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BackgroundAtopic dermatitis (AD), a chronic inflammatory skin disease, is characterized by intense itching and recurrent eczema-like lesions, requiring safer, more targeted therapies that modulate the immune response and restore skin barrier integrity. Conditioned medium derived from stem cells of human exfoliated deciduous teeth (SHED-CM) exhibits considerable immunomodulatory and regenerative effects. We evaluated the efficacy of SHED-CM in an AD-like mouse model and explored the underlying mechanisms.MethodsMice with ovalbumin (OVA)-specific allergic dermatitis (OSAD) were intravenously treated with SHED-CM, conditioned medium derived from fibroblast, or Dulbecco’s Modified Eagle Medium, followed by antigen restimulation to induce AD-like dermatitis. Disease severity was assessed macroscopically and histologically. T cell phenotypes were analyzed using immunohistochemical staining, RT-qPCR, and flow cytometry. Total serum and OVA-specific immunoglobulin E (IgE) levels were measured using ELISA. Naïve splenic CD4⁺ T cells were activated using CD3/CD28 beads in SHED-CM to evaluate their direct effects on T cell differentiation through flow cytometry. Highly expressed proteins in SHED-CM were identified using liquid chromatography–mass spectrometry, and neutralizing antibodies were used to elucidate the therapeutic mechanisms.ResultsThe treatment with SHED-CM significantly ameliorated AD-like symptoms, reduced epidermal hyperplasia, and restored filaggrin expression. SHED-CM created an anti-inflammatory microenvironment in OSAD skin and draining lymph nodes by promoting Treg differentiation and inhibiting Th2 and Th17 populations. SHED-CM markedly suppressed B cell maturation, antigen-specific IgE production, and mast cell activation. Furthermore, we found that SHED-CM directly promoted Treg differentiation from naïve CD4⁺ T cells stimulated by CD3/CD28, which was mediated, in part, through TGFβ contained in SHED-CM. SHED-CM exhibited little to no effect on in vitro Th1, Th2, and Th17 differentiation.ConclusionsSHED-CM shows significant therapeutic potential for AD-like symptom, effectively modulating immune response and enhancing skin barrier restoration. Our findings offer a new approach for developing novel, targeted therapies for inflammatory skin disorders.Supplementary InformationThe online version contains supplementary material available at 10.1186/s13287-025-04735-2.

ObjectivesThis study evaluated Biodentine (BD) after modification with 2 wt% fluorinated graphene (FG).MethodsFG was prepared using the modified Hummers’ method, where sulfuric and phosphoric acids were added to fluorinated graphite and potassium permanganate. The mixture was heated, sifted, filtered, and centrifuged to obtain FG powder. Characterization was performed using XRD, FTIR, TEM, and SEM/EDX. PH was evaluated, And Ca And F ion release were assessed by inductively coupled plasma spectroscopy and ion chromatography, at days 1,14, And 28. Cell viability was performed using the MTT Assay on pulp stem cells, while ALP assay was evaluated by a spectrophotometer. Compressive strength was evaluated by a universal testing machine. Statistical analysis was performed on the data (p ≤ 0.05).ResultsGraphene and C-F bonds of FG were confirmed in XRD and FTIR, while nanosheets were detected in TEM. SEM/EDX showed more surface roughness in modified BD-FG. pH And Ca ion release results showed significantly higher values at day 1 for modified BD-FG, with significantly higher cumulative Ca ion release. Cell viability results showed no significant difference between modified And unmodified Biodentine at days 1 And 7; however, modified BD-FG showed significantly lower values at day 3. No significant difference was observed between the two groups in ALP, while the BD-FG group showed significantly higher compressive strength.ConclusionIncorporating 2 wt% FG into BD increases ion release, hydroxyapatite formation, and mechanical properties without compromising cell viability and differentiation.Clinical relevanceThe addition of FG enhanced the bioactivity of Biodentine and improved its strength without showing cytotoxicity, making it a promising approach that needs further study.Supplementary InformationThe online version contains supplementary material available at 10.1186/s12903-025-06947-7.

Although small animal models of Fanconi anemia (FA) are useful, they do not faithfully replicate many of the clinical features seen in FA patients. We reasoned that a porcine model of FA with its similar physiology and a relatively long lifespan would produce a phenotype more similar to human FA. Targeting FANCA in domestic swine resulted in skeletal abnormalities and extreme sensitivity to interstrand DNA cross-linking agents. In addition, FANCA disruption followed by mitomycin C treatment resulted in a > 10-fold increase in chromosomal radials, a finding that is considered diagnostic for human FA. Bone marrow derived, hematopoietic progenitor cells from a FANCA null pig showed a 75% reduction in colony forming activity compared to wild type. Evaluation of steady state hematopoiesis in the peripheral blood revealed the gradual development of red cell macrocytosis and a reduction in circulating neutrophils. Targeting of FANCD2 failed to produce any biallelic animals demonstrating the loss of FANCD2 function is embryonic lethal in pigs. These results indicate that a porcine model of FANCA holds promise for the development of strategies to prevent the development of bone marrow failure and malignancies in patients with FA.

BackgroundHigh grade serous carcinoma (HGSC) is the most common and lethal subtype of ovarian cancer, yet its prognosis has remained unchanged in the past 3 decades. HGSC is known to have evolved immune evasion strategies to promote survival, but these mechanisms are not well understood. Podocalyxin (PODXL), a CD34-related sialomucin, is often expressed in HGSC patients with poor prognosis. We have recently reported that PODXL promotes the formation of compact and chemoresistant HGSC spheroids to boost their survival.MethodsIn this current study, we investigated whether PODXL may also influence HGSC spheroid susceptibility to NK cell infiltration and cytotoxicity. We co-cultured HGSC spheroids with primary human NK cells isolated from peripheral blood mononuclear cells (PBMCs) and examined the impact on these spheroids following 24, 48 and 72 h of co-culture. We first used a cell line model of HGSC spheroids employing Kuramochi cells, which express the highest level of PODXL among known HGSC cell lines. To study the impact of PODXL levels, we compared spheroids of control and PODXL knockout (PODXL-KO) cells that we have previously engineered. We then validated the data in primary cancer spheroids derived from ascites of HGSC patients that express high and low levels of PODXL.ResultsIn both the cell line and primary HGSC spheroid models, co-culture of spheroids expressing lower levels of PODXL resulted in more NK cell infiltration and cytotoxicity, while spheroids expressing higher levels of PODXL were resistant to destruction and showed more proliferation.ConclusionsCollectively, these data suggest that PODXL may play an important role in aiding immune evasion in HGSC, at least partly by conferring resistance to NK cell infiltration and the related cytotoxicity.Supplementary InformationThe online version contains supplementary material available at 10.1186/s12885-025-15108-6.

Despite recent advancements in immunotherapy, most cancer patients still struggle to achieve sustained benefits, highlighting the need for new treatment strategies. In the past, lack of available models to assess immunotherapeutic combinations hampered development. In this new study, we utilized a novel all-autologous humanized melanoma mouse model to assess the efficacy of NT-I7 (human-reagent grade, efineptakin alfa), a long-acting human IL-7. Given that NT-I7 has been shown to enhance T cell proliferation and survival in both humans and mice, we hypothesized that NT-I7 would improve the engraftment of patient immune cells and the effectiveness of anti-PD-1 therapy in our humanized melanoma model, which was reported to accurately mimic actual clinical outcome, providing more precise assessment of clinical efficacy and relevance. Our findings indicate that NT-I7 significantly enhances T cell engraftment. We discovered a synergistic effect between NT-I7 and anti-PD-1 (Pembrolizumab) that notably augments immunotherapeutic efficacy through the expansion of TCM cells and sustained cytotoxicity. In sum, our humanized model reveals that NT-I7 holds great promise as a next-generation therapy to enhance clinical responses and patient care.Supplementary InformationThe online version contains supplementary material available at 10.1038/s41598-025-20253-w.

BackgroundAir pollution, specifically fine particulate matter (PM2.5), in China is responsible for millions of excess deaths each decade. Examinations of Chinese municipalities have revealed correlations between ambient PM2.5 levels and the prevalence and severity of respiratory viral infections. Seasonal sources of ambient PM2.5 vary, with coal combustion for indoor heating significantly contributing during colder months. Due to this seasonality, we hypothesized that PM2.5 collected in Xinxiang, China would differentially alter the response to subsequent influenza A/California/04/2009 (H1N1) viral infection in a primary human nasal epithelial cell (HNEC) culture model in a seasonality-specific manner. After the PM2.5 samples were chemically analyzed, HNECs collected from males (N = 4) and females (N = 3) grown at air-liquid interface were exposed to 22 µg/cm2 of seasonal PM2.5 followed by inoculation with influenza A H1N1 at MOI = 0.001. At 2 and 24 h post infection (p.i.) we assessed transcriptional changes and basolateral release of immune and antiviral mediators.ResultsSummer and fall PM2.5 samples contained a greater organic carbon mass fraction compared to winter and spring. Winter contained the largest mass fraction of anionic components and spring the largest inorganic element mass fraction. Exposure to the seasonal PM2.5 samples without infection induced a moderate transcriptional response at 2 h, with the winter PM2.5 inducing the greatest response. The seasonal PM2.5 exposures followed by viral infection resulted in a more robust transcriptional response at 2 h p.i. with the winter, spring, and fall PM2.5 samples (but not the summer PM2.5) upregulating many inflammatory pathways. At 24 h p.i., only the spring PM2.5 sample increased inflammatory and antiviral mediator proteins in the basolateral medium, while winter PM2.5 increased these inflammatory markers in the mock infected cultures.ConclusionsSeasonal variations in PM2.5 composition during winter, spring, and fall—coinciding with influenza season—likely enhance pro-inflammatory responses to viral infection, with early inflammation contributing to worsened pathogenesis.Graphical abstract Supplementary InformationThe online version contains supplementary material available at 10.1186/s12989-025-00643-8.

ImportanceLumbar facet arthropathy (LFA) is one of the most common specific disorders associated with low back pain. Current treatments provide only symptomatic relief and are often limited in their long-term effectiveness.Objective To evaluate the safety and preliminary efficacy of intra-articular delivery of allogeneic BM-MSCs in patients with painful LFA.Design, setting, and participantsThis study was a prospective, Phase I, single-arm, open-label clinical trial. Ten patients with painful LFA were enrolled with nine completing the 2-year follow-up.InterventionsAll patients received a single intra-articular administration into two affected lumbar facet joints, with each joint injected 10 million allogeneic BM-MSCs suspended in 1 mL Lactated Ringer’s solution.Main outcomes and measures The primary endpoint was safety, as reflected by adverse events (AEs). Secondary endpoints (and the assessment tools) included low back pain (VAS), physical function (PROMIS CAT Physical Function), health status (PROMIS CATs), and the severity of facet joint degeneration on MRIs. The minimum clinically important difference (MCID) thresholds in this trial were pre-defined as a 50% reduction in low back pain (VAS), or an increase of 2.3 points on the PROMIS CAT Physical Function.ResultsThe procedures were well tolerated. There were no clinical signs of immune reaction to the allogeneic BM-MSCs. No study-related serious AEs were observed. Six patients achieved the MCID on VAS for pain at 6-, 12-, 18-, and 24-month follow-ups and five patients consistently met MCID for the PROMIS CAT Physical Function across all follow-up visits. MRIs showed reduced facet joint degeneration at one or more levels in five patients.Conclusions and relevanceThis study demonstrated a favorable safety profile for intra-articular delivery of BM-MSCs for painful LFA. Preliminary therapeutic benefits were observed, including back pain relief, improved physical function, and reduced facet joint degeneration. Further larger, randomized clinical trials are warranted to further assess its safety and efficacy. Trial Registration ClinicalTrials.gov Identifier: NCT04410731.Supplementary InformationThe online version contains supplementary material available at 10.1186/s13287-025-04674-y.

Adaptive transfer of autologous regulatory T cells (Treg), or ex vivo-generated immunomodulatory cells, has shown promise in reducing/withdrawing immunosuppression after organ transplantation. The effect of cryopreserving such cells is still unclear. This study aims to evaluate the effects of cryopreservation on the immunomodulatory functions of peripheral blood mononuclear cells (PBMCs) with or without pretreatment with red blood cell (RBC) lysate (ACK). Human PBMCs enriched from buffy coats of healthy blood donors were treated either with ACK or phosphate-buffered saline (PBS). Thereafter, a batch of the PBS-control subset was cryopreserved with 10% dimethyl sulfoxide (DMSO) and subsequently examined for phenotype, functionality, and relative gene expression. We found that ACK-treated PBMCs exhibited higher numbers of interferon gamma (IFN-γ)-producing cells when stimulated with viral peptides (p = 0.0078), indicating that ACK treatment may improve the antigen sensitivity of memory T cells. After cryopreservation, contaminated RBCs and granulocytes, cell viability, and CD4+ T-cell population decreased (p = 0.0078); IL-1β expression increased; and FoxP3 expression decreased (p = 0.0312), where the Treg population remained otherwise unchanged. Enriched Tregs from both fresh and frozen PBMCs suppressed the proliferation of anti-CD3/CD28-antibody-stimulated PBMCs equally. In conclusion, the preservation of Treg function following cryopreservation highlights its potential utility in tolerance-induction trials, providing experimental flexibility and simplified logistics. Graphical Abstract

Background: Aryl hydrocarbon receptor (AhR) is a transcription factor that is involved in the regulation of immunity. AhR inhibits T cell activation in tumors, which induces immune suppression in the blood and solid tumors. We identified effective small-molecule AhR antagonists for cancer immunotherapy. Methods: A new series of pyrazolopyrimidine derivatives was synthesized and evaluated for AhR antagonistic activity. Results: Compound 7k exhibited significant antagonistic activity against AhR in a transgenic zebrafish model. In addition, 7k exhibited good AhR antagonist activity, with a half-maximal inhibitory concentration (IC50) of 13.72 nM. Compound 7k showed a good pharmacokinetic profile with an oral bioavailability of 71.0% and a reasonable half-life of 3.77 h. Compound 7k selectively exerted anti-proliferative effects on colorectal cancer cells without affecting normal cells, concurrently suppressing the expression of AhR-related genes and the PD-1/PD-L1 signaling pathway. Compound 7k exhibited potent antitumor activity in syngeneic colorectal cancer models. Importantly, the combination of anti-PD1 and compound 7k enhanced antitumor immunity by augmenting cytotoxic T lymphocyte (CTL)-mediated activity. Conclusions: Collectively, a new pyrazolopyrimidine derivative, 7k, shows promise as a potential therapeutic agent for treating colorectal cancer.

Mesenchymal stem cells (MSCs) have emerged as a promising therapeutic approach for degenerative diseases due to their ability to modulate disease progression through paracrine mechanisms. Among various MSC sources, placenta-derived MSCs (PD-MSCs) offer significant advantages, including high proliferation capacity, reduced senescence, and low immunogenicity, making them ideal for allogeneic applications. In this study, we investigated the therapeutic effects of PD-MSC transplantation in an estrogen-deficiency-induced osteoporosis mouse model. Mice were divided into three groups: a normal control group, a non-transplanted osteoporosis group, and a PD-MSC-transplanted group. Our findings demonstrated that PD-MSC transplantation significantly improved osteoporosis-related parameters, including increased femur weight, bone volume, bone mineral density, and calcium deposition. Additionally, estrogen levels were elevated, bone formation markers were upregulated, and bone resorption markers were downregulated. PD-MSCs also reduced inflammatory cytokine levels while enhancing anti-inflammatory factors. Notably, the BMP/SMAD signaling pathway, crucial for bone formation, was significantly upregulated. These results suggest that PD-MSC transplantation effectively restores bone homeostasis by inhibiting osteoclast activity, promoting osteogenesis, and modulating inflammation. This study provides strong evidence supporting the potential of PD-MSCs as a novel therapeutic strategy for osteoporosis, offering a regenerative and anti-inflammatory approach to bone disease management.

Rheumatoid arthritis (RA) is characterized by chronic inflammation mediated by the JAK/STAT pathway. Variants in STAT4 have been associated with autoimmune susceptibility, but their functional role in RA remains unclear. The aim of this study was to genetically and functionally characterize STAT4 in RA patients with varying disease activity by analyzing two variants, mRNA expression, phosphorylated STAT4 (pSTAT4), and inflammatory cytokines (IL-12, IL-23, and IFN-γ). Sixty-three Mexican patients with RA were stratified into remission/low and moderate/high activity groups. Genotyping, STAT4 mRNA expression, pSTAT4 quantification, cytokine profiling, and treatment analyses were conducted. Patients receiving methotrexate, hydroxychloroquine, and sulfasalazine had higher IL-12 concentrations compared with those on other regimens. In remission/low activity patients, GC/GC carriers exhibited increased IL-12, PBMC levels, and anti-CCP antibodies, while GC/TT carriers in the moderate/high activity group showed distinct ESR values. Secondary analyses revealed that TT/TT carriers with STAT4 overexpression exhibited higher IFN-γ and IL-23 levels. IL-12 differences persisted among GC/GC carriers regardless of STAT4 expression status. In conclusion, these exploratory findings suggest potential interactions among STAT4 haplotypes, expression status, and treatment regimens influencing cytokine and inflammatory profiles in RA. However, due to the small subgroup sizes, the observed associations should be interpreted with caution and considered hypothesis-generating until validated in larger cohorts.

Three-dimensional (3D) hepatic tissue engineering holds great potential for liver regeneration, disease modeling, and drug screening. These applications require densely layered hepatic tissues that mimic native 3D liver architecture. However, limited oxygen supply and reduced cell viability in densely layered hepatic constructs remain key challenges. To overcome this, this study developed a photo-crosslinkable, oxygen-releasing hydrogel composed of methacrylated hyaluronic acid (MeHA) and calcium peroxide (CPO). The MeHA/CPO hydrogel exhibited favorable rheological properties and sustained oxygen release. Induced pluripotent stem cell-derived hepatocyte (iHep) sheets were cultured with or without MeHA/CPO hydrogel in single- and double-layer formats. The hydrogel enhanced structural integrity and supported the formation of a multilayer (~33 µm). Double-layered iHep sheets with MeHA/CPO showed the significantly increased expression of paracrine factors (HGF, VEGF, Alb) and improved albumin secretion without loss of hepatocyte identity (AFP, HNF4α). This oxygen-releasing system effectively alleviates hypoxic stress, supporting the structural and functional viability of multilayered iHep sheets. Our platform provides a promising approach for engineering metabolically active hepatic tissues and may serve as a foundation for 3D hepatic tissue engineering.