Search bioRxiv⌕ Search

bioRxiv · 10.1101/2023.09.27.559712

Dissolving microneedle array patches containing mesoporous silica nanoparticles of different pore sizes as a tunable sustained release platform

Abstract

Dissolving microneedle array patches (DMAPs) enable efficient and painless delivery of therapeutic molecules across the stratum corneum and into the upper layers of the skin. Furthermore, this delivery strategy can be combined with the sustained release of nanoparticles to enhance the therapeutic potential in a wide variety of pathological scenarios. Among the different types of nanoparticles that can be included in microneedle formulations, mesoporous silica nanoparticles (MSNs) of tuneable pore sizes constitute a promising tool as drug delivery systems for cargos of a wide range of molecular weights. However, the development of efficient methods to produce DMAP containing large amounts of MSNs of different pore sizes has not been reported. In this work, DMAP containing MSNs with varying pore sizes was prepared and characterized. After synthesizing and characterizing MSNs, the pore size of the nanoparticles (in the range of 3 to 13 nm for S-MSN and XL-MSN, respectively) was observed to influence the loading and release of both small and large molecules, using fluorescein and ovalbumin (OVA) as model cargos. Moreover, a new preparation method was developed to produce DMAP containing large amounts of these MSNs located mainly in the microneedle tips. The successful insertion of these DMAPs was confirmed in vitro (using Parafilm), ex vivo (using excised neonatal porcine skin) and in vivo (in the back of mice) models. The dissolution of the microneedles and deposition of the nanoparticles inside the skin were also confirmed both ex vivo and in vivo using fluorescent nanoparticles, with complete microneedle dissolution after 2 h of insertion in vivo. Through histological studies, the microneedle-delivered MSNs were found to end up inside antigen presenting cells in the skin tissue (either F4/80+ macrophages or CD11c+ dendritic cells). For this reason, the uptake and biological effect of the MSNs was evaluated in vitro in dendritic cells, showing that while smaller pore MSNs were taken up by cells more efficiently (with over 80 % of S-MSN uptake compared to ca. 55 % for XL-MSNs), the dendritic cells treated with OVA- loaded XL-MSNs underwent the largest degree of activation (inducing over 25 % of CD40 expression compared to less than 2 % for OVA-loaded S- MSNs). Finally, the immune response to OVA-loaded XL-MSNs in mice was evaluated after repeated administration either subcutaneously or through DMAP. The results of this experiment showed comparable levels of anti-ovalbumin immunoglobulin generation through both routes of administration (with significant production of OVA-specific IgG1 and IgG2b antibodies), highlighting the good potential of this delivery platform for vaccination or immunotherapy applications.

Source connections

Explore related subjects

Keep this discovery

Explore connections, maps & timelines

BibTeXRIS

Paris, J. L., Vora, L. K., Perez-Moreno, A. M., Naser, Y. A., Anjani, Q. K., Canas, J. A., Torres, M. J., Mayorga, C., Donnelly, R. F.. 2023-09-29. Dissolving microneedle array patches containing mesoporous silica nanoparticles of different pore sizes as a tunable sustained release platform. https://doi.org/10.1101/2023.09.27.559712

Cite the original work for its findings. Save a collection to share your selection of sources.

KEEP EXPLORING

Related preprints

Lipid-ASO therapeutics exhibit differential tissue targeted delivery upon systemic or local CNS administration

Antisense oligonucleotides (ASOs) are a powerful therapeutic modality, but their full potential is hindered by pharmacokinetic properties that affect tissue and cellular delivery. Lipid conjugation is increasingly used to modulate ASO's biodistribution and promote extrahepatic activity, yet lipid dependent effects on in vivo functional delivery, particularly in the central nervous system (CNS), remain less explored. Here, we performed a side by side in vivo comparison of cholesterol, palmitic acid (C16:0), docosanoic acid (C22:0), and eicosapentaenoic acid (C20:5) conjugated to a fully phosphorothioated 3 10 3 LNA gapmer ASO targeting the Malat1 long non coding RNA. Lipid-ASO conjugates were administered systemically or locally in the brain of mice and evaluated for tissue level and cellular level distribution by imaging, qPCR and single-cell RNA sequencing, simultaneously annotating cell origin and global transcriptional changes within the cell. Following systemic administration in mice, lipid conjugation improved overall multi organ efficacy compared to unconjugated ASO, but with pronounced tissue specific differences. Single cell sequencing of liver and heart transcriptomes revealed lipid dependent cellular uptake patterns and transcriptional responses distinct from administration of unconjugated ASO. After intracerebroventricular administration, selected fatty acid conjugates enhanced silencing in deep brain regions such as the striatum, whereas cholesterol conjugation impaired functional delivery despite increased CNS retention. Light-sheet microscopy showed restricted parenchymal penetration of cholesterol ASOs compared with broader but heterogeneous distribution of palmitic acid conjugate. Together, these findings demonstrate that lipid identity critically determines ASO efficacy, productive cellular uptake, and regional CNS engagement, emphasizing the need for context specific lipid design in ASO therapeutic development.

pharmacology and toxicology↗

Novel Dissymmetric Ionizable Lipid-Assembled Lipid Nanoparticles for Delivery of Ferroptosis-Related siRNA in Diabetic Treatment

Small interfering RNA (siRNA) enables precise post-transcriptional gene silencing for refractory diseases, yet its clinical translation remains limited by the lack of safe and efficient delivery vectors. Inspired by the dissymmetric alkyl chain architecture of natural membrane phospholipids, we designed and synthesized 34 novel ionizable lipids with dissymmetric hydrophobic tails and formulated them into lipid nanoparticles (LNPs). Through systematic physicochemical and biological assessments, we established clear structure-activity relationships and identified two lead LNPs (O14-LNP, H18a-LNP) with superior endosomal escape capacity, enhanced in vivo gene silencing potency, and favorable biosafety relative to the clinical benchmark MC3-LNP. In both streptozotocin-induced and spontaneous db/db type 2 diabetes (T2D) mouse models, lead LNPs delivering ferroptosis-related siRNAs effectively ameliorated glucose and lipid metabolic disorders, restored islet function, and alleviated hepatic steatosis. This study not only lays a theoretical foundation for the rational design of novel ionizable lipids, but also validates the therapeutic potential of siRNA therapy targeting ferroptosis, providing a versatile delivery platform and targeted therapeutic strategy for the treatment of T2D.

pharmacology and toxicology↗

Receptor activity modifying protein modulation of parathyroid hormone-1 receptor function and signalling.

Receptor activity-modifying proteins (RAMPs) are known to modulate the pharmacology and function of several G protein-coupled receptors (GPCRs), including the parathyroid hormone 1 receptor (PTH1R). However, the precise effects of different RAMPs on PTH1R signalling and trafficking remain poorly understood. Here we investigated the impact of RAMP2 and RAMP3 on PTH1R function using a range of PTHand PTH-related protein (PTHrP)-derived ligands. FRET imaging revealed that PTH1R preferentially interacts with RAMP2 and, to a lesser extent, RAMP3, but not RAMP1. Interestingly, RAMP3 co-expression resulted in reduced cell surface expression of PTH1R, suggesting a potential role in receptor trafficking or internalization. The presence of RAMP2 significantly enhanced PTH1R-mediated cAMP accumulation, {beta}-arrestin recruitment, and calcium signalling in response to PTH (1-34), PTHrP (1-34), PTH (1-84), and the PTH (1-17) analogue ZP2307. In contrast, RAMP3 co-expression attenuated or completely abolished those responses. We found that full-length PTHrP analogues, PTHrP (1-108) and PTHrP (1-141), exhibited lower potency and efficacy than PTHrP (1-34) in activating PTH1R. RAMP2 significantly increased potency and/or efficacy when compared to PTH1R alone cells, while RAMP3 significantly reduced these responses. Antibody-capture scintillation proximity assays demonstrated that RAMP2 differentially modulates G protein activation by PTH1R in a ligand-dependent manner, with PTH (1-34) and PTHrP (1-34) inducing distinct patterns of G protein subtype activation. These findings highlight the complex role of RAMPs in regulating PTH1R signalling and trafficking, revealing differential effects of RAMP2 and RAMP3 on receptor function. The data suggest that targeting the PTH1R/RAMP2 complex may be a promising strategy for developing novel bone anabolic therapies by leveraging biased agonism and functional selectivity. Further research using physiologically relevant models is needed to elucidate the therapeutic potential of this approach.

pharmacology and toxicology↗