Photodynamic therapy (PDT) is a minimally invasive cancer treatment strategy. Despite numerous studies confirming its potential advantages, the clinical translation of this technique remains limited owing to the impracticality of using conventional light sources and the instability of photosensitizers. This study presents a novel PDT approach that utilizes organic light-emitting diodes (OLEDs) as compact, low-thermal, wearable, and flexible light sources to activate chlorin e6 (Ce6)-loaded Pluronic nanocapsules (Ce6/Plu NCs) for anticancer therapy. Here, different types of Pluronic polymers are used and optimized to synthesize ∼120 nm-diameter Ce6/Plu NCs. Among the formulated systems, the Ce6/PP123 NCs show optimal spectral characteristics and the highest therapeutic efficacy in vitro, and efficient tumor accumulation after intravenous injection and significant antitumor efficacy upon OLED irradiation with minimal off-target toxicity in vivo. Tissue-attachable OLEDs for PDT can be placed in close proximity to tumor tissue for efficient light delivery. The combination of the Ce6/PP123 NCs with a novel wearable OLED is a promising platform for effective and safe PDT in cancer treatment.

Pulmonary fibrosis is characterized by persistent oxidative stress and excessive extracellular matrix deposition, yet effective therapeutic strategies remain limited. Here, we report a nanoengineering strategy that integrates melatonin and Nrf2 mRNA within lipid nanoparticle (LNPs) to functionally engineer mesenchymal stem cells (MSCs) for redox-targeted antifibrotic therapy. Molecular dynamics simulations and membrane fluidity analyses revealed that melatonin modulates lipid packing in cholesterol-rich LNP membranes, increasing membrane fluidity and facilitating endosomal escape, thereby improving cytosolic mRNA delivery. This membrane-level tuning enabled efficient Nrf2 expression and promoted an antioxidant phenotype in the engineered MSCs. These therapeutic LNP (TN)-engineered MSCs exhibited enhanced cytoprotective and antifibrotic activities in epithelial injury models and in a bleomycin-induced pulmonary fibrosis mouse model. The therapeutic effects were associated with suppression of epithelial-mesenchymal transition and extracellular matrix remodeling. Collectively, this study establishes melatonin-driven membrane modulation as an effective strategy to enhance LNP-mediated mRNA delivery and enable functional engineering of MSCs for redox-targeted regenerative therapy.

Intracellular delivery into suspension cells, particularly hard-to-transfect immune cells such as T- and B-lymphocytes, remains challenging. Membrane disruption-based microfluidic methods offer a carrier-free alternative but often depend on high-viscosity buffers, compromising viability and scalability. Here, we introduce a viscoelastic mechanoporation platform using a hyperbolic microfluidic channel and low-viscosity λDNA buffer for the efficient delivery of mRNA and small molecules. The system harnesses extensional strain to transiently deform cell membranes, enabling high-throughput cytosolic uptake with minimal cellular stress. Our platform achieved up to ∼17-fold enhanced mRNA delivery while maintaining >85% viability across multiple suspension cell lines. Mechanistic insights from Laurdan spectral analysis, ice incubation, and metabolic profiling revealed how membrane dynamics govern delivery outcomes. We further modulated efficiency through osmotic and cytoskeletal perturbations, demonstrating a tunable strategy for safe and effective delivery into fragile immune cells.

The clinical success of nanoparticles (NPs) with surface coating using polyethylene glycol (PEG) has been accompanied by growing concerns regarding PEG-induced hypersensitivity and immunogenicity. These limitations have spurred the search for PEG alternatives, particularly in cancer therapy, where repeated administration is common. Polyglycerol (PG), a hydrophilic and biocompatible polymer, has emerged as a promising candidate for surface coating of NPs to its antifouling properties and reduced recognition by macrophages. In this study, we developed a PG-coated liposomal nanomedicine loaded with topotecan (Tpt), a hydrophilic anticancer drug, and pheophorbide a (Pba), a hydrophobic photosensitizer. This dual-loading strategy enables two complementary therapeutic mechanisms: DNA repair inhibition by Tpt and reactive oxygen species (ROS)-mediated photodynamic therapy (PDT) by Pba under 671 nm laser irradiation. The dual drug-loaded PG-liposome (Pba/Tpt@PG) exhibited potent cytotoxicity against SCC7 tumor cells, demonstrated reduced macrophage uptake in vitro, and in vivo imaging revealed its favorable tumor accumulation. SCC7 tumor-bearing mice received intravenous injections of Pba/Tpt@PG, followed by localized laser irradiation. This treatment successfully inhibited tumor growth, reducing tumor volume to 0.58% of that in untreated controls, and outperformed single-agent treatments. No significant body weight loss or histological abnormalities were observed, indicating high biocompatibility of the materials. This study underscores the potential of dual-drug-loaded liposomes and PG-based surface modification for safe and effective combination cancer therapy.

Lipid nanoparticles (LNPs) have enabled the clinical application of RNA therapeutics, including approved mRNA vaccines and siRNA medicines. However, their predominant accumulation in liver after systemic administration and inefficient endosomal escape remain key bottlenecks for productive cytosolic delivery and gene expression. Here, we introduce a synthetic ionizable lipid with a dual pKa property that is retained in formulated LNPs. We show that after intravenous injection, these dual pKa LNPs produce lung-selective mRNA expression with higher potency than a cationic lipid-rich comparator while remaining well tolerated. Using molecular dynamics simulations, we find that protonation state can support two distinct interaction modes between the ionizable lipid and endosomal membranes, suggesting a mechanistic basis for efficient endosomal escape. Moreover, the formulation maintains robust pulmonary expression and delivers therapeutic benefit in an acute lung inflammation model. These results establish a structure-property relationship within the ionizable lipid-based LNPs investigated here and identify dual apparent pKa behavior as a promising feature for potent and tolerable lung-targeted RNA delivery.

The efficient and timely induction of chondrogenesis remains a major challenge in stem cell–based cartilage regeneration. Although mesenchymal stem cells (MSCs) have been widely explored for cartilage repair, their slow differentiation kinetics and limited efficiency of non-viral gene delivery have constrained therapeutic outcomes. To overcome these limitations, we developed a differentiation-accelerating nanoparticle (DaN) system that enables rapid and robust SOX9 mRNA expression while co-delivering the chondrogenic modulator dexamethasone (Dex) by lipid nanoparticle (LNP). The LNP formulation was systematically optimized for MSC transfection, allowing efficient SOX9 expression without interference from Dex co-loading. DaN treatment induced rapid SOX9 protein production and promoted early chondrogenic commitment in 3D MSC spheroids through the combinatorial action of SOX9 mRNA and dexamethasone, as evidenced by enhanced extracellular matrix deposition and upregulation of key chondrogenic markers. In a rat osteochondral defect model, DaN-engineered MSC spheroids significantly promoted cartilage formation, enhanced defect repair, and improved cartilage regeneration at early time points. Collectively, this study demonstrates that the simultaneous delivery of mRNA and a small-molecule modulator via LNPs can enhanced cartilage repair and enhance in vivo cartilage repair, highlighting the translational potential of mRNA-based nanotherapeutic platforms in regenerative medicine.

Ionizable lipid is a key component of lipid nanoparticle (LNP) for mRNA delivery. However, the discovery of new lipid scaffolds remains constrained by the limited availability of efficient synthetic platforms capable of generating structurally diverse libraries. Here, we report a photochemistry-based ionizable lipid library (PILL) platform that enables the rapid discovery of ionizable lipids for mRNA delivery. Using a visible-light-driven multicomponent reaction under 427 nm irradiation, 275 α-branched amine-containing ionizable lipids were generated in a one-pot process from amines, aldehydes, and boronic acids under mild conditions. This modular photochemical strategy enabled the rapid construction of a structurally diverse lipid library that was directly integrated with purification-free in vitro screening and subsequent in vivo validation. A18B8C14 was identified as a lead ionizable lipid candidate for mRNA delivery, and the optimized A18B8C14-LNP formulation exhibited substantially higher in vivo gene expression than the benchmark MC3-based LNP while maintaining a favorable safety profile. In addition, organ-specific delivery could be achieved by adjusting the A18B8C14-LNP formulation according to the selective organ targeting (SORT) strategy. Together, these results establish photochemistry as a practical platform for ionizable lipid discovery and provide a versatile framework for the development of next-generation LNP systems for nucleic acid therapeutics.

For a long time, efficient and safe gene delivery has been a key issue in gene therapy. In particular, after the Nobel Prize in Chemistry for clustered regularly interspaced short palindromic repeat (CRISPR) technology in 2020, the focus on delivery systems for genome editing has grown. In this review, we introduce the recent trends in various CRISPR delivery systems. First, we explain the impact of CRISPR in clinical settings and its history. We then focused on the physics of gene delivery systems, particularly regarding the migration of nanoparticles (NPs) under flow, cellular uptake, and formulation using microfluidics. Subsequently, various CRISPR delivery systems, both viral and non-viral, and their applications in disease therapy were introduced. Viral carriers include lentiviruses, adeno-associated viruses, and viral capsids. Exosomes, silica NPs, polymeric NPs, and lipid NPs are representative non-viral gene delivery carriers. We mainly focused on studies demonstrating promising results in animal models, not stopped at cell test considering their future potential for human application.

In the lacuna structure of cartilage, a small group of chondrocytes are contacted and surrounded by extracellular matrix. To mimic this structure, we applied tannic acid (TA)-based clustering and click chemistry-based hydrogel to our human-derived nasal septal chondrocytes (hNCs) with advantages including easy harvesting from patients, stemness, and superior differentiation capacity. We fabricated lacuna-mimic hNC clusters based on the binding ability of TA. The hNC clusters were encapsulated in a hydrogel crosslinked by click chemistry between modified polyethylene glycol (PEG) and gelatin. The hNC clusters showed high viability and improved chondrogenic differentiation in click chemistry-based hydrogel. When we injected the hNC clusters in hydrogel into a rat osteochondral defect model, they resulted in successful regeneration of cartilage in immunohistochemical staining data. These results demonstrate the potential of the lacuna-mimic structure composed of hNC clusters and click chemistry-based hydrogel in cartilage regeneration in vivo.

The direct expression of therapeutic genes in tumor tissue is a promising cancer therapy, but it has been limited by delivery issues. Herein, we fabricated optimized mRNA-encapsulating lipid nanoparticles (LNPs) and applied them to lung-targeted delivery of tumor necrosis factor-related apoptosis-inducing ligand (TRAIL) and BCL2antagonist/killer (BAK) mRNAs for lung metastasis therapy. mRNA and LNPs were optimized by in vitro and in vivo experiments to achieve the highest expression in lung metastasis tissue. The selected formulation containing luciferase mRNA generated an intense luminescence signal in the lung while using a low ratio (15 %) of cationic DOTAP, which was advantageous to reduce cytotoxicity. The TRAIL and BAK mRNAs were encapsulated in lung-targeting LNPs (LtNPs) together and delivered to lung metastasis tumor tissue. Through the in vivo transfection and generation of TRAIL and BAK, apoptosis could be induced via the extrinsic death receptor and intrinsic mitochondrial pathway simultaneously. Intravenous injections of the optimized LtNPs containing TRAIL and BAK mRNAs effectively suppressed tumors in a mouse model of A549 lung metastasis. These overall results demonstrate the promising potential of mRNA-LNPs to directly deliver therapeutic genes to tumor tissue for cancer therapy.

This study investigated the potential of mucoadhesive polymeric nanoparticles coated with polydopamine (Dopa-NPs) for inner ear drug delivery. Dopa, inspired by mussel adhesion proteins, leveraged the mucoadhesive properties of NPs and enhanced their retention within the cochlea. Dopa-NPs were compared with non-adhesive uncoated control NPs to reveal their safety and drug delivery efficiency. The safety evaluation demonstrated no toxicity during in vitro test using HEI-OC1 cells and in vivo test using mouse with auditory function assessment. When coumarin-encapsulated NPs were administered through intratympanic injection to mice, Dopa-NPs provided intense fluorescence in the inner ear compared to non-adhesive control NPs. Furthermore, dexamethasone (Dex)-encapsulated Dopa-NPs exhibited significantly higher drug concentrations in the cochlea than dexamethasone sodium phosphate and uncoated NPs. Finally, in vivo test using an ototoxicity-induced animal model showed that Dopa-NPs improved hearing protection, as indicated by the auditory brainstem response (ABR) test. In conclusion, mucoadhesive Dopa-NPs exhibit enhanced drug delivery efficiency and safety, offering a promising strategy for inner ear drug delivery and chemotherapy.

Iron oxide nanoparticles (IONPs) have been widely applied in drug delivery systems for tumor-targeting over the past decade. However, their efficacy in remote control and targeting by magnetic attraction is still limited by insufficient magnetic force. Herein, we introduce non-toxic zinc ferrite NPs (ZFNPs) with strong magnetism as enhancers for tumor-targeting. They were encapsulated in larger poly lactic-co-glycolic acid (PLGA) NPs together with the anticancer drug, SN38 resulting in ZFNP-containing magnetic NPs (ZMNPs). Upon magnetic attraction, the cellular uptake of ZMNPs increased by 15.43 times compared to treatment without a magnet. This was remarkable because the NP-encapsulating traditional IONPs (MNPs) showed only 2.47 times increase. In vivo biodistribution analysis in SCC7 tumor-bearing mice revealed that magnetic attraction enhanced the tumor accumulation of ZMNPs by 3.9 times. Furthermore, ZMNPs exhibited antitumor effects by reducing the tumor volume to 214 mm3 with magnetic attraction, whereas it was 1016.7 mm3 without it. These results demonstrate that the enhanced magnetism of ZMNPs led to improved tumor targeting, resulting in superior therapeutic outcomes compared to traditional MNPs. We expect that these small ZFNPs could be encapsulated in other types of NPs with various structures, suggesting their universal potential for enhanced tumor-targeting.

The emergence of mRNA-containing lipid nanoparticles (LNPs) during the COVID-19 pandemic has revolutionized vaccine technology and is now being explored for protein replacement therapies. This study aimed to test the hypothesis that LNP size, independent of lipid composition, critically influences their physicochemical and biological performance. To achieve this, we employed computational fluid dynamics (CFD) simulations to predict the mixing index within a microfluidic channel, identifying the flow rate conditions at which a 70

In the pharmaceutical formulation of anticancer drugs, US FDA-approved cyclodextrin (CD) and its derivates have been widely employed. Among them, hydroxypropyl β-cyclodextrin (HPCD) has attracted considerable attention owing to low toxicity, improved water solubility, and enhanced drug absorption; however, HPCD exhibits low drug loading efficiency and poor anticancer efficacy in multidrug-resistant (MDR) cancer cells. Therefore, to improve the drug loading efficiency and simultaneously overcome the MDR effects, we developed docetaxel (DTX)-loaded Pluronic®(PLU)-coated hydroxypropyl-β-cyclodextrin nanoparticles (DTX@PLU/HPCD NPs). We employed a simple nanoprecipitation technique, wherein PLU acted as a P-glycoprotein inhibitor to overcome MDR and an enhancer to achieve high drug loading. The HPCD NPs were coated with PLU (PF127, PP123, and PL81) of varying hydrophilic/lipophilic balance values, and the amount of encapsulated DTX was optimized. The DTX-loaded HPCD formulations (DTX@HPCD NP, DTX@PF127/HPCD NP, and DTX@PP123/HPCD NP) exhibited good stability and facilitated sustained release. The HPCD formulations exerted no cytotoxicity against examined cancer cell lines (HCT15 and SCC 7). DTX@PLU/HPCD NPs exerted potent anticancer effects in vitro. Notably, DTX@PP123/HPCD NPs significantly reduced the tumor volume in a mouse model. Collectively, these results indicate that PLU/HPCD NPs could function as drug carriers to alleviate toxicity and overcome MDR in various cancer models.

Directional differentiation is a key factor determining the result of stem cell therapy. Herein, we developed a polyethylenimine (PEI)-coated poly(lactic-co-glycolic) acid (PLGA) nanoparticle (mPDN) carrying both nuclear factor erythroid 2-related factor 2 (Nrf2) mRNA and dexamethasone (Dex) to human mesenchymal stem cells (hMSCs). The combination of Dex and Nrf2-mRNA delivered by mPDN promoted the osteogenic differentiation of hMSCs. In particular, Nrf2-mRNA rapidly reduced the DNA damage caused by ROS due to early and efficient gene expression at 3 h after treatment, which was not achieved in traditional pDNA systems. High and rapid transfection, effective ROS-scavenging effect, and protection of mitochondrial dynamics were observed in hMSCs after treatment with the resulting Nrf2-mPDN. Osteogenic differentiation was also observed in 3D pellets for up to 5 weeks. Finally, the effects of rapid DNA repair in hMSCs by Nrf2-mPDN and on in vivo bone regeneration were evaluated in a rat femoral bone defect model using CT. This study demonstrated the potential of an NP-based codelivery system and efficient transfection of mRNA at early stages in hMSCs for bone regeneration and stem cell therapy.

Casein micelle (CM) is a carrier of hydrophobic molecules in breast milk developed by nature. Herein, we evaluated the potential of CM for delivery of hydrophobic pheophorbide a (Pba), a photosensitizer (PS) for in vivo photodynamic therapy (PDT). CM was constructed with casein sodium salt and calcium ions, and Pba was physically loaded into the inner pockets of CM. The resulting CM-Pba was about 220.03 nm in size and showed superior stability without aggregation in aqueous conditions for 4 months. CM-Pba showed fast cellular uptake into SCC7 tumor cells and killed them by photodynamic effects upon laser irradiation. After intravenous injection into SCC7 tumor-bearing mice, CM-Pba showed about 2.0-fold higher tumor accumulation than free Pba. We observed efficient suppression of tumor growth by PDT using CM-Pba after laser irradiation, and there were no significant side effects. The overall results demonstrated that CM could be a potential drug carrier for hydrophobic PS for in vivo PDT.

An ideal hydrogel for stem cell therapy would be injectable and efficiently promote stem cell proliferation and differentiation in body. Herein, an injectable, single-component hydrogel with hyaluronic acid (HA) modified with phenylboronic acid (PBA) and spermidine (SM) is introduced. The resulting HAps (HA-PBA-SM) hydrogel is based on the reversible crosslinking between the diol and the ionized PBA, which is stabilized by the SM. It has a shear-thinning property, enabling its injection through a syringe to form a stable hydrogel inside the body. In addition, HAps hydrogel undergoes a post-injection "self-curing," which stiffens the hydrogel over time. This property allows the HAps hydrogel to meet the physical requirements for stem cell therapy in rigid tissues, such as bone, while maintaining injectability. The hydrogel enabled favorable proliferation of human mesenchymal stem cells (hMSCs) and promoted their differentiation and mineralization. After the injection of hMSCs-containing HAps into a rat femoral defect model, efficient osteogenic differentiation of hMSCs and bone regeneration is observed. The study demonstrates that simple cationic modification of PBA-based hydrogel enabled efficient gelation with shear-thinning and self-curing properties, and it would be highly useful for stem cell therapy and in vivo bone regeneration.

Although peptides notoriously have poor intrinsic pharmacokinetic properties, it is well-known that nanostructures with excellent pharmacokinetic properties can be designed. Noticing that peptide inhibitors are generally nonpolar, here, we consolidate the peptide inhibitor targeting intracellular protein-protein interactions (PPIs) as an integral part of biodegradable self-assembled depsipeptide nanostructures (SdPNs). Because the peptide inhibitor has the dual role of PPI inhibition and self-assembly in this design, problems associated with the poor pharmacokinetics of peptides and encapsulation/entrapment processes can be overcome. Optimized SdPNs displayed better tumor targeting and PPI inhibition properties than the comparable small molecule inhibitor in vivo. Kinetics of PPI inhibition for SdPNs were gradual and controllable in contrast to the rapid inhibition kinetics of the small molecule. Because SdPN is modular, any appropriate peptide inhibitor can be incorporated into the platform without concern for the poor pharmacokinetic properties of the peptide.

Directional differentiation of stem cells is a key step in stem cell therapy. In this study, we developed saponin-based nanoparticles (Ad-SNPs) containing dexamethasone (Dex) and alpha-lipoic acid (ALA) to promote osteogenic differentiation of human mesenchymal stem cells (hMSCs) and bone regeneration. The Ad-SNPs can achieve rapid cellular uptake through a pore-forming effect without cytotoxic cationic charges. They also provide extended retention in cell cytosol due to their uptake route. These properties are advantageous in efficiently supplying drugs to the hMSCs. The combination of Dex and ALA facilitated mitochondrial fusion and prevented reactive oxygen species-induced DNA damage. It also helped to preserve mitochondrial dynamics, and the efficient supply of it provided by the Ad-SNPs induced differentiation of hMSCs into osteoblasts. The Ad-SNPs showed outstanding performance in osteoblast differentiation, maturation, and mineralization in 3D culture compared with NPs without saponin and with free drugs. When Ad-SNP-treated hMSCs were tested in a rat femoral bone defect model, they showed the fastest regeneration of bones and complete repair in the shortest period among all groups. To the best of our knowledge, this study is the first application of pore-forming saponin-based NPs with rapid cellular uptake and extended retention to stem cell therapy, and we demonstrated their promising potential in bone regeneration and efficient delivery of Dex and ALA.

Biological cell membranes are a natural barrier for living cells. In the last few decades, the cell membrane has been the main hurdle in the efficient delivery of bioactive and therapeutic agents. To increase the drug efficacy of these agents, additional mediators have been considered. Cell-penetrating peptides (CPPs), a series of oligopeptides composed of mostly hydrophobic and/or positively charged side chains, can increase the interaction with the cell membrane. CPP-based delivery platforms have shown great potential for the efficient and direct cytosol delivery of various cargos, including genes, proteins, and small molecule drugs. Bypassing endocytosis allows the CPP-based delivery systems greater defense against the degradation of protein-based drugs than other drug delivery systems. However, the delivery of CPPs exhibits intrinsically non-specific targeting, which limits their medical applications. To endow CPPs with specific targeting ability, the conjugation of pH-sensitive, enzyme-specific cleavable, and multiple targeting ligands has been reported. Optimization of the length and sequence of CPPs is still needed for various drugs of different sizes and surface charges. Toxicity issues in CPP-based delivery systems should be addressed carefully before clinical use.