Bone defects pose a high risk of non-union and permanent disability, making effective bone regeneration a critical focus in the development of bone repair materials. Current research primarily emphasizes enhancing the single osteogenic function of bone repair materials, while neglecting the impact of the complex microenvironment in bone defect areas. This has resulted in the failure of many developed bone repair materials to achieve effective in vivo bone regeneration. In this study, a multifunctional near-infrared light-responsive black phosphorus (BP) bone repair scaffold was fabricated via low-temperature deposition 3D printing. In vitro characterization demonstrated that the scaffold possesses a cancellous bonelike structure, moderate compressive strength, and cytocompatibility, with the ability to promote osteogenesis under inflammatory conditions. In vivo studies further confirmed its favorable photothermal responsiveness, enabling photothermal therapy (PTT) to accelerate bone regeneration while reducing inflammation in the defect area. These findings indicate that the multifunctional BP scaffold achieves superior bone repair outcomes through synergistic effects of anti-inflammation, promotion of osteogenic differentiation, and PTT, thereby improving the success rate of defect repair. Moreover, the simple fabrication process and satisfactory therapeutic efficacy of this multifunctional BP scaffold highlight its high potential for clinical translation.

Scopoletin (Sco) is a natural coumarin compound with potent antioxidant activity; however, its poor aqueous solubility and low bioavailability significantly limit its clinical application. In this study, on the basis of density functional theory (DFT), the electronic structural characteristics of Sco were systematically analyzed via the B3LYP-D3BJ/6-311G(d,p) method. With chloromethyl-functionalized single-walled carbon nanotubes (SWCNTs (5,0)-CH2Cl) as the carrier, the adsorption interaction model between Sco and the carrier was optimized in combination with the solvation model based on density (SMD). The results of the theoretical analysis reveal that Sco forms a coplanar large π-conjugated system between its benzene ring and pyrone ring; O14-H22 and C1=O11 are identified as the key active sites of Sco; and the frontier molecular orbital (FMO) energy gap of Sco is 4.17 eV, which endows the molecule with both reactivity and chemical stability. In the adsorption interaction model, Sco is adsorbed onto the carrier surface via the synergistic effect of van der Waals forces and π-π stacking, with an adsorption energy of 0.56 eV, corresponding to exothermic physical adsorption. The charge-transfer amount is only 0.0128 e, which can prevent structural distortion of the drug molecule. After adsorption, the energy gap of the system narrows to 1.55 eV, with a considerable increase in electrical conductivity, and the electronic structure related to the core activity of Sco remains unchanged. This study theoretically elucidated the fundamental electronic structure of Sco and its interaction characteristics with functionalized carbon nanotubes (f-CNTs).

Critical limb ischemia (CLI) results from inadequate blood perfusion in the extremities, leading to accelerated muscle cell apoptosis in mild cases and lower limb claudication in severe cases. It can therefore significantly reduce the quality of life of patients. In this study, baicalin (BA), a representative flavonoid, is shown to induce myogenic differentiation thus alleviating CLI. The underlying mechanism of action of BA primarily involves the direct activation of the Wnt/β-catenin signaling pathway and the drug metabolism enzyme cytochrome P450 (Cyp1a1) in muscle cells. To address the challenging aqueous solubility of flavonoids for in vivo applications, we designed an innovative composite microneedle patch (GEL+PUE+BA MNs). This delivery strategy not only overcomes the solubility limitations of flavonoids in vivo but also promotes the repair of ischemic muscle through localized intervention. It efficiently delivers BA to the ischemic site, stimulates the associated muscle cells, and induces their differentiation into skeletal muscle. This process markedly reduces the apoptotic cells and muscle loss in the lower extremities. Our study underscores the significant therapeutic effects of GEL+PUE+BA MNs as a biomaterial for treating hindlimb ischemia and highlights its considerable potential in managing CLI.

Due to the skin's limited self-repair capacity after injury, developing effective wound dressings is crucial to accelerate wound closure and enhance tissue regeneration. In the work, a thermo-sensitive and tissue-adhesive hydrogel with an interpenetrating polymer network (QGNPI) was fabricated by using N-isopropylacrylamide (NIPAM), galactomannan, and chitin-lignin micelles. The abundant hydroxyl groups within galactomannan conferred enhanced adhesion properties to the hydrogel. To improve the stability of artesunate, it was encapsulated within the chitin-lignin micelles. Moreover, the QGNPI hydrogel exhibited excellent thermal responsiveness and strong tissue adhesion, enabling effective wound closure. It also demonstrated outstanding hemocompatibility and cytocompatibility, supporting its suitability for wound healing applications. In murine models, the QGNPI hydrogel functioned as an efficient sealant, achieving sutureless wound closure during initial healing. Moreover, it significantly accelerated tissue regeneration by reducing inflammatory cell infiltration, promoting organized collagen deposition, and stimulating robust angiogenesis. Collectively, these findings underscore the potential of QGNPI as a promising thermosensitive and adhesive hydrogel dressing for advanced wound management.

The treatment of myocardial ischemia-reperfusion injury (IRI) requires urgent improvement of mitochondrial dysfunction and sustained energy supply to restore cardiac function, but currently, there is a lack of effective strategies to meet these needs. Here, we transplanted mitochondria to treat myocardial IRI by a sequential administration approach. First, nanomotors with chemotactic target ability are modified on the surface of mitochondria to obtain engineered mitochondrial nanomotors. Then, denatured bovine serum albumin is modified outside the nanomotor, enabling mitochondria to hitchhike on activated neutrophils to accumulate in the damaged heart. During reperfusion, immediate intramyocardial injection of these mitochondria can stabilize energy supply and rescue dying cardiomyocytes from IRI. In the subsequent tissue repair stage, the mitochondria injected intravenously can achieve stepwise targeting to the damaged heart by hitchhiking on activated neutrophils and chemotactic behavior of nanomotors, thereby continuously supplementing energy to cardiomyocytes and enhancing cardiac function. In addition, in vivo results show that sequential administration reduces adverse reactions such as arrhythmia caused by high-dose mitochondrial transplantation. Compared with existing treatment methods, this design of sequential administration is a special strategy targeting the specific needs and inflammatory microenvironment of myocardial IRI, better promoting the clinical translation of mitochondrial transplantation.

For cationic nanoparticles, the spontaneous nanoparticle-protein corona formation and aggregation in biofluids can trigger unexpected biological reactions. Herein, we present a biomimetic strategy for camouflaging the cationic peptide/siRNA nanocomplex (P/Si) with single or dual proteins, which exploits the unique properties of endogenous proteins and stabilizes the cationic P/Si complex for safe and targeted delivery. An in-depth study of the P/Si protein corona (P/Si-PC) formation and protein binding was conducted. The results provided insights into the biochemical and toxicological properties of cationic nanocomplexes and the rationales for engineering biomimetic protein camouflages. Based on this, the human serum albumin (HSA) and apolipoprotein AI (Apo-AI) ranked within the top 20 abundant protein species of P/Si-PC were selected to construct biomimetic HSA-dressed P/Si (P/Si@HSA) and dual protein (HSA and Apo-AI)-dressed P/Si (P/Si@HSA_Apo), given that the dual-protein camouflage plays complementary roles in efficient delivery. A branched cationic peptide (b-HKR) was tailored for siRNA delivery, and their nanocomplexes, including the cationic P/Si and biomimetic protein-dressed P/Si, were produced by a precise microfluidic technology. The biomimetic anionic protein camouflage greatly enhanced P/Si biostability and biocompatibility, which offers a reliable strategy for overcoming the limitation of applying cationic nanoparticles in biofluids and systemic delivery.

(-)-Epigallocatechin-3-O-gallate (EGCG), one of the green tea catechins, exhibits significant antioxidant properties that play an essential role in various diseases. However, the functional role and underlying mechanism of EGCG in stimulating of hepatic stellate cells (HSCs) remain unexplored in transcriptomics sequencing studies. The present study suggests that oral administration of EGCG at a dosage of 200 mg/kg/day for a duration of four weeks exhibits significant therapeutic potential in a murine model of liver fibrosis induced by CCl4. The activation of HSCs in vitro was dose-dependently inhibited by EGCG. The sequencing analysis data reveled that EGCG exerted a regulatory effect on the calcium signal in mouse HSCs, resulting in a decrease in calcium ion concentration. Further analysis revealed that EGCG inhibited the expression of phospholipase C epsilon-1 (PLCE1) and inositol 1, 4, 5-trisphosphate (IP3) in activated mouse HSCs. Additionally, EGCG contributes to the reduction the concentration of calcium ions by regulating PLCE1. After the knockdown of PLCE1, free calcium ion concentrations decreased, resulting in the inhibition of both cell proliferation and migration. Interestingly, the expression of PLCE1 and cytosolic calcium levels were regulated by reactive oxygen species(ROS). Furthermore, our findings suggest that ROS might inhibit the expression of PLCE1 by inhibiting TFEB, a transcription activator involved in the nuclear translocation process. Our study provided novel evidence regarding the regulatory effects of EGCG on activated HSCs (aHSCs) in mice by the calcium signaling pathway, emphasizing the crucial role of PLCE1 within the calcium signaling network of HSCs. The proposition was also made that PLCE1 holds promise as a novel therapeutic target for murine liver fibrosis.

Mitochondrial transplantation is an important therapeutic strategy for restoring energy supply in patients with ischaemic heart disease (IHD); however, it is limited by the invasiveness of the transplantation method and loss of mitochondrial activity. Here we report successful mitochondrial transplantation by oral administration for IHD therapy. A nitric-oxide-releasing nanomotor is modified on the mitochondria surface to obtain nanomotorized mitochondria with chemotactic targeting ability towards damaged heart tissue due to nanomotor action. The nanomotorized mitochondria are packaged in enteric capsules to protect them from gastric acid erosion. After oral delivery the mitochondria are released in the intestine, where they are quickly absorbed by intestinal cells and secreted into the bloodstream, allowing delivery to the damaged heart tissue. The regulation of disease microenvironment by the nanomotorized mitochondria can not only achieve rapid uptake and high retention of mitochondria by damaged cardiomyocytes but also maintains high activity of the transplanted mitochondria. Furthermore, results from animal models of IHD indicate that the accumulated nanomotorized mitochondria in the damaged heart tissue can regulate cardiac metabolism at the transcriptional level, thus preventing IHD progression. This strategy has the potential to change the therapeutic strategy used to treat IHD. Ischaemic heart disease, which poses a severe threat to human health, is characterized by mitochondria damage and energy metabolism disorder. Here mitochondria were orally transplanted to the heart using nanomotors to restore energy metabolism by non-invasive administration.

Efficient neutrophil migration to infection sites plays a vital role in the body's defense against bacterial infections and natural immune responses. Neutrophils have a short lifespan and cannot be mass-cultured in vitro. Therefore, developing more stable artificial neutrophils (AN) in a controllable manner has become a research focus. However, existing AN lack chemotaxis, which is the ability to migrate toward high-signal-concentration positions in a dynamic blood- flow environment. Supplying AN with chemotaxis is key to designing AN that are more similar to natural neutrophils in terms of morphology and function. In this study, micrometer-sized, spherical, biocompatible AN are developed. These AN consist of zeolitic imidazolate framework-8 nanoparticles encapsulating two enzymes, coacervate droplet frameworks, and outer phospholipid bilayers carrying enzymes. The AN exhibit responsiveness to elevated hydrogen peroxide levels at inflammation sites, actively chemotaxing toward these sites along concentration gradients. They also demonstrate effective combat against Staphylococcus aureus infections. The capabilities of the AN are further validated through in vitro experiments and in vivo evaluations using vascular graft infection models. This study replicates natural neutrophils in terms of chemical composition, functionality, and physiological impact. It introduces new ideas for advancing the development of advanced artificial cells.

Patients who suffer from sepsis typically experience acute lung injury (ALI). Extracellular vesicles (EVs) contain miRNAs, which are potentially involved in ALI. However, strategies to screen more effective EV-miRNAs as therapeutic targets are yet to be elucidated. In this study, functional EV-miRNAs were identified based on multiomics analysis of single-cell RNA sequencing of targeted organs and serum EV (sEV) miRNA profiles in patients with sepsis. The proportions of neutrophils and macrophages were increased significantly in the lungs of mice receiving sEVs from patients with sepsis compared with healthy controls. Macrophages released more EVs than neutrophils. MiR-125a-5p delivery by sEVs to lung macrophages inhibited Tnfaip3, while miR-221-3p delivery to lung neutrophils inhibited Fos. Macrophage membrane nanoparticles (MM NPs) loaded with an miR-125a-5p inhibitor or miR-221-3p mimic attenuated the response to lipopolysaccharide (LPS)-induced ALI. Transcriptome profiling revealed that EVs derived from LPS-stimulated bone marrow-derived macrophages (BMDMs) induced oxidative stress in neutrophils. Blocking toll-like receptor, CXCR2, or TNFα signaling in neutrophils attenuated the oxidative stress induced by LPS-stimulated BMDM-EVs. This study presents a novel method to screen functional EV-miRNAs and highlights the pivotal role of macrophage-derived EVs in ALI. MM NPs, as delivery systems of key sEV-miRNA mimics or inhibitors, alleviated cellular responses observed in sepsis-induced ALI. This strategy can be used to reduce septic organ damage, particularly lung damage, by targeting EVs.

The key glycolytic enzyme phosphofructokinase (PFK) is responsible for maintaining glycolytic stability and an important energy source for activating hepatic stellate cells (HSCs). However, its regulation in activated HSCs remains unclear. Caveolin-1 (Cav1), a major constituent of caveolae, has emerged as a key target for triggering glycolysis. However, the relationship between Cav1 and glycolysis during HSC activation is not well established. In this study, Cav1 was upregulated in mouse and human fibrotic liver tissues. We concluded that HSC-specific Cav1 knockdown markedly alleviates liver injury and fibrosis. Mechanistically, Cav1 was elevated during primary mouse HSC activation, competing with SQSTM1 for the regulatory subunit of PFK liver type and inhibiting the SQSTM1-mediated autophagy-independent lysosomal degradation pathway to sustain HSC activation. We also identified the heptapeptide alamandine as a promising therapeutic agent that downregulates Cav1 protein levels via proteasomal degradation and may impair glycolysis. Our study provides evidence of the crucial role and mechanism of Cav1 in the glucose metabolic network in HSCs and highlights Cav1 as a critical therapeutic target for the treatment of liver fibrosis.

Uncontrolled and excessive fibrosis after myocardial infarction (MI) in the peri-infarct zone leads to left ventricular remodeling and deterioration of cardiac function. Inhibiting fibroblast activation during the mature phase of cardiac repair improves cardiac remodeling and function after MI. Here, we engineered a biocompatible microneedle (MN) patch using gelatin methacryloyl and loaded it with galunisertib, a transforming growth factor-beta (TGF-β)-specific inhibitor, to treat excessive cardiac fibrosis after MI. The MN patch could sustainably release galunisertib for more than 2 weeks and provide mechanical support for the fragile ventricular wall. After being applied to a rat model of MI, the galunisertib-loaded MN patch improved long-term cardiac function and reduced cardiac fibrosis by effectively inhibiting TGF-β depending on fibroblast activation. This strategy shows the potential of the MN patch as an advanced platform to locally deliver direct antifibrotic drugs to prevent myocardial fibrosis for the treatment of MI and the promotion of cardiac repair.

In the past decades, the striking development of cationic polypeptides and cell-penetrating peptides (CPPs) tailored for small interfering RNA (siRNA) delivery has been fuelled by the conception of nuclear acid therapy and precision medicine. Owing to their amino acid compositions, inherent secondary structures as well as diverse geometrical shapes, peptides or peptide-containing polymers exhibit good biodegradability, high flexibility, and bio-functional diversity as nonviral siRNA vectors. Also, a variety of noncovalent nanocomplexes could be built via self-assembling and electrostatic interactions between cationic peptides and siRNAs. Although the peptide/siRNA nanocomplex-based RNAi therapies, STP705 and MIR-19, are under clinical trials, a guideline addressing the current bottlenecks of peptide/siRNA nanocomplex delivery is in high demand for future research and development. In this review, we present strategies for improving the safety and RNAi efficacy of noncovalent peptide/siRNA nanocomplexes in the treatment of genetic disorders. Through thorough analysis of those RNAi formulations using different delivery strategies, we seek to shed light on the rationale of peptide design and modification in constructing robust siRNA delivery systems, including targeted and co-delivery systems. Based on this, we provide a timely and comprehensive understanding of how to engineer biocompatible and efficient peptide-based siRNA vectors.

A series of pH-sensitive semi-interpenetrating polymer network (semi-IPN) hydrogels poly(vinyl alcohol)/poly(hydroxypropyl methacrylate-co-methacrylic acid) (PVA/P(HPMA-co-MAA)) were synthesized by free-radical polymerization of HPMA and MAA in the presence of PVA. The physicochemical property of the obtained hydrogels was characterized by Fourier transform infrared spectroscopy, X-ray diffraction, thermogravimetric analyses and scanning electron microscopy (SEM) measurements. The SEM photograph revealed the network formation with uniform pore distribution. The swelling behavior of each hydrogel in buffer solution showed a simultaneous sensitivity to pH and ionic strength: the swelling ratio of all hydrogels was higher in neutral environment than in acidic medium; besides, equilibrium swelling ratio of the hydrogels decreases as the ionic strength increases. Insulin was loaded into the PVA/P(HPMA-co-MAA) semi-IPN hydrogel. The in vitro insulin release experiment was carried out in buffer solutions at pHs 1.2 and 6.8. Results showed that the release of entrapped insulin was inhibited at pH 1.2 but obviously increased at pH 6.8. Cell viability revealed that the hydrogels were biocompatible. After oral administration of insulin-loaded hydrogel to streptozotocin-induced diabetic rats at 75 IU/kg, a sustained reduction in blood glucose level was observed. Therefore, the semi-IPN PVA/P(HPMA-co-MAA) hydrogels are potential vehicles for oral delivery of protein drugs.

Extracellular vesicles (EVs) have emerged as important vectors of intercellular dialogue. High mobility group box protein 1 (HMGB1) is a typical damage-associated molecular pattern (DAMP) molecule, which is cytotoxic and leads to cell death and tissue injury. Whether EVs are involved in the release of HMGB1 in lipopolysaccharide (LPS)-induced acute liver injuries need more investigation. EVs were identified by transmission electron microscopy, nanoparticle tracking analysis (NTA), and western blotting. The co-localization of HMGB1, RAGE (receptor for advanced glycation end-products), EEA1, Rab5, Rab7, Lamp1 and transferrin were detected by confocal microscopy. The interaction of HMGB1 and RAGE were investigated by co-immunoprecipitation. EVs were labeled with the PKH67 and used for uptake experiments. The pyroptotic cell death was determined by FLICA 660-YVAD-FMK. The expression of NLRP3 (NOD-like receptor family pyrin domain containing 3) inflammasomes were analyzed by western-blot or immunohistochemistry. Serum HMGB1, ALT (alanine aminotransferase), AST (aspartate aminotransferase), LDH (lactate dehydrogenase) and MPO (myeloperoxidase) were measured using a commercial kit. The extracellular vesicle HMGB1 was detected in the serums of sepsis patients. Macrophages were found to contribute to HMGB1 release through the EVs. HMGB1-RAGE interactions participated in the loading of HMGB1 into the EVs. These EVs shuttled HMGB1 to target cells by transferrin-mediated endocytosis leading to hepatocyte pyroptosis by the activation of NLRP3 inflammasomes. Moreover, a positive correlation was verified between the sepsis serum EVs-HMGB1 level and clinical liver damage. This finding provides insights for the development of novel diagnostic and therapeutic strategies for acute liver injuries.

To investigate effect and mechanism of Ziqi Ruangan Decoction (ZQRGD) on hepatic fibrosis in rats. Rats were randomly assigned to blank group, model group, colchicine group, ZQRGD high-dose group, ZQRGD middle-dose group, and ZQRGD low-dose group. All groups except group A were intraperitoneally injected with 40% CCl4/olive oil for 8 weeks; group C was then given intragastric colchicine administration. Groups D, E, and F were intragastrically dosed with ZQRGD. Compared with the colchicine group, the superoxide dismutase (SOD) activity of each dose group of ZQRGD significantly increased. TNF-α and IL-6 concentration significantly decreased in each drug intervention group, while these significantly decreased in the high-dose and medium-dose ZQRGD groups. The expression of α-SMA and collagen I significantly decreased in the drug treatment group compared with the model group, as did the expression of PI3K, AKT, and mTOR. Ziqi Ruangan Decoction had a favorable anti-liver fibrosis effect and the mechanism is related to anti-oxidative stress, anti-inflammation, the inhibition of the PI3K/Akt/mTOR signaling pathway, and the inhibition of hepatic stellate cell activation.

In this research, pH-sensitive semi-interpenetrating polymer network hydrogels based on sodium carboxymethyl cellulose and poly(methacrylic acid) were synthesized using free radical polymerization and semi-interpenetrating polymer network approach for oral administration of insulin. The chemical structure and thermal stability of the hydrogels were characterized using Fourier transform infrared spectroscopy, X-ray diffraction, and thermogravimetric analysis measurements. The interior morphology was observed by scanning electron microscopy and the inner structure exhibited a porous honeycomb-like shape. The investigations on the swelling properties of hydrogels revealed their ability to response to pH value change. The in vitro release behavior of insulin was pH dependent and the release of insulin was much lower at pH 1.2 compared to pH 6.8. In vitro cytotoxicity assay indicated that the hydrogels were noncytotoxic to HeLa cells. A sustained reduction in blood glucose level was observed after oral administration of insulin-loaded hydrogel to diabetic rats at 75 IU/kg. These results indicated that the hydrogel would be a promising vehicle for oral insulin delivery systems.