Tailoring Magnetite-Nanoparticle-Based Nanocarriers for Gene Delivery: Exploiting CRISPRa Potential in Reducing Conditions.

Gene delivery has emerged as a promising alternative to conventional treatment approaches, allowing for the manipulation of gene expression through gene insertion, deletion, or alteration. However, the susceptibility of gene delivery components to degradation and challenges associated with cell penetration necessitate the use of delivery vehicles for effective functional gene delivery. Nanostructured vehicles, such as iron oxide nanoparticles (IONs) including magnetite nanoparticles (MNPs), have demonstrated significant potential for gene delivery applications due to their chemical versatility, biocompatibility, and strong magnetization. In this study, we developed an ION-based delivery vehicle capable of releasing linearized nucleic acids (tDNA) under reducing conditions in various cell cultures. As a proof of concept, we immobilized a CRISPR activation (CRISPRa) sequence to overexpress the pink1 gene on MNPs functionalized with polyethylene glycol (PEG), 3-[(2-aminoethyl)dithio]propionic acid (AEDP), and a translocating protein (OmpA). The nucleic sequence (tDNA) was modified to include a terminal thiol group and was conjugated to AEDP's terminal thiol via a disulfide exchange reaction. Leveraging the natural sensitivity of the disulfide bridge, the cargo was released under reducing conditions. Physicochemical characterizations, including thermogravimetric analysis (TGA) and Fourier-transform infrared (FTIR) spectroscopy, confirmed the correct synthesis and functionalization of the MNP-based delivery carriers. The developed nanocarriers exhibited remarkable biocompatibility, as demonstrated by the hemocompatibility, platelet aggregation, and cytocompatibility assays using primary human astrocytes, rodent astrocytes, and human fibroblast cells. Furthermore, the nanocarriers enabled efficient cargo penetration, uptake, and endosomal escape, with minimal nucleofection. A preliminary functionality test using RT-qPCR revealed that the vehicle facilitated the timely release of CRISPRa vectors, resulting in a remarkable 130-fold overexpression of pink1. We demonstrate the potential of the developed ION-based nanocarrier as a versatile and promising gene delivery vehicle with potential applications in gene therapy. The developed nanocarrier is capable of delivering any nucleic sequence (up to 8.2 kb) once it is thiolated using the methodology explained in this study. To our knowledge, this represents the first MNP-based nanocarrier capable of delivering nucleic sequences under specific reducing conditions while preserving functionality.

Functional Ability of Dental Materials After Inclusion of Nanocarriers Containing Functional Substances: A Systematic Review.

BACKGROUND: Nanocarriers are nanoparticles used to transport functional substances. OBJECTIVE: This study aimed to perform a systematic review analyzing the functional ability of dental materials after the inclusion of nanocarriers containing functional substances. MATERIALS AND METHODS: We searched PubMed, Web of Science, EMBASE, SciELO, and Science Direct from (02/03/21) to (02/06/21) without restriction on the year of publication. We included studies evaluating the incorporation of Chitosan, PEG-PCL (poly(ethylene glycol)-poly(ε-caprolactone), PLGA (poly(lactic-co-glycolic acid), and mesoporous silica into dental materials. We analyzed the risk of bias with CRIS guidelines and consolidated all analyses using Microsoft Office Excel. RESULTS: In total, we found 656 studies and included 11. The articles investigated materials to eliminate microorganisms, interference with materials' mechanical properties, elution of remineralizing substances, and reduction of oral anesthetic toxicity. Chitosan and mesoporous silica were the most prevalent nanocarriers. Among all properties, the antibacterial activity was the most analyzed. The functional ability of dental materials provided by all nanocarriers improved. CONCLUSION: The use of nanocarriers may be a useful way to produce dental materials with improved functional abilities. We registered this manuscript in the Open Science Framework platform - DOI 10.17605/OSF.IO/RP5XK, available at https://osf.io/qg49x.

All-trans retinoic acid in anticancer therapy: how nanotechnology can enhance its efficacy and resolve its drawbacks.

Introduction: All-trans retinoic acid (ATRA, tretinoin) is the main drug used in the treatment of acute promyelocytic leukemia (APL). Despite its impressive activity against APL, the same could not be clinically observed in other types of cancer. Nanotechnology can be a tool to enhance ATRA anticancer efficacy and resolve its drawbacks in APL as well as in other malignancies.Areas covered: This review covers ATRA use in APL and non-APL cancers, the problems that were found in ATRA therapy and how nanoencapsulation can aid to circumvent them. Pre-clinical results obtained with nanoencapsulated ATRA are shown as well as the two ATRA products based on nanotechnology that were clinically tested: ATRA-IV® and Apealea®.Expert opinion: ATRA presents interesting properties to be used in anticancer therapy with a notorious differentiation and antimetastatic activity. Bioavailability and resistance limitations impair the use of ATRA in non-APL cancers. Nanotechnology can circumvent these issues and provide tools to enhance its anticancer activities, such as co-loading of multiple drug and active targeting to tumor site.

Nanotheranostics through Mitochondria-targeted Delivery with Fluorescent Peptidomimetic Nanohybrids for Apoptosis Induction of Brain Cancer Cells.

Overview: Malignant brain tumors remain one of the greatest challenges faced by health professionals and scientists among the utmost lethal forms of cancer. Nanotheranostics can play a pivotal role in developing revolutionary nanoarchitectures with multifunctional and multimodal capabilities to fight cancer. Mitochondria are vital organelles to eukaryotic cells, which have been recognized as a significant target in cancer therapy where, by damaging the mitochondria, it will cause irreparable cell death or apoptosis. Methods: We designed and produced novel hybrid nanostructures comprising a fluorescent semiconductor core (AgInS2, AIS) and cysteine-modified carboxymethylcellulose (termed thiomer, CMC_Cys) conjugated with mitochondria-targeting peptides (KLA) forming a macromolecular shell for combining bioimaging and for inducing brain cancer cell (U-87 MG) death. Results: The optical and physicochemical properties of the nanoconjugates demonstrated suitability as photoluminescent nanostructures for cell bioimaging and intracellular tracking. Additionally, the results proved a remarkable killing activity towards glioblastoma cells of cysteine-bearing CMC conjugates coupled with KLA peptides through the half-maximal effective concentration values, approximately 70-fold higher compared to the conjugate analogs without Cys residues. Moreover, these thiomer-based pro-apoptotic drug nanoconjugates displayed higher lethality against U-87 MG cancer cells than doxorubicin, a model drug in chemotherapy, although extremely toxic. Remarkably, these peptidomimetic nanohybrids demonstrated a relative "protective effect" regarding healthy cells while maintaining high killing activity towards malignant brain cells. Conclusion: These findings pave the way for developing hybrid nanoarchitectures applied as targeted multifunctional platforms for simultaneous imaging and therapy against cancer while minimizing the high systemic toxicity and side-effects of conventional drugs in anticancer chemotherapy.

Fluorescent quantum dots-zika virus hybrid nanoconjugates for biolabeling, bioimaging, and tracking host-cell interactions.

The earliest possible diagnosis and understanding of the infection mechanisms play a crucial role in the outcome of fighting viral diseases. Thus, we designed and developed for the first time, novel bioconjugates made of Ag-In-S@ZnS (ZAIS) fluorescent quantum dots coupled with ZIKA virus via covalent amide bond with carboxymethylcellulose (CMC) biopolymer for labeling and bioimaging the virus-host cell interactions mechanisms through confocal laser scanning microscopy. This work offers relevant insights regarding the profile of the ZIKA virus-nanoparticle conjugates interactions with VERO cells, which can be applied as a nanoplatform to elucidate the infection mechanisms caused by this viral disease.

Dual-functional supramolecular nanohybrids of quantum dot/biopolymer/chemotherapeutic drug for bioimaging and killing brain cancer cells in vitro.

Glioblastoma (GBM) is the utmost aggressive and lethal primary brain cancer, which has a poor prognosis and remains virtually incurable. Nanomedicine with emerging disruptive nanotechnology alternatives, including designed supramolecular nanohybrids has excellent potential as multimodal tools against cancer by combining nanomaterials, biomacromolecules, and drugs. Thus, we developed and constructed for the first time quantum dot-biopolymer-drug nanohybrids based on host-guest chemistry for simultaneous bioimaging, targeting, and anti-cancer drug delivery against GBM cells in vitro. ZnS fluorescent quantum dots (ZnS-QDs) were produced using chemically modified polysaccharide, carboxymethylcellulose (CMC), as water-soluble capping ligand and biofunctional layer via a facile one-step eco-friendly aqueous colloidal process at room temperature and physiological pH. These hybrid inorganic-organic nanocolloids (ZnS@CMC) were electrostatically conjugated with doxorubicin (DOX) anti-cancer drug forming innovative supramolecular complexes (ZnS@CMC-DOX) for amalgamating bioimaging and killing cancer cells. These nanoconjugates were characterized regarding their optical and physicochemical properties combined with morphological and structural features. The cytocompatibility was evaluated by MTT assay using healthy and GBM cells. The results showed that ultra-small ZnS-QDs were expertly produced uniform nanocolloids (average size = 3.6 nm). They demonstrated photoluminescence emission within the visible range of spectra. The cell viability results in vitro showed no cytotoxicity of ZnS@CMC nanohybrids towards both cell types. In summary, the novelty of this research relies on using a nanotheranostic strategy for developing ZnS@CMC-DOX nanohybrids with supramolecular vesicle-like structures. They behaved simultaneously as active fluorescent nanoprobes and nanocarriers with modulated drug release for bioimaging and killing malignant glioma cells proving the high potential for applications in cancer nanomedicine.

Bioengineered II-VI semiconductor quantum dot-carboxymethylcellulose nanoconjugates as multifunctional fluorescent nanoprobes for bioimaging live cells.

Colloidal semiconductor quantum dots (QDs) are light-emitting ultra-small nanoparticles, which have emerged as a new class of nanoprobes with unique optical properties for bioimaging and biomedical diagnostic. However, to be used for most biomedical applications the biocompatibility and water-solubility are mandatory that can achieved through surface modification forming QD-nanoconjugates. In this study, semiconductor II-VI quantum dots of type MX (M=Cd, Pb, Zn, X=S) were directly synthesized in aqueous media and at room temperature using carboxymethylcellulose sodium salt (CMC) behaving simultaneously as stabilizing and surface biofunctional ligand. These nanoconjugates were extensively characterized using UV-visible spectroscopy, photoluminescence spectroscopy, X-ray photoelectron spectroscopy, Fourier transform infrared spectroscopy, X-ray diffraction, transmission electron microscopy, dynamic light scattering and zeta potential. The results demonstrated that the biopolymer was effective on nucleating and stabilizing the colloidal nanocrystals of CdS, ZnS, and PbS with the average diameter ranging from 2.0 to 5.0nm depending on the composition of the semiconductor core, which showed quantum-size confinement effect. These QD/polysaccharide conjugates showed luminescent activity from UV-visible to near-infrared range of the spectra under violet laser excitation. Moreover, the bioassays performed proved that these novel nanoconjugates were biocompatible and behaved as composition-dependent fluorescent nanoprobes for in vitro live cell bioimaging with very promising perspectives to be used in numerous biomedical applications and nanomedicine.

Péptidos antimicrobianos: potencialidades terapéuticas / Antimicrobial peptides: their therapeutic potential

El aumento en la incidencia de las enfermedades infecciosas en los últimos años se ha favorecido por diferentes causas. Entre estas se destacan las inmunodeficiencias adquiridas (sida, trasplantes de órganos, quimioterapia oncológica), la migración de personas que trae consigo la posibilidad de importar enfermedades hacia poblaciones susceptibles, así como el excesivo empleo de antibióticos. Debido a esta situación se ha incrementado la búsqueda de nuevos candidatos terapéuticos para el desarrollo de terapias más efectivas. En este sentido los péptidos antimicrobianos constituyen una opción promisoria, pues presentan un amplio espectro de actividad frente a varios microorganismos patógenos. Además, se encuentran ampliamente distribuidos en la naturaleza, desde organismos unicelulares hasta mamíferos. Algunos péptidos antimicrobianos ya están siendo evaluados en estudios clínicos aunque muchos de ellos no han tenido resultados favorables in vivo debido a su poca estabilidad metabólica y toxicidad, entre otros. Con el fin de optimizar estas propiedades de los péptidos antimicrobianos se han trazado diferentes estrategias como la modificación química de su estructura y la conjugación con nanopartículas magnéticas. Es por eso que este artículo tiene el objetivo de revisar las potenciales aplicaciones terapéuticas de estas moléculas, teniendo en cuenta la información publicada al respecto en MedLine, Web of Science y Scopus en los últimos años

The growing incidence of infectious disease in recent years may be attributed to several causes, among them acquired immunodeficiencies (AIDS, organ transplant, oncological chemotherapy), human migration and the consequent import of diseases into susceptible populations, and the excessive use of antibiotics. This situation has fostered the search for new therapeutic candidates for the development of more effective treatments. Antimicrobial peptides are a promising alternative in this respect, due to their broad spectrum of activity against several pathogenic microorganisms. Moreover, they are widely distributed in nature, from unicellular organisms to mammals. Some antimicrobial peptides are already being evaluated in clinical studies, though many of them have not produced any favorable results in vivo due to their low metabolic stability and their toxicity, among other factors. Several strategies have been developed to overcome the above mentioned drawbacks, among them conjugation of microbial peptides with magnetic nanoparticles and chemical modification of their structure. The present study is aimed at reviewing the potential therapeutic applications of these molecules based on information published in MedLine, the Web of Science and Scopus in recent years.

In vitro and in vivo assessment of nanotoxicity of CdS quantum dot/aminopolysaccharide bionanoconjugates.

The nanotoxicity of Cd-containing quantum dots (QDs) for biomedical applications is very controversial and not completely understood. In this study, we evaluated the cytotoxicity of surface-biofunctionalized CdS QDs with chitosan directly synthesized via aqueous route at room temperature. These core-shell CdS-chitosan nanoconjugates showed different degrees of cytotoxic responses using MTT cell proliferation assay toward three human cell cultures, human osteosarcoma cell line (SAOS), non-Hodgkin's B cell lymphoma (Toledo), and human embryonic kidney cell line (HEK293T), under three exposure times (1, 3, and 5days) and three colloidal concentrations (10nM, 50nM, and 100nM). The results clearly demonstrated that the CdS QDs, regardless to the fact that they were coated with a biocompatible aminopolysaccharide shell, induced a severe dose- and time-dependent inhibition of cell viability. In addition, the HEK293T and SAOS cell lines showed much more sensitive response compared to Toledo, which indicated that the cytotoxicity was also cell-type dependent. The exceptional resistance of Toledo cells to toxic effects of CdS nanoconjugates even at severe test conditions was assigned to specific role of B-lineage cells of the immune defense system. Remarkably, no conclusive evidence of toxicity of CdS nanoconjugates was observed in vivo using intravenous injections of CdS nanoconjugates in BALB/c mouse animal models for 30days, but localized fluorescence was detected in ex-vivo liver tissue samples. Therefore, these results prove that there is no guarantee of "risk-free" use of CdS nanoconjugates for in vivo applications, even when functionalized with biopolymer ligands, as they can pose an excessive threat due to unpredicted and uncorrelated responses under in vitro and in vivo biological assays with highly toxic cadmium ions.

Biocompatible Fluorescent Core-Shell Nanoconjugates Based on Chitosan/Bi2S3 Quantum Dots.

Bismuth sulfide (Bi2S3) is a narrow-bandgap semiconductor that is an interesting candidate for fluorescent biomarkers, thermoelectrics, photocatalysts, and photovoltaics. This study reports the synthesis and characterization of novel Bi2S3 quantum dots (QDs) functionalized using chitosan (CHI) as the capping ligands via aqueous "green" route at room temperature and ambient pressure. Transmission electron microscopy (TEM), UV-visible (UV-vis) spectroscopy, photoluminescence (PL) spectroscopy, dynamic light scattering (DLS), and zeta potential (ZP) analysis were used to characterize the hybrids made of biopolymer-functionalized Bi2S3 semiconductor nanocrystals. The results demonstrated that the CHI ligand was effective at nucleating and controlling the growth of water-soluble colloidal Bi2S3 nanoparticles. The average sizes of the Bi2S3 nanoparticles were significantly affected by the molar ratio of the precursors but less dependent on the pH of the aqueous media, leading to the formation of nanocrystals with average diameters varying from 4.2 to 6.7 nm. These surface-modified Bi2S3 nanocrystals with CHI exhibited photoluminescence in the visible spectral region. Moreover, the results of in vitro MTT (3-(4,5-dimethylthiazol-2yl)-2,5-diphenyltetrazolium bromide) assay with human osteosarcoma cells (SAOS) cell line demonstrated no cytotoxic response of the nanoconjugates.Furthermore, the results indicated that the Bi2S3 QD-CHI nanoconjugates showed HEK293T cell uptake; therefore, they can be potentially used as novel fluorescent nanoprobes for the in vitro bioimaging of cells in biomedical applications. Graphical Abstract Schematic representation of the biocompatible core-shell nanostructure of the chitosan/Bi2S3 quantum dot conjugates with photoluminescent properties.