxicity is of great concern especially for highly sensitive population groups such as pregnant women and even the developing fetus. To understand if AgNPs are taken up and cross the human placenta, we studied their translocation and accumulation in the human ex vivo placenta perfusion model by single particle ICP-MS (spICP-MS). The impact of different surface modifications on placental transfer was assessed by AgNPs with two different modifications: polyethylene glycol (AgPEG NPs) and sodium carboxylate (AgCOONa NPs). AgNPs and ionic Ag were detected in the fetal circulation in low but not negligible amounts. Slightly higher Ag translocation across the placental barrier for perfusion with AgPEG NPs and higher AgNP accumulation in placental tissue for perfusion with AgCOONa NPs were observed. Since these AgNPs are soluble in water, we tried to distinguish between the translocation of dissolved and particulate Ag. Perfusion with AgNO3 revealed the formation of Ag containing NPs in both circulations over time, of which the amount and their size in the fetal circulation were comparable to those from perfusion experiments with both AgNP types. Although we were not able to clarify whether intact AgNPs and/or Ag precipitates from dissolved Ag cross the placental barrier, our study highlights that uptake of Ag ions and/or dissolution of AgNPs in the tissue followed by re-precipitation in the fetal circulation needs to be considered as an important pathway in studies of AgNP translocation across biological barriers.
Assuntos
Nanopartículas Metálicas , Placenta/metabolismo , Prata/farmacocinética , Feminino , Humanos , Íons , Tamanho da Partícula , Gravidez , Análise EspectralRESUMO
Thermally triggered drug release from temperature-sensitive liposomes (TSL) holds great promise for cancer therapy. Different types of TSL have been designed recently for heat triggered drug release inside tumor blood vessels or after accumulation into the tumor interstitium. However, justification of drug release profiles is for far mainly based on in vitro release data. While these methods could be good enough to give early indication about the thermal sensitivity of TSL, they are still far from being optimum. This is because these methods do not take into consideration the actual adsorption of proteins onto the surface of TSL after their in vivo administration, also known as "protein corona" and the influence this could have on drug release. Therefore, in this study we compared thermal triggered drug release profile of two different types of doxorubicin encapsulated TSL; namely the lysolipid-containing TSL (LTSL) and traditional TSL (TTSL) after their in vivo recovery from the blood circulation of CD-1 mice. Ex vivo release profile at 42⯰C was then tested either in the presence of full plasma or after removal of unbound plasma proteins (i.e. protein corona coated TSL). Our data showed that the influence of the environment on drug release profile was very much dependent on the type of TSL. LTSL release profile was consistently characterized by ultrafast drug release independent on the conditions tested. On the contrary, TTSL release profile changed significantly. Doxorubicin release from in vivo recovered TTSL was slow and incomplete in the presence of unbound plasma proteins, whereas very rapid drug release was detected from in vivo recovered and purified protein corona-coated TTSL in the absence of unbound proteins. Using mass spectrometry and quantification of protein adsorption, we confirmed that this discrepancy is due to the changes in protein adsorption onto TTSL when heated in the presence of unbound proteins leading to reduction in drug release. In summary this study showed that the formation of the in vivo corona on TSL will have a dramatic impact on their release profile and is dependent on both their lipid composition and the protein content of the environment in which drug release is triggered.
Assuntos
Antibióticos Antineoplásicos/química , Doxorrubicina/química , Coroa de Proteína , Antibióticos Antineoplásicos/administração & dosagem , Proteínas Sanguíneas/química , Doxorrubicina/administração & dosagem , Liberação Controlada de Fármacos , Lipossomos , TemperaturaRESUMO
INTRODUCTION: The development of therapeutics for central nervous system (CNS) disorders is still considered a challenging area in drug development due to insufficient translocation through the blood-brain barrier (BBB). Under normal conditions, BBB restrict the penetration of more than 98% of blood-borne molecules including drugs to the CNS. However, recent research findings have proven that the nature of the BBB is altered in several neurological conditions. This complexity encourages revisiting drug delivery strategies to the CNS as this can give a wide range of opportunities for CNS drug development. AREAS COVERED: This review focuses on nanotechnology-based drug delivery platforms designed for selective recruitment into the lesioned brain by taking advantages of BBB disruption that is associated with certain neurological conditions. EXPERT OPINION: Current CNS therapeutic strategies do not fully address the pathophysiological adaptation of BBB in their design. The lack of selective delivery to the brain lesions has been the culprit behind the failure of many CNS therapeutics. This highlighted the need for smart designs of advanced drug delivery systems that take advantage of BBB structural changes in CNS diseases. Recently, promising examples have been reported in this area, however, more work is still required beyond the preclinical testing.
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Barreira Hematoencefálica/efeitos dos fármacos , Lesões Encefálicas/tratamento farmacológico , Fármacos do Sistema Nervoso Central/administração & dosagem , Sistemas de Liberação de Medicamentos/métodos , Animais , Transporte Biológico , Humanos , Nanotecnologia/métodosRESUMO
The engineering of responsive multifunctional delivery systems that combine therapeutic and diagnostic (theranostic) capabilities holds great promise and interest. We describe the design of thermosensitive liposome-nanoparticle (NP) hybrids that can modulate drug release in response to external heating stimulus. These hybrid systems were successfully engineered by the incorporation of gold, silver, and iron oxide NPs into the lipid bilayer of lysolipid-containing thermosensitive liposomes (LTSL). Structural characterization of LTSL-NP hybrids using cryo-EM and AFM revealed the incorporation of metallic NPs into the lipid membranes without compromising doxorubicin loading and retention capability. The presence of metallic NPs in the lipid bilayer reinforced bilayer retention and offered a nanoparticle concentration-dependent modulation of drug release in response to external heating. In conclusion, LTSL-NP hybrids represent a promising versatile platform based on LTSL liposomes that could further utilize the properties of the embedded NPs for multifunctional theranostic applications.
Assuntos
Doxorrubicina/química , Lipossomos/química , Nanopartículas Metálicas/química , Liberação Controlada de Fármacos/efeitos dos fármacos , Ouro/química , Temperatura Alta , Ferro/química , Bicamadas Lipídicas/química , Prata/químicaRESUMO
Nanoparticles (NPs) are instantly modified once injected in the bloodstream because of their interaction with the blood components. The spontaneous coating of NPs by proteins, once in contact with biological fluids, has been termed the 'protein corona' and it is considered to be a determinant factor for the pharmacological, toxicological and therapeutic profile of NPs. Protein exposure time is thought to greatly influence the composition of protein corona, however the dynamics of protein interactions under realistic, in vivo conditions remain unexplored. The aim of this study was to quantitatively and qualitatively investigate the time evolution of in vivo protein corona, formed onto blood circulating, clinically used, PEGylated liposomal doxorubicin. Protein adsorption profiles were determined 10 min, 1 h and 3 h post-injection of liposomes into CD-1 mice. The results demonstrated that a complex protein corona was formed as early as 10 min post-injection. Even though the total amount of protein adsorbed did not significantly change over time, the fluctuation of protein abundances observed indicated highly dynamic protein binding kinetics.
Assuntos
Circulação Sanguínea , Doxorrubicina/análogos & derivados , Nanopartículas/metabolismo , Coroa de Proteína/metabolismo , Adsorção , Animais , Disponibilidade Biológica , Doxorrubicina/sangue , Doxorrubicina/farmacocinética , Feminino , Camundongos , Nanopartículas/química , Polietilenoglicóis/farmacocinética , Coroa de Proteína/química , Fatores de Tempo , Distribuição TecidualRESUMO
In this Review, we attempt to offer a thorough description of all of the chemical components and the rationale behind the design of temperature-sensitive vesicle systems, as well as the critical pharmacological parameters that need to be combined to achieve their successful clinical translation. The focus of this Review will be predominantly on the design principles around the construction of temperature-sensitive liposomes (TSL) and their use in combination with external local hyperthermia to achieve heat-triggered drug release. The emphasis lies on the chemical components synthesized and incorporated in the design and engineering of TSL. We conclude that the development of TSL with ultrafast drug release capabilities needs to progress in parallel with vesicle pharmacokinetic profiling, imaging, and monitoring capacity and technologies for accurate temperature elevation and control. The development of heat-triggered liposome systems offer the greatest opportunity for clinical translation of the next generation, nanoscale "smart" vesicle systems of enhanced functionality, following from the successful legacy and rich clinical history from multiple earlier liposome technologies.
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Portadores de Fármacos/química , Animais , Antineoplásicos/administração & dosagem , Meios de Contraste/administração & dosagem , Doxorrubicina/administração & dosagem , Glicerofosfolipídeos/química , Humanos , Hipertermia Induzida/métodos , Lipossomos/química , Nanopartículas Metálicas/química , Peptídeos/química , Transição de Fase , TemperaturaRESUMO
The adsorption of proteins and their layering onto nanoparticle surfaces has been called the "protein corona". This dynamic process of protein adsorption has been extensively studied following in vitro incubation of many different nanoparticles with plasma proteins. However, the formation of protein corona under dynamic, in vivo conditions remains largely unexplored. Extrapolation of in vitro formed protein coronas to predict the fate and possible toxicological burden from nanoparticles in vivo is of great interest. However, complete lack of such direct comparisons for clinically used nanoparticles makes the study of in vitro and in vivo formed protein coronas of great importance. Our aim was to study the in vivo protein corona formed onto intravenously injected, clinically used liposomes, based on the composition of the PEGylated liposomal formulation that constitutes the anticancer agent Doxil. The formation of in vivo protein corona was determined after the recovery of the liposomes from the blood circulation of CD-1 mice 10 min postinjection. In comparison, in vitro protein corona was formed by the incubation of liposomes in CD-1 mouse plasma. In vivo and in vitro formed protein coronas were compared in terms of morphology, composition and cellular internalization. The protein coronas on bare (non-PEGylated) and monoclonal antibody (IgG) targeted liposomes of the same lipid composition were also comparatively investigated. A network of linear fibrillary structures constituted the in vitro formed protein corona, whereas the in vivo corona had a different morphology but did not appear to coat the liposome surface entirely. Even though the total amount of protein attached on circulating liposomes correlated with that observed from in vitro incubations, the variety of molecular species in the in vivo corona were considerably wider. Both in vitro and in vivo formed protein coronas were found to significantly reduce receptor binding and cellular internalization of antibody-conjugated liposomes; however, the in vivo corona formation did not lead to complete ablation of their targeting capability.
Assuntos
Proteínas Sanguíneas/química , Imunoglobulina G/química , Bicamadas Lipídicas/química , Lipossomos/química , Nanopartículas/química , Adsorção , Animais , Antibióticos Antineoplásicos/química , Colesterol/química , Doxorrubicina/análogos & derivados , Doxorrubicina/química , Humanos , Imunoconjugados/química , Injeções Intravenosas , Lipossomos/ultraestrutura , Células MCF-7 , Camundongos , Nanopartículas/ultraestrutura , Fosfatidilcolinas/química , Fosfatidiletanolaminas/química , Polietilenoglicóis/químicaRESUMO
Temperature-sensitive vesicles designed by inclusion of leucine zipper peptides within a lipid bilayer (Lp-Peptide hybrids) encapsulating Doxorubicin (DOX) have been reported. Intravenous administration of these constructs prolonged blood circulation kinetics and increased tumor accumulation in vivo with local mild hyperthermia. In this study, the biological activity of the DOX-loaded Lp-Peptide hybrid vesicles was further investigated at the cellular level and in vivo compared to lysolipid-containing temperature-sensitive liposomes (LTSL) and traditional temperature-sensitive liposomes. Lp-Peptide vesicles were not toxic to cell cultures at 37°C, while effective cancer cell toxicity was observed after 1 hr of heating at 42°C. The activity of Lp-Peptide vesicles in vivo was studied using two different heating protocols to obtain tumor intravascular or interstitial drug release. Lp-Peptide vesicle treatment allowing intravascular DOX release showed equally effective tumor growth retardation and survival to that of LTSL treatment. The Lp-Peptide vesicles also offered therapeutic responses using the alternative heating protocol to maximise drug release within the tumor interstitium. Matching the drug release kinetics of temperature-sensitive vesicles with the heating protocol applied is considered the most critical factor to determine therapeutic efficacy in the clinical translation of such modalities.
Assuntos
Doxorrubicina/administração & dosagem , Lipossomos , Neoplasias/tratamento farmacológico , Neoplasias/patologia , Peptídeos , Animais , Linhagem Celular Tumoral , Modelos Animais de Doenças , Doxorrubicina/farmacocinética , Doxorrubicina/toxicidade , Humanos , Zíper de Leucina , Lipossomos/química , Melanoma Experimental , Camundongos , Neoplasias/mortalidade , Peptídeos/química , Temperatura , Carga Tumoral/efeitos dos fármacos , Ensaios Antitumorais Modelo de XenoenxertoRESUMO
Indocyanine green (ICG) is an FDA-approved, strongly photo-absorbent/fluorescent probe that has been incorporated into a clinically-relevant PEGylated liposome as a flexible optoacoustic contrast agent platform. This study describes the engineering of targeted PEGylated liposome-ICG using the anti-MUC-1 "humanized" monoclonal antibody (MoAb) hCTM01 as a tumour-specific theranostic system. We aimed to visualise non-invasively the tumour accumulation of these MoAb-targeted liposomes over time in tumour-bearing mice using multispectral optoacoustic tomography (MSOT). Preferential accumulation of targeted PEGylated liposome-ICG was studied after intravenous administration in comparison to non-targeted PEGylated liposome-ICG using both fast growing (4T1) and slow growing (HT-29) MUC-1 positive tumour models. Monitoring liposomal ICG in the tumour showed that both targeted and non-targeted liposome-ICG formulations preferentially accumulated into the tumour models studied. Rapid accumulation was observed for targeted liposomes at early time points mainly in the periphery of the tumour volume suggesting binding to available MUC-1 receptors. In contrast, non-targeted PEGylated liposomes showed accumulation at the centre of the tumour at later time points. In an attempt to take this a step further, we successfully encapsulated the anticancer drug, doxorubicin (DOX) into both targeted and non-targeted PEGylated liposome-ICG. The engineering of DOX-loaded targeted ICG liposome systems present a novel platform for combined tumour-specific therapy and diagnosis. This can open new possibilities in the design of advanced image-guided cancer therapeutics.
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Anticorpos Monoclonais/imunologia , Antineoplásicos/imunologia , Antineoplásicos/farmacocinética , Doxorrubicina/administração & dosagem , Sistemas de Liberação de Medicamentos/métodos , Verde de Indocianina/análise , Polietilenoglicóis/química , Animais , Anticorpos Monoclonais/administração & dosagem , Antineoplásicos/administração & dosagem , Antineoplásicos/uso terapêutico , Linhagem Celular Tumoral , Meios de Contraste/administração & dosagem , Meios de Contraste/análise , Meios de Contraste/química , Doxorrubicina/farmacocinética , Doxorrubicina/uso terapêutico , Feminino , Fluorescência , Humanos , Verde de Indocianina/administração & dosagem , Verde de Indocianina/química , Lipossomos , Camundongos , Neoplasias/imunologia , Neoplasias/metabolismo , Técnicas Fotoacústicas , Polietilenoglicóis/administração & dosagemRESUMO
The development of actively targeted, responsive delivery vectors holds great promise for cancer therapy. Here, we investigated whether enhanced therapeutic activity of temperature sensitive liposomes (TSL) could be obtained by mild hyperthermia-triggered release of the chemotherapeutic drug doxorubicin (DOX) after hCTMO1 monoclonal antibody (anti-MUC-1) binding and uptake into cancer cells. We showed that traditional TSL (TTSL) liposome systems maintained their physicochemical and thermal properties after conjugation to hCTMO1 full IgG. Receptor-mediated cellular uptake and cytotoxic efficacy of antibody-targeted TTSL (TTSL-Ab) were investigated using 2D and 3D cell culture models. Significant enhancement in cellular uptake and cytotoxic activity after 1h of heating at 42 °C was observed for TTSL-Ab compared to non-targeted liposomes in MUC-1 over-expressing breast cancer cells (MDA-MB-435). Tissue distribution and in vivo therapeutic activity were studied using different heating protocols to explore the effect of mild hyperthermia on the tumor accumulation of targeted TTSL and their therapeutic effect. Application of local, mild hyperthermia (42°C) significantly increased the tumor accumulation of targeted TSL compared to non-targeted liposomes, associated with a moderate improvement in therapeutic activity and survival.
Assuntos
Anticorpos Monoclonais/uso terapêutico , Antineoplásicos/uso terapêutico , Hipertermia Induzida , Lipossomos/química , Neoplasias Experimentais/terapia , Administração Intravenosa , Animais , Antibióticos Antineoplásicos/administração & dosagem , Antibióticos Antineoplásicos/farmacologia , Anticorpos Monoclonais/farmacocinética , Antineoplásicos/farmacocinética , Sobrevivência Celular/efeitos dos fármacos , Doxorrubicina/administração & dosagem , Doxorrubicina/farmacologia , Feminino , Humanos , Camundongos , Camundongos Nus , Mucina-1/efeitos dos fármacos , Neoplasias Experimentais/tratamento farmacológico , Análise de Sobrevida , Temperatura , Distribuição TecidualRESUMO
The present study describes leucine zipper peptide-lipid hybrid nanoscale vesicles engineered by self-assembled anchoring of the amphiphilic peptide within the lipid bilayer. These hybrid vesicles aim to combine the advantages of traditional temperature-sensitive liposomes (TSL) with the dissociative, unfolding properties of a temperature-sensitive peptide to optimize drug release under mild hyperthermia, while improving in vivo drug retention. The secondary structure of the peptide and its thermal responsiveness after anchoring onto liposomes were studied with circular dichroism. In addition, the lipid-peptide vesicles (Lp-peptide) showed a reduction in bilayer fluidity at the inner core, as observed with DPH anisotropy studies, while the opposite effect was observed with an ANS probe, indicating peptide interactions with both the headgroup region and the hydrophobic core. A model drug molecule, doxorubicin, was successfully encapsulated in the Lp-peptide vesicles at higher than 90% efficiency following the remote loading, pH-gradient methodology. The release of doxorubicin from Lp-peptide hybrids in vitro indicated superior serum stability at physiological temperatures compared to lysolipid-containing temperature-sensitive liposomes (LTSL) without affecting the overall thermo-responsive nature of the vesicles at 42 °C. A similar stabilizing effect was observed in vivo after intravenous administration of the Lp-peptide vesicles by measuring (14)C-doxorubicin blood kinetics that also led to increased tumor accumulation after 24 h. We conclude that Lp-peptide hybrid vesicles present a promising new class of TSL that can offer previously unexplored opportunities for the development of clinically relevant mild hyperthermia-triggered therapeutic modalities.
Assuntos
Preparações de Ação Retardada/química , Doxorrubicina/administração & dosagem , Lipossomos/química , Melanoma/tratamento farmacológico , Nanocápsulas/administração & dosagem , Nanocápsulas/química , Peptídeos/química , Animais , Antineoplásicos/administração & dosagem , Antineoplásicos/química , Linhagem Celular Tumoral , Difusão , Doxorrubicina/química , Temperatura Alta , Melanoma/patologia , Camundongos , Camundongos Endogâmicos C57BL , Nanocápsulas/ultraestrutura , Tamanho da Partícula , Resultado do TratamentoRESUMO
Drug-loaded temperature-sensitive liposomes (TSL) in combination with hyperthermia (HT) have attracted considerable attention for cancer treatment. Different TSL systems have been designed with wide variations in their temperature sensitivity and drug release profile. Low temperature-sensitive liposomes (LTSL) with the capacity for ultrafast drug release, traditional temperature-sensitive (TTSL) with intermediate drug release properties and non-temperature-sensitive liposomes (NTSL) (no drug release) were dual-labeled with (3)H-cholesteryl hexadecyl ether ((3)H-CHE) lipid and loaded with (14)C-doxorubicin ((14)C-Dox). Their blood profile, serum stability, tissue distribution and tumor localization (B16F10 melanoma) were studied after intravenous administration and mild HT treatment. LTSL showed higher affinity for the liver compared to TTSL and NTSL which were uptaken mainly by spleen. Under normal conditions (no HT) Dox leakage from liposomes was expected, higher for LTSL, less for TTSL and minimal for NTSL. Localized HT did not affect the overall blood circulation or organ accumulation for all TSL studied. Since LTSL showed ultrafast Dox release kinetics at 42 °C, the highest drug accumulation in tumors was observed using this system immediately after HT, however decreased significantly after 24 h. In contrast, TTSL and NTSL showed 2-3 fold increase in both liposome and Dox levels that indicated enhanced tumor extravasation of intact Dox-loaded liposomes during the 60 min HT applications. More interestingly, high levels of drug tumor accumulation were achieved 24 h post-HT. This study offers further understanding on how the mechanisms of drug release from temperature-sensitive liposomes affect their pharmacological profile under mild hyperthermia.