Biological Barriers Research Articles

Advancements in Lipid Nanoparticle Technologies for Precision Targeting in Therapeutic Delivery

<img src=” https://s3.amazonaws.com/production.scholastica/article/121633/large/prnano_1412024bg2.jpeg?1721676909”> Lipid nanoparticles (LNPs) are at the forefront of targeted therapeutic delivery, serving as an innovative platform for precisely delivering therapeutic agents directly to specific cells or tissues. Therapeutic delivery seeks to enhance drug efficacy and safety by controlling the distribution of drugs within the body, thereby minimizing off-target effects while maximizing therapeutic impact. This review delves into advancements and innovations in LNP design to achieve unprecedented targeting precision, surmount biological barriers, and diminish off-target interactions. It encom-passes exploring targeting strategies, including passive targeting that leverages the enhanced permeability and retention effect, active targeting via receptor-ligand interactions, and stimuli-responsive methodologies for the controlled release of therapeutics. With a focus on novel tar-geting techniques, including endogenous targeting and strategic de-targeting, to enhance LNP de-livery specificity and therapeutic efficacy. This review articulates the critical role of lipid com-position, nanoparticle characterization, and surface engineering in tailoring LNPs for therapeutic applications. Through evaluating the forefront of LNP technology in drug delivery and gene therapy, this review forecasts the trajectory of nanoparticle-based therapeutics, envisioning their pivotal role in the advent of personalized medicine, thereby providing a foundation for future explorations aimed at refining and optimizing LNP systems for clinical applications.

Gold Nanoparticles: An Emerging Novel Technology for Targeted Delivery System for Site-Specific Diseases

Abstract: Gold Nanoparticles (GNPs) have emerged as a novel technology in the field of targeted delivery systems, offering promising solutions for site-specific disease treatment. These nanoparticles possess unique physicochemical properties, such as controlled size, shape, and surface chemistry, which enable precise manipulation for enhanced therapeutic efficacy. The biocompatibility and ease of functionalization of GNPs facilitate the conjugation with various biomolecules, including drugs, peptides, and nucleic acids, thereby improving their targeted delivery capabilities. Recent advancements in nanotechnology have leveraged GNPs for the treatment of a range of diseases, particularly in oncology, cardiology, and neurology. In cancer therapy, GNPs can be engineered to target tumor cells selectively, minimizing damage to healthy tissues and reducing side effects. This is achieved through the conjugation of GNPs with tumor-specific ligands, antibodies, or aptamers, which direct the nanoparticles to malignant cells, allowing for localized drug release and improved therapeutic outcomes. Moreover, GNPs exhibit remarkable potential in diagnostic imaging and photothermal therapy. Their unique optical properties, such as surface plasmon resonance, enable their use as contrast agents in imaging techniques, providing high-resolution and real-time monitoring of disease progression. In photothermal therapy, GNPs convert light energy into heat, effectively destroying targeted cells with minimal invasiveness. The development of GNP-based delivery systems also addresses significant challenges in drug resistance and bioavailability. By overcoming biological barriers and enhancing cellular uptake, GNPs improve the pharmacokinetics and pharmacodynamics of therapeutic agents. However, despite these advancements, the clinical translation of GNPs faces challenges such as potential toxicity, long-term stability, and regulatory hurdles. In conclusion, gold nanoparticles represent a cutting-edge approach in targeted delivery systems, offering significant potential for site-specific disease treatment. Continued research and innovation are essential to overcome existing challenges and fully realize the clinical applications of GNPs, ultimately revolutionizing precision medicine. Gold nanoparticles exhibit unique physicochemical and optical properties. The gold nanoparticles have advanced techniques to cure different chronic diseases. Today, gold nanoparticles are aided by photodynamic therapy and radiation therapy as drug carriers. Due to this, researchers now focus on medical sciences to treat various diseases and therapeutic applications. This review provides all the aspects of gold-based nanoparticles, methods, and their pharmacological benefits in different fields of medical sciences. We also discuss various preparation methods and their advantages in pharmaceutical formulations.

Shape Dependent Therapeutic Potential of Nanoparticulate System: Advance Approach for Drug Delivery.

Drug delivery systems rely heavily on nanoparticles because they provide a targeted and monitored release of pharmaceuticals that maximize therapeutic efficacy and minimize side effects. To maximize drug internalization, this review focuses on comprehending the interactions between biological systems and nanoparticles. The way that nanoparticles behave during cellular uptake, distribution, and retention in the body is determined by their shape. Different forms, such as mesoporous silica nanoparticles, micelles, and nanorods, each have special properties that influence how well drugs are delivered to cells and internalized. To achieve the desired particle morphology, shape-controlled nanoparticle synthesis strategies take into account variables like pH, temperatures, and reaction time. Top-down techniques entail dissolving bulk materials to produce nanoparticles, whereas bottom-up techniques enable nanostructures to self-assemble. Comprehending the interactions at the bio-nano interface is essential to surmounting biological barriers and enhancing the therapeutic efficacy of nanotechnology in drug delivery systems. In general, drug internalization and distribution are greatly influenced by the shape of nanoparticles, which presents an opportunity for tailored and efficient treatment plans in a range of medical applications.

Nano/genetically engineered cells for immunotherapy

AbstractImmunotherapy has recently emerged as a promising therapeutic modality for the treatment of various diseases such as cancer, inflammation, autoimmune diseases, and infectious diseases. Despite its potential, immunotherapy faces challenges related to delivery efficiency and off‐target toxicity of immunotherapeutic drugs. Nano drug delivery systems offer improvements in drug biodistribution and release kinetics but still suffer from shortcomings such as high immunogenicity, poor penetration across biological barriers, and insufficient tissue permeability. Targeted delivery of drugs using living cells has become an emerging strategy that can take advantage of the inherent characteristics of cells to deal with the delivery defects of nano delivery systems. Furthermore, cells themselves can be genetically engineered into cellular drugs for enhanced immunotherapy. This review provides an in‐depth exploration of cell‐derived drug carriers, detailing their biological properties, functions, and commonly used drug loading strategies. In addition, the role of genetically modified cells in immunotherapy and their synergistic therapeutic effects with drug delivery are also introduced. By summarizing the main advancements and limitations in the field, this review offers insights into the potential of cell‐based drug delivery systems to address the existing challenges in immunotherapy. The introduction to recent developments and evaluation of ongoing research will pave the way for the optimization and widespread adoption of nano/genetically engineered cells for immunotherapy.

Expanding Arsenal against Ocular Diseases through Nanoemulsion: Success So Far and the Road Ahead.

The eye is a most delicate organ protected by several complex biological barriers that are static and dynamic. The presence of these ocular barriers retards drug absorption from topically applied dosage forms at the conjunctival sac. The efficient topical delivery of the drug into the globe is more difficult to achieve and there is a need to develop a topical formulation that may reduce the use of injections and increase patient compliance with decreased frequency of administration. In the advancements of research in nanotechnology, nanoemulsions can be used as biocompatible carriers to deliver the drug to the ocular cavity. The lipophilic globules can increase the solubility of hydrophobic cargos which provides increased permeation ability and ocular bioavailability which can sustain drug release and corneal retention. Because of their small size, these formulations do not cause blurring of vision. Nanoemulsions (NEs) over the past decade have been used to treat several ocular diseases in the anterior eye segment. This review summarizes the economic burden, pathology of ocular diseases, formulation considerations for ocular formulations, and recent advances of these NEs as effective carriers for ocular drug delivery highlighting their performance in pre-clinical studies.

Terpene-based novel invasomes: pioneering cancer treatment strategies in traditional medicine.

Health care workers have faced a significant challenge because of the rise in cancer incidence around the world during the past 10 years. Among various forms of malignancy skin cancer is most common, so there is need for the creation of an efficient and safe skin cancer treatment that may offer targeted and site-specific tumor penetration, and reduce unintended systemic toxicity. Nanocarriers have thus been employed to get around the issues with traditional anti-cancer drug delivery methods. Invasomes are lipid-based nanovesicles having small amounts of terpenes and ethanol or a mixture of terpenes and penetrate the skin more effectively. Compared to other lipid nanocarriers, invasomes penetrate the skin at a substantially fasterrate. Invasomes possess a number of advantages, including improved drug effectiveness, higher compliance, patient convenience, advanced design, multifunctionality, enhanced targeting capabilities, non-invasive delivery methods, potential for combination therapies, and ability to overcome biological barriers,. These attributes position invasomes as a promising and innovative platform for thefuture of cancer treatment. The current review provides insights into invasomes, with a fresh organizational scheme and incorporates the most recent cancer research, including their composition, historical development and methods of preparation, the penetration mechanism involving effect of various formulation variables and analysis of anticancer mechanism and the application of invasomes.

Recent Advances in Photodynamic Therapy: Metal-Based Nanoparticles as Tools to Improve Cancer Therapy.

Cancer remains a significant global health challenge, with traditional therapies like surgery, chemotherapy, and radiation often accompanied by systemic toxicity and damage to healthy tissues. Despite progress in treatment, these approaches have limitations such as non-specific targeting, systemic toxicity, and resistance development in cancer cells. In recent years, nanotechnology has emerged as a revolutionary frontier in cancer therapy, offering potential solutions to these challenges. Nanoparticles, due to their unique physical and chemical properties, can carry therapeutic payloads, navigate biological barriers, and selectively target cancer cells. Metal-based nanoparticles, in particular, offer unique properties suitable for various therapeutic applications. Recent advancements have focused on the integration of metal-based nanoparticles to enhance the efficacy and precision of photodynamic therapy. Integrating nanotechnology into cancer therapy represents a paradigm shift, enabling the development of strategies with enhanced specificity and reduced off-target effects. This review aims to provide a comprehensive understanding of the pivotal role of metal-based nanoparticles in photodynamic therapy. We explore the mechanisms, biocompatibility, and applications of metal-based nanoparticles in photodynamic therapy, highlighting the challenges and the limitations in their use, as well as the combining of metal-based nanoparticles/photodynamic therapy with other strategies as a synergistic therapeutic approach for cancer treatment.

Bohemian Rhapsody of Future Drug Delivery Systems: Rational Changes Necessary for the Next Revolution.

Controlled drug delivery technology has matured for more than 70 years, starting from a twice-a-day oral formulation to 6 month long-acting injectable formulations. Further technological advances require superior formulations to treat various diseases more efficiently. Developing future formulations with practical innovations for treating existing and new diseases necessitates our continued efforts to overcome at least three main hurdles. They include (i) drug delivery with reduced side effects, (ii) long-term treatment of chronic diseases, and (iii) the overcoming of biological barriers. Such efforts start with the improved ability to accurately test drug delivery efficacy using proper controls. Future development can be aided by artificial intelligence if used properly. The next revolution of drug delivery systems will be augmented if implementation is given equal weight as discovery. Such a process can be accelerated with the systemic revamp of the research funding structure and cultivating a new generation of scientists who can think differently.

Photodynamic/photothermal pH-responsive mannose-based dimeric prodrug for synergistic overcoming the biological barriers to conquer multidrug resistance in MRSA

Methicillin-resistant Staphylococcus aureus (MRSA), recognized for its resistance, poses a formidable challenge to public health, primarily attributable to its biochemical resistant mechanisms, capacity to form protective biofilms, and capacity to evade macrophage surveillance. In response to this threat, we constructed a multifunctional pH-responsive dimeric prodrug based on mannose (MAN) and cinnamaldehyde (CMA). This prodrug, designated as MACA@indocyanine green (ICG), incorporates the photosensitizer ICG. The nanoscale properties and MAN units integrated into the MACA@ICG prodrug could promote effective binding with infectious microbes or MAN residues on mammalian cells. This specific interaction, corroborated by co-localization studies using confocal microscopy and flow cytometry, is crucial for initial bacterial binding, biofilm penetration, and macrophage-specific uptake, all of which are strategic in surmounting the biological barriers posed by MRSA. Following its targeted accumulation, MACA@ICG was demonstrated responsiveness to the acidic microenvironment of bacteria to release free CMA, a natural compound with antimicrobial properties. The drug release, synergistically coupled with the effects of photothermal therapy (PTT) and photodynamic therapy (PDT) showcased superior synergistic antibacterial efficacy both in vitro and in vivo. Therefore, MACA@ICG was verified remarkable capabilities in surmounting bacterial biological barriers and exceptional antibacterial efficiency through the combinational treatments of herbal components, PDT and PTT.

Plant‐derived exosomes: A new frontier in nano‐medicine for cancer and microbial infection therapy

AbstractExosomes, small extracellular vesicles secreted by cells, have emerged as pivotal players in cell‐to‐cell communication. Plant‐derived exosomes, in particular, are gaining attention for their potential therapeutic applications in nano‐medicine. These vesicles are naturally occurring nanoparticles that carry bioactive molecules such as proteins, lipids, and nucleic acids. Due to their biocompatibility, low toxicity, and ability to traverse biological barriers, plant‐derived exosomes present a promising alternative to synthetic nanoparticles for drug delivery, especially in cancer and microbial infection therapy. Exosomes are secreted by almost every cell and are profusely present in all living organisms, making them excellent candidates for a large spectrum of research and applications. This paper describes the highly organized and regulated biosynthesis of exosomes and the prospects of their application in cancer therapy and treatment of microbial infections.

Tumor Redox‐Responsive Minimalist B/Fe Nano‐Chains for Chemodynamically Enhanced Ferroptosis and Synergistic Boron Neutron Capture Therapy

AbstractBoron neutron capture therapy (BNCT) as a binary targeted particle radiotherapy strategy has shown potent anti‐cancer potential. However, biological barriers and restricted blood supply pose challenges in achieving adequate boron concentration within deep‐seated tumor lesions. BNCT with other anti‐cancer therapies, such as X‐ray radiotherapy and photothermal therapy, is devised to address the limitations of BNCT efficiency. However, the potential risk of organ‐accumulating toxicity and treatment complexity of dual exogenous activation hinders its development. To address this problem, newly redox‐responsive boron nano‐chains (RBNC) are reported that combine BNCT and endogenous chemodynamic therapy (CDT)‐enhanced ferroptosis. RBNC specifically activates nanoparticle size conversion (large‐to‐small) in response to GSH/H2O2 in the tumor microenvironment, releasing boron delivery agents boron quantum dots (BQD) and Fe3+. RBNC exhibits negligible systemic toxicity while demonstrating high boron accumulation at tumor. Meanwhile, the introduction of Fe3+ not only produces ·OH through reaction with H2O2, but also depletes GSH and reduces GPX4 activity in tumors, resulting in amplified intracellular oxidative stress and chemodynamically enhanced ferroptosis. Thus, the work provides a strategy to solve the problem of insufficient boron concentration and poor targeting of boron delivery agents and fill the gaps of BNCT combined with CDT and ferroptosis.

Application of Nanotechnology for Targeted Drug Delivery and Nontoxicity

Nanotechnology has applications in various fields of medicine. The health and biomedical fields can apply nanotechnology to treatment and drug delivery, enabling the targeted and controlled delivery of drugs and therapeutic compounds. Normally, the body quickly metabolizes drugs upon their entry, potentially affecting their efficiency. Additionally, drugs are often unable to specifically target cells, leading to harmful effects on healthy cells. Nanotechnology is currently being used to address these issues. Nanoparticles, which are tiny particles made up of either synthetic or semi-synthetic polymers, have introduced targeted drug delivery by allowing accurate and regulated secretion of therapeutic agents at specific activity sites. Their efficiency depends on features such as size, shape, surface, charge, and loading techniques. By utilizing their distinct attributes, nanoparticles can overcome biological barriers, improving the bioavailability of drugs and decreasing systemic toxicity. However, excessive use of nanotechnology also raises concerns about its potential nanotoxicity. The interaction between biological systems and nanoparticles can lead to hazardous effects such as genotoxicity, oxidative stress, inflammation, and neurotoxicity. Thus, it is important to examine the nanotoxicity of nanoparticles and develop various ways to diminish their toxic effects. This review aims to summarize the use of nanoparticles for drug delivery to specific sites, as well as their nanotoxicity.

Signature of click chemistry in exosome modification for cancer therapeutic

AbstractExosomes, small extracellular vesicles secreted by cells, have gained attention as potential therapeutic agents due to their natural ability to deliver biomolecules and traverse biological barriers. However, their limited targeting specificity and payload capacity necessitate modifications for improved therapeutic efficacy. Click chemistry, known for its high specificity, efficiency, and mild reaction conditions, offers an innovative solution for modifying exosomal surfaces. This technique enables precise attachment of targeting ligands, imaging agents, and therapeutic molecules, enhancing the targeting, delivery, and overall effectiveness of exosome‐based therapies. By addressing cancer heterogeneity, click chemistry‐modified exosomes can target diverse cancer cell populations within tumors, improving treatment specificity and reducing drug resistance. The development of copper‐free click chemistry, such as strain‐promoted azide‐alkyne cycloaddition (SPAAC), minimizes toxicity, ensuring biocompatibility and safety. As research progresses, this approach holds great promise for personalized and effective cancer treatment, paving the way for next‐generation therapeutics and diagnostics.

Highly Drug-Loaded Nanoaggregate Microparticles for Pulmonary Delivery of Cyclosporin A.

Nanoparticles have the advantages of improving the solubility of poorly water-soluble drugs, facilitating the drug across biological barriers, and reducing macrophage phagocytosis in pulmonary drug delivery. However, nanoparticles have a small aerodynamic particle size, which makes it difficult to achieve optimal deposition when delivered directly to the lungs. Therefore, delivering nanoparticles to the lungs effectively has become a popular research topic. Nanoaggregate microparticles were used as a pulmonary drug delivery strategy for the improvement of the bioavailability of cyclosporine A (CsA). The nanoaggregate microparticles were prepared with polyvinyl pyrrolidone (PVP) as the excipient by combining the anti-solvent method and spray drying process. The physicochemical properties, aerodynamic properties, in vivo pharmacokinetics and inhalation toxicity of nanoaggregate microparticles were systematically evaluated. The optimal nanoparticles exhibited mainly spherical shapes with the particle size and zeta potential of 180.52 nm and -19.8 mV. The nanoaggregate microparticles exhibited irregular shapes with the particle sizes of less than 1.6 µm and drug loading (DL) values higher than 70%. Formulation NM-2 as the optimal nanoaggregate microparticles was suitable for pulmonary drug delivery and probably deposited in the bronchiole and alveolar region, with FPF and MMAD values of 89.62% and 1.74 μm. In addition, inhaled NM-2 had C max and AUC0-∞ values approximately 1.7-fold and 1.8-fold higher than oral cyclosporine soft capsules (Neoral®). The inhalation toxicity study suggested that pulmonary delivery of NM-2 did not result in lung function damage, inflammatory responses, or tissue lesions. The novel nanoaggregate microparticles for pulmonary drug delivery could effectively enhance the relative bioavailability of CsA and had great potential for clinical application.

Engineered extracellular vesicle-delivered TGF-β inhibitor for attenuating osteoarthritis by targeting subchondral bone

Osteoarthritis (OA) is a disease that affects the entire joint. To treat OA, it may be beneficial to inhibit the activity of TGF-β in the subchondral bone. However, delivering drugs to the subchondral bone using conventional methods is challenging. In this study, we developed an extracellular vesicle delivery system. The utilization of macrophage-derived extracellular vesicles as a drug-carrying platform enables drugs to evade immune clearance and cross biological barriers. By incorporating targeting peptides on the surface of extracellular vesicles, the drug platform becomes targeted. The combination of these two factors results in the successful delivery of the drug to the subchondral bone. The study evaluated the stability, cytotoxicity, and bone targeting capability of the engineered extracellular vesicle platform (BT-EV-G). It also assessed the effects of BT-EV-G on the differentiation, proliferation, and migration of bone mesenchymal stem cells (BMSCs). Additionally, the researchers administered BT-EV-G to anterior cruciate ligament transection (ACLT)-induced OA mice. The results showed that BT-EV-G had low toxicity and high bone targeting ability both in vitro and in vivo. BT-EV-G can restore coupled bone remodeling in subchondral bone by inhibiting pSmad2/3-dependent TGF-β signaling. This work provides new insights into the treatment of OA.

A systematic review and meta-analysis of clinical trials assessing safety and efficacy of human extracellular vesicle-based therapy.

Nowadays, it has become clear that extracellular vesicles (EVs) are not a cellular waste disposal vesicle but are an essential part of an intercellular communication system. Besides the use of EVs in biomarker studies and diagnostics, the potential of EV-therapeutics has been seen by many. They provide unique properties for disease therapy, including strong immune-modulatory actions, the possibility of engineering, low immunogenicity, and the capability of crossing biological barriers. Proof-of-concept of EV-therapeutics for various pathologies has been achieved in preclinical studies. However, clinical trials with EVs have only been emerging slowly. Here, we aim to provide a comprehensive overview of the current state-of-the-art concerning clinical studies using EVs in human therapy. By approaching the current knowledge in a systematic manner, we were able to include 21 reports for meta-analysis of safety and evaluation of efficacy outcomes. Overall, we have shown that EV-based therapy is safe with a low incidence of serious adverse events (SAE; 0.7% (95%-CI: 0.1-5.2%), and adverse events (AE; 4.4% (95%-CI: 0.7-22.2%). Subgroup analysis showed no significant difference in SAE when comparing autologous versus allogeneic administration, as well as engineered versus non-engineered EV products. A significantly higher number of AE was seen in autologous versus allogeneic administration. However, the clinical relevance remains questionable. Evaluation of the clinical outcomes of immunostimulatory, immunosuppressive or regenerative EV-therapies indicated improvement in the majority of treated patients. Despite these promising results, data need to be approached with caution due to a high heterogeneity in the EVs manufacturing methods, study design, and reporting of (S)AE. Overall, we conclude that EV-based therapy is safe and presents a promising opportunity in therapy. More efforts are needed in the standardization and harmonization of reporting of EV isolation and characterization data as well as in the reporting of (S)AE to allow inter-study comparison.

Drug Delivery Systems based on Microneedles for Dermatological Diseases and Aesthetic Enhancement.

Microneedle (MN) devices comprise of micron-sized structures that circumvent biological barriers in a minimally invasive manner. MN research continues to grow and evolve; the technology was recently identified as one of the top ten overall emerging technologies of 2020. There is a growing interest in using such devices in cosmetology and dermatological conditions where the MNs mechanically disrupt the outer skin barrier layer, creating transient pathways that allow the passage of materials to underlying skin layers. This review aims to appraise the application of microneedle technologies in skin science, provide information on potential clinical benefits, as well as indicate possible dermatological conditions that can benefit from this technology, including autoimmunemediated inflammatory skin diseases, skin aging, hyperpigmentation, and skin tumors. A literature review was carried out to select studies that evaluated the use of microneedles to enhance drug delivery for dermatologic purposes. MN patches create temporary pathways that allow the passage of therapeutic material to deeper layers of the skin. Given their demonstrable promise in therapeutic applications it will be essential for healthcare professionals to engage with these new delivery systems as they transition to the clinic.

Exosomal microRNAs in lung cancer: a narrative review.

Exosomes are nanoscale extracellular vesicles secreted by cells, which can release bioactive macromolecules, such as microRNA (miRNA) to receptor cells. Exosomes can efficiently penetrate various biological barriers which mediate intercellular communication. MiRNA are a class of non-coding RNA that primarily regulate messenger RNA (mRNA) at the post-transcriptional level. MiRNA is abundant in exosomes, which plays an important role by being transported and released through exosomes secreted by lung cancer cells. This review aims to elucidate the roles of exosome-derived miRNAs in lung cancer. We focused on the roles of exosome-derived miRNAs in cancer occurrence and development, including angiogenesis, cell proliferation, invasion, metastasis, immune escape, drug resistance, and their clinical value as new diagnostic and prognostic markers for lung cancer. Exosomal miRNA can not only affect angiogenesis of lung cancer, induce epithelial-mesenchymal transformation, and promote reprogramming of tumor microenvironment, but also affect immune regulation and drug resistance transmission and participate in regulating lung cancer cell proliferation. Therefore, understanding the regulatory roles of exosomal miRNAs in tumor invasion and metastasis can provide new ideas for the treatment of lung cancer. Exosomal miRNA can provide some unique ideas on how to improve the efficiency of diagnosis and treatment of lung cancer in the future. Targeting tumor-specific exosomal miRNA represents a new strategy for clinical treatment of lung cancer, which can provide potential non-invasive biomarkers in the early diagnosis of lung cancer. Investigation of the involvement of exosomal miRNAs in the occurrence and progression of tumors can yield new opportunities for the clinical diagnosis and treatment of lung cancer.

Development and evaluation of self-assembling nanovaccine for immune protection against spring viremia of carp

Self-assembled peptides have become a hot topic in research on new fishery-immersed nanovaccines due to their physical and chemical properties and efficient carrying capacity. Using solid-phase synthesis, this study constructed the self-assembled nanovaccine SVG-Q11 for SVCV, assessing its safety at cellular and individual levels and comprehensively evaluating its preventive efficacy through immunization and immune challenge tests. Experimental results show that the constructed SVG-Q11 nanovaccine can be formed by self-assembly. At the cellular level, the survival rates of carp macrophages and EPC cells in each treatment group were higher than 90%. At the individual level, at the highest treatment concentration of 40 mg/L, there was no significant difference in the growth indicators and organ health of carps after immersion in SVG-Q11. The expression levels of non-specific immunity and immune-related genes in the SVG-Q11 group were significantly higher than those in the other groups. After challenge, the SVG-Q11 vaccine at the highest concentration (20 mg/L) achieved a 73% immune protection rate. The SVG-Q11 self-assembled nanovaccine can also effectively penetrate biological barriers, enhance antigen presentation, and improve immune efficacy. This study innovatively provides a theoretical basis for the research on self-assembled nanovaccines in aquaculture.

Chinese hamster ovary cell line engineering strategies for modular production of custom extracellular vesicles.

Continuously secreted by all cell types, extracellular vesicles (EVs) are small membrane-bound structures which shuttle bioactive cargo between cells across their external environment. Their central role as natural molecular messengers and ability to cross biological barriers has garnered significant attention in the use of EVs as therapeutic delivery vehicles. Still, harnessing the potential of EVs is faced with many obstacles. A cell line engineering approach can be used to exploit EVs to encapsulate a bespoke cargo of interest. However, full details regarding native EV-loading mechanisms remain under debate, making this a challenge. While Chinese hamster ovary (CHO) cells are well known to be the preferred host for recombinant therapeutic protein production, their application as an EV producer cell host has been largely overlooked. In this study, we engineered CHO DG44 cells to produce custom EVs with bespoke cargo. To this end, genetic constructs employing split green fluorescent protein technology were designed for tagging both CD81 and protein cargoes to enable EV loading via self-assembling activity. To demonstrate this, NanoLuc and mCherry were used as model reporter cargoes to validate engineered loading into EVs. Experimental findings indicated that our custom EV approach produced vesicles with up to 15-fold greater cargo compared with commonly used passive loading strategies. When applied to recipient cells, we observed a dose-dependent increase in cargo activity, suggesting successful delivery of engineered cargo via our custom CHO EVs.