The Jiangquan black pigs, a new breed of swine obtained by introducing traits from Duroc pigs into Yimeng black pigs, exhibits fast growth rates and high meat quality. To understand how daily weight gain influences muscle development in this breed, we analyzed longissimus dorsi muscle cell subpopulations from Jiangquan black pigs using snRNA and bulk RNA sequencing. Thirteen distinct cell types (e.g., muscle stem cells, satellite cells, fibroblasts) were identified, and marker genes (PAX7, MYOD, MYOG) were found to exhibit stage-specific expression during differentiation. Pseudotime analysis revealed the differentiation trajectories of these cell populations, while cell cycle analysis uncovered the higher mitotic activity in satellite cells of the fast-growth versus slow-growth groups. Furthermore, cell communication analysis highlighted the interactions between muscle cells and other cell types. Finally, intergroup analysis revealed that 2,466 and 2,597 genes were differentially expressed in muscle stem cells and muscle satellite cells, respectively. These genes were enriched in disease-related pathways. This study provides a single-cell resolution atlas of porcine muscle development, offering insights into the genetic regulation of growth and potential targets for breeding optimization.

Mechanical stimulation plays a crucial role in odontoblast differentiation. However, the underlying molecular mechanisms remain unclear. We have previously shown that hydrostatic pressure (HP) applied to stem cells from human exfoliated deciduous teeth (SHED) promotes odontoblast differentiation by translocating RUNX2 and increasing WNT16 expression through PIEZO1 signaling. In this study, we further explored the downstream signaling cascade linking PIEZO1 activation and odontoblast differentiation. HP stimulation increased the expression of odontoblast differentiation markers PANX3 and DSPP, as shown by qPCR, and enhanced Alizarin Red staining-results significantly suppressed by siRNA targeting either PIEZO1 or WNT16. RT-PCR analysis revealed that, among the two known human WNT16 isoforms, only WNT16b was expressed in SHED. qPCR demonstrated that HP-induced WNT16 expression was reduced by siPIEZO1 and further decreased by siRUNX2. Promoter analysis identified four RUNX2-binding sites within the upstream region of WNT16. A luciferase reporter assay using plasmids containing the WNT16 promoter showed that RUNX2 overexpression in HEK293 cells significantly increased luciferase activity. Moreover, HaloChIP assays with a HaloTag-RUNX2 expression vector confirmed RUNX2's binding to the WNT16 promoter. These findings suggest that PIEZO1-mediated mechanical stress promotes odontoblast differentiation through the RUNX2-dependent transcriptional activation of WNT16.

Inflammatory bowel disease (IBD) is secondary to an abnormal immune response to the microbiota. To study this, models of host-microbe interactions that represent mucosal bacterial communities and inter-patient diversity are required. Human intestinal organoids (HIOs) are an established model to investigate epithelial responses. Here, we describe a technique of culturing bacteria directly from the sites of inflammation in IBD, while simultaneously sampling host tissue. We generated HIOs from a cohort of newly diagnosed paediatric IBD patients, without confounding treatments or comorbidities, and explored their response to site-specific bacteria. A unique biobank of matched HIOs and cultured mucosa-attached bacteria was established from 27 paediatric patients. Transcriptional profiling revealed differential gene expression between control and IBD-derived organoids. We used microinjection to introduce bacteria to the apical surface of the epithelium, to determine the effect of bacteria on host epithelial cells. We measured survival and growth of bacteria within the HIOs and tested several related bacterial isolates for their impact on the epithelium. An isolate from a control patient stimulated inflammatory signalling pathways but this was not observed in response to a closely related isolate originating from an IBD patient. This study demonstrates the feasibility of isolating bacteria and generating organoids from the same biopsy tissue, to explore personalised host-microbe interactions. The microinjections, while labour-intensive, demonstrate that closely related bacteria can induce very different epithelial responses, with downstream implications for immune response. This highlights the importance of understanding host-microbe interactions in a strain- and site-specific manner and developing techniques for personalised microbiome-based therapeutics.

Keratinocytes are increasingly recognized as central regulators of cutaneous immune responses and key contributors to maintaining immune homeostasis. However, whether and how epidermal stem and progenitor cells (EPSCs) actively suppress proinflammatory signaling pathways to prevent excessive inflammation and maintain epidermal immune quiescence remains unclear. Here, we generated a conditional knockout mouse model (K14-CreERT; Supt6fl/fl) to investigate the role of SPT6, a transcription elongation factor, in epidermal and immune homeostasis. Loss of SPT6 in basal keratinocytes led to spontaneous, psoriasis-like skin inflammation, characterized by epidermal hyperplasia, immune cell infiltration, parakeratosis, and hyperkeratosis. SPT6-deficient mice also exhibited significantly delayed wound healing accompanied by impaired Wnt signaling. Moreover, single-cell RNA sequencing revealed distinct keratinocyte subpopulations with inflammatory signatures, elevated NF-κB signaling, and suppressed Wnt signaling. Mechanistically, SPT6 suppresses NF-κB signaling by binding to an enhancer of the RELA gene and preventing its positive transcriptional feedback loop. These findings support a new paradigm in which the default state of the skin may be primed for inflammation, and active suppression by factors such as SPT6 is required to maintain epidermal homeostasis. Taken together, the results of our study reveal a previously unrecognized role for SPT6 as a key regulator of epidermal immune quiescence and tissue integrity.

Selective elimination of suboptimal cells is critical for the developmental integrity of early mammalian embryogenesis. Cell competition is a non-autonomous quality control in which 'winner' cells outcompete viable but suboptimal 'loser' cells based on fitness differences. Here we investigate cell competition dynamics using mosaic mouse gastruloids, a 3D embryonic stem cell-based model of gastrulation. Introducing just two Trp53-deficient supercompetitor cells suffices to impair growth in neighbouring wild-type cells through mitochondrial apoptosis. Competition is tightly restricted to a developmental transition stage between primed pluripotency and early gastrulation and involves gene regulatory networks of pluripotency exit. Heterochronic gastruloids from developmental stage-shifted cells, EpiGastruloids, and dynamic p53-degrons reveal that both winners and losers must reside within this permissive stage, during which acute relative p53 protein levels determine competitive outcomes. These findings advance our understanding of cell fitness evaluation and establish gastruloids as a powerful 3D model for investigating developmental stage-specific cell competition in mammalian embryogenesis.

Bladder cancer is a common urogenital malignancy that remains difficult to treat, particularly due to therapeutic resistance, such as resistance to cisplatin, in which cancer stem cells (CSCs) play a central role. This study investigates the combination of 5-aminolevulinic acid-mediated photodynamic therapy (5-ALA-PDT) and berbamine as a potential multimodal treatment strategy using the bladder cancer cell lines RT112 and J82, their cisplatin-resistant variants, and generated CSC-like cells. Berbamine is a natural plant compound and was confirmed in this study to have anticancer properties by inhibiting cell migration and invasion, and by inducing apoptosis. This study also showed that berbamine enhances the accumulation of protoporphyrin IX (PpIX), the photosensitizer induced by 5-ALA. 5-ALA-PDT destroys cancer cells by stimulating PpIX via 635 nm red laser light to produce reactive oxygen species (ROS). This was found to happen in all tested cell lines, whereas berbamine could modulate the cell destruction in a concentration-dependent manner and was influenced by the specific biological characteristics of the tested cell variants. CSCs showed the strongest response to the combination therapy approach, suggesting that they may represent more vulnerable cell variants to the tested treatment. Cisplatin-resistant cell lines could also be treated successfully with 5-ALA-PDT, whereas berbamine could enhance its efficacy in the cisplatin-resistant J82 LTT. These findings suggest that the combination treatment of 5-ALA-PDT and berbamine may serve as a promising approach to overcome therapeutic resistance in bladder cancer, particularly in cisplatin-resistant and CSC-enriched tumour types.

Gastrointestinal dysfunction often precedes motor symptoms in Parkinson's disease (PD), suggesting the enteric nervous system (ENS) is central to early pathogenesis. How α-synuclein contributes to ENS dysfunction, and how inflammation modulates this, remains unclear. Here we show that Tumor Necrosis Factor alpha enhances α-synuclein accumulation in induced pluripotent stem cell-derived enteric neurons and glia, and impairs the malate-aspartate shuttle, a key pathway for mitochondrial energy production. This drives a metabolic shift toward glutamine oxidation in patient cells. This metabolic impairment reduces overall mitochondrial function, which is partially rescued by the neuroprotective compound Chicago-Sky-Blue 6B. Furthermore, transcriptomic and histological analyses of human gut tissue from inflammatory bowel disease patients reveal that inflammation-associated metabolic suppression and α-synuclein upregulation occur beyond PD, representing general hallmarks of intestinal inflammation. These findings highlight a conserved metabolic vulnerability in the ENS and establish patient-derived enteric lineages as a robust platform to model inflammatory ENS pathology.

Autism spectrum disorder (ASD) is a complex neurodevelopmental condition with limited treatment options, where neuroinflammation and gut microbiota dysbiosis are emerging as interconnected therapeutic targets. This study evaluated the therapeutic potential of non-gene-edited human chemically induced pluripotent stem cell-derived neural stem cells (hCiPSC-NSCs) in a prenatal valproic acid (VPA)-induced rat model of ASD, using a dual-pathway administration strategy (intravenous systemic delivery combined with an intracerebroventricular boost). The treatment significantly ameliorated core ASD-like behaviors, including improved sociability (increased stranger interaction time, P < 0.0001), reduced repetitive behaviors (decreased marble-burying, P < 0.0001; and self-grooming, P < 0.05), and enhanced spatial memory (shorter escape latency in the Morris water maze, P < 0.01). At the mechanistic level, hCiPSC-NSCs attenuated neuroinflammation (suppressed IL-1β, IL-6, and TNF-α; elevated IL-10, all P < 0.0001), reduced oxidative stress (restored GSH and SOD, decreased MDA and NO), diminished microglial activation in the hippocampus and cortex, and restored synaptic ultrastructure by replenishing synaptic vesicles. Furthermore, 16S rRNA sequencing revealed a rebalancing of the gut microbiota, characterized by a reduced Firmicutes/Bacteroidota ratio, enrichment of beneficial taxa like Bacteroidota and Alloprevotella, suppression of pathobionts such as Desulfovibrionales, and partial restoration of microbial diversity. These findings demonstrate that non-gene-edited hCiPSC-NSCs can simultaneously address neural pathophysiology and gut ecosystem disruption in ASD, highlighting their potential as a gut-brain axis-targeting therapy for neurodevelopmental disorders.

Acute myeloid leukemia (AML) remains a highly heterogeneous hematologic malignancy in which therapeutic resistance and disease relapse are largely driven by leukemic stem cells (LSCs). These rare, self-renewing cells possess unique biological properties that enable them to survive conventional chemotherapy and targeted therapies, thereby sustaining minimal residual disease and promoting leukemia re-emergence. LSC persistence arises from a complex and multilayered network of resistance mechanisms, including intrinsic cellular programs, adaptive molecular plasticity, and protective interactions within the bone marrow microenvironment. Intrinsic mechanisms include cellular quiescence, enhanced multidrug efflux activity, resistance to apoptosis and senescence, and activation of stress-adaptive pathways such as autophagy. In addition, LSCs exhibit remarkable metabolic and epigenetic flexibility, allowing them to rewire signaling pathways and survive therapeutic pressure. Extrinsic cues from the bone marrow niche, including stromal interactions, cytokine signaling, and metabolic support, further reinforce the survival and drug tolerance of LSCs. Together, these interconnected mechanisms create a highly resilient cellular state that limits the efficacy of current therapies. In this review, we summarize the major biological pathways that sustain LSC-mediated resistance in AML and discuss emerging therapeutic strategies aimed at selectively targeting these cells. A deeper understanding of LSC biology will be critical for the development of combination therapies capable of eradicating minimal residual disease and achieving durable remission in patients with AML.

Androgenetic alopecia (AGA) is a common hair disorder in which limited follicular drug delivery and an inflammatory and oxidative follicular microenvironment reduce topical efficacy. Herein, we developed a fast-dissolving microneedle (MN) patch of chondroitin sulfate and carboxymethyl chitosan for localized codelivery of Platycladus orientalis leaf-derived extracellular vesicles (PO-EVs) and minoxidil nanoparticles (MXD NPs). PO-EVs were separated and characterized as nanoscale vesicles and were shown to possess antioxidant, anti-inflammatory, and pro-angiogenic activities relevant to hair follicle maintenance. MXD NPs were prepared by thin-film hydration to improve the minoxidil solubility and local retention. Both were loaded into microneedles with sufficient mechanical strength that could dissolve rapidly in the skin. In a mouse model of androgenic alopecia, repeated dual-loaded MN treatment accelerated the telogen-to-anagen transition, increased hair-covered area and shaft thickness, and restored follicular morphology. Mechanistic studies showed that hair follicle stem cells were activated and proliferated, perifollicular oxidative stress and inflammation were reduced, and microvessel density around hair follicles was increased. No evident skin irritation or systemic toxicity was observed. This MN codelivery strategy improves hair regrowth by combining efficient minoxidil delivery with PO-EV-mediated microenvironment restoration and may be extended to other inflammatory/oxidative skin disorders impairing regeneration.

Habitat destruction, changing climate and other anthropogenic impacts have resulted in the recorded extinctions of hundreds of species, with many more undocumented extinctions being likely to have occurred. Approaches to conserving threatened species include protection or improvement of habitat, fenced conservation reserves, species translocations and reintroductions, elimination of environmental toxins, breeding programs in reserves or captivity, and genetic rescue and management. The latter includes storage of gametes, stem cells or embryos, to both conserve species and maintain or expand their genetic diversity. Many of these approaches require a basic knowledge of the reproductive biology of the species of interest. Such knowledge is difficult to achieve because of the astonishing diversity of species-specific reproductive strategies that have evolved. Unfortunately, for many species we simply do not have that knowledge. This report summarises key discussions from a workshop titled Reproductive Biology Research Needed for Saving our Wildlife held in Melbourne, Australia, and attended by stakeholders from zoos, wildlife organisations, universities, museums and government organisations. The workshop prioritised aspects of reproductive biology knowledge needed, how this knowledge might be obtained, and how it should be deployed. Using examples of planned and successful conservation strategies for individual species, the workshop participants considered environmental challenges, managing introduced species, captive breeding programs, challenges for assisted reproductive technologies, de-extinction science in conservation efforts, examination of reproductive steroid hormones across species, endocrine disruption, and cryopreservation of genomic diversity to assist the management of wild and captive populations. The workshop highlighted the magnitude of the issues involved and identified reproductive approaches to be used to direct future conservation efforts for saving threatened species.

The treatment of critical-size bone defect, particularly those with irregular shapes, presents a significant clinical challenge. Chitosan hydrogels, owing to their degradability, excellent biocompatibility and injectability, represent a promising carrier platform for bone tissue engineering (BTE). Bioavailable Mg2+ serves as a pivotal element in bone regeneration, promoting osteogenesis and angiogenesis to accelerate bone tissue repair. Furthermore, Gallic acid (GA) exhibits anti-inflammatory properties, modulating the immune microenvironment to create favorable conditions for bone regeneration. To control its release behavior, metal-organic frameworks (MOFs) may be constructed by leveraging the coordination interactions between Mg2+ and GA. This study constructed an injectable thermo-responsive hydrogel using chitosan and sericin as the matrix, functionalizing it by embedding Mg-GA MOF at varying loading capacities. This material system is termed CS-MOF. In vitro experiments confirmed that the Mg-GA MOF-functionalized hydrogel exerts multiple biological functions: promoting osteogenic differentiation of rat bone marrow mesenchymal stem cells (rBMSCs), inducing angiogenesis in human umbilical vein endothelial cells (HUVECs), and suppressing the inflammatory response of LPS-stimulated RAW264.7 macrophages. Using a rat critical-sized cranial defect model, we demonstrated that the 0.01% wt/vol Mg-GA MOF-functionalized hydrogel significantly enhanced new bone formation and angiogenesis compared to the MOF-free and 0.02% wt/vol MOF-loaded hydrogels. Collectively, this thermo-responsive Mg-GA MOF-functionalized hydrogel represents a promising candidate for clinical translation in bone defect repair.

Injectable biomaterials have emerged as a promising option for minimally invasive tissue repair, especially in cases with small and irregular lesions. In combination with electroconductivity and self-healing abilities, these materials provide unique benefits for neural tissue engineering. They promote adaptive defect filling, assist in post-injection structural repair, and improve bioelectrical signaling. The current study focuses on the synthesis and characterization of injectable, self-healing, electroconducting hydrogels, which are intended to serve as a potential platform for addressing small and irregular neural defects. Dynamic covalent hydrazone coupling between oxidized methacrylate sodium alginate-polypyrrole (OMA-PPy) and adipic acid dihydrazide-functionalized hyaluronic acid (HA-ADH) was employed to produce hydrogels. The hydrogels were subsequently reinforced with additional photopolymerization to ensure post-injection stability. Additionally, a laminin-derived peptide called IKVAV was covalently linked to generate biomimetic signals for enhancing neuronal cell adherence and differentiation. The hydrogels demonstrated adjustable mechanical properties (elastic modulus: 72.91 ± 10.84-103.94 ± 17.72 Pa; and compressive strength: 76.87 ± 25.06-153.60 ± 37.89 kPa) and pore size (44.86 ± 23.18-76.34 ± 59.28 μm), exceptional shear thinning behavior, and self-healing abilities (recovery up to 90.35% after six cycles), along with adequate electroconductive performance suitable for minimally invasive procedures and in-situ regeneration for neural applications. In vitro studies have shown that IKVAV-functionalized hydrogels enhance the adhesion, proliferation, and differentiation of neural stem cells (NSCs) with Tuj1 expression of 6.79 ± 0.71 in the OMA-PPy/HA-ADH/IKVAV group. In addition, the OMA-PPy/HA-ADH/IKVAV formulation demonstrated favorable biocompatibility in vivo (ISO 10993-6 reactivity score: 1.22), along with a well-regulated biodegradation profile (91.6% degradation by day 21), when tested in subcutaneous implantation models. The findings indicate that optimizing a single material characteristic is insufficient; achieving a balance among bioactive signaling, secondary network connections, and dynamic covalent bonding is essential for identifying an ideal candidate material. This versatile hydrogel platform offers an encouraging approach for future research focusing on the repair and regeneration of nerve tissues, particularly in addressing localized and small-sized nerve injuries.

Tooth extraction is a common dental procedure often accompanied by local tissue damage and alveolar bone resorption. The integration of hemostatic and regenerative materials can significantly enhance therapeutic efficacy and patient outcomes following tooth extraction. Herein, we developed dual-functional polyhedral oligomeric silsesquioxane (POSS)-functionalized carboxymethyl chitin microspheres (CMGP) via electron beam irradiation grafting. Both in vitro and in vivo studies demonstrated that CMGP possessed good biocompatibility and biodegradability. These microspheres promoted blood coagulation through robust platelet activation capacity. Notably, in a rat artery puncture model, clotting time was reduced from 555 s to 55 s, while blood loss decreased from 617 mg to 21 mg. Moreover, CMGP microspheres significantly induced M2 polarization of macrophages, demonstrating potent tissue repair-promoting properties. Furthermore, the microspheres accelerated osteogenic differentiation of mesenchymal stem cells, which is critical for bone regeneration. Therefore, POSS-functionalized radiation-grafted carboxymethyl chitin microspheres CMGP represent a promising biomaterial platform for post-extraction socket management, enabling enhanced hemostasis and tissue repair.

Mechanisms that maintain genome integrity are crucial for coordinating transcription that drives mammalian forebrain development. Neural progenitor cells and differentiating neurons in the developing forebrain sustain high transcriptional activity and metabolic demand and are therefore vulnerable to DNA damage. Transcription-coupled nucleotide excision repair is a specialized DNA repair pathway that helps to mitigate damage induced by 'bulky' adducts such as UV-induced pyrimidine dimers and monoadducts formed by reactive oxygen species. TC-NER factors, which include CSB/CSA, UVSSA-USP7, ELOF1, and STK19, coordinate and assemble the complex to initiate repair. Notably, TC-NER safeguards genome integrity and plays an essential role in neuronal differentiation, synaptogenesis, and neurogenesis. Impaired TC-NER pathway activity manifests in tissue-level pathologies, including neurodegeneration and increased susceptibility to neurological deficits. This is relevant to neurodevelopmental disorders that stem from TC-NER deficiency, such as Cockayne syndrome, Trichothiodystrophy, and Cerebro-Oculo-Facio-Skeletal syndrome. Although deficits in TC-NER have been well established as a contributor to a variety of neurodegenerative disorders, its roles in the developing forebrain across various cell types and neurodevelopmental windows remain poorly defined. In this review, we highlight recent studies investigating mechanisms linking TC-NER deficiency to forebrain developmental phenotypes and summarize knowledge gaps in the field regarding cell-type specificity, regional vulnerability, and therapeutic windows for intervention.

Intracerebral hemorrhage (ICH) is a devastating acute neurological condition with high mortality and disability. Induced pluripotent stem cell-derived neural progenitor cells (iPSC-NPCs) have been shown to promote behavioral recovery by enhancing neural connectivity and providing trophic support. As the adenosine monophosphate-activated protein kinase (AMPK)/ mammalian target of rapamycin (mTOR) signaling pathway is a key regulator of autophagy in stroke, we investigated its role in the context of iPSC-NPC transplantation for ICH.

The potential anti-obesity, anti-inflammatory, and anti-oxidative stress properties of ark shell-derived LLRLTDL (Bu1) and GYALPCDCL (Bu2) peptides were comprehensively investigated. In bone marrow-derived mesenchymal stem cells (BMMSCs), both peptides demonstrated significant anti-adipogenic effects by downregulating key adipogenic transcription factors, including peroxisome proliferator-activated receptor gamma (PPAR-γ), CCAAT/enhancer-binding protein alpha (C/EBPα), and sterol regulatory element-binding protein 1 (SREBP-1) and their downstream adipocyte-specific genes including adipocyte fatty acid-binding protein 2 (aP2), fatty acid synthase (FAS), and lipoprotein lipase (LPL). Mechanistically, Bu1 and Bu2 promoted lipolysis through the activation of AMP-activated protein kinase (AMPK) and hormone-sensitive lipase (HSL). These peptides also exhibited potent anti-oxidative stress activity by suppressing reactive oxygen species generation and activating the HO-1/Nrf2 signaling pathway, as confirmed through HO-1 siRNA silencing. In addition, Bu1 and Bu2 demonstrated robust anti-inflammatory effects by reducing pro-inflammatory cytokine production and inhibiting MAPK signaling pathways. These findings were corroborated in a high-fat diet (HFD)-induced mouse model, where oral administration of Bu1 and Bu2 resulted in significant reductions in body weight, weight gain, and adipose tissue accumulation, along with decreased expression of adipogenic transcription factors and genes while improving serum cholesterol levels, and exhibited anti-oxidative stress effects via HO-1/Nrf2 activation. Collectively, these results underline the potential of Bu1 and Bu2 as multi-target therapeutic agents against obesity and related metabolic disorders.

Myocardial ischemia/reperfusion (I/R) injury is an unsolved medical issue that is caused by additional injuries derived from reperfusion therapy for patients with acute myocardial infarction, one of the leading causes of morbidity and mortality in the world. Myocardial I/R injury causes unpredictable complications evoked by oxidative stress, endothelial dysfunction, dysregulated autophagy, apoptosis, and an imbalanced inflammatory response. Microvesicles (MVs) derived from adipose stem cells (ADSC) and egg-derived exosomes (EXOs) may confer antioxidant, anti-inflammatory, anti-apoptotic and tissue repair potential, showing potential in cardiovascular therapy. This study aims to investigate the therapeutic effects and mechanisms of MVs and EXOs on myocardial I/R injury.