Although mRNA therapy has achieved favorable outcomes, it still faces limitations such as poor stability and short duration of protein expression. In this study, we utilized a covalently closed circular RNA encoding bone morphogenetic protein-2 (BMP2 circRNA), which exhibits exceptional nuclease resistance and an extended half-life, thereby enabling sustained and efficient BMP2 protein expression. The BMP2 circRNA was encapsulated into biomimetic nanovesicles (BNVs) derived from bone marrow mesenchymal stem cells (BMSCs) using co-extrusion technology. These BNVs were then anchored onto the surface of micro-arc oxidized titanium (Ti-MAO) implants via polydopamine (PDA) adhesion, constructing a novel local gene delivery system. In vitro experiments confirmed that this system is not only efficiently internalized by cells but also evades lysosomal degradation, facilitating the sustained release of BMP2 protein. This, in turn, significantly promoted osteogenic gene expression and accelerated mineral deposition. Furthermore, in vivo animal studies demonstrated that the functionalized implant markedly enhanced bone regeneration, increasing both bone volume fraction and bone-to-implant contact. This study successfully integrated the inherent stability of circRNA with a biomimetic delivery strategy, offering an effective approach for improving implant osseointegration.

Sarcomeres are the fundamental contractile units of muscle. Despite their importance, sarcomere assembly remains poorly understood. We focused on Actn2, a protein which stabilizes the sarcomere by linking proteins to the Z-disk. During C2C12 differentiation into myoblasts, Actn2 protein levels remained constant. This finding suggested that Actn2 incorporation into the sarcomere arose from a post-translational mechanism. We hypothesized that the post-translational mechanism relied on phosphorylation. Alignment of Actn2 protein sequences from animals with three- or four-chambered hearts identified a conserved sequence, T308P309E310K311, that matches the consensus phosphorylation motif of the cell-cycle kinase CDK1. In vitro kinase assays showed that CDK1 phosphorylates Actn2 at T308. In contrast, CDK1 was unable to phosphorylate Actn2 when T308 was mutated to alanine (T308A). Using CRISPR-Cas9 gene editing, we created Actn2-T308A and phosphomimetic Actn2-T308D variants in C2C12 cells. C2C12 cells expressing Actn2-T308A differentiated rapidly and formed robust sarcomeres. However, C2C12 cells expressing Actn2-T308D failed to form organized sarcomeres. Curiously, Actn2-T308A cells were found to have less proliferative capacity than Actn2-T308D cells. Taken together, these results identify CDK1-dependent phosphorylation of Actn2-T308 as being important for sarcomere assembly. Moreover, the data also suggest a mechanism by which cell-cycle exit promotes sarcomere assembly.

The purpose of this study was to investigate the effect of mitochondrial dynamics disorder driving nucleus pulposus cell (NPC) senescence in lumbar scoliosis of aging bipedal rats under asymmetric force.

Extracellular vesicle (EV) heterogeneity is well documented but poorly defined. This is especially important in cancer where EVs serve as carriers of unique oncogenic macromolecules that can be transferred to recipient cells or targeted for liquid biopsy diagnostics. Here we employed a series of human glioma cell lines to test the content and distribution of oncogenic epidermal growth factor receptor (EGFR) including its mutant (EGFRvIII) among different subsets of tumour-derived EVs. Our results suggest that the global levels of EGFR packaged into EVs parallels its expression in parental cancer cells. However, while in glioma cells expressing high levels of EGFR (GSC83) this receptor was uniformly distributed on cellular surfaces, only a small fraction of their derived EVs contained EGFR, as documented by nano-flow cytometry, ExoView and super-resolution microscopy. Using only three protein markers (CD63, CD81 and EGFR) these single EV platforms revealed the existence of seven different EV subsets, of which four contained EGFR. Purified EGFR-positive and negative EVs contained both shared and distinctive protein markers. EGFR packaging into EVs of GSC83 cells was independent of syntenin 1 expression, but was suppressed upon treatment with pharmacological inhibitor of neutral sphingomyelinase (GW4869). Exposure of human microglial cells (HMC3) to EVs released from GW4869-treated and control glioma cells triggered distinctive changes in cellular proteome including transfer of EGFR. Overall, our results suggest that multiple pathways of EV biogenesis may operate in glioma cells resulting in formation of complex EV landscapes consisting of EGFR-positive and EGFR-negative EV subsets. This heterogeneity may have important implications for EV functions and EV-based diagnostics.

Mechanical debridement and systemic antibiotics, as current therapeutic modalities for periodontitis, suffer from limitations, including insufficient local efficacy and bacterial resistance. Herein, a dual-mode injectable hydrogel (A-CC-PBA@ZIF-8@CAPE Gel) has been designed and prepared for enhancing the therapeutic efficacy through targeted anti-infection and rectification of the dysregulated mitochondrial function of mesenchymal stem cells (MSCs) in inflammatory state. Both the in vitro antibacterial evaluation and molecular dynamics simulation confirmed the phenyl boronic acid (PBA)-modification boosted the antimicrobial efficacy against periodontal pathogenic bacteria. In addition to the targeted antibacterial efficacy, A-CC-PBA@ZIF-8@CAPE Gel exhibited remarkable capacity to scavenge ROS, ameliorate inflammatory response, relieve oxidative stress, and restore the mitochondrial function of MSCs under inflammatory conditions by activating the SIRT1/p-AMPK/PGC-1α pathway, thereby promoting the osteogenic differentiation of MSCs. In rat periodontitis model, A-CC-PBA@ZIF-8@CAPE Gel markedly reduced bone defects and ROS levels, attenuated inflammation, and restored alveolar bone height, demonstrating robust therapeutic potential in chronic inflammatory conditions. In summary, this study presents a promising therapeutic approach for periodontitis, characterized by targeted antibacterial infection, anti-inflammation, and bone regeneration.

Decellularized extracellular matrix (dECM) has been widely used as a biomimetic material for three-dimensional cell culture and tissue regeneration. Although tissue-derived ECM is the conventional source of dECM, cell-derived ECM (cECM) has emerged as an attractive alternative. Various cECM-based formulations such as powders, films, and preformed gels have been reported, but thermosensitive cECM hydrogels remain largely unexplored. Here, we report a novel method to produce a thermoreversible hydrogel exclusively made from cECM, termed CEOgel. Once in vitro-cultured umbilical cord mesenchymal stem cells were decellularized, cECM solubility was enhanced and its nanofibers were concentrated to generate CEOgel without additional factors. This fabrication strategy was applicable across multiple cell types and consistently yielded homogeneous gels with minimal donor- or batch-dependent variability. CEOgel exhibited sufficient mechanical stability for in vitro use and formed gels in vivo following injection, confirming its thermosensitive and biocompatible nature. It also served as a functional 3D matrix, supporting endothelial vascularization in microfluidic chips and the growth of colorectal cancer organoids. Proteomic profiling revealed that CEOgel incorporates a broad spectrum of proteins commonly expressed across human tissues. Additionally, we demonstrated that CEOgel properties can be tuned through transition-metal crosslinking and genetically engineering ECM-producing cells. Together, this study proposes a new class of thermoreversible cECM hydrogels that eliminate reliance on animal tissues or external cross-linkers while expanding the applicability of cECM materials and advancing conceptual diversity for ECM hydrogel design. Our findings highlight the potential of CEOgel as a regenerative biomaterial for tissue engineering and medical applications.

Castration-resistant prostate cancer is sustained by CD44+ prostate cancer stem cells (PCSCs), motivating targeted strategies that eliminate this resistant subpopulation. Here, we developed a CD44-targeted delivery system consisting of hyaluronic acid-coated selenium nanoparticles co-loaded with apigenin (HA-SeNP-Api) and evaluated its physicochemical performance, release behavior, and anti-PCSC activity. Dynamic light scattering revealed medium-dependent colloidal behavior: the hydrodynamic size in PBS (pH 7.4) increased from 169.8 nm (day 0) to 263.9 nm (72 h), remained comparatively stable in acidic PBS (pH 6.0; 223.6 to 231.8 nm), and markedly increased in PBS containing 10% FBS (192.4 to 542.5 nm). Zeta potential was strongly negative in PBS (∼-27.8 mV) but decreased in acidic PBS (∼-4.2 to -1.2 mV) and remained intermediate in PBS containing 10% FBS (∼-12.1 to -13.5 mV), consistent with medium-driven charge screening. The formulation exhibited sustained apigenin release over 72 h at pH 6.0 and pH 7.4, supported by Korsmeyer-Peppas fitting of the initial phase. In CD44+ PCSCs, HA-SeNP-Api produced the strongest cytotoxicity in the MTT assay (14.61% viability), exceeding the effects of apigenin, enzalutamide, SeNP, and non-targeted SeNP-Api (HA-). Flow cytometry confirmed pronounced apoptosis (71.67% total apoptosis) and a marked G2/M arrest (20.59%). Consistently, HA-SeNP-Api upregulated pro-apoptotic markers (BAX, CASP3, CASP8), reduced BCL2 expression, and suppressed pluripotency-associated genes (SOX2, OCT3/4, NANOG). Intracellular ROS profiling further indicated redox modulation by SeNP-containing formulations, with partial attenuation by NAC, supporting a ROS-linked contribution to the observed apoptotic response. Collectively, HA-SeNP-Api integrates CD44-mediated targeting with redox-driven stress signaling to achieve potent anti-PCSC activity, supporting further preclinical evaluation.

Multiple sclerosis (MS) is an autoimmune inflammatory disease of the central nervous system. The cause of the disease is unknown but both genetic and environmental factors are strongly involved in its pathogenesis. We derived cerebral and spinal cord organoids from induced pluripotent stem cells (iPSC) from healthy controls as well as from primary progressive MS (PPMS), secondary progressive MS (SPMS) and relapsing-remitting MS (RRMS) patients to investigate and compare oligodendrocyte differentiation and myelination capacity. In MS organoids, particularly in PPMS, we observed a decrease in p21 expression associated with a dysregulation of PAK1 and E2F1 expression. In parallel, a decrease in oligodendrocyte maturation was detected in long-term cultured cerebral and spinal cord organoids, especially in PPMS, leading to a reduced myelination capacity. Disruption of astrocyte and neuronal populations was also observed. Our findings demonstrate that in MS, inherent deficits in the p21 pathway may alter glial and neuronal cell populations and may contribute to the disease pathogenesis by reducing the capacity for myelin repair.

The Management of Myelomeningocele Study (MOMS) established that prenatal repair of myelomeningocele (MMC) improved the rate of ventriculoperitoneal shunt insertion for the treatment of hydrocephalus, compared to postnatal repair. While there was mild improvement of motor function, most children were unable to ambulate independently. Therefore, the ongoing phase 1/2a clinical trial Cellular Therapy for In Utero Repair of Myelomeningocele (CuRe trial) aims to further improve distal neurological function using placenta-derived mesenchymal stem/stromal cells seeded on an extracellular matrix (PMSC-ECM) to augment open fetal repair of MMC. Here, the authors present a video of the neurosurgical repair using the PMSC-ECM. The video can be found here: https://stream.cadmore.media/r10.3171/2026.1.FOCVID25229.

Porcine reproductive and respiratory syndrome virus (PRRSV) is a major pathogen threatening the global swine industry, with existing vaccines failing to provide broad protection due to the high viral genetic variability and immune evasion capacity. Retinoic acid (RA), a vitamin A metabolite known for immunomodulatory functions, has been reported to enhance antiviral innate immunity and suppress excessive inflammation. However, whether RA affects PRRSV infection remains unclear.

Glioma-associated microglia/macrophages (GAMs) have traditionally been described as immunosuppressive. However, within their highly heterogeneous repertoire, pro-phagocytic and cytotoxic subsets with anti-tumoral properties exist. Although macrophages (MACs) can exhibit tumor-suppressive functions, their anti-glioma properties largely remain elusive. To identify anti-glioma myeloid cell effectors, we performed directionally concatenated ligand-receptor (L-R) interactome analyses from dendritic cells (DCs) and microglia (MG) to lymphoid (CD4+T, Tregs, CD8+T, NK, and NKT) cells identified by our recent single-cell transcriptomics interrogation of tumor-associated CD45+ leukocytes from tumor brains of eighteen isocitrate dehydrogenase (IDH)-stratified glioma patients. Within DC-specific and MG-specific interactomes, we identified LGALS9, which encodes Galectin-9, as a key mediator of cell-cell interactions in IDH-wild type (IDH-wt) gliomas. Spectral cytometry, immunohistochemistry, and Western blotting analyses confirmed the abundant expression of Galectin-9 in glioma-associated MG, but not in tumor cells. Furthermore, differential gene enrichment analyses revealed transcripts associated with cellular adhesion (coronin 1A, integrin) and phagocytosis (FcγR, phospholipase D, Rab family proteins, etc.) pathways that were significantly upregulated in Galectin-9+ compared to Galectin-9- MG and MACs. Using an ex-vivo primary human microglia (pMG) and patient-derived glioma stem cells (GSCs) co-culture system, we evaluated the functional role of Galectin-9. Confocal imaging analyses of co-cultured pMG with GSCs revealed that siRNA-mediated knockdown or antibody-based neutralization of Galectin-9 significantly impaired pMG-GSC adhesion. In addition to reduced adhesion, phagocytosis of GSCs was dramatically attenuated across all Galectin-9 silenced or neutralized pMG. Altogether, our study underscores the unappreciated non-canonical role of Galectin-9 in MG as a regulator of glioma cell adhesion and phagocytosis that can be harnessed for glioblastoma immunotherapy.

Glaucoma, the leading cause of irreversible blindness worldwide, is characterized by the progressive loss of retinal ganglion cells (RGCs) and their axons. While elevated intraocular pressure (IOP) is a core risk factor, the pathogenesis of normal-tension glaucoma (NTG) remains unclear, as static IOP is within the normal range. Based on circadian fluctuations of IOP and cerebrospinal fluid pressure (CSFP), and the pressure-dependent function of the ocular glymphatic system, we propose the "dynamic trans-lamina cribrosa pressure difference (TLCPD) imbalance" hypothesis. This hypothesis posits that optic nerve damage may stem from abnormal pulse synchrony between IOP and CSFP (phase mismatch, amplitude mismatch, or abnormal frequency) rather than static TLCPD elevation alone, pending further validation. Dynamic imbalance induces RGC injury through dual mechanisms: mechanical stress on the lamina cribrosa (collagen fiber rupture, astrocyte activation) and metabolic dysfunction (ocular glymphatic clearance impairment, toxic waste accumulation), which ultimately converge on the activation of the programmed axonal degeneration (PAD) pathway-a conserved final common effector of RGC axon loss. Phase mismatch is the core pathological pattern in NTG. In contrast, high-tension primary open-angle glaucoma (POAG) is characterized mainly by amplitude mismatch and abnormal frequency, with potential coexistence and mutual influence of these mechanisms. Verifiable clinical (24-h IOP-CSFP synchronous monitoring) and animal experiments are proposed. This hypothesis may help explain unresolved clinical phenomena, provides novel diagnostic markers (a transient peak in TLCPD) and therapeutic strategies (CSFP regulation, glymphatic function enhancement, modulation of the PAD pathway), and opens new avenues for personalized glaucoma management.

Endometrial-derived hydrogels are rich in extracellular matrix proteins as well as bioactive molecules that promote cell adhesion, proliferation, and angiogenesis.

Elevated interleukin-1 beta (IL-1β) in orchitis triggers immune-mediated inflammation, disrupting spermatogenesis and impairing reproductive function.

Despite the advent of potent tyrosine kinase inhibitors (TKIs), resistance and disease persistence remain significant clinical challenges in chronic myeloid leukemia (CML). This perspective aims to synthesize concepts derived from recent advances in single-cell and multi-omics analyses, which have revealed profound heterogeneity among leukemic stem cells (LSCs). These findings augment traditional models that focus solely on clonal selection and resistance-conferring mutations. We discuss how LSCs, like normal hematopoietic stem cells (HSCs), exist in a spectrum of transcriptionally and epigenetically defined cell states, each governed by distinct gene regulatory networks (GRNs) that confer unique lineage biases and responses to therapy. Incorporating recent insights from single-cell analysis, our perspective highlights evidence for a conserved chronic phase (CP) LSC state characterized by lineage skewing, altered metabolic and environmental responsiveness, and epigenetic dysregulation, features that are likely to be underpinned by specific GRN configurations that collectively contribute to intrinsic TKI resistance. We explore how both intrinsic factors (such as germline polymorphisms and lineage bias) and extrinsic cues (including microenvironmental signals, immune interactions, and hypoxia) are likely to modulate GRN activity and LSC states, thereby affecting apoptotic thresholds, primary resistance, and the potential for treatment-free remission (TFR). Emerging data support the concept of GRNdefined LSC states at diagnosis that are predictive of TKI responses. Furthermore, multiple studies suggest that blast crisis (BC) converges on a common high-risk transcriptomic and GRN state that is agnostic to mutational diversity, and driven by polycomb and DNA methylation-dependent epigenetic reprogramming. Given that BCR::ABL1-independent mechanisms, regulated at the level of GRNs, may contribute to resistance and LSC persistence, these observations support placing greater emphasis in CML management on addressing GRN-defined cell-state vulnerabilities, with the goal of lowering the risk of blast crisis in high-risk patients and improving control of therapy-resistant CP LSCs.

Mobilized hematopoietic stem and progenitor cells (HSPCs) are essential for transplantationbased therapies, including curative gene therapies for sickle cell disease (SCD). While granulocyte colony-stimulating factor (G-CSF, filgrastim) remains the standard mobilization agent, many patients respond inadequately, and it can trigger life-threatening vaso-occlusive crises in SCD. The CXCR4 antagonist AMD3100 (plerixafor) is routinely combined with G-CSF for non- SCD settings but is ineffective as a single agent in SCD, underscoring the urgent need for alternative strategies. We previously identified 27-hydroxycholesterol (27HC) as a physiological inducer of HSPC mobilization during pregnancy. Here, we show that exogenous 27HC enhances AMD3100-induced HSPC mobilization in mice, either alone or with G-CSF. Because 27HC is metabolized by the enzyme Cyp7b1, we tested whether pharmacological Cyp7b1 inhibition could mimic this effect. Treatment with clotrimazole, an antifungal and Cyp7b1 inhibitor, significantly enhanced AMD3100-induced HSPC mobilization in wild-type, SCD, and humanized mice. Importantly, intravenous administration of voriconazole, a clinically approved systemic antifungal with Cyp7b1-binding activity, similarly augmented AMD3100-induced HSPC mobilization in wildtype and SCD mice without altering steady-state hematopoiesis. These findings establish Cyp7b1-inhibiting azoles as novel and clinically relevant enhancers of HSPC mobilization, particularly for SCD patients who cannot safely receive G-CSF but require robust HSPC yields for gene therapy.

The environmental pollutant benzo(a)pyrene (B(a)P), a representative polycyclic aromatic hydrocarbon (PAH), is a recognized carcinogen, and chronic pulmonary inflammation is closely associated with lung carcinogenesis. Although alveolar type 2 (AT2) cells are the origin of lung adenocarcinoma, the genetic and functional changes in AT2 cells and the mechanisms involved in inflammation-related lung tumorigenesis have not been elucidated. Here, C57BL/6J mice are exposed to B(a)P and the inflammatory irritant lipopolysaccharide (LPS) to establish a model of inflammation-related lung tumorigenesis. Single-cell RNA sequencing is performed on lung tissues. DNA mutations in AT2 cells are analyzed via whole-exome sequencing. The protein expression of AT2 cells in lung cancer tissue is determined by immunofluorescence staining. The results reveal that LPS promotes B(a)P-induced lung tumorigenesis; in the whole lungs of B(a)P/LPS, a decreased proportion, altered differentiation trajectory, and increased gene mutation number in AT2 cells are observed. Additionally, in B(a)P/LPS-treated lung cancer tissue, the levels of γ-H2AX DNA damage and the proliferation marker Ki67 in AT2 cells are increased, whereas the levels of differentiation markers are decreased. Single-cell RNA transcriptomics reveals that the autophagy-related genes Foxo3 and Ppp2r5, which are enriched in the PI3K-Akt pathway, and the autophagy-related genes in AT2 cells in lung cancer are decreased in the B(a)P/LPS group. Thus, chronic inflammation promotes DNA damage, gene mutation and dysfunction in AT2 cells, and decreased autophagy in AT2 cells may be an important mechanism for inflammation-related lung tumorigenesis.

Interleukin 16 (IL16) is a pleiotropic cytokine that does not belong to any interleukin family and functions as a CD4+ T-cell chemokine, a regulator of T-cell activation, and an inhibitor of human immunodeficiency virus replication in acquired immunodeficiency syndrome. Studies in the hemato-oncological disease multiple myeloma have demonstrated that IL16 plays an important role in promoting tumor cell growth and is associated with tumor burden and prognosis. However, the role of IL16 in acute myeloid leukemia (AML) remains largely unexplored.

Allogeneic hematopoietic stem cell transplantation can achieve durable HIV remission when donor cells exhibit reduced or absent C-C chemokine receptor type 5 (CCR5) function, but CCR5Δ32/Δ32 donors are rare and cord blood banks (CBBs) do not routinely screen inventories for CCR5-deficient units. We evaluated a scalable flow cytometry-based phenotyping strategy to identify cord blood units (CBUs) with intrinsically reduced CCR5 surface expression and to determine whether low-expression phenotypes enrich for CCR5Δ32 and other CCR5 variants.

The thymic medulla is essential for establishing central tolerance, orchestrating the development of a diverse yet self-tolerant T cell repertoire, and preventing autoimmunity. This process is primarily mediated through interactions between developing thymocytes and antigen-presenting cells, including thymic epithelial cells (TECs) and dendritic cells (DCs), with additional regulatory contributions from endothelial cells, mesenchymal cells, and macrophages. Despite its critical role, the complexity of late-stage thymocyte development and the dynamics of their medullary residency remain incompletely understood. Recent advances in single-cell, epigenomic, and transcriptomic technologies have begun to reveal previously unappreciated layers of cellular and molecular heterogeneity within the thymic medulla throughout life. In this review, we explore how the medulla shapes the fate of both conventional and non-conventional T cells, examine the diversity of thymocyte populations it supports, and discuss how this specialized microenvironment adapts during aging and regeneration.