Delivery Strategies Research Articles

Harnessing the Potential of Human Breast Milk to Boost Intestinal Permeability for Nanoparticles and Macromolecules.

The intricate interplay between human breast milk, nanoparticles, and macromolecules holds promise for innovative nutritional delivery strategies. Compared to bovine milk and infant formula, this study explores human breast milk's role in modulating intestinal permeability and its impact on nanoparticle and macromolecule transport. Comparative analysis with bovine milk and infant formula reveals significant elevations in permeability with human breast milk, accompanied by a decrease in transepithelial electrical resistance, suggesting enhanced paracellular transport. Mechanistically, human breast milk reduces Zonula occludens-1 levels, suggesting a regulatory role in intestinal barrier function. Through in vitro and ex vivo evaluations, we aim to understand better the mechanisms behind enhanced permeability and how human breast milk affects nanoparticle physicochemical properties, potentially modulating their behavior. Specifically, human breast milk improves the intestinal permeability of liposomes in a porcine intestinal model, with associated changes in the composition of milk proteins corona related to liposome charge. These findings underscore the unexploited potential of human breast milk in facilitating transport across the intestinal barrier, offering novel avenues for human nutritional delivery and therapeutic interventions.

Advances in IgY antibody dosage form design and delivery strategies: Current status and future perspective.

Immunoglobulin Y (IgY), a unique type of antibody found in birds, is attracting increasing attention for a broad range of biomedical applications. Rational IgY protection, dosage form design, and delivery are highly essential to transform functional IgY antibodies into desired IgY products for therapeutic and prophylactic administration. Although progress has been made in this field, it remains in the early stages, highlighting the fundamental research and development needed in this aspect of IgY technology. Hence, this article reviews the conventional and innovative IgY dosage designs and delivery strategies, emphasizes the challenges faced in various IgY delivery systems, discusses the criteria for evaluating IgY dosage form performance, and provides a comprehensive analysis of the current research status and prospects of IgY delivery strategies.

Formulation Advances in Posterior Segment Ocular Drug Delivery.

Posterior segment ocular diseases, such as diabetic retinopathy, age-related macular degeneration, and retinal vein occlusion, are leading causes of vision impairment and blindness worldwide. Effective management of these conditions remains a formidable challenge due to the unique anatomical and physiological barriers of the eye, including the blood-retinal barrier and rapid drug clearance mechanisms. To address these hurdles, nanostructured drug delivery systems are proposed to overcome ocular barriers, target the retina, and enhance permeation while ensuring controlled release. Traditional therapeutic approaches, such as intravitreal injections, pose significant drawbacks, including patient discomfort, poor compliance, and potential complications. Therefore, understanding the physiology and clearance mechanism of eye could aid in the design of novel formulations that could be noninvasive and deliver drugs to reach the target site is pivotal for effective treatment strategies. This review focuses on recent advances in formulation strategies for posterior segment ocular drug delivery, highlighting their potential to overcome these limitations. Furthermore, the potential of nanocarrier systems such as in-situ gel, niosomes, hydrogels, dendrimers, liposomes, nanoparticles, and nanoemulsions for drug delivery more effectively and selectively is explored, and supplemented with illustrative examples, figures, and tables. This review aims to provide insights into the current state of posterior segment drug delivery, emphasizing the need for interdisciplinary approaches to develop patient-centric, minimally invasive, and effective therapeutic solutions.

Natural phytochemicals reverting M2 to M1 macrophages: A novel alternative Leishmaniasis therapy.

Leishmaniasis is a tropical parasitic disease caused by the protozoan Leishmania which remains a significant global health concern with diverse clinical manifestations. Transmitted through the bite of an infected sandfly, its progression depends on the interplay between the host immune response and the parasite. The disease outcome is linked to macrophage polarisation into M1 and M2 phenotypes. M1 macrophages are pro-inflammatory and promote parasite clearance, while M2 macrophages support tissue repair and parasite survival by facilitating promastigote entry and intracellular amastigote proliferation. The review focuses on discovering novel phytochemicals that exploit the immunomodulatory properties of macrophages, which can serve as an alternative antileishmanial treatments due to their diverse chemical structures and ability to modulate immune responses. It examines the immunomodulatory effects of phytochemicals that directly or indirectly promote antileishmanial activity by influencing macrophage polarisation and cytokine secretion. They can induce M1 macrophage polarisation to directly combat leishmaniasis or suppress M2 macrophages, thereby exerting indirect antileishmanial activity by influencing the release of M1- and M2-related cytokines. & Discussion: Phytochemicals demonstrate antileishmanial effects through ROS production, M1 activation, and cytokine modulation. They regulate M1/M2-related cytokines and macrophage activity, influencing immune responses. Although their effects may be non-specific, targeted delivery strategies could overcome current therapeutic limitations, positioning phytochemicals as promising candidates for leishmaniasis treatment to counter the limitations of current medications.

Navigating Tumor Microenvironment Barriers with Nanotherapeutic Strategies for Targeting Metastasis.

Therapeutic strategy for efficiently targeting cancer cells needs an in-depth understanding of the cellular and molecular interplay in the tumor microenvironment (TME). TME comprises heterogeneous cells clustered together to translate tumor initiation, migration, and proliferation. The TME mainly comprises proliferating tumor cells, stromal cells, blood vessels, lymphatic vessels, cancer-associated fibroblasts (CAFs), extracellular matrix (ECM), and cancer stem cells (CSC). The heterogeneity and genetic evolution of metastatic tumors can substantially impact the clinical effectiveness of therapeutic agents. Therefore, the therapeutic strategy shall target TME of all metastatic stages. Since the advent of nanotechnology, smart drug delivery strategies are employed to deliver effective drug formulations directly into tumors, ensuring controlled and sustained therapeutic efficacy. The state-of-the-art nano-drug delivery systems are shown to have innocuous modes of action in targeting the metastatic players of TME. Therefore, this review provides insight into the mechanism of cancer metastasis involving invasion, intravasation, systemic transport of circulating tumor cells (CTCs), extravasation, metastatic colonization, and angiogenesis. Further, the novel perspectives associated with current nanotherapeutic strategies are highlighted on different stages of metastasis.

MicroRNAs in Parkinson's disease: From pathogenesis to diagnostics and therapeutic strategies.

Parkinson's disease (PD) is a prevalent neurodegenerative disorder characterized by pathological changes, including the loss of dopaminergic neurons and abnormal aggregation of α-synuclein (α-syn). Certain cellular and molecular events are involved; however, the origin and significance of these events remain uncertain. The discovery of microRNAs (miRNAs) predicted to play a pivotal role in various regulatory processes has emerged. Studies on the dysregulation of miRNAs in PD pathogenesis, diagnosis, and treatment have recently gained attention. This review aims to encapsulate recent research developments concerning the function of miRNAs in the pathophysiology of PD and their prospective applications as diagnostic and therapeutic biomarkers, targets, and pharmaceuticals. The most effective drug delivery approach for the treatment of PD, transnasal-cerebral drug delivery, has also been briefly described. The advantage of this delivery strategy is its capacity to bypass the blood-brain barrier, enabling direct administration of medication to the brain, which improves therapeutic efficacy and minimizes side effects.

Emerging roles of exosomes in diagnosis, prognosis, and therapeutic potential in ovarian cancer: a comprehensive review.

Ovarian cancer is a leading cause of cancer-related deaths in women, and the development of chemoresistance remains a major challenge during and after its treatment. Exosomes, small extracellular vesicles involved in intercellular communication, have emerged as potential biomarkers and therapeutic targets in ovarian cancer. This review summarizes the current literature on differences in exosomal protein/gene expression between chemosensitive and chemoresistant ovarian cancer, and the effects of exosomal modifications on chemotherapeutic response. Clinical studies have identified alterations in several exosomal components from ovarian cancer tissues and serum samples arising as a consequence of chemosensitivity, which indicates their potential usefulness as potential biomarkers for predicting the development of chemoresistance. Interventional investigations from in vitro and in vivo studies demonstrated that modulation of specific exosomal components can influence ovarian cancer cell phenotypes and individual responses to chemotherapy. Exosomal delivery of chemotherapeutic agents, such as cisplatin, has presented as a potential targeted drug delivery strategy for overcoming chemoresistance in preclinical models. In summary, this review highlights the potential for exosomal proteins and genes to be useful biomarkers for predicting chemotherapy response and being therapeutic targets for overcoming chemoresistance in ovarian cancer. However, future research is still needed to validate these findings and explore the clinical utility of exosomal biomarkers and therapeutics in ovarian cancer management. In addition, understanding the molecular mechanisms underlying exosome-mediated chemoresistance may provide valuable insights for the development of personalized therapeutic strategies, improving outcomes for patients with ovarian cancer.

Breast milk delivery of an engineered dimeric IgA protects neonates against rotavirus.

Dimeric IgA (dIgA) is the dominant antibody in many mucosal tissues. It is actively transported onto mucosal surfaces as secretory IgA (sIgA) which plays an integral role in protection against enteric pathogens, particularly in young children. Therapeutic strategies that deliver engineered, potently neutralizing antibodies directly into the infant intestine through breast milk could provide enhanced antimicrobial protection for neonates. Here, we developed a murine model of maternal protective transfer against human rotavirus (RV) using systemic administration of a dimeric IgA monoclonal antibody (mAb). First, we showed that systemically administered dIgA passively transferred into breast milk and the stomach of suckling pups in a dose-dependent manner. Next, we optimized the recombinant production of a potently RV-neutralizing, VP4-specific dIgA (mAb41) antibody. We then demonstrated that systemic administration of dIgA and IgG mAb41 in lactating dams conferred protection from RV-induced diarrhea in suckling pups, with dIgA resulting in lower diarrhea incidence from IgG. Systemic delivery of engineered antimicrobial dIgA mAbs should be considered as an effective strategy for sIgA delivery to the infant gastrointestinal tract via breast milk to increase protection against enteric pathogens.

Emerging Role of Noncovalent Interactions and Disulfide Bond Formation in the Cellular Uptake of Small Molecules and Proteins.

Intracellular delivery of proteins is an important barrier in the development of strategies to deliver functional proteins and protein therapeutics into the cells to realize their full potential in biotechnology, biomedicine, cell-based therapies, and gene editing protein systems. Most of the intracellular protein delivery strategies involve the conjugation of cell penetrating peptides to enable and enhance the permeability of plasma membrane of mammalian cells to allow proteins to enter cytosol. Small molecules conjugations such as (p-methylphenyl) glycine, pyrenebutyrate and cysteines are used for the same purpose. The molecular level interactions are governed mostly by ionic (cationic/anionic), covalent and non-covalent interactions with various molecular entities of glycocalyx matrix on plasma membrane lipid bilayer. Although the role of non-covalent interactions is not fully understood, it is intriguing to see the recent advances in the non-covalent interaction-based strategies of intracellular delivery of small molecules and proteins into mammalian cells. These are achieved by simple modification of proteins' surface with chemical moieties which can form non-covalent interactions other than hydrogen bonding. In this review, we describe the recent advances and the mechanistic aspects of intracellular delivery and role of non-covalent interactions during the cellular uptake of proteins and small molecules.

Extracellular vesicle-mediated VEGF-A mRNA delivery rescues ischaemic injury with low immunogenicity.

Lackluster results from recently completed gene therapy clinical trials of VEGF-A delivered by viral vectors have heightened the need to develop alternative delivery strategies. This study aims to demonstrate the pre-clinical efficacy and safety of extracellular vesicles (EVs) loaded with VEGF-A mRNA for the treatment of ischaemic vascular disease. After encapsulation of full-length VEGF-A mRNA into fibroblast-derived EVs via cellular nanoporation (CNP), collected VEGF-A EVs were delivered into mouse models of ischaemic injury. Target tissue delivery was verified by in situ analysis of protein and gene expression. Functional rescue was confirmed by in vivo imaging and histology. The safety of single and serial delivery was demonstrated using immune-based assays. VEGF-A EVs were generated with high mRNA content using a CNP methodology. VEGF-A EV administration demonstrated expression of exogenous VEGF-A mRNA by in situ RNA hybridization and elevated protein expression by western blot, microscopy, and enzyme-linked immunosorbent assay. Mice treated with human VEGF-A EVs after femoral or coronary artery ligation exhibited heightened neovascularization in ischaemic tissues with increased arterial perfusion and improvement in left ventricular function, respectively. Serial delivery of VEGF-EVs in injured skin showed improved wound healing with repeat administration. Importantly, as compared with adeno-associated viral and lipid nanoparticle VEGF-A gene therapy modalities, murine VEGF-A EV delivery did not trigger innate or adaptive immune responses at the injection site or systemically. This study demonstrated that VEGF-A EV therapy offers efficient, dose-dependent VEGF-A protein formation with low immunogenicity, resulting in new vessel formation in murine models of ischaemic vascular disease.

Probiotics Encapsulated via Biological Macromolecule for Neurological Therapy and Functional Food: A Review.

Probiotics are live microorganisms that confer health benefits to humans, offering significant potential for preventing and treating various diseases. Neurological disorders, driven by multifaceted factors and linked to high disability rates, have become a growing global concern, particularly in the context of an aging population. Recent studies emphasize a strong connection between dysbiosis of the gut microbiota and neurological disorders. Probiotics have emerged as promising therapeutic interventions due to their ability to modulate the gut microbiota and influence the production of key metabolites, such as short-chain fatty acids and neurotransmitters, crucial for neurological health. However, probiotic viability is often compromised, limiting their therapeutic efficacy. We propose that developing high-activity probiotic formulations, coupled with innovative delivery strategies, holds considerable promise for advancing neurological treatments. Encapsulation systems have proven effective in enhancing probiotic stability and efficacy. This review discusses advances in probiotic delivery using biological macromolecule-based encapsulation, addressing key challenges in maintaining viability during production, storage, and digestion. It also highlights emerging delivery systems, such as microencapsulation, aimed at improving stability and therapeutic effectiveness. Additionally, the review explores the potential of functional foods enriched with probiotics for neurological health. Future research should explore clinical applications of encapsulated probiotics and support the development of functional foods to enhance neurological health.

Stem Cell-Derived Extracellular Vesicle-Mediated Therapeutic Signaling in Spinal Cord Injury.

Mesenchymal stem cell-derived extracellular vesicles (MSC-EVs) have emerged as a promising therapeutic strategy for spinal cord injury (SCI). These nanosized vesicles possess unique properties such as low immunogenicity and the ability to cross biological barriers, making them ideal carriers for delivering bioactive molecules to injured tissues. MSC-EVs have been demonstrated to exert multiple beneficial effects in SCI, including reducing inflammation, promoting neuroprotection, and enhancing axonal regeneration. Recent studies have delved into the molecular mechanisms underlying MSC-EV-mediated therapeutic effects. Exosomal microRNAs (miRNAs) have been identified as key regulators of various cellular processes involved in SCI pathogenesis and repair. These miRNAs can influence inflammation, oxidative stress, and apoptosis by modulating gene expression. This review summarized the current state of MSC-EV-based therapies for SCI, highlighting the underlying mechanisms and potential clinical applications. We discussed the challenges and limitations of translating these therapies into clinical practice, such as inconsistent EV production, complex cargo composition, and the need for targeted delivery strategies. Future research should focus on optimizing EV production and characterization, identifying key therapeutic miRNAs, and developing innovative delivery systems to maximize the therapeutic potential of MSC-EVs in SCI.

Investigations of Long-Acting Formulations in Children, Adolescents, and Pregnant Women: A Systematic Review.

Background/Objectives: Long-acting and extended-release drug delivery strategies have greatly improved treatment for a variety of medical conditions. Special populations, specifically infants, children, young people, and pregnant and postpartum women, could greatly benefit from access to these strategies but are often excluded from clinical trials. We conducted a systematic review of all clinical studies involving the use of a long-acting intramuscular injection or implant in infants, children, young people, and pregnant and postpartum people. Methods: Pubmed, Embase, and Cochrane Library trials were searched. Studies published from 1980 through 2018 were included. After abstract review and duplication removal, full-text articles were obtained for further review, reviewed by two independent reviewers, and disagreements were resolved by a third reviewer. Results: a total of 101 studies of long-acting therapeutics were completed in these populations, and most (80%) of these had a sample size of <100 individuals. Therapeutics for only a small pool of indications were examined in these studies, with 72% of the studies investigating hormonal contraception or other types of hormonal treatments. Only 9.3% of the studies in children and 16.7% of the studies in pregnant people collected any pharmacokinetic (PK) data. Conclusions: Long-acting formulations may behave differently (both pharmacokinetically and pharmacodynamically) in childhood, adolescence, and pregnancy as compared to non-pregnant adulthood. Therefore, it is imperative to increase and improve upon the studies investigating long-acting formulations in order to close the knowledge gap and improve care and treatment in these special populations.

Perceptions and Use of Extended-Duration Exposure Therapy Amongst Psychologists.

Extended-duration exposure therapy, in which treatment is delivered over a single prolonged session or cluster of long-duration sessions, is a highly efficacious and efficient treatment for anxiety disorders such as specific phobias. Despite this, little is known about the use of extended-duration exposure therapy in clinical practice. In the present study we investigated the perceptions and use of extended-duration exposure therapy amongst a sample of Australian psychologists via a survey, and the Therapist Beliefs about Exposure Scale. Additionally, we compared theoretical understanding of exposure therapy, and therapy delivery strategies (using a case study vignette), between psychologists who deliver exposure therapy via an extended-duration mode, versus the more traditional multi-session mode. Extended-duration exposure therapy is widely underutilised, and this is associated with negative beliefs about exposure therapy in general, as well as several practical barriers. There were no differences in the reported theoretical mechanisms of exposure therapy between those who do and do not use extended-duration exposure therapy. However, psychologists who use extended-duration exposure therapy reported greater use of strategies with demonstrated efficacy (e.g., more intense delivery) and less use of therapy-interfering strategies (e.g., distress reduction techniques) relative to those who do not use extended-duration exposure therapy. These findings identify potential mechanisms accounting for extended-duration exposure therapy's efficiency and point to strategies that may increase the uptake of extended-duration exposure therapy in clinical practice.

Idarubicin-loaded chitosan nanobubbles to improve survival and decrease drug side effects in hepatocellular carcinoma.

Drug delivery strategies using chitosan nanobubbles (CS-NBs) could be used to reduce drug side effects and improve outcomes in hepatocellular carcinoma (HCC) treatment. To enhance their action, a targeting agent, such as the humanized anti-GPC3 antibody GC33 (condrituzumab), could be attached to their surface. Here, we investigated the use of idarubicin-loaded CS-NBs for HCC treatment and a GC33-derived minibody (that we named 4A1) to enhance CS-NB delivery. Various CS-NB formulations were prepared with or without 4A1 conjugation and idarubicin loading. CS-NBs had a positive charge and a diameter of about 360 nm. In in-vitro experiments using the HCC-like HUH7 cell line, CS-NBs showed a cytotoxic effect once loaded with idarubicin. In-vivo biodistribution in HUH7 tumor-bearing xenograft mice demonstrated that CS-NBs can accumulate in the tumor mass. This effect was enhanced by 4A1 conjugation (p = 0.0317). In HUH7 tumor-bearing xenograft mice, CS-NBs loaded with idarubicin and conjugated or not conjugated with 4A1 were both able to slow tumor growth, to increase mouse survival time compared to free idarubicin (p = 0.00044 and 0.0018, respectively) as well as to reduce drug side effects. CS-NBs loaded with idarubicin can be a useful drug delivery strategy for HCC treatment.

Constructing Activatable Photosensitizers Using Covalently Modified Mesoporous Silica.

The combination of photosensitizers (PSs) and nanomaterials is a widely used strategy to enhance PS efficacy and broaden their applicability. However, the current nanocarrier-based delivery strategies focus on conventional PSs, neglecting the critical issue of PS phototoxicity. In this study, DHUOCl-25, an activatable PS (aPS) activated by hypochlorous acid, is synthesized by combining a silicon source structure and an activation unit. DHUOCl-25 functions as a silica source for synthesizing activatable mesoporous silica nanostrctures (MSNs) using a standardized protocol, enabling the synthesis of aPS-covalently modified MSNs for a variety of biologic therapeutic applications. On one hand, the resulting nano-aPS maintains aPS functionality for antibacterial application by achieving synergistic antibacterial action via MB PDT and retained cetyltrimethylammonium bromide (CTAB) (antibacterial agent) (DHU-MSNs-2). On the other hand, the nano-aPS exhibits MSN properties for drug loading, facilitating the synergistic integration of photodynamic therapy with chemotherapy (DHU-MSNs-6) of tumors and demonstrating efficacy against the spinal metastases of lung cancer. These results validate this strategy for developing novel aPSs and expanding the application of aPSs and MSNs.

Controlled Inflammation Drives Neutrophil-Mediated Precision Drug Delivery in Heterogeneous Tumors.

Tumor heterogeneity remains a formidable obstacle in targeted cancer therapy, often leading to suboptimal treatment outcomes. This study presents an innovative approach that harnesses controlled inflammation to guide neutrophil-mediated drug delivery, effectively overcoming the limitations imposed by tumor heterogeneity. By inducing localized inflammation within tumors using lipopolysaccharide, it significantly amplify the recruitment of drug-laden neutrophils to tumor sites, irrespective of specific tumor markers. This strategy not only enhances targeted drug delivery but also triggers the release of neutrophil extracellular traps, further potentiating the anti-tumor effect. Crucially, this study demonstrates that potential systemic inflammatory responses can be effectively mitigated through neutrophil transfusion, ensuring the safety and clinical viability of this approach. In a murine breast cancer model, the method significantly impedes tumor growth compared to conventional treatments. This work offers a versatile strategy for precise drug delivery across diverse tumor types. The findings pave the way for more effective and broadly applicable cancer treatments, potentially addressing the long-standing challenge of tumor heterogeneity.

Drug delivery strategy of hemostatic drugs for intracerebral hemorrhage.

Intracerebral hemorrhage (ICH) is associated with high rates of mortality and disability, underscoring an urgent need for effective therapeutic interventions. The clinical prognosis of ICH remains limited, primarily due to the absence of targeted, precise therapeutic options. Advances in novel drug delivery platforms, including nanotechnology, gel-based systems, and exosome-mediated therapies, have shown potential in enhancing ICH management. This review delves into the pathophysiological mechanisms of ICH and provides a thorough analysis of existing treatment strategies, with an emphasis on innovative drug delivery approaches designed to address critical pathological pathways. We assess the benefits and limitations of these therapies, offering insights into future directions in ICH research and highlighting the transformative potential of next-generation drug delivery systems in improving patient outcomes.

Impact and Significance of Viral Vectors for siRNA Delivery in the Treatment of Alzheimer's Disease.

Alzheimer's disease (AD) remains a major challenge in developing effective treatments due to its complex pathophysiology, including the accumulation of amyloid-beta plaques and tau tangles. Small interfering RNA (siRNA) technology offers promise for targeted gene silencing, but effective delivery to the central nervous system remains a significant obstacle. Viral vectors have emerged as potent delivery vehicles for transporting siRNA to neural tissues. This review explores the utilization of viral vectors for siRNA delivery in AD, focusing on delivery strategies and challenges. We discuss the design and optimization of viral vectors, targeting strategies, and safety considerations. Additionally, we examine recent advancements and prospects for enhancing viral vector-mediated siRNA delivery in AD.

Focused Ultrasound and Microbubble-Mediated Delivery of CRISPR-Cas9 Ribonucleoprotein to Human Induced Pluripotent Stem Cells.

CRISPR-Cas9 ribonucleoproteins (RNPs) have been heavily considered for gene therapy due to their high on-target efficiency, rapid activity and lack of insertional mutagenesis relative to other CRISPR-Cas9 delivery formats. Genetic diseases such as hypertrophic cardiomyopathy currently lack effective treatment strategies and are prime targets for CRISPR-Cas9 gene editing technology. However, current in-vivo delivery strategies for Cas9 pose risks of unwanted immunogenic responses. This proof-of-concept study aimed to demonstrate that focused ultrasound (FUS) in combination with microbubbles can be used to deliver Cas9-sgRNA (single guide RNA) RNPs and functionally edit human induced pluripotent stem cells (hiPSCs) in-vitro, a model system that can be expanded to cardiovascular research via hiPSC-derived cardiomyocytes. Here, we first determine acoustic conditions suitable for the viable delivery of large proteins to hiPSC with clinical Definity® microbubble agents using our customized experimental platform. From here, we delivered Cas9-sgRNA RNP complexes targeting the EGFP (enhanced green fluorescent protein) gene to EGFP-expressing hiPSCs for EGFP knockout. Simultaneous acoustic cavitation detection during treatment confirmed a strong correlation between microbubble disruption and viable FUS-mediated protein delivery in hiPSCs. This study shows, for the first time, the potential for an FUS-mediated technique for targeted and precise CRISPR-Cas9 gene editing in human stem cells.