RESUMO
Tissue engineering is one of the potential fields in the domain of regenerative medicine. Engineered scaffolds are an excellent substitute for the conventional use of bone grafts as they are biocompatible, economic, and provide limitless supply with no risk of disease transmission. Gum-based scaffolds present a good scope for studying tissue-engineering models and analyzing controlled drug delivery. Uniform blending of the gums and the presence of the optimal concentration of appropriate crosslinkers are very crucial for biodegradability nature. Gum-based scaffolds containing gellan gum, xanthan gum, polyvinyl alcohol, and hydroxyapatite, cross-linked with either glutaraldehyde (GA) or sodium trimetaphosphate (STMP) were fabricated to study the efficiency of crosslinkers and were characterized for degradation profile, swelling capacity, porosity, mechanical strength, morphology, X-ray diffraction, Fourier-transform infrared, and in vitro biocompatibility. Scaffolds crosslinked with STMP exhibited higher degradation rate at Day 21 than scaffolds crosslinked with GA. However, higher compressive strength was obtained for scaffolds cross-linked with STMP signifying that they have a better ability to resist compressive forces. Superior cell viability was observed in STMP-crosslinked scaffolds. In conclusion, STMP serves as a better crosslinker in comparison to GA and can be used in the fabrication of scaffolds for bone tissue engineering.
Assuntos
Reagentes de Ligações Cruzadas , Glutaral/química , Polifosfatos/química , Alicerces Teciduais/química , Implantes Absorvíveis , Substitutos Ósseos , Sobrevivência Celular , Células Cultivadas , Humanos , Teste de Materiais , Polissacarídeos Bacterianos , Porosidade , Resistência à Tração , Engenharia Tecidual , Difração de Raios XRESUMO
The paradigm of central nervous system (CNS) drug discovery has mostly relied on traditional approaches of rodent models or cell-based in vitro models. Owing to the issues of species differences between humans and rodents, it is difficult to correlate the robustness of data for neurodevelopmental studies. With advances in the stem-cell field, 3D CNS organoids have been developed and explored owing to their resemblance to the human brain architecture and functions. Further, CNS organoids provide a unique opportunity to mimic the human brain physiology and serve as a modeling tool to study the normal versus pathological brain or the elucidation of mechanisms of neurological disorders. Here, we discuss the recent application of a CNS organoid explored for neurodevelopment disease or a screening tool for CNS drug development.
Assuntos
Encéfalo , Doenças do Sistema Nervoso Central , Avaliação Pré-Clínica de Medicamentos , Modelos Biológicos , Síndromes Neurotóxicas , Organoides , Animais , HumanosRESUMO
The blood-brain barrier (BBB) is comprised of brain microvascular endothelial central nervous system (CNS) cells, which communicate with other CNS cells (astrocytes, pericytes) and behave according to the state of the CNS, by responding against pathological environments and modulating disease progression. The BBB plays a crucial role in maintaining homeostasis in the CNS by maintaining restricted transport of toxic or harmful molecules, transport of nutrients, and removal of metabolites from the brain. Neurological disorders, such as NeuroHIV, cerebral stroke, brain tumors, and other neurodegenerative diseases increase the permeability of the BBB. While on the other hand, semipermeable nature of BBB restricts the movement of bigger molecules i.e. drugs or proteins (>500 kDa) across it, leading to minimal bioavailability of drugs in the CNS. This poses the most significant shortcoming in the development of therapeutics for CNS neurodegenerative disorders. Although the complexity of the BBB (dynamic and adaptable barrier) affects approaches of CNS drug delivery and promotes disease progression, understanding the composition and functions of BBB provides a platform for novel innovative approaches towards drug delivery to CNS. The methodical and scientific interests in the physiology and pathology of the BBB led to the development and the advancement of numerous in vitro models of the BBB. This review discusses the fundamentals of BBB structure, permeation mechanisms, an overview of all the different in-vitro BBB models with their advantages and disadvantages, and rationale of selecting penetration prediction methods towards the critical role in the development of the CNS therapeutics.
Assuntos
Barreira Hematoencefálica/metabolismo , Sistemas de Liberação de Medicamentos , Preparações Farmacêuticas/administração & dosagem , Transporte Biológico/fisiologia , Encéfalo/metabolismo , Sistema Nervoso Central/metabolismo , Humanos , Modelos Biológicos , Permeabilidade , Preparações Farmacêuticas/metabolismoRESUMO
CRISPR-Cas9/gRNA exhibits therapeutic efficacy against latent human immunodeficiency virus (HIV) genome but the delivery of this therapeutic cargo to the brain remains as a challenge. In this research, for the first time, we demonstrated magnetically guided non-invasive delivery of a nano-formulation (NF), composed of Cas9/gRNA bound with magneto-electric nanoparticles (MENPs), across the blood-brain barrier (BBB) to inhibit latent HIV-1 infection in microglial (hµglia)/HIV (HC69) cells. An optimized ac-magnetic field of 60 Oe was applied on NF to release Cas9/gRNA from MENPs surface and to facilitate NF cell uptake resulting in intracellular release and inhibition of HIV. The outcomes suggested that developed NF reduced HIV-LTR expression significantly in comparison to unbound Cas9/gRNA in HIV latent hµglia/HIV (HC69) cells. These findings were also validated qualitatively using fluorescence microscopy to assess NF efficacy against latent HIV in the microglia cells. We believe that CNS delivery of NF (CRISPR/Cas9-gRNA-MENPs) across the BBB certainly will have clinical utility as future personalized nanomedicine to manage neuroHIV/AIDS.
Assuntos
Barreira Hematoencefálica/virologia , Infecções por HIV/terapia , Infecções por HIV/virologia , HIV-1 , RNA Guia de Cinetoplastídeos/administração & dosagem , Sistemas CRISPR-Cas , Células Cultivadas , Sistemas de Liberação de Medicamentos , Edição de Genes/métodos , HIV-1/genética , Humanos , Técnicas In Vitro , Nanopartículas de Magnetita/administração & dosagem , RNA Guia de Cinetoplastídeos/genética , Latência ViralRESUMO
Nanotechnology has gained increased attention for delivering therapeutic agents effectively to the cardiovascular system. Heart targeted nanocarrier based drug delivery is a new, effective and efficacious approach for treating various cardiac related disorders such as atherosclerosis, hypertension, and myocardial infarction. Nanocarrier based drug delivery system circumvents the problems associated with conventional drug delivery systems, including their nonspecificity, severe side effects and damage to the normal cells. Modification of physicochemical properties of nanocarriers such as size, shape and surface modifications can immensely alter its invivo pharmacokinetic and pharmacodynamic data and will provide better treatment strategy. Several nanocarriers such as lipid, phospholipid nanoparticles have been developed for delivering drugs to the target sites within the heart. This review summarizes and increases the understanding of the advanced nanosized drug delivery systems for treating cardiovascular disorders with the promising use of nanotechnology.
Assuntos
Doenças Cardiovasculares/terapia , Sistemas de Liberação de Medicamentos/métodos , Nanomedicina/métodos , Nanopartículas/uso terapêutico , Doenças Cardiovasculares/patologia , HumanosRESUMO
A magnetically guided brain delivery method previously demonstrated in mice has not yet been translated for clinical applications due to the mismatch of available static magnet dimensions in relation to the human brain size and shape. To develop a human-compatible methodology, we explored magnetic resonance imaging (MRI) as a tool for the delivery of magneto-electric nanoparticles (MENPs) into the brain of a baboon, as a proof-of-concept study. MRI brain image analysis showed a reduction in T2* value at the basal ganglia, hemisphere, and vertex, thereby confirming successful MENP delivery to the brain. The observation of well-integrated morphologically healthy tissues and no blood toxicity over the study duration confirmed the biocompatibility of MENPs and the delivery procedure. Outcomes of this research present MRI-assisted delivery of MENPs to the brain as a safe and noninvasive method in larger species such as baboons and one step closer to human translation. This MENP-based nanomedicine delivery method can be used for clinical application in order to investigate effective central nervous system (CNS) therapies.
RESUMO
Neurological disorders are the biggest concern globally. Out of ~36 million human immunodeficiency virus (HIV) positive people, about 30%-60% exhibit neurological disorders, including dementia and Alzheimer's disease (AD) like pathology. In AD or AD like neurological disorders, the pathogenesis is mainly due to the abnormal accumulation of extracellular amyloid beta (Aß). In this era of antiretroviral therapy, the life span of the HIV-infected individuals has increased leading towards increased neurocognitive dysfunction in nearly 30% of HIV-infected individuals, specifically older people. Deposition of the Aß plaques in the CNS is one the major phenomenon happening in aging HIV patients. ART suppresses the viral replication, but the neurotoxic protein (Tat) is still produced and results in increased levels of Aß. Furthermore, drugs of abuse like cocaine (coc) is known to induce the HIV associated neurocognitive disorders as well as the Aß secretion. To target the Tat and coc induced Aß secretion, we propose a potent bifunctional molecule Withaferin A (WA) which may act as a neuro-protectant against Aß neurotoxicity. In this study, we show that WA reduces secreted Aß and induced neurotoxicity in amyloid precursor protein (APP)-plasmid transfected SH-SY5Y cells (SH-APP). In this study, we show that in SH-APP cells, Aß secretion is induced in the presence of HIV-1 Tat (neurotoxic) and drug of abuse coc. Our fluorescent microscopy studies show the increased concentration of Aß40 in Tat (50 ng/ml) and coc (0.1 µM) treated SH-APP cells as compared to control. Our dose optimization study show, lower concentrations (0.5-2 µM) of WA significantly reduce the Aß40 levels, without inducing cytotoxicity in the SH-APP cells. Additionally, WA reduces the Tat and cocaine induced Aß levels. Therefore, we propose that Aß aggregation is induced by the presence of Tat and coc and WA is potent in reducing the secreted Aß and induced neurotoxicity. Our study provides new opportunities for exploring the pathophysiology and targeting the neurological disorders.
RESUMO
HIV and substance abuse plays an important role in infection and disease progression. Further, the presence of persistent viral CNS reservoirs makes the complete eradication difficult. Thus, neutralizing the drug of abuse effect on HIV-1 infectivity and elimination of latently infected cells is a priority. The development of a multi-component [antiretroviral drugs (ARV), latency reactivating agents (LRA) and drug abuse antagonist (AT)] sustained release nanoformulation targeting the CNS can overcome the issues of HIV-1 cure and will help in improving the drug adherence. The novel magneto-liposomal nanoformulation (NF) was developed to load different types of drugs (LRAs, ARVs, and Meth AT) and evaluated for in-vitro and in-vivo BBB transmigration and antiviral efficacy in primary CNS cells. We established the HIV-1 latency model using human astrocyte cells (HA) and optimized the dose of LRA for latency reversal, Meth AT in in-vitro cell culture system. Further, PEGylated magneto-liposomal NF was developed, characterized for size, shape, drug loading and BBB transport in-vitro. Results showed that drug released in a sustained manner up to 10 days and able to reduce the HIV-1 infectivity up to ~40-50% (>200 pg/mL to <100 pg/mL) continuously using single NF treatment ± Meth treatment in-vitro. The magnetic treatment (0.8 T) was able to transport (15.8% ± 5.5%) NF effectively without inducing any toxic effects due to NF presence in the brain. Thus, our approach and result showed a way to eradicate HIV-1 reservoirs from the CNS and possibility to improve the therapeutic adherence to drugs in drug abusing (Meth) population. In conclusion, the developed NF can provide a better approach for the HIV-1 cure and a foundation for future HIV-1 purging strategies from the CNS using nanotechnology platform.
Assuntos
Astrócitos , Infecções por HIV/tratamento farmacológico , HIV-1/fisiologia , Nanopartículas , Transtornos Relacionados ao Uso de Substâncias/tratamento farmacológico , Latência Viral/efeitos dos fármacos , Antirretrovirais/química , Antirretrovirais/farmacocinética , Antirretrovirais/farmacologia , Astrócitos/metabolismo , Astrócitos/patologia , Astrócitos/virologia , Células Cultivadas , Infecções por HIV/metabolismo , Infecções por HIV/patologia , Humanos , Nanomedicina/métodos , Nanopartículas/química , Nanopartículas/uso terapêutico , Transtornos Relacionados ao Uso de Substâncias/metabolismo , Transtornos Relacionados ao Uso de Substâncias/patologia , Transtornos Relacionados ao Uso de Substâncias/virologiaRESUMO
This work, as a proof of principle, presents a sensitive and selective electrochemical immunosensor for Zika-virus (ZIKV)-protein detection using a functionalized interdigitated micro-electrode of gold (IDE-Au) array. A miniaturized IDE-Au immunosensing chip was prepared via immobilization of ZIKV specific envelop protein antibody (Zev-Abs) onto dithiobis(succinimidyl propionate) i.e., (DTSP) functionalized IDE-Au (electrode gap/width of 10 µm). Electrochemical impedance spectroscopy (EIS) was performed to measure the electrical response of developed sensing chip as a function of ZIKV-protein concentrations. The results of EIS studies confirmed that sensing chip detected ZIKV-protein selectively and exhibited a detection range from 10 pM to 1 nM and a detection limit of 10 pM along with a high sensitivity of 12 kΩM-1. Such developed ZIKV immune-sensing chip can be integrated with a miniaturized potentiostat (MP)-interfaced with a smartphone for rapid ZIKV-infection detection required for early stage diagnostics at point-of-care application.
Assuntos
Técnicas Biossensoriais/métodos , Espectroscopia Dielétrica/métodos , Técnicas Eletroquímicas/métodos , Proteínas Virais/análise , Zika virus/metabolismoRESUMO
Cardiovascular diseases (CVDs) are the leading cause of morbidity and mortality worldwide. Due to the significant impact of CVD on humans, there is a need to develop novel treatment modalities tailored to major classes of cardiac diseases including hypertension, coronary artery disease, cardiomyopathies, arrhythmias, valvular disease and inflammatory diseases. In this article, we discuss recent advancements regarding development of therapeutic strategies based on stem cells, aptamers, exosomes, drug-eluting and dissolvable stents, immunotherapy and nanomedicine for the treatment of CVD. We summarize current research and clinical advances in cardiovascular therapeutics, with a focus on therapies that move beyond current oral- or sublingual-based regimens. This review article provides insight into current research and future treatment strategies that hold a great relevance for future clinical practice in pursuit of improving quality of life of patients suffering from CVD.
Assuntos
Aptâmeros de Nucleotídeos/metabolismo , Doenças Cardiovasculares/terapia , Exossomos/metabolismo , Nanomedicina/métodos , Células-Tronco , Animais , Doenças Cardiovasculares/metabolismo , Doenças Cardiovasculares/patologia , HumanosRESUMO
Central nervous system (CNS) diseases are rapidly increasing globally. Currently used therapeutic agents to treat CNS diseases exhibit significant efficacy. However, the inability of these drugs to cross the blood-brain barrier (BBB) and invasiveness of the technologies to achieve localized drug delivery in disease-specific parts of the brain have thwarted pain-free and complete treatment of CNS diseases. Therefore, the safe, non-invasive, and targeted delivery of drugs to the brain using nanoparticles (NPs) is currently receiving considerable research attention. Here, we highlight advances in state-of-the-art personalized nanomedicine for the treatment of CNS diseases (with a focus on dementia), the related challenges, possible solutions, and prospects for nano-enabled personalized medicine.
Assuntos
Doenças do Sistema Nervoso Central/tratamento farmacológico , Nanomedicina , Medicina de Precisão , Animais , HumanosRESUMO
INTRODUCTION: The emergent field of nanoparticles has presented a wealth of opportunities for improving the treatment of human diseases. Recent advances have allowed for promising developments in drug delivery, diagnostics, and therapeutics. Modified delivery systems allow improved drug delivery over traditional pH, microbe, or receptor dependent models, while antibody association allows for more advanced imaging modalities. Nanoparticles have potential clinical application in the field of gastroenterology as they offer several advantages compared to the conventional treatment systems including target drug delivery, enhanced treatment efficacy, and reduced side effects. AREAS COVERED: The aim of this review article is to summarize the recent advancements in developing nanoparticle technologies to treat gastrointestinal diseases. We have covered the application of nanoparticles in various gastrointestinal disorders including inflammatory bowel disease and colorectal cancer. We also have discussed how the gut microbiota affects the nanoparticle based drug delivery in the gastrointestinal tract. EXPERT OPINION: Nanoparticles based drug delivery offers a great platform for targeted drug delivery for gastrointestinal disorders. However, it is influenced by the presence of microbiota, drug interaction with nanoparticles, and cytotoxicity of nanoparticles. With the advancements in nanoparticle technology, it may be possible to overcome these barriers leading to efficient drug delivery for gastrointestinal disorders based on nanoparticle platform.
Assuntos
Sistemas de Liberação de Medicamentos , Gastroenteropatias/tratamento farmacológico , Microbioma Gastrointestinal/fisiologia , Nanopartículas/administração & dosagem , Preparações Farmacêuticas/administração & dosagem , HumanosRESUMO
Drug abuse (e.g., methamphetamine-Meth or cocaine-Coc) is one of the major risk factors for becoming infected with HIV-1, and studies show that in combination, drug abuse and HIV-1 lead to significantly greater damage to CNS. To overcome these issues, we have developed a novel nanoformulation (NF) for drug-abusing population infected with HIV-1. In this work, a novel approach was developed for the co-encapsulation of Nelfinavir (Nel) and Rimcazole (Rico) using layer-by-layer (LbL) assembled magnetic nanoformulation for the cure of neuroAIDS. Developed NF was evaluated for blood-brain barrier (BBB) transmigration, cell uptake, cytotoxicity and efficacy (p24 assay) in HIV-1 infected primary astrocyte (HA) in presence or absence of Coc and Meth. Developed magnetic nanoformulation (NF) fabricated using the LbL approach exhibited higher amounts of drug loading (Nel and Rico) with 100% release of both the therapeutic agents in a sustained manner for 8 days. NF efficacy studies indicated a dose-dependent decrease in p24 levels in HIV-1-infected HA (~55%) compared to Coc + Meth treated (~50%). The results showed that Rico significantly subdued the effect of drugs of abuse on HIV infectivity. NF successfully transmigrated (38.8 ± 6.5%) across in vitro BBB model on the application of an external magnetic field and showed >90% of cell viability with efficient cell uptake. In conclusion, our proof of concept study revealed that sustained and concurrent release of sigma σ1 antagonist and anti-HIV drug from the developed novel sustained release NF can overcome the exacerbated effects of drugs of abuse in HIV infection and may solve the issue of medication adherence in the drug-abusing HIV-1 infected population.
Assuntos
Carbazóis/farmacocinética , Cocaína/farmacocinética , Preparações de Ação Retardada/farmacocinética , Drogas Ilícitas/farmacocinética , Metanfetamina/farmacocinética , Nelfinavir/farmacocinética , Complexo AIDS Demência/tratamento farmacológico , Astrócitos/citologia , Astrócitos/efeitos dos fármacos , Astrócitos/metabolismo , Barreira Hematoencefálica/efeitos dos fármacos , Barreira Hematoencefálica/metabolismo , Cocaína/química , Células Endoteliais/citologia , Células Endoteliais/efeitos dos fármacos , Células Endoteliais/metabolismo , Inibidores da Protease de HIV/farmacocinética , HIV-1/efeitos dos fármacos , HIV-1/crescimento & desenvolvimento , Humanos , Drogas Ilícitas/química , Imãs/química , Metanfetamina/química , Nanoestruturas/química , Fármacos Neuroprotetores/farmacocinética , Cultura Primária de Células , Abuso de Substâncias por Via Intravenosa/prevenção & controleRESUMO
In this research, we demonstrate cell uptake of magneto-electric nanoparticles (MENPs) through nanoelectroporation (NEP) using alternating current (ac)-magnetic field stimulation. Uptake of MENPs was confirmed using focused-ion-beam assisted transmission electron microscopy (FIB-TEM) and validated by a numerical simulation model. The NEP was performed in microglial (MG) brain cells, which are highly sensitive for neuro-viral infection and were selected as target for nano-neuro-therapeutics. When the ac-magnetic field optimized (60 Oe at 1 kHz), MENPs were taken up by MG cells without affecting cell health (viability > 92%). FIB-TEM analysis of porated MG cells confirmed the non-agglomerated distribution of MENPs inside the cell and no loss of their elemental and crystalline characteristics. The presented NEP method can be adopted as a part of future nanotherapeutics and nanoneurosurgery strategies where a high uptake of a nanomedicine is required for effective and timely treatment of brain diseases.
Assuntos
Encéfalo/efeitos dos fármacos , Microglia/efeitos dos fármacos , Nanopartículas/química , Linhagem Celular , Portadores de Fármacos , Eletricidade , Eletroporação/métodos , Humanos , Campos Magnéticos , Microscopia Eletrônica de Transmissão/métodos , Nanomedicina/métodosRESUMO
Magneto-plasmonic nanoparticles are one of the emerging multi-functional materials in the field of nanomedicine. Their potential for targeting and multi-modal imaging is highly attractive. In this study, magnetic core/gold shell (MNP@Au) magneto-plasmonic nanoparticles were synthesized by citrate reduction of Au ions on magnetic nanoparticle seeds. Hydrodynamic size and optical properties of magneto-plasmonic nanoparticles synthesized with the variation of Au ions and reducing agent concentrations were evaluated. The synthesized magneto-plasmonic nanoparticles exhibited superparamagnetic properties, and their magnetic properties contributed to the concentration-dependent contrast in magnetic resonance imaging (MRI). The imaging contrast from the gold shell part of the magneto-plasmonic nanoparticles was also confirmed by X-ray computed tomography (CT). The transmigration study of the magneto-plasmonic nanoparticles using an in vitro blood-brain barrier (BBB) model proved enhanced transmigration efficiency without disrupting the integrity of the BBB, and showed potential to be used for brain diseases and neurological disorders.
Assuntos
Barreira Hematoencefálica , Encéfalo/diagnóstico por imagem , Imageamento por Ressonância Magnética , Nanopartículas de Magnetita , Imagem Multimodal , Astrócitos/citologia , Encéfalo/citologia , Células Cultivadas , Células Endoteliais/citologia , Ouro , Humanos , Magnetismo , Modelos BiológicosRESUMO
Since centuries, the rapid spread and cure of infectious diseases have been a major concern to the progress and survival of humans. These diseases are a global burden and the prominent cause for worldwide deaths and disabilities. Nanomedicine has emerged as the most excellent tool to eradicate and halt their spread. Various nanoformulations (NFs) using advanced nanotechnology are in demand. Recently, hydrogel and nanogel based drug delivery devices have posed new prospects to simulate the natural intelligence of various biological systems. Owing to their unique porous interpenetrating network design, hydrophobic drug incorporation and stimulus sensitivity hydrogels owe excellent potential as targeted drug delivery systems. The present review is an attempt to highlight the recent trends of hydrogel based drug delivery systems for the delivery of therapeutic agents and diagnostics for major infectious diseases including acquired immune deficiency syndrome (AIDS), malaria, tuberculosis, influenza and ebola. Future prospects and challenges are also described.
Assuntos
Doenças Transmissíveis/tratamento farmacológico , Sistemas de Liberação de Medicamentos/tendências , Hidrogéis , Humanos , Nanomedicina/tendências , Nanopartículas/administração & dosagem , Nanopartículas/químicaRESUMO
This viewpoint is a global call to promote fundamental and applied research aiming toward designing smart nanocarriers of desired properties, novel noninvasive strategies to open the blood-brain barrier (BBB), delivery/release of single/multiple therapeutic agents across the BBB to eradicate neurohuman immunodeficiency virus (HIV), strategies for on-demand site-specific release of antiretroviral therapy, developing novel nanoformulations capable to recognize and eradicate latently infected HIV reservoirs, and developing novel smart analytical diagnostic tools to detect and monitor HIV infection. Thus, investigation of novel nanoformulations, methodologies for site-specific delivery/release, analytical methods, and diagnostic tools would be of high significance to eradicate and monitor neuroacquired immunodeficiency syndrome. Overall, these developments will certainly help to develop personalized nanomedicines to cure HIV and to develop smart HIV-monitoring analytical systems for disease management.
Assuntos
Fármacos Anti-HIV/uso terapêutico , Portadores de Fármacos/química , Sistemas de Liberação de Medicamentos/métodos , Infecções por HIV/tratamento farmacológico , Nanomedicina/métodos , Transtornos Neurocognitivos/tratamento farmacológico , Barreira Hematoencefálica/efeitos dos fármacos , Gerenciamento Clínico , Infecções por HIV/complicações , Infecções por HIV/psicologia , Humanos , Transtornos Neurocognitivos/diagnóstico , Transtornos Neurocognitivos/etiologiaRESUMO
Although the introduction of antiretroviral therapy has reduced the prevalence of severe forms of neurocognitive disorders, human immunodeficiency virus (HIV)-1-associated neurocognitive disorders were observed in 50% of HIV-infected patients globally. The blood-brain barrier is known to be impermeable to most of antiretroviral drugs. Successful delivery of antiretroviral drugs into the brain may induce an inflammatory response, which may further induce neurotoxicity. Therefore, alternate options to antiretroviral drugs for decreasing the HIV infection and neurotoxicity may help in reducing neurocognitive impairments observed in HIV-infected patients. In this study, we explored the role of magnetic nanoparticle (MNP)-bound tissue inhibitor of metalloproteinase-1 (TIMP1) protein in reducing HIV infection levels, oxidative stress, and recovering spine density in HIV-infected SK-N-MC neuroblastoma cells. We did not observe any neuronal cytotoxicity with either the free TIMP1 or MNP-bound TIMP1 used in our study. We observed significantly reduced HIV infection in both solution phase and in MNP-bound TIMP1-exposed neuronal cells. Furthermore, we also observed significantly reduced reactive oxygen species production in both the test groups compared to the neuronal cells infected with HIV alone. To observe the effect of both soluble-phase TIMP1 and MNP-bound TIMP1 on spine density in HIV-infected neuronal cells, confocal microscopy was used. We observed significant recovery of spine density in both the test groups when compared to the cells infected with HIV alone, indicting the neuroprotective effect of TIMP1. Therefore, our results suggest that the MNP-bound TIMP1 delivery method across the blood-brain barrier can be used for reducing HIV infectivity in brain tissue and neuronal toxicity in HIV-infected patients.
Assuntos
Fármacos Anti-HIV/farmacologia , Infecções por HIV/tratamento farmacológico , Nanopartículas de Magnetita , Plasticidade Neuronal/efeitos dos fármacos , Inibidor Tecidual de Metaloproteinase-1/farmacologia , Fármacos Anti-HIV/química , Fármacos Anti-HIV/farmacocinética , Barreira Hematoencefálica/efeitos dos fármacos , Encéfalo/metabolismo , Linhagem Celular , HIV-1/patogenicidade , Humanos , Magnetismo , Nanopartículas de Magnetita/química , Microscopia Confocal , Fármacos Neuroprotetores/química , Fármacos Neuroprotetores/farmacocinética , Fármacos Neuroprotetores/farmacologia , Inibidor Tecidual de Metaloproteinase-1/química , Inibidor Tecidual de Metaloproteinase-1/farmacocinéticaRESUMO
Electrochemical monitoring-on-chip (E-MoC)-based approach for rapid assessment of human immunodeficiency virus (HIV)-infection in the presence of cocaine (Coc) and specific drugs namely i.e., tenofovir (Tef), rimcazole (RA) is demonstrated here, for the first time, using electrochemical impedance spectroscopy (EIS). An in-vitro primary human astrocytes (HA) model was developed using a cultureware chip (CC, used for E-MoC) for HIV-infection, Coc exposure and treatment with anti-HIV drug i.e., Tef, and Coc antagonist i.e., RA. The charge transfer resistance (Rct) value of each CC well varies with respect to infection and treatment demonstrated highly responsive sensitivity of developed chip. The results of E-MoC, a proof-of-the concept, suggested that HIV-infection progression due to Coc ingestion and therapeutic effects of highly specific drugs are measurable on the basis of cell electrophysiology. Though, this work needs various molecular biology-based optimizations to promote this technology as an analytical tool for the rapid assessment of HIV-infection in a patient to manage HIV diseases for timely diagnosis. The presented study is based on using CNS cells and efforts are being made to perform this method using peripheral cells such as monocytes derived dendritic cells.