Periodontal regenerative therapy using recombinant human fibroblast growth factor-2 (rhFGF-2) combined with deproteinized bovine bone mineral (DBBM) has demonstrated favorable and predictable clinical outcomes for intrabony defects. However, long-term outcomes and potential challenges following such combination therapies remain poorly documented. A 40-year-old woman with generalized stage III grade C periodontitis underwent periodontal regenerative therapy using rhFGF-2 combined with DBBM for a one-wall defect at the mandibular left first molar (#19). Favorable periodontal healing was maintained for approximately 4 years. However, for tooth #19, the probing depth gradually increased at 5 years 2 months post-surgery, and the patient developed discomfort and pulpal symptoms by 6 years 2 months. Clinical examination revealed an agglomerate of DBBM particles and a palpable external root resorption lesion. The tooth was extracted due to irreversible pulpal involvement. This case illustrates that despite favorable early healing, periodontal regenerative therapy may lead to rare late-onset complications such as external root resorption. Long-term follow-up and careful consideration of biomaterial interactions are essential when employing combination regenerative approaches. Periodontal regenerative therapy using rhFGF-2 and DBBM achieved favorable clinical outcomes for 4 years before unexpected late-onset external root resorption developed. Long-term persistence and partial separation of DBBM particles, along with the response of macrophage- or osteoclast-lineage cells, are associated with a potential for delayed resorptive pathology on adjacent root surfaces. Clinicians should recognize the possibility of late complications and carefully consider biomaterial interactions when selecting regenerative strategies and informing patients. This case report describes a rare situation in which a molar initially healed well after periodontal regenerative therapy using rhFGF‐2 and a bone substitute (DBBM), but developed external root resorption more than five years later. Although the treatment first reduced inflammation and improved tissue support, the long‐term presence of graft particles, together with complex cellular responses, may be linked to delayed changes on the root surface. This case highlights that even when early healing is successful, long‐term monitoring is essential, and clinicians should carefully consider how different regenerative materials may interact over time.
Coccydynia is characterized by persistent tailbone pain that significantly impairs quality of life. Despite conservative treatments, many patients experience inadequate symptom relief. Prolotherapy, a regenerative injection technique for chronic musculoskeletal pain, shows promise; however, its efficacy for coccydynia has not been evaluated in controlled comparative studies. To the best of the authors' knowledge, this is among the first such studies. This retrospective comparative study evaluated 56 patients with primary chronic coccydynia refractory to conservative management. Twenty-nine patients received ultrasound-guided 10% dextrose prolotherapy combined with therapeutic exercises and patient education; 27 patients received exercises and education alone. Mean pain duration was 20.7 ± 17.3 months (prolotherapy) and 26.9 ± 17.3 months (control). Primary outcomes were pain intensity and functional disability at baseline, 1-month, and 3-month follow-up. Secondary outcomes comprised the Paris Functional Coccydynia Impact Questionnaire. Ultrasound-guided dextrose prolotherapy combined with therapeutic exercises demonstrated superior efficacy compared to exercises alone. The prolotherapy group achieved a 73% pain reduction at 3 months versus 26% in controls. An 82.8% clinical response rate and 37.9% complete pain resolution were observed, with a favorable safety profile. These results support prolotherapy as an effective, minimally invasive option for chronic coccydynia following failed conservative management. Tailbone pain (coccydynia) is a persistent pain at the base of the spine that gets worse with sitting or standing up. It affects daily life significantly and is more common in women. Many people do not get enough relief from standard treatments such as cushions, activity changes, and physiotherapy.This study looked at whether injecting a sugar-water solution (dextrose prolotherapy) into the tailbone area, guided by ultrasound imaging, could reduce pain and improve function in 56 patients who had tailbone pain for at least six months and had already tried standard treatments without adequate relief.Twenty-nine patients received three ultrasound-guided dextrose injections alongside a structured exercise and education program. Twenty-seven patients received the exercise and education program alone. Pain and ability to carry out daily activities were measured before treatment and at one and three months after starting treatment.Patients who received the injections had much greater reductions in pain (73%) and improvements in daily function (77%) at three months compared with those who received exercises alone (26% and 25%, respectively). The treatment was safe, with only minor and short-lived side effects such as temporary soreness at the injection site.These findings suggest that ultrasound-guided dextrose injections may be a useful and safe option for people whose tailbone pain has not responded to standard treatments.
Diabetic wound healingAC: remains challenging due to persistent inflammation, oxidative stress, and impaired macrophage polarization. Herein, a multifunctional hydrogel dressing was constructed from carboxymethyl chitosan and oxidized dextran as the dynamic network, incorporating CeO2 nanozymes for early anti-inflammatory and antioxidant effects and PLGA microspheres loaded with astragaloside IV for sustained pro-regeneration. Via Schiff base crosslinking, this hydrogel self-assembled rapidly at room temperature and adhered tightly to tissue. In vitro, the hydrogel exhibited excellent biocompatibility, potent antioxidant and anti-inflammatory activities, and promoted endothelial cell migration and angiogenesis. In a diabetic rat full-thickness wound model, this hydrogel dressing effectively reduced local inflammation, drove macrophage polarization toward the M2 phenotype, and enhanced neovascularization to accelerate wound closure. RNA sequencing further revealed that inflammatory pathways, including TNF and IL-17 signaling, were suppressed while tissue regeneration programs were activated. This stepwise therapeutic strategy offers a promising alternative for diabetic wound repair.
Cyclin-dependent kinase 4 and 6 (CDK4/6) inhibitors have significantly changed the treatment approach for hormone-receptor-positive (HR+), human epidermal growth factor receptor 2 (HER2)-negative (HER2-) breast cancer in both metastatic and high-risk early-stage settings. In combination with endocrine therapy, these agents consistently improve progression-free survival, and several phase III trials have demonstrated overall survival benefit in defined populations. Their clinical activity is supported by a well-established biologic rationale targeting dysregulated cell cycle progression, a key feature of HR+ breast cancer, which drives tumor proliferation. Despite these advances, resistance remains a clinical limitation in advanced disease. Multiple mechanisms have been identified, including loss of RB1 function; amplification of CDK6, activation of cyclin E CDK2 signaling; upregulation of bypass pathways such as PI3K, AKT, mTOR, and FGFR; and acquired alterations in estrogen receptor signaling, including ESR1 mutations. Circulating tumor DNA assays are increasingly used in clinical practice and clinical trials to detect emerging genomic alterations that may allow earlier modification of therapy based on molecular progression. The post-CDK4/6-inhibitor treatment landscape has expanded substantially and now includes treatment options such as switching CDK4/6 inhibitors, targeting the PI3K-AKT pathway in patients selected for mutation, use of oral selective estrogen receptor degraders, and incorporation of antibody-drug conjugates. Ongoing studies evaluating CDK2 inhibitors, CDK4-selective agents, immunotherapy combinations, and circulating tumor DNA (ctDNA)-guided strategies aim to further refine treatment sequencing and improve long-term outcomes in HR+/HER2- breast cancer.
Liver tissue engineering offers a promising alternative for end-stage liver disease, yet the recreation of functional vasculature remains a major bottleneck to clinical translation. Here, we developed vascularized liver organoids by integrating human induced pluripotent stem cell (iPSC)-derived hepatoblasts and endothelial cells into decellularized scaffolds functionalized with an anti-CD31 aptamer-based vascular coating agent (VCA). This facilitated spatially coordinated organization of vasculature and parenchyma. Spatial transcriptomic profiling and subsequent functional perturbation demonstrated IGF2-IGF1R-AKT/MAPK signaling as a key axis governing spatial organization and functional maturation of the liver organoids. Furthermore, exogenous IGF2 synergized with the VCA to augment the structural and functional refinement of liver organoids, which translated into markedly improved therapeutic outcomes following transplantation into a chronic liver failure mouse model. Collectively, these findings establish a comprehensive framework for generating physiologically relevant liver tissues from iPSCs and demonstrate the utility of spatial transcriptomics for uncovering regenerative mechanisms. This approach advances the feasibility of autologous, transplantable liver grafts for personalized regenerative therapy.
Infected bone defects remain a significant clinical challenge due to bacterial colonization-induced persistent inflammation, oxidative stress, and local acidification, which collectively impair bone regeneration. Conventional approaches, including antibiotic therapy and bone grafting, are often insufficient to concurrently eradicate infection and promote tissue repair. Herein, we report a multifunctional GelMA-Au@Pt@CaP (GAPCP) hydrogel that integrates second near-infrared window (NIR-II, 1000-1700 nm) photothermal antibacterial activity, nanozyme-mediated antioxidant catalysis, and programmable Ca2+/PO₄3- release to enable synergistic antibacterial and osteogenic therapy. Upon 1064 nm laser irradiation, the Au@Pt nanozyme achieves efficient bacterial ablation and biofilm disruption via photothermal effects. The calcium phosphate (CaP) layer provides responsive ion delivery to enhance osteogenic differentiation and mineralization. Concurrently, the porous Pt shell catalyzes H₂O₂ decomposition under acidic conditions, mitigating oxidative stress and hypoxia. GAPCP scavenges intracellular reactive oxygen species, promotes macrophage polarization toward an anti-inflammatory M2 phenotype, and enhances angiogenic and osteogenic activities in vitro. In an infected calvarial defect model, GAPCP combined with NIR-II irradiation achieves simultaneous antibacterial, anti-inflammatory and bone regenerative outcomes. Transcriptomic analysis reveals that these effects are associated with the coordinated regulation of inflammatory signaling, ion transport, and angiogenesis pathways. This work establishes a NIR-II-responsive multifunctional hydrogel platform that couples infection eradication with microenvironment remodeling to drive bone regeneration, providing a promising strategy for treating infected bone defects.
Injectable hydrogels represent a promising strategy for minimally invasive regenerative medicine, enabling the delivery of biomimetic microenvironments directly to irregular tissue defects. Here, we present a multi-responsive, ECM-like injectable hydrogel based on κ/λ-carrageenan, hyaluronic acid, and type I collagen, designed to combine ion-triggered gelation with ultrasound-activated piezoelectric functionality. Gelation occurs rapidly upon exposure to physiological ions, allowing straightforward injection through fine-gauge needles without the need for external crosslinkers or harsh conditions. The incorporation of submicrometric barium titanate particles confers piezoelectric responsiveness, enabling the conversion of ultrasound-induced mechanical stimuli into localised electrical cues. The resulting hydrogels exhibit mechanical properties (Young's modulus 4-9 kPa) and viscoelastic behaviour comparable to native soft tissues, alongside a highly interconnected porous architecture suitable for mass transport and cell infiltration. In vitro studies demonstrate cytocompatibility with fibroblasts, myoblasts, neuronal-like cells, and macrophages, supporting cell viability and proliferation while promoting a pro-regenerative macrophage phenotype. Preliminary ultrasound stimulation experiments confirm that piezoelectric activation does not impair cell viability, establishing a safe basis for future functional investigations. Overall, this work introduces a user-friendly, cost-effective, and multifunctional injectable hydrogel platform with potential for minimally invasive and remotely activated regenerative therapies.
Low back pain remains the leading cause of disability worldwide, with intervertebral disc degeneration representing a major biological contributor. Although cell-based therapies have shown promise in preclinical models, clinical translation has yielded modest and inconsistent outcomes. Accumulating evidence suggests that therapeutic failure reflects not only limitations in cell source or differentiation potential, but also the hostile biochemical and biomechanical microenvironment of the degenerative disc. Hypoxia, nutrient deprivation, acidity, lactate accumulation, fibrosis, senescence, inflammation, and abnormal mechanical loading collectively impair cell survival, integration, and long-term function. We performed a comprehensive review of the literature using PubMed, Web of Science, and Google Scholar, with emphasis on studies published between 2020 and 2026. Evidence was critically evaluated to examine advances in cell-based therapies for IVDD, including cell sources, mechanisms of repair, biomaterial-assisted delivery systems, microenvironment-targeted strategies, translational studies, and emerging technologies that enhance regenerative efficacy. Current evidence indicates that successful disc regeneration depends not only on selecting an appropriate therapeutic cell source but also on overcoming the biological constraints imposed by the degenerative niche. We critically compare the regenerative potential of mesenchymal stromal cells, nucleus pulposus cells, and induced pluripotent stem cell-derived therapies, highlighting their respective advantages and limitations. We further discuss how biomaterial carriers, extracellular vesicles, developmental biology-guided differentiation, genetic engineering, preconditioning approaches, and smart delivery platforms are being integrated to improve cell survival, phenotype stability, extracellular matrix restoration, and functional repair. Future success in intervertebral disc regeneration will require integrated therapeutic strategies that combine optimized cell sources with biomaterial-assisted delivery, microenvironment modulation, and precision bioengineering. Advancing these complementary approaches will be essential for achieving durable biological repair, restoring disc structure and function, and translating regenerative therapies into effective clinical treatments for patients with degenerative disc disease.
This commemorative article reflects on a research journey spanning neural development, stem cell biology, regenerative medicine, and iPSC-based drug discovery. My early work focused on RNA-mediated regulation in the nervous system, including studies on myelin basic protein gene regulation and the identification and functional characterization of the RNA-binding protein Musashi. These studies contributed to the conceptual foundation of neural stem cell biology and helped establish methods for identifying and isolating neural stem/progenitor cells, including those present in the adult human brain. Building on this foundation, my colleagues and I pursued translational research in spinal cord injury, ranging from analyses of injury pathophysiology and molecular interventions to preclinical studies using rodent and non-human primate models. These efforts ultimately led to the first-in-human clinical study of induced pluripotent stem cell-derived neural stem/progenitor cell transplantation for subacute spinal cord injury. In parallel, we developed patient-derived iPSC platforms for neurological disease modeling and drug discovery, particularly for amyotrophic lateral sclerosis, where iPSC-based screening identified Ropinirole as a therapeutic candidate and enabled reverse translational research linking cellular phenotypes with clinical responses. Looking ahead, I argue that the future of regenerative therapy will depend on the continued integration of developmental biology, stem cell science, disease modeling, rehabilitation, and clinical translation to address unmet medical needs in disorders of the central nervous system.
Cell-based therapeutics hold immense promise for clinical intervention, yet their practical translation is severely bottlenecked by suboptimal delivery kinetics, safety risks, and low patient adherence. This review provides a comprehensive overview of recent innovations in microneedle (MN)-based cell delivery systems and their applications across multiple organs. We initiate the discussion by evaluating progress in regenerative medicine, specifically targeting stem and immune-derived lineages. The article further classifies MN systems for cell therapy delivery according to material composition, fabrication techniques, and functional characteristics, with an in-depth discussion of innovative designs, highlighting their unique advantages. Additionally, we categorize and analyze recent studies on MN-based cell therapy for skin diseases, endometrial repair, cardiac regeneration, and oral disorders. Beyond summarizing current achievements, we critically examine the translational barriers that limit clinical adoption, including manufacturing scalability, storage stability, and regulatory considerations. This review provides a systematic and organ-specific overview that may help guide the design of improved and more clinically translatable cell delivery platforms.
Heart failure is a major clinical and economic burden that afflicts 60 million individuals worldwide. Guideline-directed medical therapies can slow disease progression, but they cannot restore the loss of cardiomyocytes. Over the past two decades, human pluripotent stem cell (hPSC)-based technology has emerged as a leading approach to overcome limited cardiac regenerative capacity, offering a scalable source of functional human cardiomyocytes. The field is now at a pivotal translational stage, as advances in differentiation and tissue engineering have enabled hPSC-based products to enter first-in-human clinical trials. In this review, we summarize the pathophysiological rationale for cell-based therapy in heart failure with reduced ejection fraction. Then, we examine the preclinical foundations of distinct hPSC-derived product formats, including cell suspensions, epicardial sheets, engineered heart muscle, and cardiac spheroids, each with distinct tradeoffs and translational considerations. We conclude by providing updates on ongoing and recently completed clinical trials, evaluating their safety, feasibility, and preliminary efficacy outcomes.
The development of photosensitizers with high tissue penetration and strong antimicrobial activity against pathogenic microorganisms is at the forefront of antimicrobial photodynamic therapy research. Reducing the aggregation of photosensitizers (PS) in aqueous solutions through the addition of amphiphilic polymers can enhance their ability to generate reactive oxygen species (ROS) and, consequently, improve the efficacy of photodynamic treatment. Previously, we demonstrated that amphiphilic polymers increase the singlet oxygen generation activity of porphyrin photosensitizers and certain dyes by promoting the disaggregation of hydrophobic PS molecules. In the present study, we performed a comparative investigation of photodynamic treatment of model wounds in mice using methylene blue dye and Fotoditazin as PS, both in the presence and absence of the amphiphilic polymer Pluronic F108. In addition, sodium alginate was incorporated into the photosensitizing systems to provide an additional antibacterial effect on the wound. These findings provide a theoretical basis for the application of photosensitizer-polymer systems containing sodium alginate in antimicrobial photodynamic therapy for difficult-to-treat wounds.
Frailty associated with aging, characterized by chronic inflammation and immune dysfunction, poses a significant public health challenge. Human umbilical cord-derived mesenchymal stem cells (HUC-MSCs) exhibit immunomodulatory properties; however, their mechanism of action in frail elderly populations remains unclear. Building on a prior clinical trial investigating HUC-MSCs for elderly frailty (Trial Registration: ClinicalTrials.gov, NCT04314011; PubMed ID: 38679727), this study was conducted to further elucidate the mechanism of action of HUC-MSCs. We performed a longitudinal single-cell RNA sequencing (scRNA-seq) study on peripheral blood mononuclear cells (PBMCs) from frail elderly patients before and after HUC-MSCs therapy. Immune cell dynamics were analyzed across multiple time points, and cell-cell communication networks were reconstructed. Spearman correlation analyses were performed to link immune changes with clinical frailty and physical performance metrics. HUC-MSCs induced a time-dependent recalibration of the immune system. An early, transient expansion of MAIT cells (p=0.049) was followed by a sustained reduction in B cell hyperactivity (p=0.032) and a phenotypic shift in naïve B cells. NK cell cytotoxicity was enhanced (increased GNLY), while NKT cells showed modulated stress-response signatures. Cell-cell communication was reorganized, with the EPHA/NRG pathways highlighting CD4+ TEM, MAIT, and NKT cells as central signaling hubs. Critically, these immunomodulatory changes were quantitatively associated with clinical improvement: reductions in B cell and innate immune subsets correlated with gains in grip strength and gait speed, while regulatory T cell expansion was linked to improved lower-limb function. HUC-MSCs restore immune homeostasis in aging frailty through sequential, multi-faceted immunomodulation. The association between specific immune remodeling features and functional recovery positions HUC-MSCs as a promising mechanism-based therapy for aging frailty.
Skeletal muscle function deteriorates with injury, disease, or aging. While stem cell therapies offer therapeutic potential, concerns remain regarding the safety and efficacy of transplanted cells. Satellite cells (SCs), the primary endogenous stem cells responsible for muscle repair, demonstrate safe transplantation but limited clinical effectiveness. A deeper understanding of the mechanisms regulating SC activation and differentiation is critical for improving muscle regeneration strategies. We examined the role of CSRP3 in SC-mediated muscle repair using injury models and myoblast differentiation assays. We compared its function in pigs and mice, employing genetic approaches, CSRP3 knockdown (CSRP3KD), Csrp3-/- and overexpression (CSRP3OE), to assess its impact on muscle regeneration. Additionally, we investigated the AKT-SERCA2 signaling axis and its downstream effects on calcium homeostasis, mitochondrial function, and myogenic differentiation. CSRP3 was essential for muscle regeneration and SC differentiation in pigs but not in mice. Genetic ablation of CSRP3 (CSRP3KD) impaired porcine SC differentiation and reduced regenerative capacity, while CSRP3 overexpression (CSRP3OE) enhanced these processes. Mechanistically, CSRP3 deficiency disrupted AKT-SERCA2 signaling, leading to elevated intracellular Ca2⁺, mitochondrial dysfunction, and MYOG protein degradation. Pharmacological AKT activation rescued differentiation defects in CSRP3-deficient cells. Notably, Csrp3-/- mice showed normal SC differentiation and AKT-SERCA2 activity, highlighting a species-specific regulatory role for CSRP3. Our findings identify CSRP3 as a critical regulator of porcine muscle regeneration, with minimal impact in mice. These results suggest that strategically engineered porcine SCs, through CSRP3 modulation, could improve the effectiveness of xenotransplantation-based therapies for muscle repair.
Morton's neuroma is a painful entrapment neuropathy typically affecting the third intermetatarsal space. Conventional therapies, including orthotics, corticosteroids, or surgery, often yield inconsistent outcomes. We describe a 26-year-old woman with chronic, persistent neuropathic pain due to an atypical Morton's neuroma located in the fourth intermetatarsal space. After failure of multiple conservative and infiltrative treatments, she received 2 injections of leukocyte-poor platelet-rich plasma (PRP). Pain severity (Visual Analog Scale) improved from 9/10 to 1/10, and the Foot Function Index improved by 75% at 12 months. PRP provided durable analgesia and functional recovery in this rare neuroma presentation, underscoring its regenerative and neuromodulatory potential. Our case highlights the effectiveness of PRP as a promising, minimally invasive alternative when conventional treatments fail.
Ischemic tissue perfusion remains a major clinical challenge despite VEGF's central role in angiogenesis. Limited success of VEGF-based therapies underscores the need to understand endothelial heterogeneity, as distinct subsets exhibit differential responsiveness. This study explores conserved endothelial states that govern VEGF-driven angiogenesis across human organs and evaluates their functional significance. We performed single-cell and single-nucleus RNA-sequencing (scRNA-seq/snRNA-seq) analysis of endothelial cells (ECs) from eight human tissues using GTEx data, validated findings in independent skin and heart datasets, and conducted CRISPR-mediated PLCγ2 knockout in cultured ECs followed by scRNA-seq. Analysis of 209,126 nuclei revealed a universal arterial-like endothelial subset (cluster 2) enriched across all organs, characterized by high expression of PLCγ2, a critical VEGF signaling effector. PLCγ2high cells exhibited robust angiogenic and lymphangiogenic programs, including upregulation of VEGFC, NOTCH4, and JAG1, and enrichment of pathways for vasculature development and endothelial differentiation. Cell-cell communication analysis demonstrated exclusive BMP6, IGF2, and CXCL12 signaling from PLCγ2high cells to fibroblasts, pericytes, and immune cells, positioning this subset as a central angiogenic hub. Functional validation showed that PLCγ2 knockout depleted PLCγ2high clusters, suppressed VEGF signaling, and diminished BMP6 ligand secretion, confirming its role in maintaining proangiogenic endothelial states. Ineffective targeting of the appropriate EC state may limit current therapeutic VEGF strategies, positioning PLCγ2high ECs as a clinically relevant target. We identify PLCγ2high arterial ECs as a conserved pan-organ population critical for VEGF responsiveness and vascular regeneration. These findings reimagine VEGF therapy by highlighting PLCγ2 as a critical regulator of a proangiogenic arterial-like endothelial state and a promising combinatorial target to overcome limitations of current angiogenic strategies.
Mesenchymal stem cell (MSC) therapy offers promise for treating autoimmune diseases due to its strong immunomodulatory effects. We investigated the long-term safety of a single intravenous injection with human umbilical cord blood-derived (hUCB)-MSCs in patients with rheumatoid arthritis (RA). Patients with RA who met the 2010 ACR/EULAR classification criteria and received a single intravenous infusion of hUCB-MSCs (2.5 × 107, 5.0 × 107, 1.0 × 108 cells) in a phase I trial (NCT02221258) entered this 5-year observational pilot study. Safety assessments were performed at 3, 6, and 12 months after infusion and annually thereafter. Safety endpoints included overall adverse events (AEs), serious adverse events (SAEs), and AEs of special interest. Nine patients were treated. The most common AEs were osteoarthritis (44.4%) and nasopharyngitis (44.4%). SAEs occurred in 5 patients (55.6%); a serious infection (cellulitis) occurred in 1 patient in the 1.0 × 108 group and resolved after treatment. Benign ovarian and breast tumors were reported in 2 patients 3 years post-infusion. No deaths, thromboembolism, or malignancies occurred during the follow-up period. Laboratory findings remained stable except for 1 case each of transient hypertriglyceridemia and mild eosinophilia. While DAS28 improved markedly by 3-6 months, disease activity gradually increased over 5 years, suggesting waning efficacy after a single infusion. The long-term safety profile of a single dose of intravenous hUCB-MSC in patients with RA appears acceptable; however, repeated-dose regimens may be needed for sustained disease control and further safety evaluation.
Nearly half of patients with melanoma do not respond to immune checkpoint inhibitors (CPIs) and many develop immune-related adverse events (irAEs), often forcing treatment discontinuation, and underscoring the need to predict and monitor outcomes. Responses may depend on both B cell and T cell activity. We performed high-dimensional mass cytometry profiling of coexisting peripheral B cell and key T cell states in treatment-naïve patients and healthy individuals, and paired longitudinal samples from CPI-treated patients, with clinical annotations to define immune correlates of outcomes. CPI-naive patients exhibited reduced CD19+ B cells, reduced B cell (CD21, IL-2, CXCR5) and T cell (CD38, CD27) activation markers, alongside enriched naïve (CD21lo) and double-negative (DN2)-like B cells, CD95+IL-10+plasmablasts, consistent with extrafollicular responses. Concurrently, programmed cell death protein 1 (PD-1)+ and proliferation marker protein-67 (Ki67)+ T cell expansion indicates ongoing activation with features of proliferative exhaustion. Active disease featured increased regulatory CD95 expression on B cells and expanded T follicular helper-like and activated DN (CD4-CD8-) T cells, indicating sustained antigen stimulation. Pretreatment, elevated PD-1+ T cells predicted irAEs, whereas VEGF (vascular endothelial growth factor)+TGF-β (transforming growth factor-β)+ DN T cells were enriched in patients without subsequent toxicity. Pretreatment, plasmablasts, transitional B cells, Forkhead box protein P3 (FoxP3)+ and central memory-like CD8+ T cells correlated with worse overall survival; naïve CD21lo B cells, PD-1+CD8+ T cells and CD4+follicular helper-like T cells predicted shorter event-free survival; CD4+ memory T cells predicted better prognosis, implicating dysregulated differentiation and sustained activation in adverse outcomes. On-treatment, naïve CD21hi B cells, plasmablasts, activated CD8+ and central memory-like CD4+ T cells expanded, indicating de novo humoral responses and cytotoxic T cell invigoration. On-treatment, increased class-switched memory (IgG2+) B and activated T cells predicted improved survival, while persistent naïve and DN B cells were associated with poorer outcomes. Anti-PD-1 monotherapy expanded naïve (CD21hi) B cells and global T cells. Anti-PD-1/anti-LAG-3 (lymphocyte-activation gene 3) combination contracted memory B cells. Melanoma displays aberrant peripheral B and T cell activation, maturation and exhaustion, prominent in active disease. Treatment-induced class-switched B cells and T cell invigoration predict clinical benefit and naïve/DN B cells signify resistance. Coordinated B and T cell responses, especially recurrent extrafollicular B cell and exhausted/regulatory T cell states emerge as candidate indicators of outcome.
[This corrects the article DOI: 10.3389/fimmu.2026.1796161.].
Rivaroxaban (RIV), an oxazolidinone derivative, is an oral anticoagulant used in the prevention and treatment of arterial and venous thromboembolic conditions. However, its oral administration is limited by its poor solubility and bioavailability. The current study aims to load this drug (RIV) into novel transdermal microneedle patches (MNPs) and assess the solubility-enhanced RIV inclusion complex (RIV-IC) as an alternative delivery approach. Scanning electron microscopy (SEM) confirms the presence of uniform, sharply tipped microneedles with a smooth morphology, and differential scanning calorimetry (DSC) indicates good drug-polymer compatibility. The permeation capabilities were analysed using a Franz diffusion cell across rat skin, which demonstrated efficient skin penetration and drug release (∼85% for RIV and ∼94% for RIV-IC within 90 minutes). Insertion capacity was confirmed using Parafilm and rat skin. In vivo pharmacodynamic studies in rats showed significant anticoagulant activity. Histological studies also demonstrated successful microneedle insertion through the stratum corneum without vasculature damage. Overall, these findings indicate that the RIV- and RIV-IC-loaded microneedle patches represent a promising transdermal drug delivery system with enhanced therapeutic efficacy and serve as a viable alternative to oral anticoagulants for the management of thromboembolic disorders.