International Journal of Pharmaceutics最新文献

We have previously described a model using porcine ear skin ex vivo for longitudinal studies into the disposition of macromolecules after subcutaneous injection. Since porcine skin cannot fully mimic biological responses in human skin, we now describe an ex vivo system using "full thickness" human skin. Spongiosis and epidermal detachment were the primary endpoints to evaluate skin structural integrity over a 9-day culture period. Epidermal barrier function and basal cell proliferation were monitored using expression of claudin-1 and Ki-67, respectively. Immunofluorescent staining of type I and type III procollagens and elastin after subcutaneous injection of TGF-β3, a cross-linked hyaluronic acid hydrogel, and saline solution and "no treatment" controls, showed that the model enabled visualization of changes in extracellular matrix proteins. Semi-quantitative, automated image analysis methods using multiple ROIs were evaluated to assess signal intensity and expression area of type I procollagen but displayed high inter-regional variability due to skin sample heterogeneity. Absolute quantitative methods, e.g. RT-qPCR or ELISA, which enable determination of biomarkers at either the mRNA level or the amounts of protein expressed in the sample, could be a better reporting tool. In conclusion, we successfully developed an ex vivo "full thickness" human skin model that retained viability over 9 days and which could be deployed in combination with qualitative/quantitative methods to evaluate local biological effects of subcutaneously injected biomacromolecules.

Lipid nanoparticles (LNPs) are used to encapsulate messenger ribonucleic acids (mRNAs) and enhance mRNA vaccine efficacy by producing inflammatory mediators. However, the overproduction of inflammatory mediators via LNP injection causes severe side effects, presenting a potential limitation. To resolve this issue, we developed pH-responsive dipeptide-conjugated lipid (DPL)-based LNPs (DPL-LNPs) for efficient small interfering RNA delivery with excellent biocompatibility. In detail, we optimized the dipeptide sequence and lipid-tail length of DPL, the helper-lipid compositions, and the molecular weight and lipid-tail length of the polyethylene glycol (PEG)-lipid to achieve highly efficient and safe mRNA delivery. Our results revealed that the LNPs prepared using glutamic acid (E)- and arginine (R)-conjugated DPL (DPL-ER) displayed higher protein-expression efficacy than DPL-threonine-R- and DPL-aspartic acid-R-based LNPs. Additionally, the lipid-tail length of the C22-bearing DPL-ER (DPL-ER-C22)-based LNPs displayed higher protein-expression efficacies than their C18 (DPL-ER-C18)- and C24 (DPL-ER-C24)-based LNPs. Moreover, the DPL-ER-C22-based LNPs incorporating low-lipid-tail-length phospholipids and PEG-lipids exhibited efficient protein expression. Most importantly, the injection of optimized DPL-LNPs exhibited comparable antigen-specific antibody production levels, with significantly lower inflammatory-mediator production compared with those of the commercially available LNPs. These results indicate that DPL-based LNPs (DPL-LNPs) can be deployed as highly efficient, safe carriers for mRNA delivery for developing mRNA vaccine formulations.

Given their unique phagocytic function, inflammatory site tropism, and ability to penetrate deep into the lesion sites, macrophages are considered to have promising application potential in the field of living-cell drug delivery. The methods of drug delivery using macrophages primarily include intracellular phagocytic and extracellular drug loading. Comparatively, extracellular drug loading is potential less cytotoxicity and has minimal effects on the motility and orientation of cells. In this review, we provide an overview of the methods of extracellular drug loading, and examine the effects of the different properties of nanoformulations on extracellular drug-loaded delivery systems. In addition, we assess the prospects and challenges of a self-propelled macrophage-based drug delivery system. We hope this research contributes to optimizing the design of these drug delivery systems.

The long-acting hydromorphone loaded poly-lactic-co-glycolic acid (HM-PLGA) microspheres for chronic pain management were developed and prepared using an automatic and scalable microfluidic process system in this study. The system consists of a novel designed micro-mixer for particle generation, syringe and HPLC pumps for continuous dosing, a process Raman spectrometer as process analytical technology (PAT) tool and an automation system for programmable automatic control. With the benefits of the automation and digitalization of the system, a wide range of formulation parameters was investigated for its impact on the properties of the microspheres. The particle size of the HM-PLGA microspheres was tunable with the automatic microfluidic process system. The long-acting injectable homogeneous HM-PLGA microspheres were successfully prepared with maximum drug-loading capacity of 7.71 % and drug encapsulation efficiency of 69.40 %. The physical and chemical properties were characterized using various analytical technologies. Pharmacokinetic experiments in female ICR mice confirmed prolonged exposure in plasma compared to the HM hydrochloride injection. In vivo studies in beagle dogs showed that the HM-PLGA microspheres provided sustained drug release for over 11 days. The results demonstrated the potential of the novel automatic microfluidic process system in the development and continuous manufacturing of the particle size-controllable drug loaded microspheres.

Ovarian cancer frequently develops resistance to chemotherapy, which is driven by cancer stem cells (CSCs) and an immunosuppressive tumor microenvironment (TME) shaped by tumor-associated macrophages (TAMs). This study explored the therapeutic potential of CD44-targeted, docetaxel (DTX)-loaded poly(lactic-co-glycolic acid) (PLGA) nanoparticles (CD44-PLGA-DTX NPs) in overcoming chemoresistance in ovarian cancer. A 3D spheroidal model incorporating SKOV3 ovarian cancer cells and TAMs was developed to mimic the TME for in vitro investigations. CD44-PLGA-DTX NPs exhibited significantly enhanced cellular uptake within the macrophage-embedded SKOV3 spheroids, which resulted in reduced cell viability and a reversal of chemoresistance. Cytokine profiling further revealed that the NPs promoted TAM polarization from the protumor M2 phenotype to the antitumor M1 phenotype, thus fostering an antitumor immune environment. These findings highlight the potential of CD44-targeted NPs as a dual-targeted therapeutic strategy, targeting both CSCs-driven tumor growth and TME reprogramming, thereby improving ovarian cancer treatment outcomes.

Extensive fluid loss, tissue damage, and bacterial infection are some important aspects that need to be addressed for designing ideal burn wound dressings. Hydrogel-based dressings cater to most of these functions; additionally, the incorporation of metal oxide nanoparticles (NPs) provides antibacterial properties that enhance the performance of wound dressings. We report here for the first time, how by employing different shapes of ZnO NPs, viz quasi-spherical, floral, and rods; in hydrogels made of PVA - P(AMPS) (Poly (vinyl alcohol) (PVA) - Poly (2-Acrylamido-2-Methyl Propane Sulfonic Acid)) along with g-C₃N₄, one could correlate structure-property relationships to wound healing efficiency. The incorporation of g-C3N4 was to enhance the thermo-mechanical stability of hydrogel, Maximum tensile strength of the hydrogel was obtained for 150 mg of g-C3N4 incorporated hydrogels, same amount being used for other systems studied. The impact of the incorporation of different shapes and amounts of ZnO NPs on the hydrogels has been studied and our results show maximum swelling ability (∼110 %), high moisture retention capacity (>90 %), and moderate water vapor transmission rate (82 g/m²h) for selected systems. Among these different shapes incorporated hydrogels, remarkable enhancement in tensile strength (76 %) was observed for quasi-spherical ZnO NPs incorporated hydrogels compared to bare. These hydrogels showed high cell viability (>70 %), high antibacterial activities against E. coli and S. aureus, and high wound healing efficiency (>80 %) in an in-vivo rat model, proving their potential to be used in wound dressing applications.

Developing innovative approaches to target osteomyelitis caused by polymicrobial infections remains a significant therapeutic challenge. In this study, monodispersed chitosan nanoparticles co-loaded with antibacterial (minocycline) and antifungal (voriconazole) agents were successfully prepared. Minocycline presented higher encapsulation efficiency as compared to voriconazole. Thermostability analysis suggested interactions between the co-loaded drugs within the dual-delivery system, potentially limiting voriconazole release. The dual-loaded chitosan nanoparticles exhibited significant in vitro anti-biofilm activity, achieving up to a 90% reduction in polymicrobial biofilms of S. aureus and C. albicans. Additionally, the nanoparticles showed cytocompatibility with a human osteoblast cell line. These findings highlight the potential of this dual-delivery chitosan-based nanoparticle system to address a critical gap in osteomyelitis treatment by targeting both bacterial and fungal pathogens.