Drug Delivery and Translational Research最新文献

The development of advanced topical delivery systems that enhance skin penetration and ensure controlled release of bioactives is a key focus in pharmaceutical research. Semi-solid extrusion 3D printing (SSE-3DP) has emerged as a versatile technology for fabricating topical patches, allowing precise control over internal architecture to tailor release and penetration profiles. This research investigates the integration of niacinamide (Nia) and vitamin A-palmitate (VitA) into printable gelatin-based inks, employing both pre-printing rheological assessments and post-printing structural analyses-scanning electron microscopy (SEM), micro-computed tomography (micro-CT) and mechanical properties (tension and compression). Additionally, the study evaluates how variations in patch internal design affect the release kinetics and penetration profiles of Nia and VitA, utilizing both in vitro (Franz cells and Raman Microscopy, RM) and in vivo quantitative (Confocal Raman Spectroscopy, CRS) methodologies. Results indicated a significant correlation between in vitro and in vivo data. Additionally, RM provided valuable molecular-level insights, making it an effective in vitro tool for investigating skin retention. CRS in vivo highlighted different penetration behaviors: while Nia penetration was strongly influenced by the patch design (porous vs. occlusive: 20 min, 0.074 ± 0.027 mg/cm² and 0.048 ± 0.022 mg/cm²; 40 min, 0.084 ± 0.037 mg/cm² and 0.052 ± 0.038 mg/cm²), particularly after short application times, VitA penetration was highly dependent on the integrity of the skin barrier (normal vs. slightly compromised). Notably, this work introduced the first study to apply quantitative in vivo CRS to evaluate 3D-printed topical systems, highlighting the potential of SSE-3DP as a design-driven strategy for effective and personalized topical delivery.

Laser interstitial thermal therapy (LITT) is a minimally invasive treatment for brain tumors that are recurrent or surgically inaccessible. We developed a murine model of LITT to investigate its effects on tumor burden, immune activation, and delivery of heat-activated therapeutics. We engineered a preclinical LITT system using a 1064-nm laser coupled to a 400-μm fiber-optic probe. Orthotopic gliomas were established in the right frontal cortex of BL6 mice using luciferase-transduced glioma cells. Ten days post-implantation, mice were treated with LITT (0.45 or 0.75 W). Tumor response and blood-brain barrier (BBB) disruption were assessed using bioluminescence imaging (BLI), Evans Blue dye, and histology at 3, 7, and 14 days post-treatment. Immunofluorescence (IF) staining characterized immune cell activation. The distribution of doxorubicin released from intravenously administered Thermodox^® was also evaluated. LITT disrupted the BBB, enabling Evans Blue dye and doxorubicin penetration up to 4 mm from the probe. Tumor burden was reduced by LITT, as shown by decreased hypercellularity on H&E and reduced BLI signal, while sham-treated mice showed tumor progression. A reproducible ablation zone formed at the probe site. IF revealed increased IBA1 + macrophages and T cell infiltration in LITT-treated brains. Thermodox^®-derived doxorubicin distribution correlated with thermal diffusion and matched a Fickian perfusion model. We present a reproducible preclinical model of LITT that enables investigation of tumor ablation, immune modulation, and thermally triggered drug delivery. These findings support the use of LITT as a platform for combinatorial strategies in glioma treatment.

Microneedle (MN) arrays bypass the skin's stratum corneum barrier to deliver drugs directly into the viable tissue. The skin disposition of three types of MNs-dissolvable, degradable and hydrogel-forming, fabricated using different polymers-have been evaluated post-treatment with examples of these MN arrays and then examined by confocal Raman spectroscopy and stimulated Raman scattering (SRS) microscopy. The presence of the polymers was assessed from their characteristic Raman signals. SRS image mosaics were acquired to survey and visualise larger areas of the skin surface. After MN insertion, the skin's spectrum was acquired using confocal Raman spectroscopy at the surface, and at nominal depths of 50 µm, 100 µm, and 150 µm. For dissolvable and degradable MNs, Raman signals from the constituent polymers were detectable in the skin. However, the polymer used to form the hydrogel MNs was not detectable under the experimental conditions used. SRS confirmed that the MN arrays penetrated the skin in a reasonably uniform manner. In summary, polymeric MN insertion into the skin has been visualised using confocal Raman spectroscopy and SRS microscopy. Together, these techniques have the potential to shed light on the spatial and temporal skin disposition of the constituent MN polymers used.

Microneedle patches, which are minimally invasive and have high patient compliance, offer a potential alternative for transdermal drug delivery, particularly in chronic diseases such as osteoporosis, which require consistent long-term therapy. Genistein is an osteoprotective plant-derived isoflavone that suffers from poor aqueous solubility and low bioavailability, thereby limiting its medicinal value. In this study, a genistein-loaded solid lipid nanoparticles integrated with dissolving microneedles was developed for the management of osteoporosis. SLNs were prepared using glyceryl monostearate and Tween 80, and optimized by using a Box-Behnken design. The optimised formulation of SLNs (F3) was incorporated into dissolving microneedles. The developed dissolving microneedles were studied for various characterisations, and they featured uniform geometry, sharp, pointed edges, a drug delivery profile of 69.218 ± 3.64% at 8 h. Its anti-osteoporotic studies were carried out on 6 groups of Wistar albino rats, and studies showed that significant decrease in the creatinine level and osteoclast cells; on the other hand, a substantial increase in total protein, bone weight, hardness, thickness, and trabecular composition. A skin irritation test, which was performed, manifested the biocompatible, dermatologically safe quality of the developed dissolving microneedles. This microneedle platform, coupled with an optimised SLNs carrier, ensures a non-invasive controlled transdermal delivery, maximising genistein's osteogenic potency for the effective management of osteoporosis.

With nearly half of all pregnancies occurring unintended, effective and acceptable contraceptive options remain a global necessity. Though contraceptives with extended durations of action reduce the need for strict daily adherence, thus enhancing compliance and reliability, only one of these methods - the copper intrauterine device (IUD) - prevents pregnancy without administering exogenous hormones. Herein, we demonstrate co-delivery of two nonhormonal contraceptive agents, lactic acid (LA) and glycerol monolaurate (GML), using a next-generation 3D-printed intravaginal ring (IVR). Through alterations in ring properties and drug loading, a range of LA and GML release rates were achieved in vitro, demonstrating the flexibility of the platform technology. Rings elicited sustained release of LA and GML at target release rates over 30 days or longer. Additionally, these studies explored how drugs with different physiochemical properties interact within the IVR matrix and further elucidated IVR drug loading and release mechanisms.

Human microbiota is now recognized as a fundamental organ of the body. In its healthy state, it fulfills essential local and systemic functions, whereas dysbiosis disrupts these roles and can contribute to disease. Although numerous studies have examined the relationship between microbiota and cancer, often revealing conflicting mechanisms and outcomes, this work has focused almost exclusively on the gut, leaving the lung microbiota largely unexplored. In this project, a ferrocifen compound was selected as an anticancer agent for lung cancer therapy. We found that lung microbiota actively degraded the ferrocifen. To prevent this degradation, the antibacterial peptide buforin II was synthesized, purified, and characterized. After confirming its antimicrobial activity, it was covalently conjugated to the ferrocifen, yielding an amphiphilic bioconjugate. This prodrug was subsequently formulated into self-assembled structures to enhance ferrocifen solubility and bioavailability. The resulting self-assemblies were evaluated in an orthotopic murine model of lung cancer and administered via nebulization to assess their therapeutic efficacy. A significant reduction in tumor progression and an improved predicted survival in mice were obtained. Together, these findings highlight the capacity of the lung microbiota to interfere with anticancer therapies and underscore the importance of considering this flora when designing treatment strategies for lung cancer.

Leveraging the intrinsic platelet-cancer cell crosstalk through platelet membrane-coated nanosystems offers a promising avenue for targeted drug delivery, particularly against triple-negative breast cancer, the most aggressive highly metastatic breast cancer subtype. In parallel, repurposing the antiparasitic ivermectin (Ivm) for anticancer applications is hindered by poor solubility and high toxicity, restricting its parenteral administration. In this study, ivermectin nanocrystals (Ivm-NC) were first developed to enhance drug solubility and systemic delivery. Afterwards, platelet-membrane was employed for realizing platelet-mimetic-camouflaged PMV/Ivm-NC for active-targeted tumor homing, immune evasion and higher biocompatibility. The innovative PMV/Ivm-NC presented optimum particle-size and zeta-potential, while exhibiting a sustained release pattern. Successful coating and retention of platelet membrane proteins was confirmed by SDS-PAGE profiling and immunocytochemistry for the platelet-membrane-protein P-selectin. In vitro studies for PMV/Ivm-NC demonstrated higher selective-cytotoxicity (IC₅₀ 2.89 ± 0.38 µg/mL) and anti-migratory potential on MDA-MB-231 cells, and cytocompatibility on normal human fibroblasts vs. uncoated Ivm-NC. In 4T1-tumor-bearing BALB/c mice, PMV-functionalization fostered preferential tumor-homing and reduced off-target distribution, compared to uncoated-NC. In addition, PMV/Ivm-NC secured pronounced tumor-growth inhibition, down-regulation of oncogenic markers (VEGF and cyclin D1), upregulation of pro-apoptotic Bax and caspase-3, and enhanced immune-infiltration of CD4⁺ and CD8⁺ T-cells, suggesting Ivm-induced immunogenic cell death. Histological evaluation confirmed higher tumor-necrosis and lower mitotic-count, as well as a notable lung-antimetastatic activity. Serum biochemistry and histopathology confirmed favorable biocompatibility. Together, our findings highlight PMV/Ivm-NC as a promising biomimetic-camouflaged nanoplatform for harnessing Ivm repurposed anticancer immunotherapy and reducing possible toxicity with selective, active targeting of triple negative breast cancer.

Intraoperatively applied local drug delivery systems (LDDS) offer a means of overcoming blood-brain barrier (BBB) impermeability. However, there is a paucity of LDDS development for paediatric tumours arising in the posterior fossa. Here we demonstrate applicability of an LDDS against medulloblastoma group 3 (G3 MB) and atypical teratoid/rhabdoid tumours (AT/RT), neoplasms associated with poor prognoses. A poly(ethyleneglycol)-poly(caprolactone)-poly(ethyleneglycol) (PECE) hydrogel loaded with chemotherapeutics identified as effective against primary G3 MB and AT/RT in vitro, was prepared as an injectable, biodegradable formulation. CHIR99021 (glycogen synthase kinase-3 inhibitor), ribavirin (guanosine analogue) and PG545 (heparanase inhibitor) were chosen based upon an inability to traverse the BBB. The hydrogel alone was well-tolerated, and drug-loaded hydrogel achieved > 1-month therapeutic release. Orthotopic xenograft studies against G3 MB and AT/RT indicated good tolerability to combined CHIR99021 and PG545 or combined CHIR99021 and ribavirin loaded loaded LDDS respectively. Median survival of AT/RT arms receiving XRT alone was comparable to CHIR99021- and ribavirin-loaded LDDS, with long-term survivors observed only in the latter arm, demonstrating a significant survival benefit. LDDS against cerebellar tumours using PECE offers a promising therapeutic alternative and the possibility of circumventing radiation-induced adverse effects for children impacted by these diseases.

Mucosal vaccination generates protective immune responses directly at the primary site of STI infection. However, the delivery of nanoparticles is hindered by the mucus barrier at these mucosal surfaces. Due to this interference, research on mucosal administration of self-amplifying (sa)-mRNA encapsulated in lipid nanoparticles (LNP) is currently limited and inconsistent. Some progress has been reported for nasal mRNA vaccination. However, for STIs, protective immune responses are required at the urogenital tract, which is achieved through intravaginal or intranasal administration. Therefore, in this research, we aimed to determine whether an sa-mRNA-LNP reporter vaccine could be effectively administered mucosally, evaluating its potential as a novel platform for STI vaccination. The sa-mRNA luciferase construct was encapsulated in two LNP formulations. In vitro studies demonstrated that these formulations maintained their potency after being sprayed with different sprayers and exposed to different mucus solutions, except for a human cervicovaginal simulant. Next, pigs received 15 µg of the sa-mRNA intravaginally and intranasally through a mucosal spray or injection. The mucosal spray resulted in expression and uptake only at the vaginal mucosa, whereas injection of the formulations resulted in expression at both mucosal sites. However, expression after spraying in the vaginal mucosa disappeared by day 4 post-administration. No differences were observed between both LNP formulations. These findings demonstrate that sa-mRNA can be used for mucosal administration, and expression can be achieved in a more relevant animal model. However, additional research is needed to develop more suitable particles for these complex environments.

Neurodegenerative diseases are increasingly significant causes of mortality and morbidity worldwide, particularly among the elderly. Despite their widespread prevalence, effective treatment options remain inadequate. A significant challenge contributing to this therapeutic gap is the impermeability of the blood-brain barrier to many drugs. Thus, developing new strategies to bypass this barrier and deliver therapeutic agents to the central nervous system (CNS) is crucial. The intranasal (IN) route has emerged as a promising approach in animal models of neurodegenerative diseases. This method of administration is gaining attention as a viable alternative for delivering various pharmacological agents, including proteins, miRNA, and oligonucleotides, to the CNS. It offers advantages over oral and intravenous routes. However, translating IN formulations from preclinical models to clinical practice presents several challenges. Assessing the adequacy of current clinical trials in evaluating IN delivery efficacy is crucial. Furthermore, the introduction of novel formulations such as nanoparticles sparks excitement for enhancing the effectiveness of IN drug administration compared to traditional free drug solutions. This review summarizes recent advancements in delivering therapeutic molecules to the CNS to treat neurodegenerative diseases. We explore critical strategies to overcome the blood-brain barrier obstacle, focusing on recent progress using the IN route as a potential avenue for effective neurodegenerative disease therapies. Additionally, we will delve into the preclinical studies that have provided the basis for the clinical trials conducted.