Photothermal therapy (PTT) leverages light-absorbing nanoparticles (NPs) to convert light energy into localized heat, providing a targeted and minimally invasive approach for tumor ablation. Despite its promise, PTT faces several limitations, chiefly the poor tissue penetration depth and the risk of cancer recurrence due to incomplete tumor eradication. Recent breakthroughs, however, reveal that carefully engineered NPs can trigger immunogenic cell death (ICD) within an optimal thermal range. This process releases a cascade of damageassociated molecular patterns (DAMPs), activating dendritic cells (DCs) and priming tumor-specific T-cell immunity. By converting immunologically “cold” tumors into “hot” ones, this strategy opens the door for synergistic combination therapies with immunotherapy. This review outlines cutting-edge progress across precious-metal, organic, and hybrid photothermal nanomaterials, emphasizing how PTT-triggered ICD works at the molecular level and highlighting key strategies to enhance synergy with other treatments, such as immunotherapy, radiotherapy (RT), and chemotherapy. It also explores future directions and the challenges that remain in improving ICD efficiency, overcoming immunosuppressive tumor microenvironments (TME), and translating these strategies into clinical practice.
Background: Insufficient drug targeting and the immunosuppressive tumor microenvironment are key factors contributing to the high mortality rate of hepatocellular carcinoma (HCC). The targeted cavitation effect mediated by low-intensity pulsed ultrasound (LIPUS) provides a promising strategy to address these challenges. Although Piezo1 is known to be dysregulated in multiple cancer types, it remains unclear whether ultrasound stimulation can activate Piezo1 and trigger downstream cell death and immune activation. Methods: Perfluoropentane (PFP)@R-Lip was prepared to assess its combined effects with LIPUS on proliferation, migration, invasion, apoptosis, and ferroptosis in HCC cells. The specific Piezo1 inhibitor GsMTx4 was used to verify the role of Piezo1 activation and immunogenic cell death (ICD) induction. A tumor-bearing mouse model was established to validate the in vivo antitumor efficacy of LIPUS combined with PFP@R-Lip. Student’s t-test, one-way ANOVA test, and two-way ANOVA test were used. Results: PFP@R-Lip was successfully prepared and exhibited enhanced targeting ability to HCC cells. The combination of LIPUS and PFP@R-Lip inhibited the proliferation, migration, invasion, and cell cycle of HCC cells. Mechanistically, the combination activated the Piezo1 channel, leading to intracellular calcium overload, mitochondrial membrane potential (MMP) depolarization, and reactive oxygen species (ROS) accumulation. This cascade induced both apoptosis and ferroptosis, ultimately triggering ICD. These effects were partially reversed by pretreatment with GsMTx4. Conclusions: PFP@R-Lip with ultrasound imaging properties and tumor-targeting specificity was developed. LIPUS combined with PFP@R-Lip was verified to induce intracellular calcium overload and synergistically trigger apoptosis and ferroptosis to prompt ICD in HCC via Piezo1 activation.
Background: Repairing tracheal defects remains a significant challenge in tracheal surgery. Previous attempts using traditional tissueengineered tracheal scaffolds in large animal models have largely failed due to inflammation and insufficient mechanical properties. Methods: In this study, we propose a novel approach that uses cell sheet technology to construct scaffold-free cartilage sheets in vitro. These sheets are stacked and then implanted into goats for further maturation, resulting in large neocartilage tissue with mechanical properties comparable to native tracheal cartilage. We evaluated three methods of tracheal defect repair in goat models using the stacked cartilage sheets: (1) In situ repair with stacked cartilage sheets without vascular pedicle; (2) In situ repair with stacked cartilage sheets and preservation of the vascular pedicle; (3) In situ repair with stacked cartilage sheets, preservation of the vascular pedicle, and T-tube insertion. Results: Airway stability was successfully restored in all animals, with all goats surviving until the end of the experiment without surgery-related complications. Goat 1 exhibited significant granulation tissue hyperplasia compared to goats 2 and 3. The airway morphology was best maintained in goat 3, which showed the highest degree of re-epithelialization, followed by moderate reepithelialization in goat 2, and minimal in goat 1. Conclusions: These results suggest that stacked cartilage sheets are a viable option for tracheal repair, with vascular pedicle preservation and T-tube insertion enhancing therapeutic outcomes. Our study represents the first successful use of scaffold-free cartilage sheets for tracheal repair and provides a theoretical foundation for applying this technology in tracheal reconstruction.
Background: Endometriosis (EMs) is a prevalent gynecological inflammatory disorder characterized by ectopic endometrial tissue growth and a high-ROS microenvironment. In this study, mesoporous silica (mSiO2) was employed to load the COX2 inhibitor celecoxib (CXB), followed by Fe3+-epigallocatechin gallate (EGCG) self-assembly to form a metal-polyphenol coating. The efficacy and biosafety of the resulting nanoparticles (SC@FEG) combined with photothermal therapy (PTT) were systematically investigated, aiming to achieve precise treatment for EMs. Methods: SC@FEG was characterized by dynamic light scattering (DLS), transmission electron microscopy (TEM), and UV-visible spectroscopy, and its ROS-responsive release and photothermal performance were examined. In ectopic endometrial stromal cells (eESCs), the effects of SC@FEG combined with near-infrared (NIR) irradiation were assessed via CCK-8, Calcein-AM/PI staining, Transwell assays, and Western blotting. Moreover, an EMs mouse model was established, and lesion-targeted accumulation and heating effects were monitored by infrared thermography. Apoptosis and fibrosis of ectopic lesions were examined by TUNEL and Masson staining, while anti-inflammatory efficacy was evaluated by Western blotting and ELISA. Biosafety was further assessed by hemolysis testing, histology, and serum biochemistry. Results: SC@FEG displayed stable physicochemical properties, efficient ROS-responsive release, and excellent photothermal conversion. In vitro, SC@FEG with NIR irradiation markedly suppressed eESCs’ proliferation, migration, and invasion, while reducing COX2 expression. In vivo, SC@FEG accumulated in lesions, induced local hyperthermia under laser irradiation, inhibited lesion growth, promoted apoptosis, alleviated fibrosis, and markedly reduced systemic inflammation, without systemic toxicity. Conclusions: SC@FEG represents a multifunctional nanoplatform that combines photothermal ablation and anti-inflammatory drug delivery, providing safe and precise therapy for EMs.
The prevention and treatment of tumors have become a major public health priority. While surgery and chemoradiotherapy remain the cornerstone of current therapeutic strategies, they are accompanied by notable toxic side effects, which severely compromise patients’ quality of life. Flavonoids with precisely characterized molecular structures have demonstrated clinical utility in cancer treatment. Nevertheless, their therapeutic potential is hampered by inherent limitations such as poor aqueous solubility, suboptimal absorption, and chemical instability, ultimately resulting in diminished bioavailability. Recently, the synergistic combination of flavonoid compounds with nanocarrier-based delivery systems has gained prominence as a research focus, owing to its capacity for controlled release of bioactive components and site-specific targeting of tumor tissues. This study examines five flavonoids currently employed in clinical settings, reviewing the latest advancements and future directions in flavonoid-based drug delivery systems (including nanoparticles, hydrogels, and scaffolds) for anticancer applications. Additionally, we review the therapeutic applications of these five flavonoids in other prevalent diseases, proposing novel strategies to overcome the limitations of free drugs and improve therapeutic outcomes in disease management.
Background: Tuberculosis (TB), an infectious disease caused by Mycobacterium tuberculosis (M.tb), remains a formidable global public health threat. The currently available Bacillus Calmette-Guérin (BCG) vaccine, while effective in children, provides inadequate protection for adults, highlighting an urgent need for novel vaccine development. Methods: This study constructed an inhalable biomimetic nanoparticle vaccine, designated ECQ@PS-LPs. The vaccine utilizes liposomes that mimic natural pulmonary surfactant as a carrier to co-encapsulate two key TB antigens, ESAT-6 and CFP-10, along with the immunoadjuvant QS-21. Following intranasal administration in mice, immune responses and protective efficacy were evaluated. Using one-way ANOVA with post-hoc testing, statistical significance was defined as *p < 0.05, **p < 0.01 and ***p < 0.001. Results: The ECQ@PS-LPs vaccine effectively breached the alveolar surfactant barrier, significantly enhancing uptake by antigen-presenting cells. This triggered a robust multidimensional immune response, including potent cellular immunity (characterized by Th1 and Th17 responses), systemic humoral immunity (indicated by IgG production), and strong mucosal immunity (evidenced by sIgA). Furthermore, the vaccine induced durable immune memory, marked by the generation of tissue-resident memory T cells (TRM) and central memory T cells (TCM). In a M.tb challenge model, ECQ@PS-LPs conferred a level of protection comparable to the BCG vaccine and elicited a superior mucosal immune response. The vaccine formulation also demonstrated a simple preparation process and favorable biological safety profile. Conclusions: ECQ@PS-LPs represents a promising inhalable candidate vaccine for TB. It effectively induces comprehensive and persistent immune protection, particularly at the mucosal site, offering a novel and strategic direction for preventing TB in adults.
Osteogenic differentiation of mesenchymal stem cells (MSCs) serves as the cornerstone of bone tissue engineering and regenerative medicine. Traditional biochemical induction methods exhibit limitations, whereas physical stimulation—as a non-invasive, precise, and controllable regulatory approach—demonstrates significant potential in guiding osteogenic differentiation of MSCs. This review comprehensively examines the biological mechanisms by which diverse physical stimuli (including mechanical forces, matrix properties, electromagnetic fields, low-intensity ultrasound, and photobiomodulation (PBM)) promote osteogenic differentiation in bone marrowderived MSCs (BMSCs), analyzes parameter optimization strategies for multi-modal physical stimulation, and envisions the broad application prospects of intelligent and dynamic biomaterial systems in bone regeneration and tissue repair. Finally, this review proposes key directions for future research, emphasizing the importance of multifactorial synergistic regulation, intelligent precision interventions, development of non-invasive techniques, and clinical translation, aiming to provide theoretical foundations and novel insights for designing next-generation efficient and safe bone regeneration strategies.
Background: Chemokine (C-C motif) ligand 2 (CCL2) enhances bone regeneration when integrated into a composite scaffold and show promise for clinical applications in persistent bone defects. Methods: A composite biodegradable scaffold was fabricated using 3D bioprinting with a polylactic acid-glycolic acid copolymer (PLGA) and β-tricalcium phosphate (β-TCP). Gelatin methacrylate (GelMA) hydrogel was used as carrier of CCL2 to enable prolonged release. The scaffolds were immersed in GelMA solution with or without CCL2. In vitro experiments were conducted to analyze the kinetics of CCL2 release and its influence on the proliferation and osteogenic differentiation of rat bone marrow mesenchymal stem cells (BMSC). Scaffolds PLGA/β-TCP (PT), GelMA/PLGA/β-TCP (GPT), and CCL2-conjugated GelMA/PLGA/β-TCP (CGPT) were implanted into a rat femoral defect model (n = 8) and analyzed to evaluate bone mass regeneration at 4 and 8 weeks. Results: CGPT exhibited a prolonged release period lasting 30 days. In vitro, CCL2 significantly promoted BMSCs proliferation (p < 0.05) and the formation of mineralized nodules, and it markedly increased tube formation in HUVECs. In the femoral defect model, bone mineral density and volume fraction in the CGPT group increased compared to the controls (PT, GPT, and control) after implantation. Histological analysis revealed enhanced new bone formation, and integration of the scaffold with surrounding tissue in this group. The expression of osteogenic markers (runt-related transcription factor 2 (Runx2), osteocalcin (OCN), and osteopontin (OPN)) and the angiogenic factor vascular endothelial growth factor (VEGF) significantly increased at the defect site within the CGPT group. Conclusions: Scaffolds made from a composite of GelMA-infused 3D-printed PLGA/β-TCP incorporated with CCL2 greatly enhance osteogenesis and angiogenesis to facilitate bone healing.
Background: Obtaining sufficient chondrocytes by monolayer expansion in vitro is used for articular cartilage tissue engineering. However, chondrocytes lose their chondrogenic phenotype after monolayer expansion via mitochondrial dysfunction-induced senescence. Adipose-derived stem cell mitochondrial transfer (ADSC-MT) improves senescent cell function. We hypothesise that ADSC-MT improves the chondrogenic phenotype of senescent chondrocytes. Methods: After monolayer expansion in vitro, chondrocytes were subjected to ADSC-MT. Cell senescence was evaluated via analysis of p16 and p21 expression and senescence-associated β-galactosidase (SA-β-gal) staining. The chondrogenic phenotype was evaluated by measuring collagen type II (Col-II) and collagen type I (Col-I) levels. Oxidative stress was assessed by determining the mitochondrial superoxide and 8-hydroxydeoxyguanosine (8-OHdG) levels. Mitochondrial dysfunction was assessed by determining the mitochondrial membrane potential (MMP) and PGC-1α levels. Finally, SOD-2, SIRT-1, SIRT-3, TFAM, MFN-1, MFN-2, OPA-1, PINK-1 and Parkin levels were used to assess mitochondrial quality control (MQC). Results: ADSC-MT-recipient chondrocytes exhibited alleviated senescence with decreased p16 and p21 expression and SA-β-gal staining. The increased Col-II and decreased Col-I expression indicated that the chondrogenic phenotype of the chondrocytes was restored. Decreased mitochondrial superoxide and 8-OHdG levels indicated alleviated oxidative stress. The increased MMP indicated alleviation of mitochondrial dysfunction. For MQC, SOD‑2, PGC‑1α, TFAM, SIRT-1, and SIRT-3 were upregulated, indicating that antioxidant defences and mitochondrial biogenesis in MQC were increased in ADSC‑MT–recipient chondrocytes. PINK‑1 and Parkin were downregulated, suggesting that damaged mitochondria were reduced through mitophagy. In contrast, MFN‑1, MFN‑2, and OPA‑1 were not changed, indicating that mitochondrial dynamics were not affected. Conclusions: ADSC-MT improves the chondrogenic phenotype of senescent chondrocytes by ameliorating mitochondrial dysfunction.
Background: Huperzine A (HupA), a potent and selective acetylcholinesterase inhibitor with neuroprotective properties, faces significant clinical challenges in Alzheimer’s disease (AD) management. Its amphiphilic nature results in poor encapsulation efficiency within conventional hydrophobic polyesters or hydrophilic hydrogels, while its requirement for repeated dosing leads to systemic toxicity and poor patient compliance. Methods: We developed a novel hyaluronic acid (HA)-based biphasic delivery platform. This system utilized a HA gel matrix to encapsulate HupA-loaded polyhydroxyalkanoate nanoparticles (HupA@(NP/Gel)). Its sustained-release efficacy and therapeutic efficacy were evaluated in AD mice through a single administration, followed by behavioral, biochemical, and histopathological analyses. Results: The HupA@(NP/Gel) platform achieved complete drug encapsulation and successfully extended HupA release for over 20 days, matching the therapeutic requirements for AD. This sustained delivery translated to therapeutic efficacy in vivo: it significantly ameliorated core AD pathologies, including reduced Aβ deposition and restored cholinergic function. Moreover, the treatment conferred comprehensive neuroprotection by suppressing glial activation and neuroinflammation, enhancing neuronal survival, and preserving synaptic integrity. Crucially, these multifaceted benefits resulted in improved spatial memory and reduced anxiety/depressionlike behaviors. Conclusions: Our study demonstrates that the HupA@(NP/Gel) system can serve as an effective and translatable strategy for AD intervention. By effectively overcoming the key hurdles of HupA delivery, this work establishes a translatable sustained-delivery strategy that highlights carbohydrate polymer technology’s role in neurologic therapeutics.
The article titled “Biomimetic Neuropeptide Y/collagen I/β-tricalcium Phosphate Scaffold Mediated Macrophage Polarization and Vascularization for Bone Regeneration” was published in European Cells & Materials, Volume 55, pages 1–16. Fig. 3 of the originally published article was incorrect. The correct version of Fig. 3 is provided below. This correction does not affect the results or conclusions of the article. The underlying data remain unchanged and continue to fully support the original conclusions of the study.Publisher’s Note: The relevant content in the original manuscript contained inaccuracies due to the authors’ oversight. Following a subsequent review, the authors proactively requested revisions, and the corresponding corrections have been made accordingly.Editor’s Note: The Editor-in-Chief responsible for this correction was Martin Stoddart.
As a critical subclass of extracellular vesicles, exosomes have emerged as a research focus in tissue regeneration and precision therapy because of their unique molecular delivery and intercellular communication capabilities. This article systematically reviews the biogenesis, isolation, and characterization of exosomes and their role in advancing tissue engineering applications. Their multifaceted regulatory roles in bone/cartilage repair, neural regeneration, wound healing, and cardiovascular regeneration, including antiapoptotic, proangiogenic, immunomodulatory, and antifibrotic mechanisms, are highlighted. The innovations of this work lie in (1) the comprehensive analysis of engineered exosome strategies—such as surface modification, cargo-loading optimization, and synergistic integration with biomaterials—to overcome the limitations of traditional delivery systems; (2) the proposal of the dual regulatory potential of exosomes in cancer immunotherapy and autoimmune diseases, offering novel insights for clinical translation; and (3) the envisioning of future directions by integrating artificial intelligence (AI) and three-dimensional (3D) bioprinting to advance scalable production and precision design of exosome-based therapies. This article further addresses current challenges (e.g., heterogeneity, standardization, and safety) and emphasizes interdisciplinary collaboration to bridge the gap between fundamental research and clinical translation. This review provides a theoretical framework and technical foresight for advancing regenerative medicine and precision therapeutics.
Objective: The load-bearing structures of the subchondral bone undergo alterations in osteoarthritis (OA) joints and exhibit distinct bone remodelling properties. This study examined the pathological features and cellular components of the subchondral bone plate (SCBP) and subchondral cancellous bone (SCCB) in OA-affected regions of human knee joints. Methods: Tibial plateaus were obtained from patients with varus knee OA (n = 42; women: n = 22, aged 57–87 years; men: n = 20, aged 59–82 years). Osteochondral specimens were collected from OA lesion sites in the medial compartment (OA region) and paired control sites in the lateral compartment (C region). Bone mineral density (BMD) was evaluated using micro-computed tomography, osterix+ osteoprogenitors, cathepsin K (CTSK)+ osteoclasts, and F4/80+ macrophages were quantified by immunohistochemistry, and correlations between cellular components were analysed by sex and region. Results: The OA SCBP had a significantly higher BMD than did the C region. In male patients, more F4/80+ macrophages were present in the SCBP C region than in the OA region. Female OA SCCB samples showed an increased number of CTSK+ osteoclasts. In both sexes, compared with the C region, the OA SCCB contained more CTSK+ osteoclasts and macrophages. Positive correlations between macrophage and osteoprogenitor densities were observed in most subchondral bone regions, except in male OA samples. Conclusions: Region-specific differences in cellular components were identified in the OA subchondral bone. Asynchronous remodelling responses were noted between the SCBP and SCCB. These findings provide detailed insights into OA pathology and can inform future therapeutic strategies.
Background: Osteoporosis, a metabolic disorder with reduced bone density and high fracture risk, can be alleviated by ginsenoside Rg3 (GRg3), which promotes human bone marrow-derived mesenchymal stem cell (hBMSC) osteogenic differentiation via multiple pathways. However, GRg3’s clinical use is limited by hydrophobicity, short half-life, and degradation susceptibility. Methods: This study developed a novel nanocarrier using biodegradable poly (3-hydroxybutyrate-co-3-hydroxyvalerate-co-3-hydroxyhexanoate) (PBVHx) (the latest terpolymer in polyhydroxyalkanoates) doped with a small amount of poly (lactic-co-glycolic acid)-polyethylenimine conjugate (PLGA-PEI) to encapsulate GRg3 (GRg3-PBVHx-based nanoparticles (PNPs)). GRg3-PNPs (1%, 5%, 10% loadings) were spherical, with sizes 105–160 nm (increasing with GRg3) and stable dispersibility. Results: Encapsulation efficiency was ∼ 89% for 1% and 5% groups, but 38.21% for 10% due to leakage. In vitro, GRg3 released sustainably over 20 days, synchronized with degradation. Cellular experiments showed efficient uptake by hBMSCs. 5% and 10% GRg3-PNPs enhanced cell viability; the 5% group exhibited the strongest osteogenic efficacy, with increased alkaline phosphatase (ALP) activity, calcium deposition, upregulated pro-osteogenic markers (collagen type I (COL-1), osteocalcin (OCN), osteopontin (OPN), runt-related transcription factor 2 (RUNX2)), and downregulated inhibitory markers (matrix Gla protein (MGP), osteoprotegerin (OPG)). Conclusions: This nanosystem, with 5% GRg3-PNPs as the optimal formulation, efficiently delivers poorly soluble GRg3 and shows promise for traditional Chinese medicine (TCM)-based osteoporosis treatment and bone regeneration.
Background: Endometrial cancer (EC) is a prevalent gynecological malignancy with limited therapeutic options due to drug resistance and systemic toxicity. Exosomes have emerged as promising targeted drug delivery vehicles, but their application in EC remains underexplored. Thrombospondin-1 (THBS1), a key regulator of extracellular matrix remodeling and transforming growth factor β (TGF-β) signaling, may offer a novel therapeutic strategy for EC. Methods: Differentially expressed genes were identified from the public transcriptomic datasets and subjected to GO and KEGG enrichment analyses. Exosomes derived from Human embryonic kidney 293 (HEK293) cells overexpressing THBS1 were isolated and characterized by Western blotting, transmission electron microscopy, and nanoparticle tracking analysis. Their effects on EC cells were assessed using Cell Counting Kit-8, 5-ethynyl-2’-deoxyuridine incorporation, colony formation, wound healing, and cell invasion assays. TGF-β pathway involvement was assessed using the agonist SRI-011381. In vivo experiments were conducted to evaluate tumor growth, epithelial–mesenchymal transition (EMT) marker expression, and treatment safety. Results: THBS1 expression was downregulated in the EC cells. The restoration of THBS1 expression through exosome delivery inhibited cell proliferation, migration, and invasion. THBS1-overexpressing exosomes attenuated the activation of the TGF-β/Smad pathway and modulated EMT-related markers. The inhibitory effects of THBS1-exosomes were reversed by SRI-011381. In vivo, THBS1-exosome treatment markedly suppressed tumor growth, enhanced E-cadherin expression, reduced vimentin levels, and exhibited no observable toxicity. Conclusions: Exosomes overexpressing THBS1 effectively suppressed EC progression by targeting the TGF-β/Smad signaling axis and EMT. These findings support the potential of THBS1-enriched exosomes as a novel, safe, and targeted therapeutic modality for future EC.
Background: Currently, the clinical treatment of severe bone defects remains a major challenge. Gelatin methacryloyl (GelMA) hydrogels with extracellular matrix (ECM)-like properties are commonly used materials for bone defect repair. However, due to the lack of osteogenic activity, researchers have considered using tissue engineering methods to address this issue. Methods: A composite hydrogel scaffold was fabricated by incorporating acryloylated polyethylene glycol N-hydroxysuccinimide (AC-PEG-NHS)-modified osteogenic growth peptide (OGP)(10-14) (AC-PEG-OGP(10-14)) and primary osteoblasts into a GelMA matrix. The hydrogel scaffold was characterized using scanning electron microscopy (SEM), mechanical testing, and the bicinchoninic acid (BCA) protein assay. In vitro, the effects of the scaffold on the differentiation of osteoblasts and bone marrow-derived macrophages (BMMs) were evaluated. In vivo, its role in bone defect repair was assessed using a rat model. Results: The results demonstrated that the AC-PEG-OGP(10-14)-loaded hydrogel scaffold significantly enhanced the mechanical properties and slowed the degradation rate based on the GelMA scaffold, and it could also achieve the controlled release of the osteogenic peptide OGP(10-14). In vitro and in vivo experiments showed that the material exhibited excellent biocompatibility and osteogenic mineralization properties. It inhibited osteoclast formation, thereby significantly promoting the repair of mouse femoral bone defects. The underlying mechanism is closely related to the activation of the Wnt/β-catenin signaling pathway. Conclusions: AC-PEG-OGP(10-14)-loaded hydrogel scaffold offers an effective solution for repairing bone defects and significantly enhances bone regeneration.
Background: Post-surgical biofilm infections present a major clinical challenge due to their exceptional tolerance to antibiotics and the physical barrier of extracellular polymeric substance (EPS), calling for innovative non-antibiotic therapeutic strategies. Methods: We engineered a synergistic platform by constructing curcumin-loaded iron-based metal-organic framework (MIL@Cur) nanoparticles and incorporating them into a dissolvable hyaluronic acid-based hydrogel to fabricate composite microneedles (MIL@Cur microneedle (FCMN)). The system was characterized for its physicochemical properties and evaluated for antibacterial efficacy in vitro and in a murine methicillin-resistant staphylococcus aureus (MRSA)-infected wound model. Results: The MIL@Cur nanoparticles demonstrated well-defined morphology, high photothermal conversion efficiency (reaching >50°C under laser irradiation), and pH-responsive drug release. In vitro, MIL@Cur with laser irradiation achieved synergistic bacterial eradication through photothermal therapy and ironoverload-induced chemodynamic therapy (CDT), while also disrupting pre-formed biofilms and inhibiting new biofilm formation via quorum sensing (QS) suppression. The FCMN patch exhibited excellent mechanical strength and efficient transdermal delivery. In vivo, the FCMN + Laser group showed accelerated wound closure, ∼ 2-log reduction in bacterial load, enhanced collagen deposition and angiogenesis, and no systemic toxicity. Conclusions: This microneedle-mediated platform effectively combines multiple antimicrobial modalities, providing a powerful and translatable strategy for treating stubborn biofilm infections and promoting wound repair.
Background: Bone grafting is the primary clinical intervention for bone defects. β-tricalcium phosphate (β-TCP) is an absorbable ceramic for its excellent biocompatibility and bioactivity. Neuropeptide Y (NPY) participates in bone homeostasis and vascular regeneration. This research explored the TCP/collagen I (Col)/NPY scaffolds, which controlled release of NPY, on macrophage polarization during bone repair. Methods: The scaffold was characterized by scanning electron microscopy (SEM) and Fourier-transform infrared (FTIR) spectrometry. The cumulative NPY release from the TCP/Col/NPY scaffold was tested by an enzyme-linked immunosorbent assay. Biocompatibility of the TCP/Col/NPY scaffold was evaluated using cell counting kit (CCK)-8 and calcein-acetoxymethyl ester (AM)/propidium iodide (PI). Angiogenic activity was detected by scratch and tube-formation assays with human umbilical vein endothelial cells (HUVECs). Osteogenic differentiation was detected by alkaline phosphatase (ALP) staining. Flow cytometry and immunofluorescence staining were used to evaluate RAW264.7 polarization. In vivo, bone-defect repair was evaluated using micro-computed tomography scans, hematoxylin and eosin (H&E) staining, Masson and immunohistochemical staining. Results: The SEM images disclosed an interconnected pore structure. FTIR of TCP/Col/NPY scaffolds showed a characteristic peak of NPY. The TCP/Col/NPY scaffold exhibited favorable biocompatibility with bone marrow mesenchymal stem cells and promoted the migration and angioenesis of HUVECs. RAW264.7 upregulated cluster of differentiation-206 (CD206) in the TCP/Col/NPY group (p < 0.05). In vivo, the TCP/Col/NPY scaffold promoted the repair of cranial defects. H&E and Masson revealed that new bone formation in TCP/Col/NPY group was significantly higher (p < 0.05). Runx2, osteocalcin, platelet-derived growth factor-BB and CD206/CD80 expression were higher in the TCP/Col/NPY group (p < 0.05). Conclusions: TCP/Col/NPY scaffolds significantly promoted the repair of critical bone defects by modulating macrophage polarization toward the M2 phenotype and enhancing vascular regeneration.
Background: Rheumatoid arthritis (RA) disproportionately affects postmenopausal women and manifests as accelerated cartilage and bone erosion driven by hyperactive osteoclasts (OCs). Oestrogen deficiency exacerbates OC activity, while the Eph receptor interacting protein B2 (EphrinB2)/Eph receptor B4 (EphB4) signalling pathway acts as a critical negative regulator of OC differentiation by suppressing the cellular oncogene c-Fos (c-Fos)/nuclear factor of activated T cells cytoplasmic 1 (NFATc1) transcription cascade. The traditional Chinese medicine (TCM) Yishenjuanbi pill (YSJB) enhances EphrinB2-mediated bone protection and demonstrates superior therapeutic efficacy in an ovariectomized collagen-induced arthritis (OVX + CIA) rat model compared with its effects in CIA rats, although its effects on osteoclasts (OCs) remain incompletely characterized. Methods: Bone marrow-derived macrophages (BMMs) and splenic monocytes were employed to elucidate the EphrinB2-dependent mechanisms through which YSJB inhibits OC differentiation. Rat serum from the Control, OVX, CIA, OVX + CIA, OVX + CIA + oestradiol valerate (EV), and OVX + CIA + YSJB groups was prepared and added during osteoclast induction. Quantitative reverse transcription PCR (RT-qPCR), tartrate-resistant acid phosphatase (TRAP) staining, and bone resorption assays were performed on both cell types. Results: YSJB reversed the ovariectomy + CIA-induced upregulation of the osteoclastogenic factors c-Fos, transcription factor c-Jun (c-Jun), Nfatc1, and receptor activator of nuclear factor-κb (Rank) while restoring encoding EphrinB2 (Efnb2) expression (n = 3). Crucially, Efnb2 knockdown abolished the protective effects of YSJB, restoring pathological OC activity and increasing gene expression levels (n = 3). Conclusions: Our findings suggest that by upregulating EphrinB2, YSJB serum inhibits osteoclastogenesis in both BMMs and splenic monocytes.
Preclinical surgical studies in rodent models play a pivotal role in elucidating the mechanisms of bone repair and regeneration. Similar to clinical practice, intraoperative and postoperative complications in preclinical studies significantly affect treatment outcomes and may even alter the overall experimental results. Well-established laboratory infrastructure and experimental procedures can reduce the risk of adverse events, such as intraoperative technical errors and postoperative infection. However, these aspects are often underreported or overlooked in the literature. In this study, fracture fixation in rats is presented as an example to highlight the essential but often unreported details, such as virtual surgical planning, preoperative rehearsal, disinfection protocols, intraoperative management, postoperative support, multidisciplinary collaboration, and research documentation. Supplementary and alternative solutions are also proposed for laboratories with limited resources. By applying virtual planning and rehearsal with three-dimensional (3D)-printed samples, the authors have performed 108 consecutive external fixation procedures for rat femoral fractures since 2020, with no intraoperative dropouts attributable to surgical technique errors. The operative time stabilized at 45.6 ± 3.8 minutes (mean ± standard deviation), reflecting low variability and a reproducible workflow. Systematic implementation of these strategies helps prevent postoperative complications, enhances animal welfare, and improves the reproducibility and translational potential of preclinical research.
