RESUMEN
The COVID-19 pandemic involving SARS-CoV-2 has raised interest in using antimicrobial lipid formulations to inhibit viral entry into their host cells or to inactivate them. Lipids are a part of the innate defense mechanism against pathogens. Here, we evaluated the use of nano-monocaprin (NMC) in inhibiting enveloped (phi6) and unenveloped (MS2) bacteriophages. NMC was prepared using the sonochemistry technique. Size and morphology analysis revealed the formation of ~ 8.4 ± 0.2-nm NMC as measured by dynamic light scattering. We compared the antiviral activity of NMC with molecular monocaprin (MMC) at 0.5 mM and 2 mM concentrations against phi6, which we used as a surrogate for SARS-CoV-2. The synthesized NMC exhibited 50% higher antiviral activity against phi6 than MMC at pH 7 using plaque assay. NMC inactivated phi6 stronger at pH 4 than at pH 7. To determine if NMC is toxic to mammalian cells, we used MTS assay to assess its IC50 for HPDE and HeLa cell lines, which were ~ 203 and 221 µM, respectively. NMC may be used for prophylactic application either as a drop or spray since many viruses enter the human body through the mucosal lining of the nose, eyes, and lungs.(AU)
Asunto(s)
Humanos , Coronavirus Relacionado al Síndrome Respiratorio Agudo Severo , Pandemias , Bacteriófagos , Lípidos , Infecciones por Coronavirus/epidemiología , Antiinfecciosos , Microbiología , Técnicas MicrobiológicasRESUMEN
Photodynamic therapy (PDT) is a promising anticancer noninvasive technique that relies on the generation of reactive oxygen species (ROS). Unfortunately, PDT still has many limitations, including the resistance developed by cancer cells to the cytotoxic effect of ROS. Autophagy, which is a stress response mechanism, has been reported as a cellular pathway that reduces cell death following PDT. Recent studies have demonstrated that PDT in combination with other therapies can eliminate anticancer resistance. However, combination therapy is usually challenged by the differences in the pharmacokinetics of the drugs. Nanomaterials are excellent delivery systems for the efficient codelivery of two or more therapeutic agents. In this work, we report on the use of polysilsesquioxane (PSilQ) nanoparticles for the codelivery of chlorin-e6 (Ce6) and an autophagy inhibitor for early- or late-stage autophagy. Our results, obtained from a reactive oxygen species (ROS) generation assay and apoptosis and autophagy flux analyses, demonstrate that the reduced autophagy flux mediated by the combination approach afforded an increase in the phototherapeutic efficacy of Ce6-PSilQ nanoparticles. We envision that the promising results in the use of multimodal Ce6-PSilQ material as a codelivery system against cancer pave the way for its future application with other clinically relevant combinations.
RESUMEN
The COVID-19 pandemic involving SARS-CoV-2 has raised interest in using antimicrobial lipid formulations to inhibit viral entry into their host cells or to inactivate them. Lipids are a part of the innate defense mechanism against pathogens. Here, we evaluated the use of nano-monocaprin (NMC) in inhibiting enveloped (phi6) and unenveloped (MS2) bacteriophages. NMC was prepared using the sonochemistry technique. Size and morphology analysis revealed the formation of ~ 8.4 ± 0.2-nm NMC as measured by dynamic light scattering. We compared the antiviral activity of NMC with molecular monocaprin (MMC) at 0.5 mM and 2 mM concentrations against phi6, which we used as a surrogate for SARS-CoV-2. The synthesized NMC exhibited 50% higher antiviral activity against phi6 than MMC at pH 7 using plaque assay. NMC inactivated phi6 stronger at pH 4 than at pH 7. To determine if NMC is toxic to mammalian cells, we used MTS assay to assess its IC50 for HPDE and HeLa cell lines, which were ~ 203 and 221 µM, respectively. NMC may be used for prophylactic application either as a drop or spray since many viruses enter the human body through the mucosal lining of the nose, eyes, and lungs.
Asunto(s)
Antivirales , COVID-19 , Animales , Humanos , Antivirales/farmacología , SARS-CoV-2 , Células HeLa , Pandemias , MamíferosRESUMEN
The use of nanoparticle-based materials to improve the efficacy of photodynamic therapy (PDT) to treat cancer has been a burgeoning field of research in recent years. Polysilsesquioxane (PSilQ) nanoparticles with remarkable features, such as high loading of photosensitizers, biodegradability, surface tunability, and biocompatibility, have been used for the treatment of cancer in vitro and in vivo using PDT. The PSilQ platform typically shows an enhanced PDT performance following a cell death mechanism similar to the parent photosensitizer. Ferroptosis is a new cell death mechanism recently associated with PDT that has not been investigated using PSilQ nanoparticles. Herein, we synthesized a protoporphyrin IX (PpIX)-based PSilQ platform (PpIX-PSilQ NPs) to study the cell death pathways, with special focus on ferroptosis, during PDT in vitro. Our data obtained from different assays that analyzed Annexin V binding, glutathione peroxidase activity, and lipid peroxidation demonstrate that the cell death in PDT using PpIX-PSilQ NPs is regulated by apoptosis and ferroptosis. These results can provide alternative approaches in designing PDT strategies to enhance therapeutic response in conditions stymied by apoptosis resistance.
RESUMEN
Programmable nucleic acid nanoparticles (NANPs) with precisely controlled functional compositions can regulate the conditional activation of various biological pathways and responses in human cells. However, the intracellular delivery of NANPs alone is hindered by their susceptibility to nuclease activity and inefficient crossing of biological membranes. In this work, we optimized the internalization and therapeutic performance of several representative NANPs delivered with mesoporous silica nanoparticles (MSNPs) tailored for efficient electrostatic association with NANPs. We compared the immunostimulatory properties of different NA-MS-NP complexes formed with globular, planar, and fibrous NANPs and demonstrated the maximum immunostimulation for globular NANPs. As a proof of concept, we assessed the specific gene silencing by NA-MS-NP complexes functionalized with siRNA targeting green fluorescent protein expressed in triple-negative human breast cancer cells. We showed that the fibrous NANPs have the highest silencing efficiency when compared to globular or planar counterparts. Finally, we confirmed the multimodal ability of MSNPs to co-deliver a chemotherapy drug, doxorubicin, and NANPs targeting apoptosis regulator gene BCL2 in triple-negative breast cancer and melanoma cell lines. Overall, the combination of NANPs and MSNPs may become a new promising approach to efficiently treat cancer and other diseases via the simultaneous targeting of various pathways.
Asunto(s)
Portadores de Fármacos/química , Nanopartículas/química , Ácidos Nucleicos/química , Dióxido de Silicio/química , Línea Celular Tumoral , Supervivencia Celular/efectos de los fármacos , Doxorrubicina/química , Doxorrubicina/farmacología , Humanos , Porosidad , Proteínas Proto-Oncogénicas c-bcl-2/antagonistas & inhibidores , Proteínas Proto-Oncogénicas c-bcl-2/genética , Proteínas Proto-Oncogénicas c-bcl-2/metabolismo , Interferencia de ARN , ARN Interferente Pequeño/química , ARN Interferente Pequeño/metabolismoRESUMEN
Chronic liver dysfunction often begins with hepatic fibrosis. A pivotal event in the progression of liver fibrosis and cirrhosis is hepatic stellate cell (HSC) activation and secretion of extracellular matrix proteins, including tenascin-C (TnC). TnC is often chosen as a therapeutic target for treatment of liver disease. TnC is minimally detected in healthy tissue, but is transiently expressed during tissue injury, and plays a critical role in fibrogenesis and tumorigenesis. siRNA therapy is a promising alternative to knock-down proteins relevant for fibrosis therapy. This study describes the application of a functionalized mesoporous silica nanoparticles (MSNs) for the efficient transport and delivery of siTnC in HSCs. Silencing experiments in HSCs demonstrate the effective reduction of TnC mRNA and protein levels. In addition, attenuation of TnC expression due to the cellular uptake and release of siTnC from MSNs resulted in decreases of inflammatory cytokine levels and hepatocyte migration. We envision this siTnC-MSN platform as a promising alternative to evaluate siRNA therapy of chronic liver disease in preclinical trials.
Asunto(s)
Silenciador del Gen , Células Estrelladas Hepáticas/citología , Nanopartículas/química , Tenascina/genética , Animales , Movimiento Celular , Supervivencia Celular , Progresión de la Enfermedad , Matriz Extracelular/metabolismo , Proteínas de la Matriz Extracelular/genética , Regulación de la Expresión Génica , Hepatocitos/citología , Humanos , Inflamación , Hígado/citología , Cirrosis Hepática/genética , Masculino , Ratones , Ratones Endogámicos C57BL , Nanomedicina , ARN Interferente Pequeño/metabolismo , Ratas , Ratas Sprague-Dawley , Dióxido de Silicio/químicaRESUMEN
Multifunctional hybrid nanoparticles are being developed to carry a wide variety of therapeutic and imaging agents for multiple biomedical applications. Polysilsesquioxane (PSilQ) nanoparticles are a promising hybrid platform with numerous advantages to be used as a delivery system. In this report, we demonstrate the ability of a stimuli-responsive PSilQ-based platform to transport and deliver simultaneously protoporphyrin IX, curcumin, and RNA interference inducers inside human cells. This multimodal delivery system shows a synergistic performance for the combined phototherapy and chemotherapy of triple-negative breast cancer and can be used for efficient transfection of therapeutic nucleic acids. The current work represents the first report of using the PSilQ platform for the combined phototherapy and chemotherapy and gene delivery.