Drug Delivery and Translational Research最新文献

SARS-CoV-2 continues to circulate globally, with persistent hospitalizations, despite a successful global vaccination strategy. We have developed highly conserved, antiviral short interfering (si)RNA and demonstrated in vivo antiviral efficacy following intranasal treatment of mice with naked siRNA. To enhance antiviral efficacy and siRNA protection, in this study we investigated the use of LNP packaging to improve delivery and efficacy. We examined three clinically approved lipid nanoparticle (LNP) formulations that mimic the compositions of Alnylam's Onpattro (MC3), Moderna's Spikevax (SM-102), and Pfizer-BioNTech's Comirnaty (ALC-0315) RNA-based therapeutics, to identify the optimal formulation for antiviral siRNA therapeutic respiratory delivery and antiviral efficacy. All LNP formulations assessed showed successful delivery of siRNA to respiratory cells in vitro and provided effective silencing of siRNA targeted SARS-CoV-2 genes. However, the MC3-based LNP-siRNA (MC3 LNP-siRNA) treatment elicited the least off-target immune activation, with no induction of interferon stimulated genes. Additionally, the MC3 LNP-siRNA remained effective when administered 24 h post-infection, significantly reducing viral RNA levels in vitro. Chemical modification of siRNA with 2'‑O‑methyl incorporation further attenuated immune activation, without compromising efficacy. In vivo intranasal delivery of MC3 LNP-siRNA was generally well tolerated, with no adverse effects on body weight or pulmonary function at therapeutic doses, although mild pulmonary leukocyte infiltration was observed at higher or repeated doses. Our study demonstrates that LNP-encapsulated and chemically modified siRNAs can provide an effective and mutation-resilient antiviral strategy. This study compares clinically relevant LNP formulations for siRNA delivery to the respiratory tract, demonstrating that MC3-based LNPs offer a promising platform for safe and effective RNA therapeutic delivery.

Vitamin B12 (cyanocobalamin) deficiency is a prevalent global health issue which has been historically treated with oral supplements or intramuscular injections. In case of pernicious anemia, there is poor absorption of Vitamin B12 in the intestine, so injection is a typical treatment option. However, injections are painful and require a trained medical professional, which can often lead to the patient visiting clinics, thereby leading to patient non-compliance. In order to overcome these disadvantages, this research investigates an alternative, minimally invasive, and patient-friendly self-administration option, transdermal delivery via dissolvable microneedle (MN) patch. We fabricate a poly (methyl vinyl ether/maleic acid) (PMVE/MA) based MN patch, a biocompatible material, for the rapid delivery of cyanocobalamin. An in-house developed polydimethylsiloxane (PDMS) mold was used to prepare the MN patch. The MN patches were characterized by optical microscopy and scanning electron microscopy (SEM) to determine MN dimensions and morphology. Mechanical strength and skin penetration ability of the MNs were assessed through a universal testing machine (UTM) and a parafilm layer model. The findings suggest that 20% PMVE/MA formulation offered sufficient mechanical strength for skin penetration and more than 90% insertion efficiency which is total number of holes created by all the MNs. Dissolution of the needles was achieved within 45 s in agarose phantom and within 120 s in porcine skin. In vitro Franz diffusion experiments revealed continuous release, achieving a cumulative release mass of ~ 229 µg of vitamin B12 within 210 min. UV-Vis and FTIR analysis confirmed excellent B12 stability in the MN patch up to 60 days. The reported observations emphasize the advantages and the translational potential of PMVE/MA-based dissolvable MN patch as a safe, effective, and user-friendly system for the transdermal delivery of vitamin B12.

Long COVID has been increasingly linked to chronic inflammatory skin conditions driven by cytokine overproduction. Topical tacrolimus, a calcineurin inhibitor, is commonly used to manage such conditions due to its immunosuppressive properties. However, due to poor dermal penetration, tacrolimus oftens produce to suboptimal efficacy and adverse effects such as local irritation and burning sensation. Effective management of chronic inflammatory skin conditions linked to long COVID necessitates targeted, controlled drug delivery into deeper skin layers to modulate excessive cytokine production and attenuate localized inflammation. This study explores fibroblast-derived small extracellular vesicles (sEVs) as a new controlled delivery vehicle for tacrolimus. The sEVs were isolated using sucrose-cushioned density ultracentrifugation and characterized by TEM, NTA, Dot blot, and MicroBCA assay, confirming their successful isolation and purity. Tacrolimus was encapsulated into sEVs via sonication, with successful drug loading confirmed by morphological and physicochemical characterization. The resulting Tac-sEVs exhibited an encapsulation efficiency of 79.19% ± 0.01. Franz diffusion studies revealed a rapid initial release within the first 10 h, followed by sustained higher release over time. Tape-stripping demonstrated significantly deeper dermal penetration of tacrolimus loaded sEVs (Tac-sEVs) compared with commercial tacrolimus ointment and free drug. Both tacrolimus and Tac-sEVs downregulated IFN-γ, GCS-F, IL-2, and IL-4 expression, indicating potent suppression of SARS-CoV-2 spike glycoprotein-induced cytokine overproduction. PKH-26 fluorescence labelling confirmed efficient cellular uptake, while cytotoxicity assays (Alamar Blue, CCK-8) showed high cell viability for both formulations. In summary, these results position Tac-sEVs as a safe and promising therapeutic platform for cytokine-driven inflammatory skin diseases associated with long COVID, meriting further clinical investigation.

Within this study, thiolated and alkenylated β-cyclodextrins were developed as novel mucoadhesive excipients capable of interacting with mucosal surfaces and prolonging the residence time of incorporated drugs at absorption sites, thereby potentially enhancing their bioavailability. For this purpose, cysteine-, cysteamine-, allylcarbamate-, and methacrylate-functionalized oligosaccharides were synthesized, and the resulting structures were identified by ¹H NMR spectroscopy, Ellman's and disulfide bond tests, and iodometry. The thiol and double bond content, as well as the stability of these functional groups at 37 °C, over 4 h, were evaluated. Alkenylated β-cyclodextrins showed significantly higher stability in aqueous solution compared to the thiolated products with oxidative sensitivity. Furthermore, the cytotoxicity and rheological, mucodiffusive, and mucoadhesive properties of the derivatives were assessed to elucidate their potential as multifunctional excipients for mucosal drug delivery. The cytotoxicity study confirmed that all derivatives were non-toxic within 4 h of incubation. Rheological measurements showed that β-CD highly modified with allylcarbamate exhibited a 15-fold increase in dynamic viscosity after incubating with intestinal mucus compared to the native β-CD. Regarding mucopenetration, β-CD cysteine was able to penetrate the mucus by more than 3% per 5 cm in 24 h. The results on porcine intestine revealed the superiority of thiolated derivatives in mucoadhesion, with at least 98% of the samples remaining on the intestinal tissue after 3 h of rinsing. For alkenylated β-CD with a similar degree of modification, low mucoadhesiveness was detected, but it was increased significantly with the degree of modification. The current study compares various mucoadhesive approaches and demonstrates that thiolated derivatives are more effective than alkenylated derivatives for mucosal drug delivery, but are less stable in physiological fluids.

Albumin-based nanoparticles (NPs) are typically synthesized by harsh conditions-based methods that limit their application in clinics and can seriously damage the entrapped drug and even their base material. Despite the potential of the use of chitosan (CS) as stabilizing agent by adapting the ionic gelation method or by adding CS as a coating to albumin NPs generated by desolvation, the influential factors of these methods have not yet been studied. In this article, these synthesis approaches have been optimized by a 2-step DoE-based methodology (a screening process with fractional designs plus a response surface methodology using central composite designs). The application of the ion gelation method to produce albumin-based NPs generates sizes from 66 to 1017 nm, PDI (polydispersity index) values of 0.3-0.6 and surface charges (ZP) from neutral to positive (> 20 mV). The fitted models of the responses depend on four factors (albumin and CS concentration, CS pH and CS:albumin mass ratio). On the other hand, the modification of the desolvation method using CS as a stabilizing coating generates 37-1305 nm NPs, with PDI between 0.4 and 0.7 and highly positive ZP (20-40 mV). In this case, the approximate models for the responses depend on four main effects (albumin and CS concentration, pH of CS and albumin:EtOH volume ratio). Furthermore, in this work the best combinations of factors and levels that allow minimizing PDI and obtaining the minimum and maximum expected values for mean size and ZP of NPs were determined for both synthesis methods. Focusing on the minimum possible PDI, the predicted values for the ion gelation- and desolvation-based methods are 0.363 and 0.341, respectively, which are achieved with values of [BSA] (mg/ml), [CS] (mg/ml), CS pH and CS:BSA or BSA:EtOH ratios (mL:mL) of {2.3,1.4,2.2,1:7.3} and {10,0.5,1.8,1:1}, respectively. These optimized conditions yield acceptable size and ZP values for the ion gelation-based (27.7 nm; 16.4 mV) and optimal values for the desolvation-based (146.2 nm; 29.5 mV).

Hair loss is a prevalent dermatological disorder that significantly affects quality of life. Among FDA approved treatments, topically applied minoxidil is commonly used. Nevertheless, its low water solubility necessitates use of irritating solvents which hinder its clinical administration. Since these solvents usually cause adverse reactions, nanotechnology-based carriers have emerged as promising strategies for topical delivery of minoxidil. Thus, we prepared topical delivery nanocarriers, i.e., chitosan-coated nanocapsules (ChiNCs), to increase percutaneous delivery through intrinsic characteristics of chitosan and to increase encapsulation of minoxidil into the nanocarriers. ChiNCs were prepared through simple nanoprecipitation and encapsulated in high amounts of minoxidil. The minoxidil-loaded ChiNCs (MXD@ChiNCs) were evaluated for their physicochemical characteristics, colloidal stability, release profile, skin penetration, cytotoxicity, proliferation, hair growth via animal, and histological analyses. MXD@ChiNCs were successfully loaded with 50 wt.% of MXD and maintained good stability. Additionally, release profile of MXD from MXD@ChiNCs exhibited sustained release behavior and improved skin penetration. Particularly, ChiNCs showed no cytotoxicity in skin-related cell lines and did not influence cell proliferation. MXD@ChiNCs demonstrated dose-dependent therapeutic efficacy and considerably boosted hair regrowth compared to other MXD formulations, as evidenced by macroscopic and histological results. These observations highlight potential of MXD@ChiNCs as topical delivery system for alopecia.

Obesity is not only a direct cause of health problems, but is also closely related to the incidence rate of a variety of noncommunicable diseases. Intervention for obesity requires high self-discipline and compliance from individuals, which often makes weight loss less smooth and prone to rebound. In this study, we prepared a dissolving microneedle (MN) loaded with compound traditional Chinese medicine (TCM). The base material was composed of a mixture of polyvinyl alcohol (PVA) and polyvinyl pyrrolidone (PVP), which met the requirements of solubility and sufficient mechanical properties. For the drug, we chose the Huatanqushi formula, which has a clear fat-reducing effect in clinical practice and is food and medicine homology. The results showed that the MN can penetrate the stratum corneum, rapidly release drugs in the dermis of the abdominal wall, alleviate the inflammatory state of local adipose tissue, regulate systemic lipid metabolism, and ultimately achieve weight loss. This MN patch is suitable for busy office workers to use conveniently, avoiding the discomfort of oral medication and the drawbacks of a large dosage and long medication time. It can effectively intervene in obesity with a minimal effect on the body.

Triple-negative breast cancer (TNBC) is highly aggressive with limited treatment options, and resistance to doxorubicin (DOX) further compromises outcomes. Cannabinoids such as cannabichromene (CBC) and cannabidiol (CBD) possess anticancer properties, but their combined effects in resistant TNBC remain unexplored. This study evaluated the antitumor efficacy of a CBC + CBD combination against DOX-resistant (DOX-RT) TNBC using in vitro, in vivo, and pharmacokinetic models. Cytotoxicity was assessed in DOX-RT MDA-MB-231 cells using 2D and 3D assays, with synergy confirmed by combination index (CI) analysis. Cell cycle and invasion assays were performed. Xenograft studies were conducted in BALB/c nude mice bearing DOX-RT tumors treated intraperitoneally with CBC (10 mg/kg), CBD (20 mg/kg), or CBC + CBD. Pharmacokinetics were evaluated in rats, complemented by GastroPlus™ simulations. CBC + CBD synergistically inhibited cell growth induced G0/G1 arrest, and reduced invasiveness by ~ 55% in a Transwell Matrigel invasion assay. In xenografts, combination therapy reduced tumor volume by two-folds compared to single treatments and fourfolds versus control. Western blotting revealed downregulation of MEK/ERK, PI3K/AKT/mTOR, Cyclin D1, CDK6, SOD2, and NF-κB. Pharmacokinetic studies showed co-administration increased Cmax and AUC without altering Tmax, supported by simulations predicting enhanced jejunal absorption. CBC + CBD co-therapy demonstrates synergistic efficacy against resistant TNBC by inhibiting oncogenic pathways and enhancing systemic exposure. This first study of its kind highlights CBC + CBD as a promising strategy to overcome DOX resistance in TNBC.

There is a poor response of prostate cancer to immunotherapies because of dysfunctions in the tumor microenvironment (TME) characteristics, like abnormal vasculature structure, stiffened stroma, increased interstitial fluid pressure (IFP), and regions of hypoxia. However, the existing computational modelings are unable to tackle this issue, as they are based on two-dimensional (2D) geometry that ignores TME properties (like TME biphasic composition) and interaction between cancer and immune cells. To address this knowledge gap, this paper offers a patient-specific multiphysics model for prostate cancer. The proposed model is based on three-dimensional (3D) geometry obtained from magnetic resonance imaging (MRI) and combines three complementary approaches to normalizing the tumor microenvironment: vascular normalization via anti-angiogenic therapy, stromal normalization via extracellular matrix softening, and immune checkpoint blockade. One important new aspect of this work is that new nanoparticle delivery models have been developed for 20-100 nm nanoparticles (NP) delivering immunotherapy agents. These equations explicitly incorporate interactions between the components of the TME and directly account for mechanical stress induced by tumor growth, enabling mathematical modeling of physical TME changes and their subsequent impact on the dynamics of immune cells (such as cytotoxic T cells (CD8 + T cells), regulatory T cells (Treg), and pro-inflammatory macrophages (M1-like)/anti-inflammatory macrophages (M2-like) and cancer cells. This capability is absent in previous models. The other important novelty is that for the first time in a prostate cancer model, factors for vascular and stromal normalization and immunotherapy have been incorporated in a 3D geometry. The parameters of this model have been optimized based on literature and preclinical trial data related to immunology and tumors. The sensitivity analysis has confirmed that all therapeutic factors, optimized vascular function (functional vessel density increases from 43 to 112 cm²/cm³), reduced stromal solid stress (decrease in shear modulus from 10.4 to 6.1 kPa), as well as a 70% reduction in IFP (from 1471 to 441 Pa), in combination contribute to a 30% increase in accumulation of nanoparticles in the tumor, 60% increase in the ratio of CD8 + /Tregs, a 45% decrease in the ratio of M1/M2 macrophages, a 15% reduction in the tumor hypoxia gradient, and a 40% decrease in the size of the tumor within 50 days. This model can thus provide a clinically applicable tool for predicting the efficacy of nano-immunotherapy in prostate cancer. Experimental confirmation is required to better evaluate NP toxicity.

Glaucoma is a leading cause of irreversible blindness, with current treatment strategies focusing on reducing the intraocular pressure (IOP). The Durysta^® (bimatoprost) intracameral implant offers a sustained-release alternative to conventional eye drops by continuously delivering medication directly into the anterior chamber (AC). This study employs advanced computational modeling using ANSYS Fluent to investigate the interaction between aqueous humor (AH) flow and drug distribution following Durysta^® implantation. Drug transport is modeled using the unsteady convection diffusion equation, and concentration profiles are analyzed at different times: 10 min, 30 min, 1 h, 1 day, 15 days, 30 days, and 60 days. The results show that AH circulation significantly influences by Durysta^® implant, with flow-driven mixing facilitating its distribution across ocular tissues. In the early phase, high concentration zones appear near the implant, while long-term simulations demonstrate sustained and uniform drug distribution throughout the AC. These findings advance the understanding of sustained intraocular drug delivery and establish a computational framework to guide the optimization of future implant designs, aiming to improve therapeutic outcomes in glaucoma management.