Diabetic ulcers (DUs), a severe complication of diabetes, are characterized by impaired wound healing and contribute significantly to morbidity and mortality. A key pathological driver is the persistent accumulation of neutrophil extracellular traps (NETs), which extend inflammation and tissue damage; however, appropriate therapeutic strategies to resolve NETs remain underdeveloped. We engineered a self-assembled nanocomplex, O/DNase-I, through structural and functional integration of oligomerized epigallocatechin gallate (OEGCG) and deoxyribonuclease-I (DNase-I). Its functionality was systematically evaluated in vitro and in a diabetic murine wound model using molecular and histological analyses. The O/DNase-I nanocomplex simultaneously eliminates existing NETs via DNase-I-mediated DNA hydrolysis and suppresses further NET formation through OEGCG. This synergistic action robustly cleared NETs, mitigated pro-inflammatory signaling, and critically, promoted a reparative immune microenvironment by driving M2 macrophage polarization, ultimately accelerating diabetic wound closure in vivo. This study not only validates O/DNase-I as a potent therapeutic approach for diabetic wound management but also establishes a novel supramolecular strategy for targeting dysregulated inflammation, with broad potential applications in other NET-associated pathologies.

BACKGROUND:Transdermal drug delivery (TDD) offers a convenient administration for treating local or systemic diseases. However, the dense "brick-mortar" structure of the stratum corneum hinders the skin permeation of most of bioactive molecules, which highly constrained the application of TDD. METHODS:This study analyzed stratum corneum disruption and enhanced permeation by gently rubbing calcium hydrogen phosphate (Chp) particles of varying sizes on skin. Enhanced anesthetic efficacy of Chp-containing microemulsion gel (MG) loaded with lidocaine-tetracaine eutectic was evaluated in guinea pigs using a needle-prick model. RESULTS:Using Chp as dermabrasion particle could effectively disrupt the barrier of stratum corneum after applying gently rubbing on the skin, which resulted in increasing skin permeation of both FITC, FITC-Dextran4000 and MG loaded with lidocaine-tetracaine eutectic. The enhanced skin permeation depended not only on the increase of particle size and amount of Chp in the MG, but also on the increase of rubbing pressure and duration after being applied to the skin, finally shortening the onset time and improving the efficacy of local anesthetics. CONCLUSIONS:The incorporation of Chp into the topical formulation, followed by rubbing them on the skin with appropriate pressure and a certain duration, can significantly disrupt the stratum corneum, enhance drug permeation through the skin, and shorten the onset time of local anesthetics. This study provided a potential strategy for improving the skin permeation with well tolerance, which could be further used to expand the range of bioactive molecules for TDD.

Lipid nanoparticles (LNPs) have emerged as a powerful platform for mRNA vaccine delivery, with ionizable lipids playing a pivotal role in enhancing cellular uptake and endosomal escape, thereby improving therapeutic efficacy. In this study, we designed and synthesized 12 novel ionizable lipids via molecular hybridization, incorporating a hydroxyalkylamine headgroup, an ester linkage, and a cis-double bond tail. These lipids were then formulated into LNPs with helper lipids and eGFP or LUC mRNA. Through cell transfection and in vivo imaging experiments, the O2-N5-OLE LNP was identified with superior mRNA delivery capabilities, particularly in immune cells. Subsequently, it was employed to encapsulate the tumor model antigen ovalbumin (OVA) mRNA vaccine, and its preventive effects against mouse melanoma were evaluated. The results demonstrated that O2-N5-OLE LNP@OVA mRNA exhibits superior tumor suppression. Additionally, the OVA mRNA delivered by O2-N5-OLE LNP significantly elevated serum OVA-IgG and IFN-γ levels and activated dendritic cells and CD8+ T lymphocytes, indicating that the vaccine effectively activated both humoral and cellular immunity. Coupled with the excellent biocompatibility of the O2-N5-OLE LNP, these findings highlight its remarkable delivery efficacy and provide a strong experimental foundation for its potential clinical application.

Mitochondria-targeted drug delivery is a promising strategy to potentiate anti-tumor efficacy. Current mitochondria-targeting nano-delivery strategies are mainly based on modification of lipophilic cation or targeting peptides. Due to endo/lysosomal capture and size limitation from mitochondrial outer membrane pores, neither approach guarantees efficient mitochondrial entry of the modified nanosystems. Herein, an ultrasmall hydrophobic cationic graphene oxide (GO-ODA) based nanoplatform is developed for cascading of endo/lysosomal escape and precise inner mitochondrial membrane (IMM) targeting. The functional cascading originates from the strong Brownian motion and high edge density of the ultrasmall hydrophobic cationic GO-ODA, which increases the frequency of edge-induced scratching and lipid extraction against bio-membranes during its random movement. Photosensitizers (PSs) with different charges are used as model therapeutics and loaded onto GO-ODA surface, followed by co-encapsulated within hyaluronic acid based amphiphile (IPHD) to form IPHD/GO-ODA@PSs nanoparticles. The PSs loading dose-optimized GO-ODA@PSs are validated to feature membrane-interactive properties for penetration, and prominently accumulate to IMM with inherent electronegativity. Upon laser irradiation, GO-ODA@PSs induce severe mitochondrial dysfunction both in vitro and in vivo, and thus not only potentiate tumor inhibition, but also significantly enhance tumor immunogenicity through activating the cGAS-STING pathway and GSDME-mediated pyroptosis. It ultimately elicits a strong systemic antitumor immune response.

With the rapid development of human-machine interfaces, the demand for next-generation patches with versatile functionality is increasing to overcome the limitations of conventional patches. This study presents an ultrathin multifunctional patch consisting of two main layers-a bioadhesive layer copolymerized with thioctic acid and sulfobetaine methacrylate (PTA-SBMA) and a top-protective layer made of porous and breathable styrene-ethylene/butylene-styrene. The PTA-SBMA layer demonstrated robust adhesion properties (382.08 +/- 7.32 J m-2 on porcine skin) and could be easily detached (13.70 +/- 3.46 J m-2, resulting in a switching ratio of 27.89) in a mixed solution of glutathione (GSH) and sodium bicarbonate. The patch exhibited biocompatibility and antifouling properties (3.33 +/- 5.77 CFU/cm2), making it suitable for personalized health monitoring and wound healing. Through masking and spraying, the patch achieved precise drug application on unique wound shapes. Silver nanowires (AgNWs) were used as a conductive layer to reduce the noise caused by skin deformation during health monitoring. The resistance of the patch remained stable even under strain because of the Ag-S bonding between the PTA-SBMA adhesive and AgNW. These features improve the acquisition and analysis of physiological signals during varying and complex motions. This dual superiority of adhesion and breathability made the m-patch a promising material for biomedical and bioelectronic applications.

Proteolysis targeting chimeras (PROTACs) hold immense promise for targeted protein degradation; however, challenges such as off-target effects, poor drug-likeness properties, and the "hook effect" remain. This study introduces Nano-Click-formed PROTACs (Nano-CLIPTACs) for precise tumor protein degradation in vivo. Traditional PROTACs with high molecular weight were first divided into two smaller druglike precursors capable of self-assembling to form functional PROTACs through a bioorthogonal reaction. Then, optimal CLIPTACs precursors (W4 and Z2) were encapsulated individually into cyclic RGDfC-peptide-modified liposomes to prepare Nano-CLIPTACs, enabling tumor-targeted delivery and subsequent in situ self-assembly to form PROTACs WZ42 within tumor cells. The degradation abilities of Nano-CLIPTACs in vitro and in vivo were further verified using a key oncology target, anaplastic lymphoma kinase (ALK), validating the safety, efficacy and "anti-hook effect" of this strategy. Overall, Nano-CLIPTACs represent a critical step towards the clinical translation of PROTACs technology for precise targeted anti-cancer therapies.

The accumulation of atherosclerosis plaques within arterial walls leads to cardiovascular events. Lipid-laden macrophages, known as foam cells play a pivotal role in atherosclerotic plaque progression by disrupting cholesterol homeostasis and facilitating inflammation. This study presents a rational and multivalent nanoplatform (siTTENPs) for atherosclerosis treatment. siTTENPs can form electrostatic complexes with the nucleic acid siTRPM2, thereby reducing oxidized low-density lipoprotein (oxLDL) uptake by foam cells and alleviating inflammation. Concurrently, β-cyclodextrin (β-CD) modified siTTENPs facilitate cholesterol clearance, further re-establishing lipid homeostasis. The nanometer size and S2P peptide (CRTLLTVRKC) modification endow these particles with specific targeting capabilities toward lesional macrophages, thereby enhancing their anti-atherosclerotic efficacy. Consequently, the siTTENPs delivery system effectively inhibits pathological cholesterol internalization while simultaneously promoting cholesterol efflux mechanisms and reducing inflammation. This therapeutic intervention leads to significant regression of atherosclerotic plaque. This study introduces an innovative therapeutic strategy aimed at improving cholesterol homeostasis, with promising implications for the treatment of atherosclerosis.

Islet-antigen-specific tolerization is a key goal of experimental immunotherapies for type 1 diabetes. mRNA-based vaccines have demonstrated the feasibility of RNA delivery in inducing antigen tolerance in autoimmune diseases. In this study, mRNA vaccine, encoded tandem glutamic acid decarboxylase 65 (GAD65) epitopes and cholera toxin B subunit (CTB-GADIII), prepared by an in vitro transcription (IVT) system and encapsulated with lipid nanoparticles (LNP), was intramuscularly administered to non-obese diabetic (NOD) and cyclophosphamide (Cy)-NOD mice respectively. The results showed that the mRNA vaccines significantly reduced the incidence rate of type 1 diabetes, delayed the disease progression, improved glucose tolerance, and protected pancreatic morphology and function compared with the controls. Meanwhile, the vaccines also reduced the levels of autoantibodies to glutamic acid decarboxylase (GADA) and insulin (IAA) in the serum. Furthermore, the proportion of CD4+ T helper cell subsets was modulated in the spleen of mice treated with mRNA vaccines, in correspondence with the increased levels of IL-10 and TGF-β in serum, suggesting the possible mechanism of immune tolerance. This study provides experimental evidence for the application of mRNA vaccines encoding self-antigens in the prevention or treatment of type 1 diabetes.

Mild photothermal therapy (PTT) shows the potential for chemosensitization by tumor-localized P-glycoprotein (P-gp) modulation. However, conventional mild PTT struggles with real-time uniform temperature control, obscuring the temperature-performance relationship and resulting in thermal damage. Besides, the time-performance relationship and the underlying mechanism of mild PTT-mediated P-gp reversal remains elusive. Herein, we developed a temperature self-limiting lipid nanosystem (RFE@PD) that integrated a reversible organic heat generator (metal-phenolic complexes) and metal chelator (deferiprone, DFP) encapsulated phase change material. Upon NIR irradiation, RFE@PD released DFP for blocking ligand-metal charge transfer to self-limit temperature below 45 degrees C, and rapidly reduced P-gp within 3 h via Ubiquitin-proteasome degradation. Consequently, the DOXHCl-loaded thermo-chemotherapeutic lipid nanosystem (RFE@PD-DOX) led to dramatically improved drug accumulation and 5-fold chemosensitization in MCF-7/ADR tumor models by synchronizing P-gp reversal and drug pulse liberation, achieving a tumor inhibition ratio of 82.42%. This lipid nanosystem integrated with "intrinsic temperature-control" and "temperature-responsive pulse release" casts new light on MDR tumor therapy.

Neutrophil extracellular traps (NETs) play a crucial role in the formation of vulnerable plaques and the development of atherosclerosis. Alleviating the pathological process of atherosclerosis by efficiently targeting neutrophils and inhibiting the activity of neutrophil elastase to inhibit NETs is relatively unexplored and is considered a novel therapeutic strategy with clinical significance. Sivelestat (SVT) is a second-generation competitive inhibitor of neutrophil elastase with high specificity. However, therapeutic effect of SVT on atherosclerosis is restricted because of the poor half-life and the lack of specific targeting. In this study, we construct a plaque-targeting and neutrophil-hitchhiking liposome (cRGD-SVT-Lipo) to improve the efficacy of SVT in vivo by modifying the cRGD peptide onto SVT loaded liposome, which was based on the interaction between cRGD peptide and integrin ανβ3 on the surface of cells in blood and plaque, including epithelial cell, macrophage and neutrophils. The cRGD-SVT-Lipo could actively tend to or hitchhike neutrophils in situ to reach atherosclerotic plaque, which resulted in enhanced atherosclerotic plaque delivery. The cRGD-SVT-Lipo could also reduce plaque area, stabilize plaque, and ultimately alleviate atherosclerosis progression through efficiently inhibiting the activity of neutrophil elastase in atherosclerotic plaque. Therefore, this study provides a basis and targeting strategy for the treatment of neutrophil-related diseases. STATEMENT OF SIGNIFICANCE: Neutrophil extracellular traps (NETs)-inhibiting is a prospective therapeutic approach for atherosclerosis but has received little attention. The NETs can be inhibited by elastase-restraining. In this work, an intriguing system that delivers Sivelestat (SVT), a predominantly used neutrophil elastase inhibitor with poor targeting capability, is designed to provide the drug with plaque-targeting and neutrophil-hitchhiking capability. The result suggests that this system can effectively hinder the formation of NETs and delay the progression of atherosclerosis.

Checkpoint blockade immunotherapy (CBI) have exhibited remarkable benefits for cancer therapy. However, the low responsivity of CBI hinders its application in treatment of bladder cancer. Ferroptosis shows potential for increasing the responsivity of CBI by inducing immunogenic cell death (ICD) process. Herein, we developed a mitochondrial-targeted liposome loaded with brequinar (BQR) (BQR@MLipo) for enhancing the mitochondrial-related ferroptosis in bladder cancer in situ. It could be found that BQR@MLipo could selectively accumulate into mitochondria and inactivate dihydroorotate dehydrogenase (DHODH), which induced extensive mitochondrial lipid peroxidation and ROS, finally triggering ferroptosis of bladder cancer cells to boost the release of intracellular damage-associated molecular patterns (DAMPs) such as calreticulin (CRT), adenosine triphosphate (ATP), high mobility group box 1 (HMGB1). In addition, BQR@MLipo further promoted the release of mtDNA into the cytoplasm to activate the cGAS-STING pathway for the secretion of IFN-β, which would increase the cross-presentation of antigens by dendritic cells and macrophage phagocytosis. Furthermore, the in vivo studies revealed that BQR@MLipo could remarkably accumulate into the bladder tumor and successfully initiate the infiltration of CD8+ T cells into tumor microenvironment for enabling efficient CBI to inhibit bladder tumor growth. Therefore, BQR@MLipo may represent a clinically promising modality for enhancing CBI in bladder tumor.

Atherosclerotic cardiovascular diseases remain the leading causes of morbidity and mortality worldwide. Cholesterol crystals in atherosclerotic plaques play an essential role in atherosclerosis progression. However, no clinical drugs have been used for removing cholesterol crystals from plaque to counter atherosclerosis. Previous studies identified the hydrophobic domain of lipid bilayer in liposomes acted as sinks for solubilizing hydrophobic cholesterol. Moreover, adjusting the composition of the lipid bilayer in liposomes can enhance its hydrophobic molecule loading capacity. Therefore, in this study, ginsenosides Rb1 (Rb1), one of main active components of ginseng which has a similar structure to cholesterol, is anchored into soy phospholipids bilayer with its hydrophobic region to prepare nano-sponge-like liposomes (Rb1-LPs), aiming to amplify the solubilization of cholesterol in lipid bilayer. For targeting delivery to atherosclerotic plaques, Annexin V (AnxV), a protein that can specifically recognize phosphatidylserine upregulated in atherosclerotic plaques, is applied to decorate the surface of Rb1-LPs by click reaction to obtain the final preparation of AnxV-Rb1-LPs. The in vitro studies showed that incorporating Rb1 into lipid bilayer remarkably increased the affinity of the lipid bilayer to free cholesterol and the solubilization of cholesterol crystals. Additionally, nano-sponge-like liposomes could efficiently reduce the accumulation of cholesterol crystals and improve cholesterol efflux, finally inhibiting inflammation and apoptosis in cholesterol-laden cells. Furthermore, AnxV-Rb1-LPs could efficiently accumulate in atherosclerotic plaques after intravenous injection, exert nano-sponge-like functions to remove intra- and extracellular cholesterol crystals, ultimately alleviating inflammation and apoptosis in atherosclerotic plaques for antiatherosclerosis. Therefore, AnxV-Rb1-LPs provide a potential strategy for removing cholesterol crystals in atherosclerotic plaques and can be further utilized in other diseases with excessive cholesterol accumulation.

Immune cells stand as a critical component of the immune system to maintain the internal environment homeostasis. The dysfunction of immune cells can result in various life-threatening diseases, including refractory infection, diabetes, cardiovascular disease, and cancer. Therefore, strategies to standardize or even enhance the function of immune cells are critical. Recently, nanotechnology has been highly researched and extensively applied for enhancing the cytoplasmic delivery of bioactive molecules to immune cells, providing efficient approaches to correct in vivo and in vitro dysfunction of immune cells. This review focuses on the technologies and challenges involved in improving endo-lysosomal escape, cytoplasmic release and organelle targeted delivery of different bioactive molecules in immune cells. Furthermore, it will elaborate on the broader vision of applying nanotechnology for treating immune cell-related diseases and constructing immune therapies and cytopharmaceuticals as potential treatments for diseases.

Abstract: Cardiovascular disease is responsible for the largest number of deaths worldwide, and atherosclerosis is the primary cause. Apoptotic cell accumulation in atherosclerotic plaques leads to necrotic core formation and plaque rupture. Emerging findings show that the progression of atherosclerosis appears to suppress the elimination of apoptotic cells. Mechanistically, the reduced edibility of apoptotic cells, insufficient phagocytic capacity of phagocytes, downregulation of bridging molecules, and dysfunction in the polarization of macrophages lead to impaired efferocytosis in atherosclerotic plaques. This review focuses on the characteristics of efferocytosis in plaques and the therapeutic strategies aimed at promoting efferocytosis in atherosclerosis, which would provide novel insights for the development of antiatherosclerotic drugs based on efferocytosis.

Tumor metastasis is directly correlated to poor prognosis and high mortality. Circulating tumor cells (CTCs) play a pivotal role in metastatic cascades, of which CTC clusters is highly metastatic compared to single CTCs. Although platelets and neutrophils within the bloodstream could further exacerbate the pro-metastatic effect of single CTCs, the influence of platelets and neutrophils on CTC clusters mediated metastasis remains unclear. In this study, a pro-metastatic complex composed of CTC clusters, platelets and neutrophils, namely circulating tumor microemboli (CTM), was identified in vivo among different metastatic tumor, which was demonstrated with highly upregulation of hypoxia-inducible factor-1α (HIF-1α). While knock-out of HIF-1α or therapeutically downregulating of HIF-1α via HIF-1α inhibitor (BAY87-2243)-loaded neutrophil cyto-pharmaceuticals (PNEs) could efficiently restrain CTM mediated lung metastasis. The underlying mechanism of metastasis inhibition was attributed to the downregulation of HIF-1α-associated PD-L1, which would enhance immune response for inhibiting metastatic cells. Thus, our work here illustrates that hypoxia was an essential factor in promoting CTM colonization in lung. More importantly, we provide a promising strategy by targeted downregulation of HIF-1α in CTM via neutrophil cyto-pharmaceuticals for treatment of CTM mediated metastasis.

Naringenin (NGN) can be used to inhibit the progression of nonalcoholic fatty liver disease (NAFLD) in mice, but its poor water solubility limits its applications. Nanostructured lipid carriers (NLCs) have recently attracted much attention in the field of nanodrug delivery systems because they increase the drug loading capacity and impressively enhance the solubility of indissolvable drugs. Herein, a thin-film dispersion method was used to prepare naringenin-loaded nanostructured lipid carriers (NGN-NLCs). These NGN-NLCs have a narrow size distribution of 171.9 ±2.0 nm, a high drug loading capacity of 23.7 ± 0.3%, a high encapsulation efficiency of 99.9 ± 0.0% and a drug release rate of 86.2 ± 0.4%. NGN- NLCs elevated the pharmacokinetic parameters (Cmax and AUC0→t) of NGN, accelerated NGN transepithelial transport in MDCK cells and intestinal absorption in the jejunum and ileum, and reduced hepatic lipid accumulation in an oleic acid (OA) plus lipopolysaccharide (LPS)-induced lipid deposition cell model in primary hepatocytes and in a methionine/choline deficient (MCD) diet-induced NAFLD mouse model. A detailed study of the mechanism showed that this NLC formulation elevated the drug release rate in simulated intestinal solutions in vitro, the transepithelial transport in MDCK cells, the oral absorption in mice and the ex vivo intestinal absorption of NGN. Thus, NGN-NLCs significantly enhanced the inhibitory effects of NGN on MCD diet induced mouse NAFLD.

Mitochondrion is an important organelle in the cell, playing critical roles in various biological processes, such as calcium homoeostasis, cellular stress response, cellular metabolism and cellular apoptosis.Thus, mitochondrion is also a promising therapeutic target for solving current clinical issues and preventing various human diseases such as diabetes, Alzheimer's disease, cardiovascular disorders, and cancer.In this review, firstly, we aimed at the function and influence of mitochondria in diseases.Furthermore, the targeting strategies to mitochondrion based on the highly negative potential of mitochondrial membrane and mitochondrial import proteins were also discussed for developing more efficient treatments for mitochondrial diseases.

动脉粥样硬化是导致心脑血管疾病的主要诱因.主动脉血管内皮下斑块的形成是动脉粥样硬化的病理特征,而在斑块中胆固醇结晶的出现不仅会加剧斑块炎症反应,还会造成斑块去稳定和破裂,最终导致心脑血管事件.因此,清除斑块胆固醇结晶将有利于动脉粥样硬化的归转.本文综述了斑块中胆固醇结晶的形成过程和特点、胆固醇结晶促进动脉粥样硬化的作用机制以及清除斑块胆固醇结晶的策略,为开发以斑块胆固醇结晶为靶点的抗动脉粥样硬化药物提供理论依据和参考.

Limb and CNS expressed 1 like (LIX1L) is over-expressed in several types of tumors. However, the function of LIX1L in glucose metabolism and hepatocellular carcinoma (HCC) progression remains elusive. Here we report that LIX1L is over-expressed in human HCC tissues, which predicts unfavorable prognosis. LIX1L deficiency in vivo significantly attenuated liver cancer initiation in mice. Functional studies indicated that LIX1L overexpression elevated proliferation, migratory, invasive capacities of HCC cells in vitro, and promoted liver cancer growth and metastasis in vivo. LIX1L knockdown up-regulated fructose-1,6-bisphosphatase (FBP1) expression to reduce glucose consumption as well as lactate production. Mechanistically, LIX1L increased miR-21-3p expression, which targeted and suppressed FBP1, thereby promoting HCC growth and metastasis. MiR-21-3p inhibitor could abrogate LIX1L induced enhancement of cell migration, invasion, and glucose metabolism. Inhibition of miR-21-3p suppressed tumor growth in an orthotopic tumor model. Our results establish LIX1L as a critical driver of hepatocarcinogenesis and HCC progression, with implications for prognosis and treatment.

Multidrug resistance (MDR) due to P-glycoprotein (P-gp) overexpression is a major obstacle to successful leukemia chemotherapy. The combination of anticancer chemotherapy with a chemosensitizer of P-gp inhibitor is promising to overcome MDR, generate synergistic effects, and maximize the treatment effect. Herein, we co-encapsulated a chemotherapeutic drug of mitoxantrone (MTO) and a P-gp inhibitor of β-elemene (βE) in solid lipid nanoparticles (MTO/βE-SLNs) for reversing MDR in leukemia. The MTO/βE-SLNs with about 120 nm particle size possessed good colloidal stability and sustained release behavior. For the cellular uptake study, doxorubicin (DOX) was used as a fluorescence probe to construct SLNs. The results revealed that MTO/βE-SLNs could be effectively internalized by both K562/DOX and K562 cells through the pathway of caveolate-mediated endocytosis. Under the optimized combination ratio of MTO and βE, the in vitro cytotoxicity study indicated that MTO/βE-SLNs showed a better antitumor efficacy in both K562/DOX and K562 cells than other MTO formulations. The enhanced cytotoxicity of MTO/βE-SLNs was due to the increased cellular uptake and blockage of intracellular ATP production and P-gp efflux by βE. More importantly, the in vivo studies revealed that MTO/βE-SLNs could significantly prolong the circulation time and increase plasma half-life of both MTO and βE, accumulate into tumor and exhibit a much higher anti-leukemia effect with MDR than other MTO formulations. These findings suggest MTO/βE-SLNs as a potential combined therapeutic strategy for overcoming MDR in leukemia.