Search Results (378)

Diabetic wounds are characterized by persistent hyperglycemia, excessive ROS accumulation, sustained inflammation, and impaired angiogenesis, yet current treatments remain suboptimal. To address these challenges, we developed a mild heat stimulating and microenvironment reprogramming hydrogel (termed C-4-N) via a green synthetic strategy. L-Arginine (L-Arg) triggered the spontaneous self-polymerization of protocatechuic aldehyde (PA) into poly (protocatechuic aldehyde) (PPA) nanoparticles, onto which ginsenoside Compound K (CK) was subsequently loaded, yielding CK/L-Arg/PPA nanoparticles. These nanoparticles were then uniformly embedded into a dynamic disulfide network composed of α-lipoic acid (LA)-modified chitosan (CS-LA) and 4-arm-PEG-SH under UV irradiation without toxic photo-initiators, forming the C-4-N hydrogel. The C-4-N hydrogel reprogrammed the diabetic wound microenvironment through three synergistic mechanisms, lowering blood glucose and scavenging ROS via the coordinated actions of LA, CK and PPA, promoting M1-to-M2 macrophage polarization via downregulation of pro-inflammatory cytokines (TNF-α, IL-6) and upregulation of anti-inflammatory cytokines (IL-10, TGF-β1), further amplified by mild photothermal stimulation of 40–43 °C. In a diabetic rat model, the C-4-N hydrogel achieved a near-complete wound closure rate of 99.49 ± 0.10% on day 13 upon mild photothermal stimulation, accompanied by enhanced re-epithelialization, organized collagen deposition, vascular maturation, and systemic glucose regulation. In summary, this green synthesized, mild heat-stimulating hydrogel establishes a synergistic microenvironment reprogramming paradigm for chronic diabetic wound managements. Full article

Background: Macrophage pyroptosis contributes substantially to sepsis-induced lung injury, yet effective therapeutic strategies remain limited. This study aimed to determine the protective effects of meloxicam, a non-steroidal anti-inflammatory drug, and the underlying mechanisms in this context. Methods:In vivo, CLP mice were treated with meloxicam (20 mg/kg). In vitro, LPS-primed macrophages were stimulated with ATP or nigericin in the presence or absence of meloxicam. Levels of pyroptosis-associated proteins (cleaved Caspase-1, mature IL-1β, GSDMD-NT), NLRP3 inflammasome assembly, and the CBP/TXNIP/p38 signaling axis were assessed by Western blot. Mitochondrial membrane potential (ΔΨm) and intracellular ROS were measured. Overexpression of COX-2, TXNIP, and CBP was also performed. Results: Meloxicam significantly improved survival, reduced lung injury, and suppressed pyroptosis-associated proteins in CLP mice. In vitro, meloxicam dose-dependently enhanced macrophage viability and reduced LDH, IL-1β, and IL-18 release. The protective effects of meloxicam were mediated by inhibition of NLRP3 inflammasome priming and assembly, disruption of NLRP3-ASC-pro-Caspase-1 complex formation, and suppression of ASC oligomerization. Meloxicam also inhibited the CBP/TXNIP/p38 axis, an effect reversed by TXNIP or CBP overexpression. Furthermore, meloxicam restored ΔΨm and reduced ROS accumulation; these effects were abrogated by the ROS inducer imiquimod. Importantly, the anti-pyroptotic effects of meloxicam were independent of COX-2 inhibition. Conclusions: These findings expand the pharmacological profile of meloxicam and support its repurposing as a therapeutic agent for sepsis-associated lung injury. Full article

Tuberculosis (TB) treatment is severely hampered by the rise in multi-drug-resistant strains and the prevalence of drug-induced toxicities. Host-Directed Therapies (HDTs) have emerged as a promising strategy to overcome these challenges by modulating innate immunity and circumventing Mycobacterium tuberculosis (Mtb) evasion mechanisms. A hallmark of Mtb pathogenesis is the arrest of phagosome maturation and the induction of host cell necrosis over protective apoptosis. In this study, we investigated the potential HDT effects of α-Lipoic acid (α-LA), a well-known antioxidant and metabolic cofactor, within an in vitro model of Mtb-infected THP-1 macrophages. Our findings indicate that α-LA treatment modulates the macrophage redox state and selectively promotes apoptosis in infected cells without increasing necrotic lysis. Furthermore, α-LA administration led to a significant, dose-dependent restoration of phagolysosome acidification, effectively reversing the maturation blockade imposed by Mtb. Notably, this enhanced acidification inversely correlated with intracellular bacterial survival. These results suggest that α-LA might act as a multifaceted HDT agent capable of restoring both host-protective cell death and phagosomal microbicidal mechanisms. Given its established safety profile and its ability to complement standard anti-TB drugs like Bedaquiline (BDQ), α-LA represents a highly promising candidate for adjunct therapy to improve TB treatment outcomes and mitigate the impact of antibiotic resistance. Full article

Myeloid malignancies encompass a heterogeneous group of haematological disorders, primarily including myelodysplastic syndromes (MDSs) and myeloproliferative neoplasms (MPNs). MDS is characterised by defective myeloid cell maturation, while MPNs involve the pathological overproduction of myeloid lineage cells. In the absence of timely diagnosis and effective clinical intervention, both entities carry a substantial risk of progression to acute myeloid leukaemia (AML). Although allogeneic haematopoietic stem cell transplantation remains the only potentially curative therapy, its application is frequently constrained by patient-related factors such as advanced age and comorbid conditions. While currently, hypomethylating agent therapy (azacitidine and decitabine) is mainly used in high-risk MDS patients, and ruxolitinib is primarily used in symptomatic primary myelofibrosis (PMF-MPN), their clinical efficacy remains suboptimal. More recently, focus has turned toward the role of the tumour microenvironment (TME) in disease pathogenesis and whether therapeutically targeting the TME, either alone or in combination with conventional therapy, could present a new treatment option. Emerging evidence underscores the significant influence of TME components, particularly macrophages and T cells, in modulating immune responses and shaping the leukaemic niche to either facilitate or hinder malignant progression. In response, a new generation of immune checkpoint inhibitors are being developed to target the TME, including PD-1/CTLA-4 blockers, macrophage-directed agents including CD47 inhibitors, and T cell-targeting checkpoint inhibitors such as TIM-1 and LAG-3. This review will describe the functional role of key TME constituents in the progression of myeloid malignancies and explore the current landscape and future potential of advanced cellular and molecular immunotherapies in the treatment of these disorders. Full article

Pancreatic ductal adenocarcinoma (PDAC) remains highly lethal and largely refractory to immune checkpoint inhibition because limited antigen-specific priming, myeloid suppression, dense desmoplasia, and abnormal vasculature enforce immune exclusion. CD40 links CD4+ T-cell help through CD40L/CD154 to antigen-presenting-cell (APC) licensing and CD8+ T-cell priming, making CD40 agonism a rational strategy to stimulate antitumor immunity in PDAC. CD40 is expressed on APCs and has also been reported on subsets of PDAC tumor cells, cancer-associated fibroblasts, and endothelial cells, indicating that CD40 agonists may affect immune activation, stromal/vascular remodeling, and context-dependent tumor-cell-intrinsic signaling. TRAF-dependent CD40 signaling activates canonical and non-canonical NF-kB, MAPK, and PI3K/AKT pathways, promoting APC maturation, IL-12-associated Th1 programming, macrophage repolarization, and matrix remodeling; tumor-intrinsic effects remain more variable, ranging from apoptotic to pro-survival programs. Clinically, CD40 agonists have shown pharmacodynamic immune engagement and manageable toxicity, mainly in combinations with chemotherapy, checkpoint inhibitors, and vaccine platforms, but efficacy remains inconsistent, and randomized validation is incomplete. Baseline CD40 expression has not reliably predicted benefit. Future development should prioritize spatially resolved tumor-immune profiling, longitudinal pharmacodynamic biomarkers, optimized sequencing, and agent-specific dosing strategies. This review integrates CD40 expression, signaling, and clinical evidence in PDAC to support more rational, biomarker-guided development of CD40-directed immunotherapy. Full article

Testicular development and spermatogenesis are critical for male reproduction, but their molecular mechanisms in Dezhou donkeys remain understudied. This study used single-cell RNA sequencing (scRNA-seq) to analyze testicular tissues from Dezhou donkeys at juvenile (2 months), pre-pubertal (12 months), and mature (24 months) stages. A total of 24,606 high-quality cells were profiled, constructing a comprehensive single-cell transcriptional atlas. Unsupervised clustering identified nine major cell types: three germ cell subtypes (spermatogonia, spermatocytes, spermatids) and six somatic cell subtypes (Leydig cells, Sertoli cells, peritubular muscle cells, macrophages, endothelial cells, T cells). Key marker genes (AMH, TNP1, UTF1, ZMYND10) were validated by immunofluorescence. Pseudotemporal trajectory analysis revealed sequential germ cell differentiation (spermatogonia → spermatocytes → spermatids) and Sertoli cell maturation (immature → mature), while Leydig cells and peritubular muscle cells shared common progenitors. CellChat analysis identified critical ligand–receptor pairs in BMP, IGF, WNT, and FSH pathways, which regulate testicular development. This study provides the comprehensive single-cell transcriptional map of Dezhou donkey testicular development, elucidating key molecular mechanisms of germ and somatic cell maturation. The findings offer valuable insights into donkey reproductive biology, supporting breeding improvement and male infertility research. Full article

Chamaecrista nomame (Siebold) H. Ohashi (C. nomame), a leguminous plant traditionally consumed in East Asia, contains diverse bioactive phytochemicals, but whether its activities act convergently under obesity-related pathological conditions remains unclear. This study investigated the anti-inflammatory, anti-obesity, and insulin-sensitizing effects of a 40% ethanol extract of C. nomame (ECNE) and its marker compound luteolin in lipopolysaccharide (LPS)-stimulated RAW 264.7 macrophages, differentiating and mature 3T3-L1 adipocytes, and tumor necrosis factor-α (TNF-α)-induced insulin-resistant adipocytes. In LPS-stimulated macrophages, ECNE and luteolin reduced nitric oxide and pro-inflammatory cytokine (TNF-α, interleukin (IL)-6, IL-1β) production, accompanied by suppression of nuclear factor-κB and mitogen-activated protein kinase signaling. In differentiating adipocytes, both reduced lipid accumulation and downregulated peroxisome proliferator-activated receptor γ, CCAAT/enhancer-binding protein α, and adipocyte protein 2. In mature adipocytes, they enhanced insulin-stimulated glucose uptake and Akt phosphorylation. In TNF-α-challenged adipocytes, pretreatment partially restored glucose uptake and Akt phosphorylation while attenuating IL-6 and monocyte chemoattractant protein-1 production. ECNE exerted effects equal to or greater than those of luteolin at equivalent luteolin-based concentrations, indicating contributions from additional phenolic constituents. These findings support ECNE as a multifunctional natural resource against obesity-associated inflammation and insulin resistance. Full article

Earthworm-derived bioactive compounds are emerging as promising pharmaceutical agents; however, the immunomodulatory effects of Eisenia fetida proteins at different developmental stages remain unclear. This study evaluated, for the first time, the stage-dependent immunomodulatory activity of E. fetida protein extracts in RAW 264.7 macrophages. Soluble proteins isolated from juvenile, mature, and senescent worms were lyophilized and tested for their effects on cell viability, phagocytic activity, nitric oxide (NO), reactive oxygen species (ROS), and inflammatory gene expression. Amino acid profiling and Western blot analysis were additionally performed to investigate biochemical composition and signaling mechanisms. Mature-stage extracts exhibited the highest protein yield, minimal cytotoxicity, enhanced macrophage phagocytosis, and significant suppression of LPS-induced NO, ROS, and proinflammatory cytokines. In contrast, juvenile-stage extracts showed moderate immunomodulatory activity, whereas senescent-stage extracts induced oxidative stress and inflammatory responses. Western blot analysis demonstrated that mature-stage proteins strongly inhibited phosphorylation of NF-κB and MAPK signaling proteins, including p65, IκBα, p38, ERK1/2, and JNK, while senescent-stage extracts maintained elevated pathway activation. Amino acid analysis further revealed enriched immunologically relevant amino acids in mature-stage extracts. These findings demonstrate that developmental stage strongly influences the biological activity of E. fetida proteins and highlight mature-stage extracts as promising natural immunomodulatory agents. Full article

Radiotherapy (RT) is a cornerstone of cancer treatment, traditionally recognized for its direct cytotoxic effects via DNA damage. However, emerging evidence highlights RT as a profound modulator of the tumor microenvironment (TME), acting as a “double-edged sword” that greatly influences the success of immune checkpoint inhibitors (ICIs). On the one hand, RT acts like an in situ vaccine, causing immunogenic cell death and activating the cGAS-STING pathway, which leads to dendritic cell maturation, T-cell infiltration, and reactive PD-L1 expression. This effect can turn “cold” tumors into “hot” ones, making them more responsive to immune checkpoint blockade. On the other hand, RT can lead to resistance to ICIs by promoting an immunosuppressive environment, recruiting regulatory T cells, M2 macrophages, and myeloid-derived suppressor cells. This review analyzes the mechanisms behind this immunological duality and assesses how parameters such as dose, fractionation, and particle type (e.g., carbon ion versus photon therapy) can be optimized to enhance immune activation. Lastly, we discuss future strategies that focus on innate immunity and tumor metabolism, showing how targeting nutrient depletion and ferroptosis can break down immunosuppressive barriers and position RT as an essential component of precision immuno-oncology. Full article

Mesenchymal stem/stromal cells (MSCs) are multipotent progenitor cells with potent immunomodulatory properties, making them attractive candidates for treating inflammatory and autoimmune diseases. A key mediator of MSC-induced immunosuppression is programmed death-ligand 1 (PD-L1), a checkpoint molecule that interacts with PD-1 on immune cells to regulate immune responses and promote tolerance. This review synthesizes current evidence on the role of PD-L1 expression in MSCs, emphasizing its effects on both the innate and adaptive immune systems, its therapeutic potential, and its utility as a biomarker for MSC potency and clinical efficacy. We examine how PD-L1 modulates T cell activation, dendritic cell maturation, macrophage polarization, and cytokine profiles, including its role in exosomal contexts. Additionally, we highlight its synergistic interactions with other immune checkpoints and discuss its dual function as both a therapeutic effector and a dynamic biomarker. Finally, we explore its relevance in clinical contexts such as autoimmune diseases, graft-versus-host disease, sepsis, and transplantation and conclude with a discussion of challenges and future directions in harnessing PD-L1 for MSC-based therapies. Full article

Background: Impaired angiogenesis and persistent inflammation are hallmarks of chronic diabetic wounds. Extracellular vesicles derived from dental pulp stem cells (DPSC-EVs) represent a promising cell-free therapy for tissue repair; however, their clinical translation is hindered by suboptimal yields and attenuated bioactivity associated with conventional two-dimensional (2D) culture. This study investigated whether a biomimetic three-dimensional (3D) fibrin/gelatin hydrogel system could optimize the therapeutic potency of DPSC-EVs for diabetic wound healing. Methods: DPSCs were encapsulated within 3D fibrin/gelatin scaffolds, followed by comprehensive characterization of cell viability and morphology. 3D-EVs and 2D-EVs were isolated via ultracentrifugation and validated by transmission electron microscopy and nanoparticle tracking analysis. The pro-angiogenic capacity of 3D-EVs was evaluated using human umbilical vein endothelial cells (HUVECs) under high-glucose (HG) stress. Additionally, the immunomodulatory effects were assessed by monitoring macrophage polarization in lipopolysaccharide-stimulated RAW 264.7 cells. The therapeutic efficacy was further validated in vivo using a streptozotocin (STZ)-induced diabetic mouse model with full-thickness cutaneous wounds. Results: The 3D fibrin/gelatin hydrogel provided a supportive microenvironment that significantly augmented the secretory productivity of DPSCs. Compared to 2D-EVs, 3D-EVs exhibited superior functional resilience in restoring HUVEC migration and tube formation under HG-induced oxidative stress. Furthermore, 3D-EVs effectively orchestrated the macrophage transition from a pro-inflammatory M1 phenotype toward an anti-inflammatory M2 phenotype, thereby modulating the immune microenvironment. In vivo, topical administration of 3D-EVs markedly accelerated wound closure, promoted re-epithelialization, and enhanced microvascular density and collagen maturation in diabetic mice. Conclusions: Our findings demonstrate that the 3D fibrin/gelatin culture system effectively primes the therapeutic profile of DPSC-EVs. These engineered vesicles accelerate diabetic wound healing by synergistically promoting angiogenesis and resolving chronic inflammation, offering a robust and potent cell-free strategy for the management of chronic diabetic ulcers. Full article

Fetal skeletal muscle development involves coordinated interactions among myogenic, stromal, vascular, and immune compartments, yet the cellular and molecular programs guiding tissue maturation remain incompletely understood. To address this, we generated a high-resolution single-cell atlas of fetal female goat skeletal muscle and performed trajectory analysis, transcription factor activity profiling, and intercellular communication mapping. Unsupervised clustering identified RUNX2 mesenchymal progenitors, fibro-adipogenic progenitors (FAPs), myofibroblasts, endothelial cells, macrophages, differentiating myocytes, and mature skeletal muscle fibers, revealing a heterogeneous ecosystem in which stromal populations support myogenic progression and vascular and immune cells contribute to tissue organization. Pseudotime analysis traced a maturation continuum from differentiation-competent myocytes to contractile fibers, marked by sequential activation of extracellular matrix remodeling, cytoskeletal stabilization, and sarcomere assembly. KEGG and GO enrichment highlighted stage-specific engagement of ErbB, Hedgehog, and Hippo signaling, as well as cell cycle and ubiquitin-mediated proteolysis pathways, linking proliferation, differentiation, and structural maturation. Transcription factor profiling revealed early-stage proliferative and morphogenetically permissive states driven by E2F4/5, HMGA2, and HAND2, transitioning to late-stage differentiation, ECM remodeling, and tissue stabilization orchestrated by CEBPB, CREB3L1, ELK1, and E2F2. Cell–cell communication analysis showed a developmental redistribution of signaling authority, from ECM-driven, progenitor-centered networks to modular, structurally stabilized interactions. These findings define the cellular, transcriptional, and signaling framework orchestrating fetal skeletal muscle maturation. Full article

Peripheral T-cell lymphomas (PTCLs) are malignancies of mature T cells with a poor prognosis. Most PTCL cases express follicular T-helper (T_FH) cell antigens and are classified as T_FH cell lymphoma (TFHL). Contact-dependent signaling between CD40 and its ligand, CD154, is essential for immune functions. CD154 is expressed by activated T cells, while CD40 is found on B cells, follicular and other dendritic cells, macrophages, and stromal cells. Although the CD40–CD154 crosstalk is a key costimulatory pathway in immune responses, data on its role in PTCLs are limited. To explore the role of the CD40–CD154 axis in TFHLs, we conducted an in-depth immunomorphological study of 111 PTCL cases, including 93 TFHL cases. We found that neoplastic T cells in TFHL are consistently CD154-positive. The CD154 expression increased in histologically advanced cases and correlated with the extent of CD40 positivity. We showed that CD154-positive neoplastic T cells recapitulate the intranodal migration of normal TFH cells, disrupting and remodeling each functional compartment, thereby explaining the disease-related immune dysfunction. Our findings indicate that pathological CD40–CD154 interaction is a potential driver mechanism in TFHL and offers a promising target for future therapies. Full article

The cardiac extracellular matrix (ECM) is a dynamic, mechanically active network continuously shaped and interpreted by cardiomyocytes, fibroblasts, and macrophages. Interdependent mechanosensing, force transmission, and ECM remodeling functions create multicellular feedback loops that control tissue stiffness, alignment, maturation, and fibrotic remodeling. Together, these biomechanical processes create reciprocal signaling pathways in which cellular behavior modifies the ECM while the ECM’s mechanics concurrently shape cellular phenotype and function. This review explores cell–ECM mechanoreciprocity, a physiologic framework that unifies cell-sensing mechanotransduction, mechano-electrical coupling, and ECM-based biochemical signaling with cell-driven ECM remodeling. We propose three interconnected feedback loops that integrate biochemical and mechanical cues across cell types: load amplification, structural alignment, and immune regulation. We discuss how advanced two- and three-dimensional engineered cardiac systems incorporating tunable and dynamic mechanical cues can be used to model these interactions. We address the limitations of existing experimental platforms and the need for better models to fully recapitulate in vivo complexities. Understanding and recreating these reciprocal mechanical interactions will provide essential frameworks for disease modeling and therapeutic development while reducing reliance on in vivo studies. Full article

Myocardial infarction (MI) triggers a robust inflammatory response that is essential for tissue repair but, when excessive or prolonged, drives pathological cardiac remodelling and heart failure. Colony-stimulating factor 2 (CSF2) signalling has been implicated in driving pro-inflammatory macrophage activation post-MI. Here, we investigated the role of macrophage-specific CSF2 receptor alpha (CSF2RA) signalling in post-MI remodelling using a tamoxifen-inducible genetic mouse model and permanent coronary artery ligation. Macrophage-specific Csf2ra deficiency significantly improved left ventricular systolic function post-MI without altering cardiac fibrosis burden. Functional improvement was associated with enhanced collagen scar maturation, characterised by an increased proportion of mature collagen fibres, and with accumulation of anti-inflammatory, pro-reparative macrophages within the infarct. These macrophage changes were accompanied by increased fibroblast density, consistent with altered macrophage–fibroblast crosstalk. Collectively, these findings identify macrophage-intrinsic CSF2RA signalling as a critical regulator of inflammatory resolution and scar maturation after MI and provide mechanistic support for the rationale of selective CSF2RA inhibition as a therapeutic strategy to limit adverse cardiac remodelling and improve post-infarction recovery. Full article