The application of conductive substrates in anaerobic digestion has garnered growing interest due to their potential to enhance wastewater treatment performance. However, the initial attachment mechanisms governing distinct colonization across substrates remain unclear. In this study, the most commonly used conductive materials from previous research (biochar, activated carbon, magnetite, and graphite) were employed to investigate the adhesion and biofilm formation characteristics of a typical electron acceptor, Methanosarcina barkeri. Results indicate that adsorption is governed primarily by Lewis acid-base interactions between acidic functional groups on the activated carbon surface and methanochondroitin in the cellular outer layer. During early colonization, cell attachment to activated carbon was 2.3, 3.3 and 6.3 times higher than biochar, magnetite and graphite, respectively. This mechanism was corroborated by substrate modification experiments demonstrating that removing surface acidic functional groups reduced attachment capacity by 57.6%, whereas enriching carboxyl groups increased it by 116.8%. Theoretical calculations revealed that this enhancement arises from an attractive interaction energy of approximately -15 kcal/mol between methanochondroitin and carboxyl groups. Consequently, this advantage in early colonization promoted the preferential formation of a structurally mature, extracellular polymeric substances-rich biofilm on the activated carbon surface, ensuring mechanical stability under fluid shear stress and providing substantial material support for efficient electron transfer and storage. This study provides a predictive framework for substrate selection, in which tailoring surface functional-group density to match specific microbial biochemical motifs enables the development of high-performance systems, shifting the field from empirical trial and error to rational design.
Poly(methyl methacrylate) (PMMA) remains a clinical standard in orthopedic procedures due to its reliable primary fixation. However, its inherent lack of bioactivity and poor osseointegration limit long-term success. While bioactive additives can improve biological response, they frequently compromise structural integrity, challenging the effective design of load-bearing implants. This study introduces calcium methacrylic phyllosilicate (CaMP) as a novel bioactive comonomer, designed to induce osteogenic activity in PMMA bone cements. Chemical characterization confirms the successful synthesis of CaMP, which is covalently incorporated into the PMMA matrix via radical copolymerization (up to 15 wt%) without compromising the structural integrity of the bone cements. Surface analysis after biomimetic mineralization reveals a significant increase in the calcium and phosphorus content on the surface of the samples, demonstrating induced calcium nucleation. Deconvolution of Raman and XPS spectra confirms the template-driven upregulation of the calcium phosphate mineralization, promoting the transition from amorphous phases into poorly crystalline apatite-like deposits. Moreover, the hybrid bone cements elicit sustained calcium release governed by Fickian diffusion, which validates the ability of the material to induce osteogenic differentiation, endogenous biomimetic mineralization, and long-term apatitic crystal maturation. Consequently, the in situ functionalization of PMMA bone cements via CaMP copolymerization provides a robust pathway for developing mechanically stable, bioactive hybrid implants for load-bearing applications.
Triple-negative breast cancer (TNBC) remains one of the most aggressive and therapeutically challenging breast cancer subtypes due to the lack of effective treatments, high recurrence rates, and poor clinical outcomes. Curcumin (Cur) is a natural anticancer agent; however, its clinical translation is limited by poor aqueous solubility, low stability, and insufficient tumor selectivity. In this study, Cur/carboxymethyl-β-cyclodextrin (CM-βCD) supramolecular assemblies were developed using different formulation approaches. Then, GE11 was covalently coupled to the carboxyl groups of CM-βCD via post-assembly functionalization. For this purpose, the formulation exhibiting the highest Cur loading (16.95%) was selected to construct EGFR-targeted supramolecular delivery systems. GE11-functionalized Cur/CM-βCD supramolecular assemblies significantly enhanced Cur cytotoxicity, reducing IC50 values by ~1.7-fold in MCF-7 cells and up to 2.2-fold in MDA-MB-231 TNBC cells compared to free Cur at 48 h. Quantitative uptake studies revealed a ~3.25-fold higher cellular internalization in MDA-MB-231 cells relative to MCF-7 cells. Moreover, GE11-Cur/CM-βCD markedly suppressed proliferation, evidenced by a 3.29-fold reduction in PCNA-positive cells, and induced apoptosis, with a 5.1-fold increase in TUNEL-positive cells. Overall, the findings suggest that GE11-functionalized Cur/CM-βCD is a promising targeted strategy to overcome Cur limitations and address TNBC therapy needs.
The pursuit of effective anti-aging interventions remains a significant challenge in dermatological research. Ellagic acid (EA), a natural polyphenol with demonstrated anti-skin-aging properties, faces limitations in therapeutic application due to poor percutaneous absorption. To address this challenge, we developed TPGS-functionalized liposomes (EA-TPGS-L) through systematic optimization using a Box-Behnken design, achieving an encapsulation efficiency of 94.19% and an optimal particle size of 152.50 nm. MD simulations revealed that TPGS enhances skin permeation by spontaneously inserting its hydrophobic domain into the stratum corneum, thereby disrupting the ordered lipid packing and specifically reducing the free energy barrier within the hydrophobic core of the bilayer. The optimized EA-TPGS-L formulation exhibited comprehensive anti-aging effects, including potent antioxidant activity with greater radical scavenging capacity compared to free EA and EA-L, significant inhibition of key aging-related enzymes, and effective cellular protection evidenced by a reduction in lipid peroxidation. Furthermore, skin-on-a-chip evaluations demonstrated the formulation's ability to promote tissue remodeling, showing 84.11% recovery of type I collagen while effectively regulating oxidative stress markers. These collective findings establish TPGS-modified liposomes as a promising transdermal delivery platform that overcomes the skin barrier while amplifying the anti-aging efficacy of natural polyphenols like EA, offering significant potential for advanced dermatological and cosmeceutical applications.
Multidrug-resistance (MDR) remains a formidable obstacle in the treatment of breast cancer, primarily due to the overexpression of P-glycoprotein (P-gp) efflux pumps. The objective of this study is to develop a copper-based nanotherapeutic agent capable of simultaneously inhibiting drug efflux and inducing a novel form of cell death, cuproptosis, to overcome MDR. The nanoparticle known as DOX-P-gp ASO-Cu2 + NPs has been engineered to facilitate the delivery of doxorubicin (Dox), P-gp antisense oligonucleotide (P-gp ASO), and copper ions into cancer cells. The incorporation of ASO effectively silenced P-gp expression, significantly enhancing the retention of Dox within the cells. Consequently, the IC50 value of Dox in the nanoparticle formulation was reduced by 17.5-fold compared to free Dox in MCF-7/ADM cells. This synergistic strategy not only restored chemosensitivity but also triggered cuproptosis, as evidenced by mitochondrial dysfunction, downregulation of LIAS/FDX1, and DLAT oligomerization. The results of drug-resistant breast cancer xenograft model further demonstrated superior tumor suppression with the nanoformulation compared to monotherapy. This integrated strategy, which involves the inhibition of drug efflux and the induction of cuproptosis, offers a promising approach to eradicating multidrug-resistant breast cancer.
Diverse living systems and organisms harness the positional and directional distribution of solutes to drive essential functions. Developing engineered platforms that can mimic these complex spatial domains is crucial for achieving more effective operational biotech workstations. To this end, we first demonstrate that oil confined within the nanocavities of mesoporous thin films serves as reservoirs for hydrophobic drug release when immersed in aqueous media, modulating cell behavior in local environments. We further show that the spontaneous interaction of the mesoporous architecture with discrete liquid volumes (droplets) enables radial release as function of volume and incubation time, evidenced by using cell staining as readout of the generation of programmable spatiotemporal action patterns. This approach takes advantage of mesoporous system features coupled with liquid-liquid dynamics to achieve vectorial biological effects, thereby providing a biomimetic platform that emulates natural settings in advanced biotechnological applications.
Stimuli- responsive nanocarriers must retain their therapeutic cargo in the body yet release it efficiently at disease sites. Achieving both properties in a simple, stable, and adjustable system remains challenging. In this study, we developed a pH- responsive polymer- lipid hybrid nanovesicle, PC7ALV, by incorporating poly(2- hexamethyleneiminoethyl methacrylate) (PC7A) into dimyristoylphosphatidylcholine (DMPC) bilayers. We investigated whether PC7A domains within the membrane could enhance stability under physiological pH conditions and promote cargo release under mildly acidic conditions. The novelty of this approach lies in using PC7A as a structural component of the membrane that responds to pH changes, rather than as a conventional self- assembling polymer carrier. Using a combination of techniques, including dynamic light scattering, zeta-potential analysis, electron microscopy, fluorescence colocalization, Förster resonance energy transfer (FRET), and Laurdan membrane-fluidity measurements, we demonstrated that PC7A integrates with the lipid bilayer and forms small polymer-rich regions at pH 7.4. This organization enhances the membrane's order and stability. Upon decreasing the pH to 6.4, PC7A becomes protonated and undergoes disassembly; the association between the polymer and lipid weakens, FRET decreases, vesicle shape changes, and the membrane becomes more permeable and less stable, thereby facilitating cargo release. For the 1:1 lipid-to-PC7A formulation, approximately 16% of FAM-labeled ovalbumin was released at pH 7.4, whereas release increased to approximately 48% at pH 6.4 after 12 h. Increasing the PC7A content accelerated cargo release under acidic conditions while further improving vesicle stability at physiological pH. In MDA-MB-231 cells, PC7ALVs resulted in higher intracellular fluorescence and increased paclitaxel-mediated cytotoxicity under acidic conditions compared with a non-pH-responsive model vesicle. These findings demonstrate that incorporation of PC7A into the membrane enables pH-dependent control over polymer- lipid vesicle organization, vesicle stability, and molecular transport. PC7ALVs provide a proof-of-concept for designing pH-responsive nanocarriers that preserve cargo integrity during circulation while promoting release under acidic conditions. Further research with other models is warranted to understand how this system works and to assess its broader use, stability, and safety.
Osteosarcoma, the most common primary malignant bone tumor in adolescents, has a poor prognosis due to high rates of metastasis, recurrence, and chemoresistance, necessitating novel treatments. Herein, a biomimetic nanoplatform CFCM/VK3 was engineered to exploit synergistic iron-copper interference against osteosarcoma. This platform comprises an optimized CuFe2O4 nanoparticle core with superior Fenton-like activity, glutathione depletion capacity, and near-infrared photothermal properties, combined with vitamin K3 (VK3) as an endogenous H2O2 generator specifically activated by tumor-overexpressed quinone oxidoreductase 1 (NQO1). The core is cloaked with K7M2 osteosarcoma cell membranes to enable homologous targeting and immune evasion. After tumor-specific uptake, the acidic microenvironment activates VK3 to generate H2O2 and liberates iron and copper ions. This initiates a self-reinforcing cascade wherein iron-driven oxidative stress impairs mitochondrial ATP synthesis, and the resultant energy shortage traps copper inside cells, thereby amplifying the toxicity of both ions. Critically, the cooperative action of iron and copper simultaneously activates two regulated cell death pathways: iron‑dependent lipid peroxidation leads to ferroptosis and copper‑induced aggregation of lipoylated proteins triggers cuproptosis. Near-infrared irradiation further accelerates these catalytic reactions and provides photothermal ablation. The CFCM/VK3 nanoplatform demonstrates significant tumor suppression and excellent biosafety in an osteosarcoma mouse model, establishing a new paradigm of metal-ion interference therapy that leverages the mutual dependency between iron and copper for self-amplifying antitumor effects.
Repeated intravitreal injections of anti-vascular endothelial growth factor agents for choroidal neovascularization (CNV) impose significant patient burden, increase the risk of ocular complications, and lead to poor compliance. To address these limitations, we developed a transscleral, sustained-release platform aiming to enable single-administration combination therapy targeting multiple pathological pathways while reducing injection frequency and avoiding intravitreal puncture. Cationic solid lipid nanoparticles (Res/Conb-SLNs) were fabricated via thin-film ultrasonication and electrostatic adsorption to co-encapsulate resveratrol (Res) and conbercept (Conb). These were integrated into hyaluronic acid-based annular dissolvable microneedles (Res/Conb-SLNs@MN, 500 μm height) designed to target the suprachoroidal space (SCS) while avoiding the optical axis. The system achieved biphasic sustained release over 48 h (>80% Res, >90% Conb) and demonstrated rapid microneedle dissolution (<30 s) with complete micropore closure within 24 h. Enhanced cellular uptake, potent antioxidant activity, and significant inhibition of endothelial cell migration/tube formation were observed. Intraocular pharmacokinetic experiments confirm that this Res/Conb-SLNs@MN achieved SCS-targeted accumulation with significantly extended posterior segment retention. In a laser-induced rat CNV model, Res/Conb-SLNs@MN delivered therapeutic agents efficiently to the retina-choroid region, significantly suppressing CNV leakage, reducing neovascularization, and attenuating inflammatory cell infiltration. No changes in intraocular pressure or retinal histology were observed over 42 days. Moreover, the safety of repeated microneedle administration was good. This annular microneedle-nanocomposite system enables transscleral, sustained, and targeted co-delivery to the posterior segment without the need for intravitreal puncture, offering a transformative strategy to replace frequent intravitreal injections with a synergistic, patient-compliant approach for neovascular retinal diseases.
Achieving coordinated inflammation resolution, oxidative stress control, and angiogenesis remains a key goal for skin wound repair. The plant-derived C-glycosyl flavonoid swertisin has known anti-inflammatory and antioxidant effects, but its role in skin wound repair remains untested. Herein, we loaded swertisin into a sodium alginate/silk sericin composite hydrogel and tested it in a mouse full-thickness skin wound model. The hydrogel released swertisin steadily over 7 days without burst release and maintained a moist, cytocompatible wound environment. In vitro, it promoted fibroblast migration and proliferation. In vivo, it accelerated wound closure and achieved near-complete re-epithelialization by day 10, with improved collagen deposition and extracellular matrix organization. Specifically, the treatment reduced NF-κB p65 nuclear translocation, lowered pro-inflammatory cytokines (IL-6, IL-1β, TNF-α), and increased angiogenic markers CD31 and VEGF. It also upregulated Nrf2 and CD206, reduced iNOS, and moderately modulated TGF-β1, indicating coordinated regulation of antioxidant defense, macrophage polarization, and matrix remodeling. No adverse effects were detected in major organs. Notably, this study provides the first evidence that locally delivered swertisin, released from a natural polymer-based hydrogel, can promote full-thickness skin wound repair by dampening inflammation, restoring redox balance, and boosting angiogenesis, offering a promising approach for wound management.
Idiopathic pulmonary fibrosis (IPF) lacks therapies that reverse disease progression, partly due to persistent senescent alveolar epithelial type II cells (SAEC II). We report a synergistic senolytic strategy combining mitochondrial calcium overload with AKAP1 silencing. An acid-responsive nanoplatform (CCMD@Lipo-Apt) comprising a CaCO3 core co-loaded with curcumin (CUR), Mn2+, and an AKAP1-targeting DNAzyme, enveloped by an L1CAM aptamer-modified lipid bilayer, enables SAEC II-specific homing. Upon lysosomal acidification, CaCO3 degrades to release Ca2+, CUR (mobilizing ER Ca2+), Mn2+, and the DNAzyme. Mn2+ simultaneously activates DNAzyme to knockdown AKAP1 (sensitizing mitochondria to calcium overload) and directly activates caspase-8. This triple mechanism collapses mitochondrial membrane potential and triggers robust apoptosis exclusively in SAEC II. In a bleomycin-induced mouse model, CCMD@Lipo-Apt clears SAEC II, attenuates fibrosis, and restores lung function without overt toxicity. This work establishes a synthetic-lethal axis for targeted IPF therapy.
Acute lung injury (ALI) is a common critical illness driven by uncontrolled pulmonary inflammation, with hyperactivated macrophages sustaining pro-inflammatory cytokine release. Activation of liver X receptor α (LXRα) can suppress inflammatory gene expression. However, clinical translation of LXRα agonists such as T0901317 (T09) is limited by off-target effects, particularly hepatotoxicity. Here, we developed a biomimetic controlled-release platform (AB/NPs@T09) using mesenchymal stem cell (MSC)-derived apoptotic bodies (ABs) as natural carriers for T09. The structural properties of ABs facilitated macrophage uptake via the efferocytosis pathway. Inhalation administration of AB/NPs@T09 enabled dual organ-cell targeting, increasing pulmonary drug accumulation while markedly reducing liver distribution. In an ALI mouse model, inhaled AB/NPs@T09 at a low dose (1 mg/kg) achieved anti-inflammatory efficacy comparable to that of high-dose intraperitoneal injection (10 mg/kg), without inducing hepatotoxicity. The biomimetic platform presented here not only offers a new avenue for therapeutic intervention in clinical ALI, but also establishes a generalizable strategy for repurposing potent yet toxic small molecules in inflammatory diseases.
Chitosan is widely employed as a hemostatic material for hemorrhage control due to its excellent biocompatibility, and intrinsic cationic property that promotes red blood cell aggregation. However, the insufficient stability under high-pressure arterial flow and the poor mechanical strength of the formed clots can lead to a high risk of secondary bleeding. Here, machine learning-guided hierarchical sized chitosan hemostatic particles (CSHP) with electrostatic-hydrophobic synergy were fabricated through multistage particle size distributions and lauric acid loading. The CSHP combined physical plugging (large, medium, and small particles for anchoring, bridging, and dense sealing) with electrostatic attraction and hydrophobic interactions to form a compact clot network that resisted arterial flow independent of systemic coagulation activation. In vitro coagulation tests showed that CSHP-3 reduced whole blood clotting time by 33.3% compared to LA-free CS. Rheological measurements showed that CSHP-3 formed blood clots exhibited a storage modulus of 1566 Pa, which was 51.3% higher than that of the commercial product CELOX™ (1035 Pa) and 6 times that of the blank group (263 Pa). Moreover, in a Bama minipig femoral artery injury model, CSHP-3 achieved successful hemostasis within 180 s. This work paves the way for more stable, more effective hemorrhage control and faster clinical translation in emergency management of deep arterial wounds.
