The COVID-19 pandemic, caused by SARS-CoV-2, has underscored the urgency of understanding viral entry mechanisms to develop effective therapeutic strategies. SARS-CoV-2 primarily exploits angiotensin-converting enzyme 2 (ACE2) as its entry receptor and relies on the serine protease TMPRSS2 to prime its spike protein, enabling membrane fusion and infection. Traditionally, TMPRSS2 has been described as a cell surface protein, but our study reveals that in human lung epithelial cells, TMPRSS2 is largely absent from the plasma membrane and instead resides intracellularly. We show that TMPRSS2 is secreted together with ACE2 in extracellular vesicles (EVs) from lung epithelial cells, which are subsequently taken up by non-epithelial cells, specifically alveolar macrophages, endothelial cells, and pericytes, that do not express TMPRSS2 or ACE2 mRNAs under homeostatic conditions. This EV uptake deposits ACE2 and TMPRSS2 protein onto recipient cells, equipping them for SARS-CoV-2 entry. By transferring these viral entry proteins, EVs expand the spectrum of susceptible cell types in the lung, offering a new explanation for how the virus can infect diverse cell populations and cause widespread tissue damage. Identifying EVs as vehicles for delivering functional ACE2 and TMPRSS2 across cell types reveals a previously unrecognized pathway of viral entry with important implications for not only COVID-19 pathogenesis but also for other viral infections that exploit similar entry mechanisms. These findings open new avenues for therapeutic intervention aimed at disrupting EV-mediated protein transfer, potentially limiting viral dissemination and severity, and may also represent a generalizable mechanism exploited by other viral pathogens, highlighting the potential relevance of EV-mediated protein transfer beyond SARS-CoV-2.

Hematologic malignancies are a group of malignant diseases originating from hematopoietic stem cells or the lymphatic system, mainly including leukemia, lymphoma, myeloma and so on. These diseases are characterized by their high heterogeneity, rapid disease progression and poor prognosis. Glycosylation is one of the most common post-translational modifications. In recent years, it has been found that abnormal glycosylation plays an important role in the genesis, development and treatment of hematologic malignancies. Aberrant glycosylation has been demonstrated to exert a significant influence on the progression of disease, impacting various biological processes including tumor cell signaling, the tumor microenvironment, cellular recognition, and immune evasion. This review synthesizes recent advances in how dysregulated glycosylation drives the biology of leukemia, lymphoma, myeloma, and other types of malignancies. Additionally, it discusses the potential value of glycosylation products that can be used as biomarkers for disease diagnosis and prognosis.

Epithelial cell adhesion molecule (EpCAM) is a tumor-associated antigen that marks pluripotent embryonic stem cells (ESCs). Regulation of Epcam expression yields a spatiotemporal patterning during embryogenesis that is thoroughly mimicked in a 3D model of spontaneous differentiation of embryoid bodies (EBs). Here, we present a role of EpCAM in exit from pluripotency of murine ESCs (mESCs) to establish cardiomyocytes in EBs. Comparative transcriptomic analysis of wildtype and Epcam-knockout mESCs at strategic time points of spontaneous differentiation uncovered molecular deficiencies of Epcam-knockout ESCs in "Wnt signaling" and "Heart development". Multi-level bioinformatic analyses revealed central lineage-defining transcription factors Eomes, Foxa2, and Gata6 as differentially expressed genes (DEGs) that are misregulated in Epcam-knockout mESCs. Gene expression association of Epcam with Eomes, Foxa2, and Gata6 was prominent at day three of spontaneous differentiation, representing primitive streak formation in EBs. Interrogation of public single-cell RNA sequencing (scRNAseq) datasets supported a co-expression of Epcam and Eomes at early stages of murine embryogenesis in epiblast, primitive streak, nascent mesoderm, extraembryonic ectoderm and endoderm. Newly generated scRNAseq of wildtype mESCs in spontaneous differentiation delineated the formation of epiblast, primitive streak, endo- and mesoderm cells, and cardiomyocytes. Expression and pseudotime analysis positioned Epcam expression slightly ahead of Eomes at the transition of early to late primitive streak, along with rising Wnt signaling. Accordingly, conditional re-expression of Epcam or Eomes but not of Foxa2 or Gata6 complemented differentiation defects of Epcam-knockouts and confirmed an involvement of Wnt signaling in the EpCAM-dependent activation of Eomes. Hence, defective exit of pluripotency in Epcam-deficient ESCs is linked to Eomes regulation via Wnt signaling.

Tissue engineering, particularly utilizing mesenchymal stem cells (MSCs), represents a rapidly advancing strategy for craniofacial bone regeneration. While traditional autologous bone grafting remains the standard, its widespread application is severely limited by donor site morbidity, prolonged recovery, and inconsistent results.

The generation of hepatocyte-like cells (HLCs) from human pluripotent stem cells (hPSCs) holds great promise for drug discovery and cell-based therapy for liver disease. However, current differentiation protocols are complicated and unstable, and the underlying gene regulatory mechanisms of hepatic differentiation remain incompletely defined. Here, we developed a machine learning-based artificial intelligence (AI) tool using phase-contrast images of hepatic progenitor cells (HPCs), which are essential for generating HLCs. The AI tool significantly improves the success rate of hepatic differentiation without the need for immunostaining or lineage tracing. By optimizing the methodology, we achieved an impressive purity of 90-95% for HLCs derived from hPSCs, aided by the AI algorithm. Through further investigating transcriptomes and epigenomic changes, we discovered the pivotal roles of NR5A2 and AP-1 transcription factors in regulating the maturation of hepatocytes. Single-cell RNA sequencing (scRNA-seq) demonstrated the upregulation of NR5A2 and AP-1 during hepatic differentiation. Importantly, mutagenesis and tumorigenesis assays confirmed the safety of this modified hepatic differentiation protocol. This work highlights the potential of combining AI algorithm and computational genomics to facilitate development of lineage differentiation and molecular mechanism study.

The fallopian tube microenvironment supports gamete transport, fertilization and early embryo development. Any disturbances in this microenvironment can lead to fertilization failure, infertility or ectopic pregnancy. In this study, we systematically investigated the effects of clomiphene citrate (CC) on human PAX8-positive fallopian tube secretory epithelial cells (hPFTSECs) to assess CC-induced alterations in the tubal microenvironment. Human fallopian tube tissues obtained from women undergoing postpartum tubectomy were enzymatically digested, and the isolated hPFTSECs were cultured with CC. CC exposure reduced hPFTSEC viability, clonogenicity, proliferation and organoid forming efficiency while inducing apoptosis, DNA damage, and senescence in a dose-dependent manner. Further, delayed cell-cycle progression and impaired DNA replication were observed in CC-exposed hPFTSECs. CC when administered to adult female Swiss albino mice, both single-dose (25, 50, and 100 mg/kg; intraperitoneally) and multiple-dose (10 mg/kg; intraperitoneally for four consecutive days) exposure caused several oviductal abnormalities, including epithelial disorganization, loss of ciliation, dysplasia, and hyperplasia. Our findings reveal that CC induces a spectrum of cytotoxic, genotoxic, and structural alterations in the fallopian tube epithelium, with potential implications for tubal function impairment and disruption of the optimal microenvironment essential for fertilization and early embryo development, which might negatively impact overall reproductive outcomes.

Chimerism-based strategies remain promising for tolerance induction in solid organ and vascularized composite allograft (VCA) transplantation. This study aimed to develop a novel, less toxic chimeric cell therapy to prolong allograft survival and reduce the need for lifelong immunosuppression. Di-chimeric cells (DCC) were created via polyethylene glycol (PEG)-mediated ex vivo fusion of allogeneic hematopoietic stem cells (HSC) and mesenchymal stem cells (MSC) derived from August Copenhagen Irish (ACI) and Lewis rats. Twenty-four fully major histocompatibility complex (MHC)-mismatched groin flap VCAs were transplanted from ACI rat major histocompatibility complex (rat MHC) (RT1a) donors to Lewis (RT11) recipients under a 7-day immunosuppressive protocol of anti-αβTCR antibody and tacrolimus, combined with four different cell therapies of n = 6/group: Group 1, saline control; Group 2, MSC; Group 3, HSC/HSC DCC; and Group 4, HSC/MSC DCC. DCC were delivered via the intraosseous injection. DCC phenotype was confirmed by flow cytometry (FC). Graft rejection was evaluated macroscopically. A single DCC dose significantly prolonged VCA survival, with the best results in Group 4 (94 ± 1.65 days), followed by Group 3 (66 ± 1.24 days), Group 2 (45.5 ± 4.08 days), and Group 1 (38 ± 4.29 days). This study confirmed immunomodulatory and tolerogenic properties of DCC, supporting VCA transplantation.

Antisense oligonucleotides (ASOs) have emerged as a powerful therapeutic modality for targeting disease‑associated RNAs. However, most clinical ASOs rely on chemical modifications to enhance stability and pharmacokinetic properties. In this study, we developed a series of pH‑responsive hairpin‑structured ASO prodrugs based on the i‑motif. By systematically varying loop size, loop position, and stem length, we obtained prodrugs with distinct structural stability and release kinetics. Notably, the R-series ASO prodrugs demonstrated the highest stability among all designed sequences, a property attributable to their largest loop structure. Among these, the construct with a 5‑base‑pair stem exhibited the optimal balance between stability and acid‑triggered release efficiency. In SK-BE (2) cells, these ASO prodrugs, particularly the R3‑5 and R5‑5 sequences, effectively silenced MYCN expression and induced apoptosis. This work provides a rational structure‑activity design strategy for tumor‑microenvironment‑responsive nucleic acid therapeutics and establishes a reference for further structural optimization.

Capping materials are critical for vital pulp therapy in endodontic treatment, whereas the currently available ones remain inadequate in meeting the multiple requirements of the complex tissue defects. Herein, we report an injectable and self-setting calcium polyphosphate coacervate composite (polyP-Ca-CS) that exploits an acid neutralization mechanism based on the coacervate and chitosan, engineered for direct pulp capping. The material leverages polyP-Ca coacervate (formed through liquid-liquid phase separation) as an injectable matrix and chitosan that initiates setting via acid neutralization, enhances mechanical strength, and confers antibacterial properties. polyP-Ca-CS sets into a rigid solid under both aqueous and anhydrous conditions without significant exothermic reaction or volume change-features that are critical for clinical reliability. Mechanistic investigations reveal that neutralization of the coacervate's intrinsic acidity drives the setting, thereby advancing fundamental understanding of setting mechanisms in polyphosphate-based materials. In vitro, polyP-Ca-CS significantly boosts ATP production, mitochondrial function, cell migration, metabolic activity and odontogenic differentiation of dental pulp stem cells (DPSCs). In a rabbit pulp exposure model, it effectively induces reparative dentin formation and preserves pulp vitality, performing comparably to commercial bioceramics. This work presents a bioenergetic-active biomaterial that meets the complex requirements of vital pulp therapy and offers a promising alternative for regenerative endodontics. STATEMENT OF SIGNIFICANCE: This work presents an application of a polyphosphate-based coacervate system in vital pulp therapy, opening an avenue for bioactive dental biomaterials. Besides, it elucidates that setting of the calcium polyphosphate coacervate is driven by acid neutralization, advancing fundamental insights into inorganic coacervate chemistry. The prepared calcium polyphosphate coacervate composite not only meets stringent physical requirements but also activates cellular energy metabolism. Additionally, this work also demonstrates that chitosan-a normal biopolymer-can serve as an setting initiator for inorganic coacervate systems, while simultaneously enhancing mechanical integrity and conferring antibacterial activity.

Enzalutamide resistance is a major clinical bottleneck limiting the therapeutic efficacy of castration-resistant prostate cancer (CRPC). Enzalutamide treatment can exacerbate tumor hypoxia, which is a critical contributor to the development of its resistance. The traditional Chinese formula Guben Qingyuan Herbal Medicine (GBQY), coupled with enzalutamide, has demonstrated potential in delaying enzalutamide resistance. Nevertheless, its interaction with the tumor microenvironment (TME) remains insufficiently investigated.

Bosma arhinia microphthalmia syndrome (BAMS) is a rare congenital disorder characterized by the absence of a nose, along with eye and reproductive anomalies. BAMS is caused by heterozygous missense variants in SMCHD1, an epigenetic regulator. Despite uncovering its genetic basis, the cellular basis of the disease has remained elusive. The embryonic development of the nose involves direct contributions from both the neural crest and cranial placodes. Here, we differentiated patient-derived induced pluripotent stem cells toward the cranial placode lineage and found that they exhibited significant differentiation defects. Combined transcriptome and DNA methylome analyses further revealed dysregulation in cell adhesion but without overt apoptosis. Together our research suggests that BAMS is caused by impaired differentiation to cranial placode cells and changes in cell adhesion, offering new insights into the cellular pathology of this enigmatic syndrome.

Corneal injuries remain a major cause of visual impairment, where inflammation, infection, and stromal scarring severely hinder functional regeneration. Here, we propose a multifunctional hydrogel concept that integrates structural biomimicry with immunoregulatory and antimicrobial therapy to achieve scar-free corneal healing. The injectable hydrogel is constructed from type I collagen and oxidized dextran via dynamic Schiff-base crosslinking, incorporating poly(lactic-co-glycolic acid)-poly(ethylene glycol) (PLGA-PEG) nanomicelles encapsulating magnolol (Mag-NMs) and bone marrow-derived mesenchymal stem cells (BMSCs). This composite system exhibits excellent transparency, tunable viscoelasticity, and enzymatic biodegradability, forming a biocompatible microenvironment that supports cell integration and drug release. Mag-NMs endow potent antibacterial and anti-angiogenic properties while modulating macrophage polarization toward an anti-inflammatory M2 phenotype. Meanwhile, embedded BMSCs further promote epithelial repair and suppress fibrotic remodeling. In a rabbit corneal injury model, the OC/Mag-NMs/BMSCs hydrogel accelerated re-epithelialization, restored optical clarity, and markedly reduced inflammatory (IFN-γ, CD4, CD68) and fibrotic responses without systemic toxicity. This multi-targeting strategy demonstrates that coupling small-molecule nano-delivery with stem-cell therapy within a biomimetic scaffold enables comprehensive regulation of infection, inflammation, and fibrosis, offering a promising direction for translational corneal regeneration.

Psoriasis is a chronic immune-mediated inflammatory disease with systemic manifestations beyond the skin, yet the role of subcutaneous adipose tissue (SAT) in disease biology and therapeutic response remains poorly understood. Here, we investigated inflammatory features of SAT in psoriasis and the effects of dimethyl fumarate (DMF) on this compartment. Six adults with moderate-to-severe plaque psoriasis received oral DMF for 24 weeks and were clinically evaluated measuring the Psoriasis Area and Severity Index (PASI), showing a consistent reduction in disease severity during treatment. Publicly available spatial transcriptomic data were analysed to profile inflammatory signatures in SAT clusters of psoriatic versus healthy skin. Bulk RNA sequencing was performed on SAT biopsies obtained from psoriatic plaques before and after DMF treatment in four patients. Complementary in vitro models using murine 3T3-L1 adipocytes and human adipocytes differentiated from mesenchymal stem cells were exposed to pro-inflammatory cytokines or macrophage-conditioned media (CM) with or without DMF to assess effects on inflammatory gene expression and NF-κB signalling. Spatial transcriptomics identified enrichment of inflammation-related pathways in SAT beneath psoriatic lesions. DMF treatment was associated with reduced expression of inflammatory mediators and with a shift in SAT transcriptional profile toward patterns observed in healthy tissue. In vitro, DMF significantly attenuated cytokine- and CM-induced adipocyte activation and reduced NF-κB phosphorylation in both murine and human adipocyte models. These data provide integrated clinical and experimental evidence that DMF treatment is associated with reduced disease activity and attenuation of inflammatory signalling within psoriatic SAT, supporting adipose tissue as a potentially modifiable inflammatory compartment in psoriasis.

Pineapple (Ananas comosus (L.) Merr.) from Tac Cau, Vietnam, represents a valuable source of bioactive compounds, particularly in its peel, a traditionally used ethnomedicinal material. This study evaluated the antioxidant and anti-melanogenesis activities of methanol and ethanol extracts from different pineapple parts (peel, leaf, flesh, and stem) and further characterised metabolites in the ethyl acetate peel fraction. Among all samples, the methanolic peel extract exhibited the highest phenolic and flavonoid contents and demonstrated the strongest radical scavenging, reducing power, and tyrosinase inhibitory effects, which correlated with significant suppression of melanin synthesis in B16F10 cells. The ethyl acetate fraction showed enhanced bioactivity, and LC-MS/MS analysis identified caffeic acid, p-coumaric acid, chlorogenic acid, and related phenylpropanoid derivatives as key constituents. These findings substantiate traditional uses of pineapple peel and highlight its potential for functional food and cosmeceutical development while promoting sustainable utilisation of local agro-resources.

Articular cartilage injuries pose a major clinical obstacle due to their inability to regenerate contributed by cartilages' intrinsic properties and close association with osteoarthritis and progressive joint degeneration. Cartilage damage may be a consequence of acute trauma, repeated mechanical overload or age-related degenerative processes which often leads to chronic pain, joint dysfunction and a deterioration in the quality of life of patients.Established treatments such as; conservative management, intra-articular drug administrations and surgical cartilage repair typically provide relief. However, it's important to note that these treatments rarely lead to complete, permanent regeneration of natural hyaline cartilage. Recently, regenerative medicine has been paying significant attention to stem cell therapies. It aims to support cartilage repair while simultaneously impacting the intra-articular environment. It's safe to say that these approaches are increasingly being considered as potential therapeutic methods. Between the various cell populations, mesenchymal cells have gained particular attention due to their ability to promote chondrogenic differentiation, immunomodulatory properties, and paracrine effects.There is growing evidence suggesting that stem cells effects can be mediated not only by direct source replacement but are also contributed by the secretion of bioactive factors that influence physical processes, cartilage metabolism, and endogenous repair mechanisms. This narrative review aims to concisely summarize and critically evaluate novel evidence and scientific data on the biological repair mechanisms, clinical outcomes and safety assessment of stem cell-based therapies used to treat articular cartilage repair.

Oncofertility has emerged as a critical interdisciplinary field addressing the reproductive challenges faced by cancer patients, particularly those undergoing chemotherapy. While chemotherapeutic agents remain indispensable in cancer therapy, their gonadotoxic effects frequently result in diminished ovarian reserve, impaired spermatogenesis, and long-term infertility. Exosomes - small extracellular vesicles enriched with nucleic acids, proteins, and lipids - are increasingly recognized as key mediators of intercellular communication in both pathological and regenerative contexts. Recent evidence suggests that chemotherapy alters exosome cargo, thereby amplifying cellular stress responses, oxidative damage, and bystander effects in gonadal tissues. Conversely, exosomes derived from mesenchymal stem cells, induced pluripotent stem cells (iPSCs), and other regenerative sources demonstrate the ability to restore ovarian and testicular function by reducing apoptosis, enhancing angiogenesis, and supporting germ cell survival. This dual role positions exosomes as both contributors to reproductive toxicity and promising therapeutic agents in fertility preservation strategies. However, clinical translation remains hindered by challenges including source heterogeneity, isolation methods, safety concerns, and regulatory barriers. This review highlights the emerging roles of exosomes in chemotherapy-induced reproductive damage, explores their regenerative potential, and outlines future directions for their integration into oncofertility practice.

Multiple biomarkers have been proposed to identify cancer stem cells in tongue squamous cell carcinoma. This study evaluated ALDH1A1, an ALDH1 subtype implicated in head and neck cancer stem cells, and examined its association with histopathological features (depth of invasion, worst pattern of invasion, perineural invasion, grade, inflammation, TNM stage) and prognostic outcomes in tongue tumors.

While Fgf and Hippo-Yap signaling are fundamental for proper development, homeostasis, and disease, their crosstalk remains largely unknown. Here, we identified that Yap and Taz, canonical Hippo effectors, function as noncanonical effectors of Fgf signaling to maintain the proper function of neural crest (NC) lineages. NC cells are a multipotent stem cell population during vertebrate embryogenesis that contribute to numerous structures and diverse cell lineages, including craniofacial and cardiac tissues, neurons, and suture mesenchymal cells (SMCs), a specified cell population required for cranial bone growth and repair. We observed that activation of Fgf signaling in NC cells and NC-derived SMCs inhibited osteogenesis while simultaneously enhancing stemness and proliferation. Interestingly, these effects were reversed by inhibition of either Yap/Taz or phosphorylated Erk1/2 (pErk1/2). Mechanistically, Fgf signaling promotes the interaction of Yap and pErk1/2, increasing the chromatin occupancy of Yap at genes regulating stemness, proliferation, and osteogenesis. We further show that pERK1/2 phosphorylates YAP at the noncanonical S128 site, enhancing YAP's nuclear localization. This mechanism is conserved across mouse and human cells and is active in Apert syndrome-associated FGF gain-of-function models, revealing a previously unrecognized FGF-YAP axis in stem cell regulation.

The infrapatellar fat pad (IFP) is a rich source of mesenchymal stem cells (MSCs) with dual contributions from adipose and synovial tissues. The heterogeneity of IFP-derived MSCs and the lack of standardized isolation protocols, however, hinder consistent therapeutic outcomes. This study aimed to optimize collagenase-based isolation protocols for IFP-MSCs, with a focus on the effects of enzyme concentration and treatment duration on tissue digestion, cell origin, viability, and functional properties.

Prolonged glucocorticoid exposure leads to oxidative stress, mitochondrial damage and impaired myogenesis reducing the overall health of the skeletal muscles. Dexamethasone (dex), a synthetic glucocorticoid, induces proteolysis and inflammation by disrupting cellular energetics and mitochondrial function. Vitamin B3 (vit B3), an NAD+ precursor, is known to be a natural antioxidant and anti-inflammatory compound. This study investigates the protective role of vit B3 against dex-induced skeletal muscle damage, focusing on mitochondrial homeostasis and the IKK/FoxO3a signalling axis. C2C12 myoblasts were treated with dex (200 µM) and/or vit B3 (1 mM). Oxidative stress, mitochondrial potential and DNA damage was evaluated using DCFDA, JC1, and γH2AX immunostaining, respectively. Gene expression analysis was performed to assess the mitochondrial fission/fusion and the extent of electron transport chain (ETC) gene expression. Protein expression of inflammatory (IKKα/β, NFκB) and atrophy markers were analysed using immunoblotting and flow cytometry. The extent of myogenic differentiation was evaluated using MyoD and MyHC1 immunostaining along with measurement of the morphometric parameters. Vit B3 treatment significantly enhanced C2C12 viability and reduced dex-induced ROS production while restoring Nrf2 expression. It prevented DNA damage and preserved mitochondrial membrane potential. The results also implicated increased mitochondrial fusion upon vit B3 treatment as seen by the elevated gene expression of Mfn1, Mfn2 and Opa1 and decreased fission as observed by the reduced expression of Fis1 and Drp1. The NADH levels were also seen to be rescued by vit B3 supplementation which translates to better energy production by the electron transport system. Additionally, vit B3 was observed to suppress inflammation and prevent muscle proteolysis by modulating an IKK/FoxO3a axis. Finally, vit B3 was able to improve differentiation as seen by the levels of MyoD and MyHC1 expression in the cells. Vit B3 acts in a multifaceted manner and reduces dex-induced skeletal muscle atrophy which is primarily a result of reduced oxidative stress and restored mitochondrial homeostasis. These findings highlight vit B3 as a potential therapeutic and nutritional supplement for maintaining the skeletal muscle health under myopathic conditions.