Nanomed (Brazil)

This thesis explores a range of strategies aimed at enhancing RNA delivery beyond the liver through the use of lipid nanoparticles (LNPs). A major focus of the work is on mitigating the rapid clearance of LNPs by the reticuloendothelial system (RES), which is attempted by employing RES blockade strategies aiming to extend LNP circulation time and enhance their accumulation in tumors. Additionally, the study investigates the functionalization of LNP surfaces with targeting ligands, a strategy designed to improve the specificity of RNA delivery to prostate cancer cells in vivo. By engineering LNPs to recognize and bind selectively to cancer cell receptors, this approach aims to maximize therapeutic efficacy while minimizing off-target effects. Another key focus of this thesis is the exploration of alternative administration routes to extend the applicability of LNP-mediated RNA delivery to different disease contexts. Specifically, in this thesis we examine intramyocardial administration of various LNP formulations as a promising strategy for treating heart diseases. By directly injecting LNPs into cardiac tissue, the study evaluates the efficiency of LNP uptake and RNA expression within the heart. In this thesis we evaluate LNP-based delivery systems for both small interfering RNA (siRNA) and messenger RNA (mRNA) in diverse in vivo animal models. This includes a thorough assessment of administration routes, biodistribution, gene silencing or expression efficiency, and the immune responses triggered by these nanoparticles. Special attention is given to the associated inflammation and toxicity profiles of different LNP formulations, providing critical insights into their safety and translational potential. Finally, the research explores the rapidly expanding field of extracellular vesicles as alternative drug delivery systems (DDS) to synthetic lipid-based nanoparticles. Over the past decade, extracellular vesicles have gained significant interest due to their innate biocompatibility, lower immunogenicity, and potential for cell-specific targeting. By comparing these naturally derived vesicles with synthetic LNPs, the study aims to elucidate their respective advantages and limitations, ultimately contributing to the broader effort of optimizing RNA-based therapeutic strategies for a range of medical applications.

The production of electrochemical devices and systems using additive manufacturing technology, particularly three‐dimensional (3D) printing, has proven to be highly promising. This work reports the development of 3D‐printed electrochemical sensors for the determination of the antibiotic ciprofloxacin (CIP). To achieve this, a lab‐made conductive filament composed of carbon black (CB) and polylactic acid (PLA) was produced and utilized in the fabrication of the sensors. Additionally, an electrochemical cell was constructed using a nonconductive filament, resulting in a miniaturized and entirely additively manufactured platform. The characterization of the proposed CB–PLA sensor was carried out using scanning electron microscopy and electrochemical techniques. The proposed sensor has shown a linear range of 1.0–12.5 µmol L−1, with a sensitivity of 3.77 µA µmol−1 L, and limits of detection and quantification of 0.3 and 0.9 µmol L−1 for CIP, respectively. Regarding the analysis of the samples (tap water and synthetic urine), it was observed recovery values close to 100% for all samples. Thus, the 3D‐printed electrochemical device presents itself as a high‐potential alternative for CIP drug control, with the possibility of being used in the field and point of care.

Lung cancer remains a challenge within the realm of oncology. Characterized by late-stage diagnosis and resistance to conventional treatments, the currently available therapeutic strategies encompass surgery, radiotherapy, chemotherapy, immunotherapy, and biological therapy; however, overall patient survival remains suboptimal. Nanotechnology has ushered in a new era by offering innovative nanomaterials with the potential to precisely target cancer cells while sparing healthy tissues. It holds the potential to reshape the landscape of cancer management, offering hope for patients and clinicians. The assessment of these nanotechnologies follows a rigorous evaluation process similar to that applied to chemical drugs, which includes considerations of their pharmacokinetics, pharmacodynamics, toxicology, and clinical effectiveness. However, because of the characteristics of nanoparticles, standard toxicological tests require modifications to accommodate their unique characteristics. Effective therapeutic strategies demand a profound understanding of the disease and consideration of clinical outcomes, physicochemical attributes of nanomaterials, nanobiointeractions, nanotoxicity, and regulatory compliance to ensure patient safety. This review explores the promise of nanomedicine in lung cancer treatment by capitalizing on its unique physicochemical properties. We address the multifaceted challenges of lung cancer and its tumor microenvironment and provide an overview of recent developments in nanoplatforms for early diagnosis and treatment that can enhance patient outcomes and overall quality of life.

The production of electrochemical devices and systems using additive manufacturing technology, particularly three‐dimensional (3D) printing, has proven to be highly promising. This work reports the development of 3D‐printed electrochemical sensors for the determination of the antibiotic ciprofloxacin (CIP). To achieve this, a lab‐made conductive filament composed of carbon black (CB) and polylactic acid (PLA) was produced and utilized in the fabrication of the sensors. Additionally, an electrochemical cell was constructed using a nonconductive filament, resulting in a miniaturized and entirely additively manufactured platform. The characterization of the proposed CB–PLA sensor was carried out using scanning electron microscopy and electrochemical techniques. The proposed sensor has shown a linear range of 1.0–12.5 µmol L−1, with a sensitivity of 3.77 µA µmol−1 L, and limits of detection and quantification of 0.3 and 0.9 µmol L−1 for CIP, respectively. Regarding the analysis of the samples (tap water and synthetic urine), it was observed recovery values close to 100% for all samples. Thus, the 3D‐printed electrochemical device presents itself as a high‐potential alternative for CIP drug control, with the possibility of being used in the field and point of care.

This study focuses on the development and implementation of a computational model based on a two-level system framework, employing a kinetic Monte Carlo approach, to simulate the behavior of magnetic nanoparticles in fluid environments under alternating magnetic field excitation. The primary goal is to develop a tool to be used in the investigation of the impact of different nanoparticle properties on the efficiency of magnetic hyperthermia. A crucial aspect of this investigation is the rigorous validation of the computational model through comprehensive comparisons with experimental observations varying particle size, concentration, and magnetic field amplitude and frequency, ensuring the reliability and accuracy of the model in capturing the complex dynamics of nanoparticles in suspension and its influence on hyperthermia efficiency. Our findings underscore the potential of this computational model as a predictive tool for estimating Specific Loss Power (SLP) metrics across various systems subjected to different applied magnetic fields. By delineating the relationship between the nanoparticle’s characteristics and hyperthermia efficiency, this study contributes to advancing the fundamental understanding and predictive capabilities of magnetic nanoparticle-based hyperthermia therapies.

Herein, we present the first 3D-printed electrochemical portable biodevice for the detection of monkeypox virus (MKPV). The electrochemical device consists of two biosensors: an immunosensor and a genosensor specifically designed for the detection of the protein A29 and a target DNA of MKPV, respectively. The electrodes were manufactured using lab-made ultraflexible conductive filaments composed of carbon black, recycled PLA from coffee pods, and castor oil as a plasticizer. The sensors created through 3D printing technology exhibited good reproducibility and repeatability of analytical responses. Furthermore, both the immunosensor and genosensor demonstrated excellent MKPV detection capabilities, with a linear range from 0.01 to 1.0 μmol L-1 for the antigen and 0.1 to 20.0 μmol L-1 for the DNA target. The biosensors achieved limits of detection of 2.7 and 29 nmol L-1 for the immunosensor and genosensor, respectively. Interference tests conducted with the biosensors demonstrated their selectivity for MKPV. Moreover, analyses of fortified human serum samples showed recoveries close to 100%, confirming the absence of significant matrix effects for MKPV analysis. Therefore, the 3D-printed multiplex device represents a viable and highly promising alternative for on-site, portable, and rapid point-of-care MKPV monitoring.

Viruses are infectious agents that cause various diseases worldwide. The recent COVID-19 pandemic has shown the need for rapid and reliable tests to confirm viral infections, aiming at the rapid isolation, treatment, and identification of high-incidence regions. Rapid antigen tests based on lateral flow immunochromatography have proven to be very useful. However, they are not accurate in patients with low viral loadings. The gold standard test is RT-PCR, which identifies parts of the viral genome by detecting specific DNA or RNA sequences. RT-PCR or similar tests such as RT-LAMP involve several steps for sample preparation and amplification of target sequences, require trained personnel to be performed, and can be time-consuming and expensive, limiting their point-of-care application. Biosensors are promising analytical devices for detecting nucleic acids, mainly RNA from viruses, offering advantages such as rapid results, high sensitivity, and low cost compared with the RT-PCR test since the amplification of target sequences is not necessary. Recently, several biosensors have been developed to detect RNA viruses without sequence amplification. Here, we present a review on the design and technology of amplification-free biosensors for the detection of viral RNA as an alternative for diagnosing infectious diseases. The challenges and advances for the point-of-care electrochemical, electrical, and optical biosensors will be addressed.