pH-responsive drug delivery systems, which enable site-specific drug release and reduce systemic toxicity, offer a promising strategy to exploit the acidic tumor microenvironment. We previously demonstrated the effectiveness of poly(ethylene glycol)-poly[(benzyl-L-aspartate)-co-(N-(3-aminopropyl)imidazole-L-aspartamide)] (PEG-PABI) as a pH-sensitive nanocarrier for anticancer therapy. The purpose of this study is to optimize PEG-PABI for enhanced pH-sensitive targeting and controlled drug release under acidic conditions. To optimize this design, we synthesized two structural variants of PEG-PABI, namely, linear PEG-PABI (AB type, 5 and 10 kDa) and branched PEG-(PABI)2 (AB2 type, 10 kDa), where two PABI arms are conjugated to a single PEG backbone, by tuning the polymer architecture and molecular weight. All PABI variants were investigated using molecular dynamics simulations. The simulations revealed that extended PABI chains enhanced intermolecular interactions and improved nanoparticle stability. The branched PEG-(PABI)2, which mimics phospholipid amphiphilicity, showed superior colloidal stability. All variants maintained their pH responsiveness, thereby enabling drug release under acidic conditions. Doxorubicin-loaded nanocarriers showed efficient drug encapsulation and potent anticancer effects both in vitro and in vivo. Among these nanocarriers, the branched PEG-(PABI)2 displayed the most favorable performance, with enhanced colloidal stability, efficient endosomal escape, and increased tumor accumulation. Overall, the PEG-PABI system maintained strong pH responsiveness. Notably, the structure of branched PEG-(PABI)2 resembles that of polymersomes, with dual PABI arms and a central PEG mimicking amphiphilic bilayers. These findings highlight the promise of branched PEG-(PABI)2 as an advanced system for pH-sensitive and tumor-specific drug delivery.

Sepsis is a complex and heterogeneous inflammatory and immune response, causing multi-organ dysfunction and accounting for approximately 20% of all global deaths. Currently, antibiotics and organ-supportive therapy are the major available supportive treatments for sepsis, which may be due to multiple pathways of disease progression that make them highly evasive. Besides, Food and Drug Administration-approved drugs, cells, or biologics for sepsis treatment are unavailable, and preclinical data do not seem to reduce mortality in clinical trials, indicating the need for an alternative therapeutic approach. Here, we introduce bioengineered cell-particle hybrid spheroids (HS), composed of mesenchymal stem cells (MSCs) and metformin-loaded poly (lactic-co-glycolic acid) microspheres (Met-MS) as MSCargo, which exhibit synergistic and enhanced antioxidants, antiinflammatory, and immunomodulatory activities. We demonstrated that MSCargo significantly enhances cell viability and functionality of hepatocytes, thereby protecting them from oxidative stress and inflammatory cytokine-induced damage. Furthermore, MSCargo post-transplantation via the hepatic portal vein remarkably reduced sepsis-associated organ dysfunction, including liver, kidney, and lungs through the modulation of TLR4/ NF-kB signaling pathways as well as enhancing hepatic regeneration. In summary, MSCargo holds a promising therapeutic alternative for treating sepsis as well as other inflammatory diseases by integrating immune modulation, anti-inflammatory, and regenerative effects.

Alendronate, marketed under the brand name Fosamax (R), is a bisphosphonate that has been actively used in the clinical setting for the treatment of bone disease. However, the low intestinal permeability and bioavailability of alendronate significantly limit its therapeutic effectiveness. We developed alendronate/permeation enhancer co-loaded solid lipid nanoparticles (SLNs) composed of Compritol (R) 888 ATO or GelotTM 64, Span 20, and F108, and demonstrated that the SLN formulation improved drug intestinal permeability with an excellent Papp value. Salcaprozate sodium (SNAC) was selected as the permeation enhancer because it enhanced drug transport across Caco-2 cell monolayers. Through the X-ray diffraction and differential scanning calorimetry, it was confirmed that both drugs were encapsulated in SLNs. Moreover, the alendronate/SNAC co-loaded SLNs exhibited a sustained-release profile, offering the potential to extend the therapeutic effects and reduce the dosing frequency, which is crucial for long-term osteoporosis treatment. Subsequently, the freeze-drying process improved the storage stability of SLNs, increasing their commercialization potential. This study demonstrates how SLNs and permeability enhancers can be used to bypass the drawbacks of the drug with poor permeability.

Intraportal islet transplantation is an effective beta-cell replacement therapy for restoring insulin production in patients with type 1 diabetes. However, transplanted islets are rapidly lost due to a strong immediate inflammatory reaction, termed as instant blood-mediated inflammatory reaction (IBMIR). Current preventive therapies for IBMIR face systemic side effects such as bleeding risks. Although surface islet modification with biocompatible polymers or antithrombotic molecules shows promise, the limited anchoring of these molecules on the islet surface, caused by low conjugation efficiency and complex procedures in current methods, often lead to suboptimal outcomes. In this study, a novel islet conjugation platform is presented for effectively delivering heparin (HEP) for overcoming IBMIR. The islet surface is first conjugated with chitosan microparticles (CSMP) via electrostatic interaction. HEP is then absorbed into the conjugated CSMP with a high amount (4.4 ng IEQ-1). The conjugation system is stable and highly biocompatible with cell viability and functionality in vitro. Importantly, intraportal transplantation of the HEP/CSMP-islets significantly reduces coagulation and complement activation and improves blood glucose levels, thereby achieving a higher survival rate. This strategy can provide a versatile platform to deliver various therapeutic agents, including bioactive anti-inflammatory cytokines and growth factors, leading to enhanced islet transplantation outcomes.

Antigen‐presenting cells (APCs), particularly dendritic cells (DCs), play key roles in activating T cells for enhanced immune response in cancer immunotherapy. In cancer progression, an immunosuppressive tumor microenvironment (TME) is gradually developed, shielding tumor cells from immune surveillance. One of the defects created by the TME is the presence of dysfunctional DCs, which triggers failures in antigen recognition, processing, and presentation to T cells, inducing the impairment of anti‐tumor immune responses. The demand for ex vivo T cell activation and expansion by the replacement of autologous DCs is imperative in adoptive cell therapy (ACT) due to the limited availability and the laborious isolation of natural DCs. Therefore, the fabrication of artificial APCs (aAPCs) mimicking the function of natural DCs holds promise for cancer immunotherapy, especially in ACT. This review concentrates on the design of aAPCs using the principles of cell signaling for the immunological synapse: T cell receptor (TCR)‐specific activation (signal 1), co‐stimulatory signal (signal 2), and cytokine‐mediated signal (signal 3). Particularly, the customization of size, shape, stiffness, density, and mobility of ligands, as well as the dimension of activating engagers for the optimization of aAPCs, is also discussed.

Ligand-functionalized particulates have received considerable attention for targeted drug delivery. Previous studies have reported several methods for functionalizing various molecules on the surfaces of nano- and micro- particulates which involve multiple steps, reaction buffers and chemicals, and long reaction times. Here, we report a single-step method that uses catechol chemistry to rapidly functionalize ligands on the surfaces of polymeric nanoparticles (NPs). We synthesized dopamine-conjugated poly (ethylene-alt-maleic alt- maleic acid) (D-PEMA) by performing a nucleophilic addition reaction between dopamine and poly (ethylene-alt-maleic alt-maleic anhydride) (PEMAnh). We then used D-PEMA as a stabilizer while preparing NPs which led to the construction of NPs that were non-adhesive per se but provided active sites for conjugating multiple different ligands in an alkaline buffer via Michael's addition and Schiff's base substitution reactions. The ligand-functionalized NPs were nontoxic and effectively internalized by both primary and cancer cells. In addition, NPs prepared using the modified stabilizer conjugated rapidly on the surfaces of pancreatic islets in alkaline conditions. Our study provides evidence that the modified stabilizer is versatile and has the potential for drug delivery applications and cell surface modifications.

Recently, nanovaccine-based immunotherapy has been robustly investigated due to its potential in governing the immune response and generating long-term protective immunity. However, the presentation of a tumor peptide-major histocompatibility complex to T lymphocytes is still a challenge that needs to be addressed for eliciting potent antitumor immunity. Type 1 conventional dendritic cell (cDC1) subset is of particular interest due to its pivotal contribution in the cross-presentation of exogenous antigens to CD8+ T cells. Here, the DC-derived nanovaccine (denoted as Si9GM) selectively targets cDC1s with marginal loss of premature antigen release for effective stimulator of interferon genes (STING)-mediated antigen cross-presentation. Bone marrow dendritic cell (BMDC)-derived membranes, conjugated to cDC1-specific antibody (αCLEC9A) and binding to tumor peptide (OVA257-264), are coated onto dendrimer-like polyethylenimine (PEI)-grafted silica nanoparticles. Distinct molecular weight-cargos (αCLEC9A-OVA257-264 conjugates and 2′3′-cGAMP STING agonists) are loaded in hierarchical center-radial pores that enables lysosome escape for potent antigen-cross presentation and activates interferon type I, respectively. Impressively, Si9GM vaccination leads to the upregulation of cytotoxic T cells, a reduction in tumor regulatory T cells (Tregs), M1/M2 macrophage polarization, and immune response that synergizes with αPD-1 immune checkpoint blockade. This nanovaccine fulfills a dual role for both direct T cell activation as an artificial antigen-presenting cell and DC subset maturation, indicating its utility in clinical therapy and precision medicine.

Idiopathic pulmonary fibrosis (IPF) is a chronic inflammatory and fibrotic response-driven lung disease that is difficult to cure because it manifests excessive profibrotic cytokines (e.g., TGF-β), activated myofibroblasts, and accumulated extracellular matrix (ECM). In an attempt to develop an inhalation formulation with enhanced antifibrotic efficacy, we sought to fabricate unique aerosolizable inhaled microgels (μGel) that contain nintedanib-poly(lactic-co-glycolic acid) (PLGA) nanoparticles (NPs; n-PN) and pirfenidone-liposomes (p-LP). The aero-μGel was ∼12 μm, resisted phagocytosis by alveolar macrophages in vitro and in vivo, and protected inner-entrapped n-PN and p-LP. The n-PN/p-LP@aero-μGel caused enhanced/extended antifibrotic efficacy in a bleomycin-induced pulmonary fibrosis mouse presumably due to prolonged lung residence. Consequently, the results obtained by intratracheal aerosol insufflation of our n-PN/p-LP@aero-μGel twice a week were much better than those by as many as seven doses of single or mixed applications of n-PN or p-LP. The antifibrotic/pharmacokinetic results for the n-PN/p-LP@aero-μGel included reduced fibrosis progression, restored lung physiological functions, deactivated myofibroblasts, inhibited TGF-β progression, and suppressed ECM component production (collagen I and α-SMA) along with prolonged lung retention time. We believe that our n-PN/p-LP@aero-μGel increased the local availability of both nintedanib and pirfenidone due to evasion of alveolar macrophage phagocytosis and prolonged lung retention with reduced systemic distribution. Through this approach, our inhalation formulation subsequently attenuated fibrosis progression and improved lung function. Importantly, these results hold profound implications in the therapeutic potential of our n-PN/p-LP@aero-μGel to serve as a clinically promising platform, providing significant advancements for improved treatment of many respiratory diseases including IFP.

Chemodynamic therapy (CDT) has emerged as a novel approach to overcome cancer resistance and enhance anticancer efficacy. Despite the considerable effort devoted to current chemodynamic therapeutic agents, developing efficient delivery systems to induce ferroptosis remains demanding due to their limited efficacy and lack of selectivity. Herein, an iron-based single-atom upconversion photocatalyst (UmFe-OA@hPM) mimicking natural horseradish peroxidases has been developed. This nanoformulation not only targets tumors via the existence of a hybrid platelet membrane (hPM) coating but also generates excessive hydroxyl radicals in response to both tumor microenvironment and external laser irradiation. This nanoenzyme overcomes the low tissue penetration of UV light, which sensitizes the iron-doped graphitic carbon nitride network, attributed to the unique anti-Stokes shift from infrared to UV displayed by upconversion nanoparticles. Together with an increase in intracellular polyunsaturated fatty acid accumulation induced by oleanolic acid (OA), lipid peroxidation is significantly elevated, leading to the enhancement of CDT. UmFe-OA@hPM is demonstrated to induce significant ferroptosis in vitro, superior antitumor efficacy in breast cancer mouse models, and suppression of metastasis status when incorporated with an immune checkpoint blockade. These findings provide a potential strategy for developing a precisely controlled CDT to deal with aggressive cancers, especially in combination with immunotherapy. UmFe-OA@hPM is an iron-based single-atom upconversion photocatalyst mimicking horseradish peroxidase. It targets tumors with a hybrid platelet membrane coating and generates hydroxyl radicals in response to tumor microenvironment and near-infrared irradiation via an upconversion process that sensitizes the g-C3N4-Fe network. Coupled with the increased production of polyunsaturated fatty acids induced by oleanolic acid, UmFe-OA@hPM demonstrates potent antitumor efficacy and metastasis suppression in breast cancer models. image

The precise and selective modification of the silica shell on the anisotropic structures of gold nanorods (AuNRs) is crucial for advancing their applications in areas such as catalysis, sensing, and directional self-assembly. However, a limitation of existing methods to synthesize patched silica shells on the sides of AuNRs is that they require the introduction of polymeric blocking ligands. Herein, we present a novel method for synthesizing polymer-free, side-silica-patched AuNRs (PF/side-SiO2 AuNRs). Utilizing sodium iodide as the key agent, we achieved a yield of approximately 88%. The underlying mechanism involves the curvature-induced, tip-selective chemisorption of iodide, which reduces the charge attraction between negatively charged silica precursors and the AuNR surface. A comparative study of the localized surface plasmon resonance and surface-enhanced Raman scattering (SERS) properties of PF/side-SiO2 AuNRs with those of conventional silica-coated AuNRs was conducted. The SERS signals of the PF/side-SiO2 AuNRs intensified, whereas interference by the polymeric ligand signals was absent to expose the areas near the hotspots. An efficient technique for synthesizing anisotropic silica-coated AuNRs for various applications is presented herein, and our findings offer mechanistic insights related to the core materials and anisotropic deposition of other oxides.

Purpose Our goal was to increase the targetability of our nanoparticles to specific targets to increase their penetration into the tumor and to enhance immunotherapy with ketoconazole (KTZ), BMS-202, and indocyanine green (ICG)-induced photothermal therapy (PTT) for anticancer effects. Methods We prepared liposomes through a thin film formation process in which hydrophobic drugs BMS-202 and KTZ were embedded in a lipid bilayer, and ICG was embedded inside the liposomes as a hydrophilic drug. In the process, the cell membranes of specific target cancer cells were fused to increase penetration and targeting. The resulting nanoparticles were evaluated in vitro and in vivo to confirm their anticancer effects. Results Numerous experiments have demonstrated the properties of BMS-202/ICG/KTZ-loaded hybrid liposomes. The results of the in vitro/in vivo experiments confirmed successful enhancement of the targeting ability of the nanoparticles prepared by fusing with the cell membranes of specific cancer cells to increase their targetability and penetration. In addition, although the drug alone did not show significant cytotoxicity to 4T1 cancer cells by MTT assay, liposomes containing both KTZ and BMS-202 showed a significant effect on secondary tumor suppression in animal experiments compared to liposomes containing only a single drug, confirming the effectiveness of the enhanced immunotherapy. Conclusion Our study showed that exosome inhibition and immune checkpoint blockade act synergistically in cancer immunotherapy.

Pharmacological research has expanded to the nanoscale level with advanced imaging technologies, enabling the analysis of drug distribution at the cellular organelle level. These advances in research techniques have contributed to the targeting of cellular organelles to address the fundamental causes of diseases. Beyond navigating the hurdles of reaching lesion tissues upon administration and identifying target cells within these tissues, controlling drug accumulation at the organelle level is the most refined method of disease management. This approach opens new avenues for the development of more potent therapeutic strategies by delving into the intricate roles and interplay of cellular organelles. Thus, organelle-targeted approaches help overcome the limitations of conventional therapies by precisely regulating functionally compartmentalized spaces based on their environment. This review discusses the basic concepts of organelle targeting research and proposes strategies to target diseases arising from organelle dysfunction. We also address the current challenges faced by organelle targeting and explore future research directions.

Mesenchymal stem cell (MSC)-based therapies show great potential in treating various diseases. However, control of the fate of injected cells needs to be improved. In this work, we developed an efficient methodology for modulating chondrogenic differentiation of MSCs. We fabricated heterospheroids with two sustained-release depots, a quaternized chitosan microsphere (QCS-MP) and a poly (lactic-co-glycolic acid) microsphere (PLGAMP). The results show that heterospheroids composed of 1 x 104 to 5 x 104 MSCs formed rapidly during incubation in methylcellulose medium and maintained high cell viability in long-term culture. The MPs were uniformly distributed in the heterospheroids, as shown by confocal laser scanning microscopy. Incorporation of transforming growth factor beta 3 into QCS-MPs and of dexamethasone into PLGA-MPs significantly promoted the expression of chondrogenic genes and high accumulation of glycosaminoglycan in heterospheroids. Changes in crucial metabolites in the dual drug depot-engineered heterospheroids were also evaluated using 1H NMRbased metabolomics analysis to verify their successful chondrogenic differentiation. Our heterospheroid fabrication platform could be used in tissue engineering to study the effects of various therapeutic agents on stem cell fate.

Mushrooms contain chitin-glucan complex (CGC), a natural copolymer of chitin and glucan, and nanofibrillation enhances its applicability. Here, a novel method was used to fabricate chitin-glucan nanofibers (CGNFs) from white button mushrooms. The first stage was to pretreat the raw mushroom using hot water and alkali to remove water-soluble glucans and alkali-soluble proteins, respectively, producing a CGC amenable to nanofibrillation. The second stage was nanofibrillation via esterification using acidic deep eutectic solvents (DESs) and subsequent ultrasonication. Five choline chloride-based DESs containing mono-or dicarboxylic acid were tested for the CGC esterification. DESs with strong dicarboxylic acids expedited nanofibrillation by homogeneously dispersing the solid CGC, swelling CGC fibrils, and facilitating acidity-dependent esterification leading to steric and electrostatic repulsions. One CGNF, namely CGNF_CCMnA, was characterized: it contained chitin and glucan at an approximate ratio of 8:2 and exhibited desirable properties as nanomaterials, including small diameter (11 nm) and high colloidal (zeta potential <-30 mV above pH 5.8) and thermal stability (T-m, 315 degree celsius). CGNF_CCMnA was tested for the adsorption to methylene blue, revealing a maximum adsorption capacity of 82.58 mg/g. The proposed approach is an efficient and readily applicable method to fabricate various mushroom-derived safe CGNFs and to produce related nanomaterials.

This study aims to enhance the solubility of Olaparib, classified as biopharmaceutical classification system (BCS) class IV due to its low solubility and bioavailability using a solid self-nanoemulsifying drug delivery system (S-SNEDDS). For this purpose, SNEDDS formulations were created using Capmul MCM as the oil, Tween 80 as the surfactant, and PEG 400 as the co-surfactant. The SNEDDS formulation containing olaparib (OLS-352), selected as the optimal formulation, showed a mean droplet size of 87.0 ± 0.4 nm and drug content of 5.53 ± 0.09

A novel nanoparticle screening technique was established to mostly enhance the aqueous solubility and oral bioavailability of aceclofenac using nanoparticle systems. Among the polymers investigated, sodium carboxymethylcellulose (Na-CMC) showed the greatest increase in drug solubility. Utilizing spray-drying technique, the solvent-evaporated solid dispersion (SESD), surface-attached solid dispersion (SASD), and solvent-wetted solid dispersion (SWSD) were prepared using aceclofenac and Na-CMC at a weight ratio of 1:1 in 50 % ethanol, distilled water, and ethanol, respectively. Using Na-CMC as a solid carrier, an aceclofenac-loaded liquid self-emulsifying drug delivery system was spray-dried and fluid-bed granulated together with microcrystalline cellulose, producing a solid self-nanoemulsifying drug delivery system (SNEDDS) and solid self-nanoemulsifying granule system (SNEGS), respectively. Their physicochemical properties and preclinical assessments in rats were performed. All nanoparticles exhibited very different properties, including morphology, crystallinity, and size. As a result, they significantly enhanced the solubility, dissolution, and oral bioavailability in the following order: SNEDDS ≥ SNEGS > SESD ≥ SASD ≥ SWSD. Based on our screening technique, the SNEDDS was selected as the optimal nanoparticle with the highest bioavailability of aceclofenac. Thus, our nanoparticle screening technique should be an excellent guideline for solubilization research to improve the solubility and bioavailability of many poorly water-soluble bioactive materials.

While spray-drying has been widely utilized to improve the bioavailability of poorly water-soluble drugs, the outcomes often exhibit suboptimal particle size distribution and large particle sizes, limiting their effectiveness. In this study, we introduce electrostatic spraying as an advanced technology tailored for poorly water-soluble drugs, enabling the fabrication of nanoparticles with fine and uniform particle size distribution. Regorafenib (1 g), as a model drug, copovidone (5 g), and sodium dodecyl sulfate (0.1 g) were dissolved in 200 ml ethanol and subjected to conventional-spray-dryer and electrostatic spray dryer. The electrostatic spray-dried nanoparticles (ESDN) showed smaller particle sizes with better uniformity compared to conventional spray-dried nanoparticles (CSDN). ESDN demonstrated significantly enhanced solubility and rapid release in water. In vitro studies revealed that ESDN induced apoptosis in HCT-116 cells to a greater extent, exhibiting superior cytotoxicity compared to CSDN. Furthermore, ESDN substantially improved oral bioavailability and antitumor efficacy compared to CSDN. These findings suggest that ESD shows potential in developing enhanced drug delivery systems for poorly water-soluble drugs, effectively addressing the limitations associated with CSD methods.