Qihuang Biwen Formula (QHBWF), derived from the classic Yupingfeng Powder, is traditionally used for immune regulation. While clinically applied, its mechanism of action in ulcerative colitis (UC) remains unclear.

Intervertebral disc degeneration (IVDD) is a leading cause of low back and leg pain and is primarily driven by inflammation-induced apoptosis of nucleus pulposus (NP) cells. Bone marrow mesenchymal stem cell-derived exosomes (BMSC-exos) have shown strong therapeutic potential for degenerative diseases. Sirtuin 6 (SIRT6), a key regulator of aging and inflammation, has been closely associated with IVDD progression. Therefore, we hypothesized that BMSC-exos might alleviate IVDD by promoting SIRT6-mediated deacetylation of high mobility group box 1 (HMGB1), thereby suppressing inflammation.

Spinal cord injury (SCI) can result in irreversible neurological deficits, such as limb paralysis and dysfunctions of urination and defecation. Currently, there remains a dearth of effective therapeutic strategies to fully reverse the detrimental consequences of SCI. Stem cell transplantation, particularly adipose-derived stem cells (ASCs) transplantation, shows great promise in treating SCI. However, the low survival rate of transplanted cells and limited neural differentiation impede its long-term clinical effectiveness. In this study, platelet lysate-rich plasma (PLP) was utilized as the primary matrix component. Through sodium alginate cross-linking, a platelet lysate-rich plasma hydrogel (PLPH) with a micron-scale macroporous structure was fabricated. By precisely modulating the component ratios, PLPH was engineered to possess a low Young's modulus (approximately 1000 Pa), which is comparable to that of spinal cord tissue, along with characteristics such as a low swelling ratio, high porosity, and slow degradation. In vitro, PLPH facilitated the proliferation of ASCs and offered neuroprotection under oxidative stress conditions. Moreover, it promoted axonal growth in neuronal cells. In a mouse model of spinal cord injury, PLPH-loaded ASCs induced M2 polarization of microglia/macrophages at the injury site by activating the signal transducer and activator of transcription 6 (STAT6) pathway, thereby suppressing the inflammatory response. Additionally, PLPH promoted the survival and neural differentiation of loaded ASCs in vivo, and also recruited neural progenitor cells, induced early differentiation of neurons and oligodendrocytes, inhibited glial scar formation, promoted axonal and myelin regeneration, and ultimately enhanced neurological functional recovery. As a hydrogel material derived from natural biological sources, PLPH holds great potential in providing novel alternatives for clinical stem cell transplantation therapies.

Inflammation-associated alveolar bone defects, such as periodontitis, remain a major clinical challenge, as persistent inflammatory microenvironments severely impair bone regeneration and tissue repair. The cathepsin family plays a significant role in inflammatory responses and the clinicopathological process of periodontitis. Studies have shown that the cathepsin K inhibitor, odanacatib (ODN), can repair inflammatory bone defects. However, its administration via oral or local delivery fails to maintain sustained drug concentrations within the periodontal pocket. Here, we report a gel sustained-release system that coordinates immunomodulation with osteogenic induction to overcome inflammation-impaired bone regeneration. We utilized the emulsification-solvent evaporation method to prepare ODN-loaded poly(lactic-co-glycolic acid) (PLGA) microspheres (ODN-MS). These microspheres were thoroughly mixed with gelatin methacryloyl (GelMA) to form a gel composite sustained-release system (ODN-MS-Gel). The ODN-MS-Gel prepared has excellent photopolymerization properties and can achieve sustained release of ODN for a long time, thus enabling local and targeted drug delivery within the periodontal pocket. In vitro, ODN-MS-Gel not only promoted the proliferation, adhesion, and migration of BMSCs but also induced macrophage polarization toward the M2 phenotype, demonstrating a significant ability to promote osteogenic differentiation under inflammatory conditions. Furthermore, this study also distinguished the differences between the two situations of continuous release and non-continuous release and revealed the potential role of macrophage immune regulation in conjunction with stem cell osteogenic differentiation. In vivo, ODN-MS-Gel gradually degraded over time, exhibiting excellent long-term biocompatibility. By using a rat periodontitis model, it was demonstrated that ODN-MS-Gel could reduce inflammatory expression in periodontal tissues under inflammatory conditions and repair the alveolar bone defect to 77% of its normal height. In conclusion, ODN-MS-Gel achieves anti-inflammatory and bone repair effects with a single drug, representing a promising biomaterial for treating inflammatory alveolar bone defects.

Neuroendocrine tumors (NETs) secrete vasoactive hormones that promote hemodynamic instability. This study investigated whether NETs alter vascular responsiveness using a murine model. J:Nu mice received intrasplenic injections of BON-1 NET cells (BON-1, n = 21) or PBS (VEH, n = 21) and monitored for 10 weeks. Liver metastases were identified by histological analysis, chromogranin A expression, and presence of ALDH1-positive cancer stem cells. Vasomotor function of isolated mesenteric arteries was assessed to acetylcholine, serotonin, vasopressin, and endothelin-1, and with nitric oxide synthase inhibition (L-NAME), 5-HT receptor blockade, or serotonin incubation. BON-1 mice exhibited impaired vasodilation to acetylcholine and serotonin compared with VEH (p < 0.05), specifically in males (p < 0.001). L-NAME and 5-HT1b/d receptor blockade attenuated vasodilatory responses to serotonin only in VEH mice. Serotonin incubation reduced vasopressin-mediated vasoconstriction (p < 0.0001). These findings indicate that NET metastases are associated with impaired serotonin-mediated vasodilation, which is associated with reduced nitric oxide bioavailability, altered 5-HT1b/d receptor action, and sex-dependent impairment of endothelium-dependent vasodilation. Elevated serotonin levels may further compromise vasopressin-mediated vasoconstriction. The presence of NET metastases alters vascular responsiveness, which may contribute to hemodynamic instability.

The engineering three-dimensional (3D) cartilage tissue from mesenchymal stem cells is often obstructed by diffusion limitations, uncontrolled signal delivery, and hypertrophic differentiation following chondrogenesis. We herein report a 3D cartilaginous construct by encapsulation of spheroids of human umbilical cord-derived MSCs (hUCSCs) within in Gelatin methacryloyl hydrogels where the mineral-coated fibers (MFs) were integrated within the spheroid for localized ion delivery, thereby alleviating diffusion limitations. Through the intrinsic properties of hUCSCs, this system achieved robust chondrogenesis while minimizing hypertrophic progression. MFs led to a greater than threefold upregulation in chondrogenic gene expression and enhanced deposition of chondrogenic extracellular matrix in hUCSC spheroids, without concomitant increases in hypertrophic markers or matrix components. Comparative analysis revealed that hUCSCs exhibited superior chondrogenic potential and reduced hypertrophic gene expression relative to human bone marrow-derived MSCs. These findings highlight the potential of the MFs-incorporated composite spheroids-laden hydrogels as a novel biomimetic strategy for stable cartilage biofabrication, as they selectively promote hUCSC chondrogenic differentiation while mitigating hypertrophic maturation in a controlled 3D microenvironment.

Mesenchymal stem cells have great therapeutic potential due to their antiinflammatory, anti-apoptotic and pro-angiogenic effects through their secretome. Wharton’s gelatin is one of the best sources of mesenchymal stromal stem cells; their secretome is rich in bioactive molecules with the ability to maintain cellular and tissue homeostasis for the treatment of pathologies such as inflammation, skin wounds, neurodegenerative disorders, and diabetes.

Aging of the male reproductive system is characterized by declining fertility, with epididymal dysfunction being a critical yet poorly understood contributor. Through a multimodal analysis in non-human primates that integrated histology and transcriptomics, we delineated a coherent epididymal aging phenotype encompassing epithelial senescence, chronic inflammation, fibrosis, and functional decline. Single-nucleus transcriptomics revealed principal cells (PCs) as the predominant and most transcriptionally perturbed epithelial cell type. Within PCs, the longevity-associated transcription factor FOXO1 was markedly downregulated with age. Functional studies in human epididymal epithelial cells demonstrated that FOXO1 deficiency drives cellular senescence. Mechanistically, FOXO1 transcriptionally activates LHX1, and this axis is essential for counteracting senescence. Furthermore, intervention with senescence-resistant mesenchymal progenitor cells or their exosomes mitigated epididymal aging phenotypes and restored FOXO1 expression in vivo and in vitro. Our study establishes the FOXO1-LHX1 axis as a key protective pathway against primate epididymal aging, providing mechanistic insights and potential therapeutic targets for preserving male reproductive health.

Recombinant insulin, used to treat diabetes, has an enormous impact on diabetes management. However, this essential hormone is prone to aggregation during transportation and storage, reducing its efficacy in glycemic control. To prevent insulin aggregation, here we repurposed an antifungal drug called Nystatin (Nys) identified through a virtual screening of 5500 FDA-approved drugs. Nys effectively inhibited the aggregation of insulin under physiological conditions. Nys-treated insulin was completely soluble, retained its native secondary structure, and was nontoxic to HEK293T cells. Notably, this drug completely abrogated fibrillation of fast-acting insulin formulations Aspart and Lispro, as well as slow-acting insulin, Glargine. The ability of insulin formulations to activate the insulin receptor was maintained after Nys treatment. The efficacy of Nys-treated formulations in maintaining glycemic control was further validated in a diabetic rat model. Thus, this FDA-approved drug has the potential to be used in commercial insulin formulations to maintain stability and activity while preventing its aggregation for an extended period.

Photobiomodulation (PBM) is a non invasive technique that utilizes light to modulate cellular processes and promote tissue regeneration. In the context of regenerative therapies, PBM has emerged as a promising approach for enhancing the differentiation of adipose-derived stem cells into neurospheres. This study aimed to investigate the effects of PBM on neurosphere growth, mitochondrial function, and cellular differentiation, focusing on 825 and 525 nm wavelengths and at 5 and 10 J/cm2 fluences. Our results demonstrate that PBM modulates neurosphere size and growth kinetics with distinct effects observed at different wavelength and fluence combinations. Notably, 825 nm at 5 J/cm2 promoted larger neurospheres with slower growth rates, while 525 nm at 10 J/cm2 induced smaller, rapidly differentiating clusters. We also observed wavelength-dependent effects on mitochondrial function and cellular differentiation, accompanied by increased gene expression of β-tubulin III and NeuN levels by Day 11. These findings highlight the multifaceted impact of PBM on cellular behavior, promoting differentiation and modulating mitochondrial function. Our results suggest that PBM has potential as a non invasive approach for stem cell-based therapies aimed at neural repair and regeneration and warrant further investigation into its mechanisms and applications.

Neural crest-derived cells offer valuable opportunities to dissect mechanisms of cell fate specification and differentiation and the underpinnings of cell type diversification over evolutionary time. Particularly useful for such analyses are pigment cells of ectothermic vertebrates that arise from neural crest cells or via latent neural crest-derived stem cells. Among these are white cells, leucophores, present in a variety of species that contribute to patterns on the body or ornamentation on the fins. To better understand developmental and evolutionary origins of these cells, we examined leucophores harboring deposits of yellow/orange carotenoids-xantholeucophores-of zebrafish and leucophores of white cloud minnow. We show that white phenotypes of both cell types require sepiapterin reductase and an accumulation of pale and colorless pteridines. We further demonstrate that xantholeucophores of zebrafish develop from yellow, sepiapterin-rich xanthophore-like cells and that this transition requires both gap junctional communication and the aquaglyceroporin/peroxiporin channel Aquaporin 3, revealing similarities and differences in differentiation and patterning compared to pigment cells on the body. These findings identify xantholeucophores of zebrafish and leucophores of white cloud minnow as distinct developmentally, genetically, and biochemically from other white cells of zebrafish-melanoleucophores-that develop directly from melanophores and depend on guanine crystals, as well as white cells of medaka fish and anemonefish. Our results highlight remarkable convergences and parallelisms in the acquisition of white cell phenotypes within and between phylogenetic lineages and identify this as a rich system for enquiries into the evolutionary individuation of novel cell types.

Ependymal cells in the adult ventricular-subventricular zone are increasingly recognized for functions extending beyond cerebrospinal fluid dynamics; however, their identity and functional specialization remain incompletely understood. While ependymal cells (EP) have been implicated in interactions with the stem cell niche and the vasculature, their role in repair processes following neural injury remains elusive. In this study, we employed region-specific single-cell transcriptomics of the subventricular zone (SVZ) and ipsilesional peri-infarct territory in mice to identify a distinct subpopulation of GFAP+ FOXF2+ EP that selectively expand within the SVZ after neural injury. Notably, these cells were absent from other brain regions. Immunohistochemical validation revealed characteristic ependymal features, including typical pinwheel architecture and expression of Foxj1 and β-catenin. Furthermore, the absence of the proliferation marker Ki67 and the resistance of this subpopulation to Ara-C-mediated ablation indicate that these cells do not possess proliferative properties. Conditional deletion of Foxf2 in GFAP+ cells led to impaired endothelial junction integrity and increased blood-brain barrier (BBB) permeability. In contrast, overexpression of Foxf2 via GFAP promoter-driven adeno-associated virus delivery enhanced vascular repair and facilitated functional recovery. Mechanistically, these GFAP+ FOXF2+ EP secrete exosomal DLL4, which was associated with enhanced NOTCH pathway activity and restoration of BBB function. While the mechanism linking this limited cell population to the broad reparative effect, particularly the complete signaling amplification cascade, remains to be fully elucidated, these findings identify a subset of EP that contributes to BBB repair.

The aim of this White Paper is to establish a foundational framework for research, technological development, and regulation in the emerging field of stem cell-based embryo models (SCBEMs). These models, generated from Pluripotent Stem Cells, are designed to recapitulate essential events in early stages of human development. They have the potential to illuminate the early stages of embryo development and implantation and hold promise as an avenue to address global health challenges, including infertility and pregnancy loss, congenital, neonatal and adult conditions, and the need for organ transplants. While SCBEMs are not a substitute for human embryos, their tractability for large-scale analysis and their abilities to model the earliest stages of embryonic development suggest that they will have a significant impact on reproductive biology and regenerative medicine. But SCBEMs do not just raise novel scientific questions; they pose ethical and legal questions that need to be addressed. The paper stems from a meeting of a core group of researchers that met at the Institut Pasteur in Paris in November 2024 and represents the views of an extended group that has worked to elaborate the documents as a consensus for the field. Here, we provide a framework to guide research in this new field. We do this by summarizing the state of the science, assessing current SCBEM research in relation to its primary future applications and addressing the need for continued ethical and regulatory oversight associated with this new field.

Cell therapy for neurodegenerative diseases (NDs) is considered a promising strategy to halt disease progression. Currently, most clinically applied cells are derived from two-dimensional (2D) cultures. However, 2D-cultured mesenchymal stem cells (MSCs) are prone to aging and functional deterioration after multiple passages, and the availability of neural precursor cells for cell replacement therapy remains limited. In contrast, three-dimensional (3D) cell cultures have garnered significant attention due to their unique 3D spatial interactions. The unique spatial architecture of 3D culture not only enhances cell-cell and cell-extracellular matrix (ECM) interactions in MSC spheroids, thereby preserving MSCs properties, but also facilitates developmental processes of brain organoids derived from pluripotent stem cells, including embryogenesis, morphogenesis, and organogenesis. This review highlights the therapeutic ability of 3D-cultured MSC spheroids and brain organoids for NDs and summarizes advanced engineering platforms for their production. Future research should integrate the strengths of both technologies by establishing standardized quality control systems and scalable production processes to harness the microenvironmental modulation capacity of MSC spheroids and the precise cell replacement ability of brain organoids, ultimately advancing personalized therapies for NDs.

This study aims to investigate the effect of exosomes derived from olfactory mucosa mesenchymal stem cells (OM-MSCs-Exo) on microglial polarization and its potential therapeutic role in Alzheimer's disease (AD). OM-MSCs-Exo were isolated and purified from the mice olfactory mucosa, followed by phenotypic characterization. Proteins transferred by OM-MSCs-Exo were screened using proteomic analysis. The AD model was established in microglial cells and mice with Aβ1-42. Immunofluorescence and biochemical assays were employed to assess the impact of OM-MSCs-Exo and its secreted protein FGFR1 on microglial polarization. Protein-protein interactions and immunoprecipitation were used to identify the target proteins of FGFR1 in microglial cells. Additionally, the effects of OM-MSCs-Exo-induced microglial polarization on neuronal inflammation and cognitive function in mice were evaluated. OM-MSCs-Exo were successfully isolated and purified. FGFR1 was significantly upregulated in OM-MSCs-Exo compared to OM-MSCs. Aβ1-42 induced M1 polarization and suppressed M2 polarization of microglia, which was reversed by OM-MSCs-Exo. FGFR1 overexpression in OM-MSCs-Exo further enhanced M2 polarization in microglial cells. Phospholipase C gamma 1 (PLCγ1) was identified as the target of FGFR1, and knocking down PLCγ1 reversed the effects of FGFR1-overexpressing OM-MSCs-Exo. OM-MSCs-Exo alleviated cognitive decline and neuroinflammation in AD mice, with FGFR1 overexpression further enhancing these effects. OM-MSCs-Exo promote M2 polarization of microglia in AD mice through the FGFR1/PLCγ1 pathway, alleviating neuronal inflammation and cognitive dysfunction.

Spinal cord injury (SCI) is a major traumatic insult that leads to significant motor and sensory impairments. Following SCI, an inhibitory microenvironment develops, accompanied by substantial neuronal loss. A single therapeutic approach may not achieve substantial functional recovery. Therefore, strategies aimed at addressing the inhibitory environment and enhancing cellular engraftment are crucial for optimal recovery. In this study, we investigated the efficacy of combining an injectable fibrin hydrogel with subventricular zone-derived neural stem cells (SVZ-NSCs) and CT04, a RhoA inhibitor, in a rhesus monkey model of SCI. After characterizing SVZ-NSCs and optimizing the CT04 dosage, we administered the combination therapy during the sub-acute phase of SCI. Our results indicated that this combination treatment significantly improved neuronal differentiation and integration of grafted NSCs and promoted axonal regeneration. Compared to cell transplantation or CT04 treatment alone, animals receiving the combination therapy exhibited superior functional recovery. These findings highlight a promising approach for treating SCI by combining therapies to enhance NSC transplantation and axonal repair using an advanced injectable hydrogel.

The main aim of the present study is to fabricate a 3D scaffold loaded with bioglass nanofibers to orchestrate robust bone regeneration via enhanced osteogenesis and mineralization. We applied a sol-gel method to fabricate the bioglass nanofibers and incorporated them into 3-D hydrogel (1%, 5%, and 10% wt.%). The fabricated scaffolds were characterized to determine their morphology, surface functional groups, porosity, swelling capacity, biodegradation profile, cytocompatibility, and hemocompatibility. Finally, the scaffolds were implanted in critical-size bone-defect models in the rabbit calvaria. The results showed that the calcination of precursor nanofibers to fabricate bioglass nanofibers reduced the nanofibers' diameter from around 981 nm to around 566 nm. Hydrogels containing 0%, 1%, and 5% BGNFs demonstrated no cytotoxicity, with viability percentages of 113%, 105.7%, and 112%, respectively. The 10% BGNF formulation exhibited a minor decrease in vitality (84.3%). Notably, the scaffolds promoted osteogenesis to a high degree, as evidenced by a concentration-dependent increase in mineral deposition under Alizarin Red staining. The animal studies showed that scaffold implantation supported new bone formation, as evidenced by histological analysis and micro-CT imaging. A denser, more interconnected bony network with superior mineralization (BMD) was the result of a remarkable rise in bone volume (BV/TV), trabecular thickness (Tb.Th), and trabecular number (Tb.N), as well as a decrease in trabecular separation (Tb.Sp). The most significant outcomes were achieved when stem cells were combined with the BGNF scaffold. In summary, these results demonstrate that the bioactive nanocomposite scaffold is an excellent starting point for functional bone repair.

Patients with relapsed/refractory T-cell acute lymphoblastic leukemia (R/R T-ALL) have very poor prognosis. CAR T-cell therapy is being explored in these patients with encouraging results. Specifically, CD1a-directed CAR T-cells are being explored in clinical trials to treat cortical T-ALL (CD1a+), but there are no data on outcome in this subgroup of patients before this therapy. This retrospective, observational study aimed to describe the characteristics and outcomes of patients with R/R CD1a + T-ALL. We included pediatric and adult patients diagnosed with R/R CD1a + T-ALL in 25 sites of Spain between March 2006 and May 2022. The primary outcome of the study was overall survival (OS). Forty-three patients, 28 adults and 15 children, with R/R CD1a + T-ALL were included. Median (range) age at inclusion was 24 years (5–57), and 82.5% were male. After a median (range) follow-up of 7.0 years (5.1–13.6), 6 (14.0%) patients were alive and in complete remission (CR) and 37 (86.0%) had died. Five-year OS was 16% (95% CI; 7–29%). Bone marrow relapse, an interval < 12 months between first CR and relapse, and lack of allogeneic hematopoietic stem cell transplantation during salvage treatment were associated with worse OS. Our results confirm that patients with R/R CD1a + T-ALL have a very poor prognosis, highlighting the need for new treatment alternatives in these patients.

Standard preclinical human tumor models lack a human tumor stroma. However, as stroma contributes to therapeutic resistance, the lack of human stroma may make current models less stringent for testing new therapies. To address this, using patient-derived tumor cells, patient-derived cancer-associated mesenchymal stem/progenitor cells, and human endothelial cells, we created a human stroma-patient-derived xenograft (HS-PDX) tumor model. HS-PDX, compared to the standard PDX model, demonstrates greater resistance to targeted therapy and chemotherapy and better reflect patient response to therapy. Furthermore, HS-PDX can be grown in mice with humanized bone marrow to create humanized immune stroma patient-derived xenograft (HIS-PDX) models. The HIS-PDX model contains human connective tissues, vascular and immune cell infiltrates. RNA sequencing analysis demonstrated a 94-96% correlation with primary human tumor. Using this model, we demonstrate the impact of human tumor stroma on recruitment of TAMs and tumor immune exclusion to impact to response to immunologic therapy. We show an immunosuppressive role for human tumor stroma and that this model can be used to identify immunotherapeutic combinations to overcome stroma-mediated immunosuppression. Combined, our data confirm a critical role for human stroma in therapeutic response and indicate that HIS-PDX can be an important tool for preclinical drug testing.