Hyperbaric oxygen therapy (HBOT) enhances wound healing by promoting angiogenesis and reducing hypoxia. However, the role of air-breaks-intermittent exposures to ambient air during HBOT-remains unclear. We investigated the effects of air-breaks on HBOT-mediated wound healing, particularly in combination with adipose-derived stromal cells (ASCs). Full-thickness wounds were created in C57BL/6 mice (n = 36) and assigned to control, HBOT (1 h/day, 2 ATA for 11 days), or HBOT with a 10-min air-break groups. In a second experiment, we evaluated ASC treatment combined with HBOT and air-breaks. Wound healing was assessed via gross examination, histology and gene expression analysis of collagen type 1 alpha 1 (Col1a1), hypoxia-inducible factor 1 alpha (Hif1a) and tumour necrosis factor (Tnf-α). Compared with HBOT alone, air-breaks significantly improved wound closure, epithelial regeneration and collagen deposition (p < 0.05). Gene analysis showed higher Col1a1 expression and lower Hif1a and Tnf-α levels in the air-break group. In ASC-treated wounds, air-breaks further accelerated healing, enhancing collagen synthesis and reducing hypoxia and inflammation. These findings suggest that incorporating air-breaks into HBOT protocols improves wound healing outcomes, both generally and in ASC-based therapies, by modulating collagen production, hypoxia and inflammation, and could optimise HBOT efficacy, particularly in cell-based regenerative therapies.

Osteoporotic bone defects, characterized by chronic inflammation and impaired osteogenesis, pose a formidable challenge for functional bone regeneration. Conventional scaffolds lack effective regulation of the inflammatory microenvironment and fail to coordinate anti-inflammatory and osteogenic signals, thereby limiting their ability to couple inflammation resolution with new bone formation. Here, we developed a "Brake-Drive Osteo System", a spatiotemporally programmed biomaterial integrating quercetin-loaded nanovesicles and teriparatide-loaded nucleic acid frameworks within a calcium phosphate scaffold (BCP-T/N@Q/V). This design enables a sequential therapeutic cascade-quercetin first "disengages the inflammatory brake", alleviating microenvironmental resistance to osteogenesis, while teriparatide subsequently "activates the osteogenic drive", promoting bone regeneration. It effectively reprogrammed macrophages toward a pro-regenerative phenotype and mitigated inflammatory stress, establishing an immunologically permissive microenvironment. This osteoimmune modulation significantly enhanced the osteogenic commitment and maturation of osteoporotic bone marrow mesenchymal stem cells. In a rat osteoporotic femoral condyle defect model, the scaffold achieved accelerated, structurally integrated bone regeneration, underscoring its translational potential. Collectively, this "Brake-Drive Osteo System" provides a sequential strategy that couples inflammation resolution with osteogenesis for effective osteoporotic bone regeneration.

Human embryos undergo pivotal morphogenetic remodelling shortly after implantation. The understanding of this crucial stage is severely impeded by the scarcity of embryonic samples and ethical constraints. Pluripotent stem cells with the competence for somatic and germline differentiation serve as in vitro models of epiblast. In this study, we established human formative pluripotent stem cell-like cells (hfPSC-LCs) from naïve human embryonic stem cells (hESCs), conventional hESCs, human induced pluripotent stem cells (hiPSCs), as well as human blastocysts using the three-dimensional (3D) Matrigel culture system. Similar to pre-gastrula stage epiblast, hfPSC-LCs self-organise into self-renewing colonies with an apical lumen and exhibit several hallmarks of formative pluripotency, consistent with the properties observed in mouse fPSCs. Notably, single cells of hfPSC-LCs could differentiate into amnion-like precursor cells (hALPCs) which are transcriptionally and morphologically similar to the bona fide amnion. Meanwhile, hfPSC-LCs directly respond to primordial germ cell (PGC) induction signals, generating PGC-like cells (PGCLCs) either as single-cell aggregates or intact colonies, with an efficiency of approximately 50%. Chromatin accessibility analysis revealed that the differentiation capacity of hfPSC-LCs for gametes and amnion lineages might correlate with the accessible chromatin architecture of PGC and amnion associated genes. Loss of 3D-Matrigel niche disrupts formative pluripotency in both mouse and human, manifesting as downregulated formative markers and compromised differentiation capacity. Collectively, our findings establish hfPSC-LCs as a 3D model for investigating formative pluripotency of humans, thereby probably addressing a critical gap in the understanding of human pluripotency transitions.

Aspiration pneumonia (AP) is a prevalent and life-threatening pulmonary disease resulting from repeated aspiration of exogenous materials such as food or gastric contents. However, most existing animal models simulate only acute injury and fail to reproduce the chronic pathological features observed in patients. This study aimed to establish a stable and clinically relevant chronic AP mouse model.

Human induced pluripotent stem cells (hiPSCs) may acquire genomic alterations during reprogramming and culture, which poses significant risks for clinical applications. Current detection methods, such as karyotyping analysis, often fail to identify critical submicroscopic variations. This highlights an urgent need for comprehensive genomic surveillance strategies.

Neurodegenerative diseases are characterized by progressive neuron loss and brain atrophy. While conventional studies focused on neuronal death as the primary cause of these diseases, accumulating evidence suggests that impaired neurogenesis, particularly the dysfunction of adult neural stem cells (NSCs), may also contribute significantly to disease pathogenesis. Adult neurogenesis occurs primarily in two adult NSC niches. These specialized niches are enriched with complex cytokine networks, neuronal activity, and non-cellular components such as the extracellular matrix. Understanding the regulation of NSCs in the adult brain and how their dysregulation exacerbates neurodegeneration provides novel insights into therapeutic strategies. This review proposes that dysfunction of the NSC microenvironment, rather than neuronal death alone, may drive neurodegeneration, and that restoring this microenvironment offers a novel research direction of stem cell-based therapies.

The role of inflammation-induced myeloid-biased hematopoiesis in driving resistance to immune checkpoint blockade (ICB) is recognized, yet the intricate mechanisms through which tumors orchestrate it are not fully defined.

Short stature (SS) is a common growth disorder with multiple aetiologies. Variants in the MMP13 gene can result in varying degrees of SS, typically accompanied by pronounced skeletal abnormalities. This study aimed to investigate the genetic basis of SS in a family lacking significant imaging abnormalities and elucidate the underlying pathogenic mechanism.

Steroid-refractory acute graft-versus-host disease (SR-aGVHD) is the predominant cause of morbidity and mortality after allogeneic hematopoietic stem cell transplantation (allo-HSCT), with gastrointestinal involvement (SR-GI-aGVHD) remaining a key obstacle. We conducted a multicenter, single-arm, pivotal trial to assess the efficacy and safety of hUC-MSC PLEB001, a human umbilical cord-derived mesenchymal stromal cells (MSCs) product, plus anti-CD25 monoclonal antibody as second-line therapy for SR-GI-aGVHD.

Clinical responses to mesenchymal stromal cell (MSC) therapies remain variable because MSCs are often treated as uniform biologics despite donor-programmed differences. Evidence indicates that developmental maturity (fetal vs. adult) and biological sex (female vs. male) bias the MSC secretome and downstream signalling (NF-κB, PI3K/AKT-ERK, TGF-β/Smad, Wnt/β-catenin), potentially shaping anti-inflammatory, angiogenic, anti-fibrotic, and regenerative functions.

Strontium (Sr)-based nanofibers have gained great attention in biomedical and tissue engineering applications due to their unique ability to combine nanoscale structural features with the biological activity of Sr ions (Sr2+). Nanofibers offer a versatile platform to harness these properties owing to their high surface area, tunable porosity, and mechanical strength. The incorporation of Sr2+ ions further enhances their bio-functionality and offers a cost-effective alternative to growth factor-based strategies. Sr2+ ions could stimulate the production of growth factors such as vascular endothelial growth factor (VEGF) and basic fibroblast growth factor (bFGF), thereby promoting neovascularization, while also enhancing osteogenesis by mimicking calcium's physiological role, inducing mesenchymal stem cell differentiation, and stimulating extracellular matrix mineralization. This review summarizes recent advances in the fabrication techniques such as electrospinning, assisted-electrospinning, and non-electrospinning, including the design, control composition, morphology, and functionality of Sr-based nanofibers. The mechanisms governing Sr2+ ions interactions with cells and tissues are discussed, along with in vitro and in vivo biological outcomes. Our bibliometric analysis shows that Sr-based nanofibers have been most extensively investigated in bone tissue engineering, followed by applications in drug delivery and tumor therapy, with fewer studies exploring skin and cartilage regeneration. This review highlights the advantages and disadvantages of every fabrication strategy, discusses biomedical applications of Sr-based nanofibers, and outlines challenges and future directions for their clinical translation.

Telomeres are protective DNA caps at chromosome ends that prevent cells from mistakenly recognizing them as broken DNA. These structures are safeguarded by a protein complex called Shelterin, particularly through the TRF2 protein encoded by Trf2. Surprisingly, in mouse embryonic stem cells, TRF2 is not essential for telomere protection, suggesting that other mechanisms compensate for its loss. Here we show that a cellular quality control system called nonsense-mediated mRNA decay (NMD), which normally eliminates defective RNA molecules, plays an unexpected role in maintaining telomere integrity in pluripotent cells. Through a genome-wide genetic screen, we discovered that NMD is essential for cell survival when TRF2 is absent. NMD accomplishes this by degrading an aberrant form of the messenger RNA encoded by Trf1, which produces the TRF1 protein, another Shelterin component. Without NMD, this aberrant RNA produces a truncated, harmful version of TRF1 that interferes with normal telomere protection. Our findings reveal that embryonic stem cells use a unique strategy for chromosome end protection, linking RNA quality control to genome stability in a previously unrecognized way.

Advances in transcriptomic technologies have progressively transformed the questions we can ask and answer about muscle stem cells (MuSCs) during aging. Early microarray and bulk RNA sequencing studies established foundational population-level signatures of aged MuSCs, including attenuation of myogenic and metabolic programs as well as induction of inflammatory and stress-associated transcription. However, these averaged readouts obscured cell-to-cell variability and rare functional states. The transition to single-cell and single-nucleus RNA sequencing marked a turning point by resolving MuSC heterogeneity and revealing that MuSC aging is not purely stochastic. Instead, aged MuSC pools show reproducible changes in state composition, delayed or altered myogenic lineage progression, and selective vulnerability of specific functional subsets. Emerging spatial transcriptomic approaches, although still limited by sensitivity and cell-type discrimination in muscle, are beginning to place these MuSC states into their native tissue context, directly linking transcriptional states, niche organization, and age-associated remodeling. In parallel, integrative multi-omic designs that pair transcriptomics with chromatin accessibility and metabolic measurements have strengthened mechanistic connections among age-associated gene programs, epigenetic remodeling, and metabolic state shifts. Finally, computational frameworks - including trajectory inference, dynamic modeling, and machine learning - are increasingly applied to high-dimensional transcriptomic data to predict aging trajectories and identify candidate rejuvenation targets. In this Perspective, we trace the evolution of transcriptomic technologies through the lens of MuSC aging and highlight how increasing resolution has reframed core models of MuSC decline and plasticity.

Cystinosis is a systemic lysosomal storage disease resulting from mutations in the CTNS gene encoding the lysosomal cystine transporter cystinosin, leading to cystine accumulation in all organs. Despite cystinosin's ubiquitous expression, renal Fanconi syndrome (FS) is the first clinical manifestation of cystinosis, which is not prevented by cystine reduction therapy with cysteamine. Here, we report a novel interaction of cystinosin and sodium/hydrogen (Na+/H+) exchanger proteins in the endosomes of yeast and mammalian cells. NHE3 is a major absorptive sodium transporter at the apical membrane of proximal tubular cells (PTCs). Cystinosin is required for the correct subcellular localization and trafficking of NHE3 and for sodium uptake. Introducing CTNS successfully rescues these defects in CTNS- deficient PTCs, whereas CTNS-LKG, encoding the lysosomal and plasma membrane isoform of cystinosin, did not. NHE3 mislocalization was confirmed in Ctns-/- mice and cystinosis patient kidney. Transplantation of wild-type hematopoietic stem and progenitor cells in Ctns-/- mice restores NHE3 expression at the brush border membrane and improves FS-related phenotypes. This study uncovers an evolutionary conserved novel role of cystinosin in NHE3 trafficking, offering insights into FS pathogenesis and potential new therapeutic avenues.

DNA replication timing (RT) often correlates with transcription during cell fate transitions, yet notable exceptions indicate a complex relationship. Using a reductionist system in mouse embryonic stem cells, we manipulate transcriptional length and strength at a single locus upstream of the silent, late-replicating Pleiotrophin (Ptn) gene. Small reporter genes driven by two of four promoters advance RT, whereas all promoters advance RT when driving the 96-kb endogenous Ptn gene. Inducible transcription of Ptn, but not the reporter, triggers a rapid and reversible RT advance, providing a system to manipulate RT independent of differentiation. Strikingly, deletion of the Ptn promoter and enhancers abolishes transcription yet does not prevent the developmental RT switch to early replication during neural differentiation. These findings, supported by parallel genome-wide analyses during differentiation, demonstrate that transcriptional elongation can causally advance RT in a rate-dependent and context-specific manner, but that transcription is neither necessary nor sufficient for RT advancement. Our results provide a solid empirical base with which to re-evaluate decades of seemingly contradictory literature.

The Drosophila midgut epithelium undergoes nutrient-dependent growth regulated by the InR-Akt-TOR signaling pathway, though the downstream transducers that coordinate this response remain incompletely defined. We demonstrate that hipk is selectively expressed in intestinal stem cells (ISCs) and their immediate progeny, enteroblasts (EBs), the transient precursors to the absorptive enterocytes (ECs) that form the bulk of the gut epithelium. Hipk expression is dynamically regulated by nutritional status and requires active InR-Akt-TOR signaling; notably, ectopic activation of this pathway is sufficient to induce hipk expression even under nutrient-restricted conditions. Through genetic analysis, we show that Hipk promotes ISC proliferation while simultaneously directing lineage specification toward the EB fate, thereby facilitating expansion of the absorptive epithelium in response to nutrient availability. These findings establish Hipk as a critical nutrient-responsive effector that couples insulin signaling to both stem cell division and lineage commitment in the adult intestine.

A reduction in the number of neurons and an increased susceptibility to apoptosis are one of the characteristic features observed in Down syndrome (DS). However, the multifaceted actions of chromosomal aneuploidy hinder the elucidation of the underlying mechanism. Here, using neurons and astrocytes differentiated from patient-derived induced pluripotent stem cells (iPSCs), we aimed to clarify the neuropathology of DS by focusing on aneuploidy-associated stress and the dosage effects of specific genes. Human chromosomal trisomies 13, 18, and 21 exert a stress response on neurons, resulting in the accumulation of protein aggregates. In addition, DYRK1A overdosage in trisomy 21 astrocytes causes intrinsic activation of the NLRP3 inflammasome, which leads to the release of inflammatory cytokines. These dual actions reciprocally interact and enhance neuronal apoptosis in trisomy 21. Notably, correction of DYRK1A copy number successfully rescued apoptotic neural death in combination with a chemical chaperone treatment. Our study provides insights into the neuropathological mechanism of DS and its potential therapeutic strategy.