Cell mechanics, elasticity and viscoelasticity, are key markers of biological states like cancer. Atomic force microscopy (AFM) is ideal for such studies, but its low throughput limits large-scale use. Two solutions exist: automation for higher throughput, or high-density measurements for richer data. The latter enables machine learning (ML)-based classification, with viscoelastic parameters offering unique insights beyond static measures like Young's modulus.

The global aging crisis presents significant challenges in treating age-related skeletal disorders, characterized by impaired bone regeneration due to senescent microenvironment dysfunction. While anti-senescence drugs have shown promising therapeutic potential, their clinical use is hindered by severe side effects and limited regenerative efficacy. Biomaterials-based anti-senescence strategies offer a promising alternative. This study is the first to demonstrate that silicate bioceramics possess not only osteogenic properties but also intrinsic anti-senescence capabilities, laying the foundation for the concept of "Silicate Anti-Senescence (SAS)". Using hardystonite (Ca2ZnSi2O7, ZnCS), one of the classic silicate-bioceramics, we reveal its biphasic biological function-delaying cellular senescence and promoting bone formation-through various formulations, including porous scaffolds, ionic extracts, and particle suspensions. In vitro, ZnCS effectively delays senescence of bone marrow mesenchymal stem cells (BMSCs) and enhances osteogenic differentiation. In vivo, ZnCS 3D scaffolds remodel the senescent microenvironment and accelerate osteoporotic bone regeneration, while oral administration of ZnCS particles exhibits significant anti-osteoporotic effects. Mechanistically, Zn2+ and SiO32- ions released from ZnCS exert synergistic effects that converge on the PI3K-AKT-SIRT1 signaling axis, thereby attenuating senescence-associated programs and reinforcing osteogenic gene networks. Notably, compared to classic anti-senescence drugs (dasatinib and quercetin), ZnCS shows superior performance compared to the control groups in both anti-senescence and osteogenesis. The study provides new insights into developing anti-senescence strategies for age-related skeletal diseases with silicate-based bioceramics.

Autologous hematopoietic stem cell transplantation (AHSCT) is an established therapy for diffuse progressive systemic sclerosis (DpSSc), improving progression-free and overall survival compared with cyclophosphamide; however, relapse occurs in ∼12-17% of patients. Chimeric antigen receptor T cell (CAR-T) therapy offers a novel approach to deplete autoreactive B cells. This study aimed to assess the feasibility, efficacy, and safety of anti-CD19 CAR-T therapy as a rescue treatment in patients with SSc relapse following AHSCT.

Previous studies have shown that the lung tissues of preterm infants with bronchopulmonary dysplasia (BPD) are infiltrated by pro-inflammatory macrophages. The Wnt5a/frizzled-5/CaMKII signaling pathway plays a critical role in regulating macrophage activation. The antimicrobial peptide LL37, an important paracrine factor secreted by cord blood mesenchymal stem cells, modulates macrophages. Lower LL37 levels are associated with a higher risk of BPD. However, whether LL37 protects against inflammation-induced lung injury and its underlying mechanisms remain unclear.

Zinc is a component of the antioxidant defense system. Its functions protecting biological systems from oxidation are exerted at multiple levels including competing with redox active metals for binding sites, dynamically interacting with thiol groups and inducing metallothionein (MT) expression, regulating oxidant production, and increasing antioxidant defenses in part via NRF2 modulation. Zinc also directly and indirectly modulates redox regulated signaling cascades. Zinc deficits can affect not only the capacity of cells to defend against oxidative challenges but also alter redox signaling that modulate key cellular processes. Zinc is essential at different stages of development given its capacity to regulate key participating processes, e.g. cell proliferation, differentiation and survival. In the developing brain, the adverse consequences of a decrease in zinc availability depend on the severity and the timing of the deficiency. While gestational severe zinc deficiency causes teratogenesis in the brain and several other organs, mild zinc deficiency has significant deleterious consequences on the neural stem cell pool, neurogenesis, oligodendrogenesis, and astrogliogenesis in the offspring. Alterations in neuron, oligodendrocyte and astrocyte number, neuronal specification and myelination associated with zinc deficits in early development persist into adulthood, affecting behavior and motor performance. This review will focus on the role of zinc on brain development and on the interconnection between zinc and the redox tone in shaping different windows of neurodevelopment.

Bone marrow organoids (BMOs) are three-dimensional cell culture models that recapitulate key structural and functional features of the bone marrow (BM) niche. BMOs offer important advantages in hematopoietic research by modeling key aspects of human hematopoiesis compared to classical in vitro two- and three-dimensional cellular models including bioreactors, BM-on-a-chip platforms, 2D models or BM ossicles by better recreating the three-dimensional architecture, cellular heterogeneity, and spatial organization of the BM microenvironment. They offer a scalable and cost-effective alternative to animal models and reduce the need for animal experiments. Induced pluripotent stem cell (iPSC)-derived BMOs can be generated from a patient's own cells, enabling personalized disease modeling and drug testing and are highly amenable to gene editing technologies allowing precise modifications to study gene function or model diseases. Recent landmark studies from Christoph Klein and Abdullah Khan have established protocols for the generation of BMOs and demonstrated their applications in disease modeling. Here, we review the critical steps in BMO generation, their structural/ functional validation and discuss how BMOs can be applied to model inflammatory responses, rare genetic bone marrow failure syndromes, and multiple myeloma. These advances demonstrate BMOs' growing potential as powerful tools in hematopoietic research and will pave the way for further innovation and increasingly refined systems in future studies.

Regenerative medicine increasingly relies on therapeutic cells such as mesenchymal stem cells (MSCs), induced pluripotent stem cells (iPSCs), and engineered cellular constructs to repair and restore damaged tissues. However, clinical translation is constrained by challenges in maintaining viability, ensuring precise localization, achieving durable engraftment of transplanted cells, and producing enough clinically relevant cells at scale. Microfluidic technologies are emerging as transformative tools to address these barriers by enabling precise manipulation of fluids, biomaterials, and cells at the microscale. In the context of therapeutic cell delivery, these platforms can improve early retention and engraftment compared with conventional needle injection, tighten control over delivered cell dose, preserve viability under defined shear conditions, enable site-specific placement of cell-laden carriers, and support immunoisolating or immunomodulatory architectures that enhance immune safety. These platforms provide controlled microenvironments that mimic native tissue architecture, regulate biochemical and mechanical cues, and support scalable production of cell-laden carriers. Advances in microfabrication, from soft lithography and thermoplastics to 3D printing and hydrogel integration, have expanded device versatility, while embedded sensors allow real-time monitoring of cell state, metabolism, and differentiation. Beyond single-cell delivery, microfluidics facilitates encapsulation, co-culture, and organoid assembly, enabling multicellular systems with physiologically relevant interactions. Coupled with CRISPR genome editing and synthetic biology, these platforms allow the engineering of "smart" therapeutic cells with enhanced regenerative and immunomodulatory functions. Applications extend to microfluidic sorting for stem cell purification, controlled differentiation, and advanced manufacturing of immune cell therapies such as Chimeric Antigen Receptor (CAR)-T cells, alongside exosome-based strategies for precision delivery. Despite promising progress, obstacles remain in regulatory standardization, large-scale manufacturing, and integration with clinical workflows. This review highlights state-of-the-art microfluidic approaches for controlled delivery of stem cells and engineered cells, emphasizing how these systems impact key delivery metrics such as retention, dose control, shear resilience, spatial targeting, and immune interfaces to advance precision and personalized regenerative medicine.

Ovarian aging, which progresses faster than overall organismal aging, represents a major public health challenge, profoundly impacting female reproductive health and accelerating societal aging. The traditional Chinese medicine formula Zuogui Pill (ZGP) has shown great potential in delaying ovarian aging, warranting further investigation and development.

Dormant cancer stem cells (CSCs) are the root cause of the drug resistance and metastatic processes of malignant tumors, but an in-depth analysis of their biological mechanisms is needed. Dormant CSCs are in the G0 phase of the cell cycle and are characterized by enhanced autophagic activity, a stable genomic structure and strong plasticity. Recently, several new specific markers of dormant CSCs, such as p27, CD13, QSOX1, Survivin, GPD1 and BEX2, have been identified, which offer hope for targeted therapy. In addition, epigenetic modifications such as DNA methylation and histone modifications have been reported to regulate the transition between the quiescent and proliferative states of dormant CSCs. From a clinical perspective, keeping cancer stem cells in a dormant state is helpful for preventing tumor recurrence and metastasis. To this end, clarifying the potential mechanisms and molecular regulation of cancer stem cell dormancy is vital. Here, in this review, we examine recent significant findings regarding tumor stem cell dormancy in both experimental and human disease models, emphasizing the underlying molecular mechanisms, regulatory processes, experimental models, and prospective research directions aimed at advancing this field and enhancing clinical translation.

Homologous recombination (HR) is crucial for maintaining genomic integrity and is tightly regulated, yet the role of ubiquitin-dependent degradation in HR proteins remains poorly understood. Through high-throughput screening for compounds that modulate the DNA replication stress response, we identify ML367 and its derivative, UNI418. Kinase profiling and detail molecular analyses reveal that UNI418 inhibits PIKfyve and PIP5K1C, reducing inositol hexaphosphate (IP6) levels and triggering Cul4A-dependent degradation of RAD51, CtIP, and CHK1. Further analysis identifies WDR5 as a DCAF protein that facilitates Cul4A-mediated proteolysis of RAD51 and CHK1. Functionally, UNI418 suppresses HR, enhances tumor sensitivity to PARP inhibitors (PARPis), and re-sensitizes PARPi-resistant tumor cells in both in vitro and in vivo xenograft models. These findings reveal a Cul4A-WDR5-dependent proteolysis pathway regulating HR protein stability via phosphatidyl inositol signaling. This mechanism offers a promising therapeutic strategy for overcoming PARPi resistance and improving combinatorial cancer treatment strategies.

T-cell acute lymphoblastic leukemia (T-ALL) is an aggressive malignancy of T-cell precursors. Although the survival rates have improved with the use of intensive chemotherapy, the emergence of relapse as well as treatment-related morbidity and mortality remain major challenges. Novel treatment approaches include the inhibition of anti-apoptotic regulators or cellular immunotherapies. Here, we analyzed the sensitivity of T-ALL to inhibitors of BCL-2 (venetoclax), BCL-XL (A1331852), MCL-1 (AZD5991) and dual inhibition of BCL-2/BCL-XL (AZD4320) and evaluated their combination effects with natural killer (NK) cells. While only early T-cell precursor (ETP) ALL was sensitive to BCL-2 inhibition, MCL-1 inhibition alone was not effective in most cell lines and patient-derived xenograft (PDX) samples. For BCL-XL and dual BCL-2/BCL-XL inhibition, we observed heterogeneous sensitivities, which were associated with anti-apoptotic dependencies on the respective BCL-2 family members as assessed by BH3-profiling. Moreover, we identified functional shifts in anti-apoptotic dependencies upon exposure to AZD4320 or AZD5991 alone and synergistic effects when both inhibitors were combined with each other, allowing cell death induction in resistant samples. We then explored the potential use of apoptosis-inducing drugs as sensitizers for immunotherapy. Therefore, we investigated the potential of NK cell-mediated killing in T-ALL and found heterogeneous sensitivity, with some cell lines showing responses even at low effector-to-target (E:T) ratios. Importantly, NK cell-mediated killing could be further enhanced by combining NK cells with AZD4320, proposing this combination as a potential effective treatment. Taken together, we demonstrated promising potential of BH3-mimetics and NK cells for the treatment of T-ALL alone and in combination, warranting further preclinical and potential clinical evaluation.

Microplastics (MPs) are emerging pollutants that are associated with many diseases including atherosclerosis, inflammatory bowel disease (IBD), and Alzheimer's. The oral cavity is the primary point for the uptake of MPs by human, where MPs could pose risks to oral and even system health. Various polymer-based materials have been used as dental materials in oral treatment, however, the assessment of MPs in dental treatments remains limited and the processes and mechanisms by which MPs affect human health through the oral route are elusive. Here, we report the assessment of the risks and sources of MPs in dental clinics, the establishment of the relationship between MPs and oral inflammatory disorders, and also the elucidation of underlying mechanisms. Our results showed that commonly used therapeutic dental materials could generate MPs in dental clinics with proportions significantly higher than those in office areas and outdoors. As a representative, polymethyl methacrylate (PMMA) MPs showed significant toxicity to human oral keratinocytes (HOK), human periodontal ligament stem cells (hPDLCs), and THP-1-derived macrophages. Mechanistic investigations of apoptosis and inflammation processes revealed that MPs could lead to mitochondrial stress and autophagy and trigger the Notch signaling pathway and the JAK-STAT signaling pathway. Mice experiments showed that prolonged high-dose MPs exposure induced periodontal inflammatory reactions and even led to inflammatory bone resorption. This study provides a scientific basis for the oral health risks by MPs in dental practice and addresses the need for the development of dental materials with higher biocompatibility and environmentally sustainability.

This study investigates the therapeutic effects of ultrasound-guided transplantation of human umbilical cord mesenchymal stem cells (HUC-MSCs) on nerve injury and neuronal recovery in chronic constriction injury (CCI) rats. Behavioral analysis showed that HUC-MSCs treatment significantly improved Paw Mechanical Withdrawal Threshold (PMWT) and Thermal Paw Withdrawal Latency (TWL), with faster recovery in the HUC-MSCs groups. Histological analysis revealed enhanced repair and regeneration, reduced inflammation, and improved results in the dual-treatment group (CCI + U2). Ultrasound and histological examination demonstrated significant nerve fiber repair, reduced inflammation, and enhanced myelin recovery, especially in the CCI + U2 group. TMT-based proteomics identified differential protein expression in the spinal dorsal horn (SDH), confirming molecular changes associated with neuronal recovery. Analysis of microglial polarization in the SDH, and macrophage polarization in the dorsal root ganglion (DRG) and sciatic nerve (SN), showed that HUC-MSCs promoted the M2 phenotype, reducing inflammation and enhancing repair. HUC-MSCs also modulated the NLRP3 inflammasome, lowering NLRP3, ASC, caspase-1, and IL-1β levels. The study further explored HUC-MSCs-derived exosomes' effects on microglial polarization and NLRP3 activity, regulated by miR-223-3p to promote anti-inflammatory and neuroprotective effects, suggesting the potential of HUC-MSCs and exosomes in nerve regeneration and neuroinflammation reduction after SN injury.

Ameloblastoma is a benign odontogenic tumor with an aggressive growth phenotype orchestrated by a complex and heterogeneous tumor microenvironment. This review addresses how tumor cells, cancer-associated fibroblasts, mesenchymal stem cells, endothelial cells, and immune cells interact with non-cellular elements especially the extracellular matrix and hypoxic niches to drive invasive growth and recurrence. Several genetic changes associated with ameloblastoma activate mitogen-activated protein kinase (MAPK), Hedgehog (HH), Wnt/β-catenin, and less commonly PI3K/AKT signaling pathways. These pathways increase matrix-degrading enzymes such as matrix metalloproteinases and heparanase and reorganize collagen to create paths for local spread of ameloblastoma cells. Hypoxic niches in ameloblastoma stabilize hypoxia-inducible factor (HIF-1)α and activate vascular endothelial growth factor (VEGF) thereby linking low oxygen tension to new blood vessel growth within the microenvironment. Crosstalk between ameloblastoma epithelium and stroma through interleukin-6, transforming growth factor (TGF)-β, and connective tissue growth factor (CTGF) activates a positive feedback loops that stiffen the extracellular matrix and promote collective invasion. Within the encompassing jaw bone, a higher receptor activator of nuclear factor kappa-Β ligand/osteoprotegerin (RANKL/OPG) ratio and parathyroid hormone-related protein (PTHrP) level stimulate osteoclastogenesis, which accounts for the characteristic osteolysis displayed by ameloblastoma. Additionally, PD-L1 expression in ameloblastoma weakens T-cell activity in spite of the high population of M1 macrophages at the tumor leading edge. Collectively, coordinated interplay of these molecular processes define the invasive and aggressive growth phenotypes of ameloblastoma. Opportunities abound for development of targeted therapies for management of ameloblastoma. Potential candidates are inhibitors of BRAF/MEK and smoothened (SMO) gene/HH pathways, interruption of the TGF-β-Cancer-associated fibroblast axis, anti-angiogenic strategies, immune checkpoint blockade, and RANKL-directed therapy.

Background and objectives Atherosclerosis is a chronic disease marked by the build up of lipids and inflammatory cells in arterial walls, leading to vessel narrowing and increasing the risk of serious complications like heart attack and stroke. Recent findings suggest that microRNAs (miRNAs) serve as key regulators in the mechanisms driving atherosclerotic disease. However, the expression levels and functional roles of miRNA-133a-3p and miRNA-124-3p in atherosclerosis remain incompletely understood. The aim of this study was to determine the relationship between the expression levels of miR-124-3p and miR-133a-3p, and the phenotypic changes of S100A4-positive vascular smooth muscle cells in atherosclerosis. Methods We collected tissue samples from 25 patients with atherosclerosis who underwent coronary artery bypass graft surgery. IMA tissues were used as controls; atherosclerotic aortic tissues as cases. Expression levels of miRNAs were assessed using reverse transcription polymerase chain reaction (RT-PCR). Tissue samples underwent immunohistochemical staining with S100A4 protein to evaluate cellular and structural characteristics. Results A marked decrease in the expression of miR-133a-3p and miR-124-3p was observed in the atherosclerosis group compared to the control group, and both differences were statistically significant (P=0). Additionally, an increase in S100A4 protein immunoreactivity was detected in the atherosclerosis group. Interpretations and conclusions The downregulation of miRNA-133a-3p and miRNA-124-3p in atherosclerotic tissues, along with the observed increase in S100A4 protein immunoreactivity, suggests that these two miRNAs may play a role in the regulation of inflammatory endothelial phenotypes. Therefore, the interaction between miRNA-133a-3p, miRNA-124-3p, and S100A4 protein may help elucidate a potential mechanism underlying the prevention of atherosclerosis.

Kidney size is sex-dimorphic and regulated by androgens in adult humans and mice. However, the effects of developmental androgen deficiency on kidneys remain elusive. We hypothesized that androgens program future kidney growth during fetal development. Male mice lacking the main testosterone-producing enzyme HSD17B3 had reduced testosterone at embryonic day 15.5, but the concentrations increased by E18.5, creating a short time window of androgen deficiency resulting in reduced kidney size in adult males. In male Hsd17b3-/- kidneys, nephron development was qualitatively normal, but the number of glomeruli and proliferation of proximal tubules were reduced, as was proximal tubule size in adults. Testosterone supplementation at E14.5-17.5 normalized the renal size in adult Hsd17b3-/- males. Our data suggest that androgen receptor and HNF4A jointly regulate IGFBP5, putatively influencing FOXO1 and mTOR signaling to promote male-specific kidney growth in the fetal period. In conclusion, we have identified a novel developmental programming effect on male kidneys, where fetal androgen deficiency reduces kidney growth and androgen responsiveness in adult males.

Mesenchymal stem cells (MSC) represent the main stromal component of the bone marrow (BM) niche and are crucial to maintain hematopoietic tissue homeostasis. MSC exhibits extraordinary and multiple properties. In terms of expanding potential and differentiation capacity, Wharton jelly MSC (WJ-MSC), derived from the umbilical cord, was described as being greater and more performing than MSC from BM or other sources. WJ-MSC mimics the hematopoietic niche and supports hematopoietic stem cells (HSC) expansion ex vivo. This study aimed to evaluate the effects of human WJ-MSC cocultured with HSC in a B-cell differentiation protocol. Remarkably, results highlight WJ-MSC use as a preferable feeder layer to efficiently support HSC commitment toward the B-lineage. Over 11 days of HSC coculture with WJ-MSC, B-cell genes (E2A, RAG1, RAG2, etc.) expression patterns and B-cell markers (CD19, immunoglobulin chain, etc.) acquisition were evidenced. WJ-MSc were also able to unlock the B-lineage differentiation blockade of the acute lymphoblastic leukemia cell line Nalm16. This model might provide a new strategy to support ex vivo B-cell differentiation using the powerful properties of WJ-MSC. This study implements a new approach to improve understanding of B-leukemogenesis and B-cell acute lymphoblastic leukemia (B-ALL) pathophysiology.

The objective of the study was to compare the role of Cryopreserved human amniotic membrane (cHAM) for early pulp biomineralisation in comparison to Biodentine using the tooth culture model. This study was conducted in two stages. The first stage aimed to establish the tooth culture model and assess the viability of pulp cells at 28 and 50 days. In the second stage, pulp capping was performed in the tooth culture model using cHAM and Biodentine. A total of 20 freshly extracted teeth (for orthodontic reasons) were used for pulp capping with either cHAM or Biodentine. The samples were evaluated for biomineralisation at 14 and 28 days using micro-computed tomography (micro-CT) and histological analysis. Morphometric data from micro-CT scans were analysed using the Chi-square test. The viability of pulp cells in the tooth culture model was 66.1% at 28 days and 38.4% at 50 days. Pulp capping with cHAM resulted in a thicker and larger volume of mineralisation at 28 days (720 μm, 0.7 mm3) compared to Biodentine (630 μm, 0.46 mm3). Histological analysis revealed that the dentine formed in the HAM group was homogeneous and continuous, while the Biodentine group showed discontinuous foci of mineralisation in the newly formed hard tissue. Within the study's limitations, it is observed that cHAM induces enhanced early pulpal mineralisation compared to Biodentine.

Human embryonic stem cell (hESC) can be differentiated into definitive endoderm (DE) through multiple branching lineage choices. Although different DE differentiation methods have been established, there are still several limitations, such as the yield of heterogeneous cell populations containing undifferentiated or non-DE cells. Therefore, this study aimed to suppress the alternate fates at branch points and establish a robust and highly efficient differentiation protocol for hESC-derived hepatocytes (hESC-Heps). We developed a two-step DE induction protocol. First, hESCs were treated with a GSK-3α/β inhibitor and an mTOR inhibitor combined with TGF-β activation to generate an anterior primitive streak (progenitor to endoderm). Subsequently, a BMP inhibitor combined with TGF-β activation was used to abolish the mesoderm lineage. The resulting DE cells were further differentiated into hESC-Heps to evaluate their functionality. By regulating the branching lineage choices, we established an efficient two-step method that yielded up to 96% DE cells with minimal expression of pluripotency and mesodermal markers. Notably, this method reduced the dosage of Activin A, which makes it cost-effective for future applications. The derived hESC-Heps exhibited mature hepatocyte characteristics, including glycogen storage, indocyanine green uptake, and cytochrome P450 activity. Additionally, these cells demonstrated robust liver-specific functions such as sensitive innate immune responses and permissiveness to hepatitis B virus infection. In summary, we developed a novel and cost-effective method that achieves high-purity DE by precisely modulating cell fate decisions in the early stages. The derived hESC-Heps can serve as a model for further studies, such as host-virus interaction and hepatotoxicity testing.