Over the past two decades there have been remarkable advances in stem cell biology, bioengineering, and lung regenerative research, transforming our understanding of pulmonary biology from development to repair, and disease. Strategies using endogenous lung progenitor cells, pluripotent stem cell technologies, and engineered tissue platforms have become central tools for interrogating lung biology. Major breakthroughs have included the identification of diverse cell populations that coordinate lung homeostasis and repair, facilitated by the extensive adoption of single cell, multiomic and spatialomics approaches. Simultaneous progress in biomaterials, organoid systems, decellularized lung scaffolds, and lung-on-chip platforms has uncovered how extracellular matrix composition, mechanical forces, and tissue architecture contribute to the regulation of cell fate and function. These advances have enabled increasingly physiologically relevant in vitro, and ex vivo models while informing tissue engineering strategies aimed ultimately at functional lung replacement. Translation toward the clinic has advanced through both cell-based and cell-free therapeutic strategies. Early efforts focused largely on mesenchymal stromal cell-based approaches and extracellular vesicles, which have demonstrated safety and context-dependent efficacy in inflammatory lung diseases, alongside emerging preclinical evidence of functional engraftment of induced pluripotent stem cell-derived lung lineages. The past twenty years of progress, captured at the 20th Anniversary Stem Cells, Cell Therapies, and Bioengineering in Lung Biology and Diseases Conference, highlights the power of interdisciplinary collaboration in advancing lung regeneration from foundational discovery toward therapeutic reality.
Particulate matter (PM) in the air, classified as PM2.5 and PM10, enters the body through the nose and mouth during breathing and reaches the lungs. PM is linked to respiratory damage, lung cancer, and the activation of immune system cells, including inflammation and metastasis. Previously, our work group showed that PM upregulates the expression of adhesion molecules and their ligands in lung cancer cells (A549), favoring their adhesion to monocytes; however, its role in cancer progression remains unclear. Therefore, this study examined whether PM exposure promotes a pro-migratory behavior and cell death in A549 cells and monocytes (U937). PM uptake was detected in cells by transmission electron microscopy (TEM) and flow cytometry; cell migration was measured by wound-healing and transwell assays in cocultures of U937 and A549 cells; and cell death was measured by Annexin-V-FLUOS/propidium iodide-mediated flow cytometry. In A549 cells, both PM exposures led to cellular uptake, morphological changes, and increased cell migration. In cocultures of U937 and A549 cells, treatment of both cells with 10 μg/cm2 of PM2.5 and PM10 resulted in the most significant increase in monocyte migration, and PM10 had the maximum effect compared to PM2.5. High PM concentrations (30 μg/cm2) induced necrosis in A549 cells, with rates of 17.7% for PM2.5 and 63.3% for PM10. Therefore, these results suggest that PM exposure in lung cancer cells and monocytes could promote an inflammatory environment and tumor cell progression.
NK cells are promising candidates for adoptive cell therapy; however, their proliferative capacity and functional persistence may be limited. Genetic modification with hTERT enhances their proliferative potential, while co-expression of the iCASP9 suicide gene provides a safety mechanism based on late-stage apoptosis induction by chemical dimerizer (CID). Whether hTERT overexpression interferes with iCasp9-mediated cell death remains unclear, and the non-canonical functions of telomerase in this context are poorly understood. This study served a dual purpose: to assess the efficacy of the iCasp9 "suicide switch" in NK cells, and to investigate a non-canonical role of telomerase in NK cell-mediated evasion from cell death. Here, we demonstrate that hTERT-modified NK cells exhibit significant resistance to CID-induced apoptosis, an effect independent of telomerase catalytic activity, as confirmed using a dominant-negative hTERT (DN-hTERT) mutant. Transcriptomic profiling revealed that both CID-resistant iCasp9-NK cells and hTERT-iCasp9-NK cells share common gene expression signatures: upregulation of cell cycle-associated genes and downregulation of splicing-related factors, including HNRNPH1 and SNRPD3, accompanied by shared patterns of alternative splicing. Among apoptosis-related transcripts, BIRC3, which encodes c-IAP-2, a direct inhibitor of caspase 9, was consistently elevated in both "resistant" and "survived" NK cells. However, shRNA-mediated knockdown of BIRC3 failed to restore sensitivity to CID, indicating that BIRC3 upregulation is not the unique determinant of resistance and suggesting involvement of additional compensatory pathways. Overall, our findings define specific transcriptional signatures associated with evasion of NK cells from iCasp9-mediated apoptosis, implying the contribution of cell cycle progression, enhanced anti-apoptotic signaling, and alterations in splicing regulation, and highlighting the complex role of non-canonical hTERT functions in these adaptations. In the rational design of next-generation gene-modified NK cell therapies with improved safety and persistence, the uncovered insights should be considered.
IgG4-related disease (IgG4-RD) is a systemic fibroinflammatory disorder characterised by tissue infiltration of IgG4+ plasma cells, yet the precise roles of B cells remain unclear. This study aimed to characterise the longitudinal immune dynamics in IgG4-RD during B-cell depletion therapy to identify pathogenic cell subsets and specific biomarkers associated with relapse. We performed a longitudinal multiomics analysis on 119 peripheral blood samples from patients with IgG4-RD undergoing rituximab treatment, spanning active, remission, and relapse phases. Integrated single-cell transcriptomics, B-cell receptor (BCR) sequencing, and serum proteomics were performed on matched blood and tissue samples from submandibular gland and lymph nodes, with Sjögren's syndrome and healthy individuals as controls. We identified a previously unrecognised, disease-specific IgG4+ plasma cell subset characterised by aberrant expression of the neuropeptide NMU and highly skewed IGHV1-69 usage. Circulating follicular helper T cells (CD4+Tfh) displayed signatures consistent with chronic antigen stimulation and type 2 immunity. Furthermore, markedly expanded double-negative T cells (dnT) highly expressed IL10, suggesting their involvement in immune regulation and isotype class switching. Longitudinal analysis following B-cell depletion emphasised the reconstitution dynamics of pathogenic plasma cells, furthermore highlighting the concurrent shifts in CD4+Tfh and dnT that mirrored disease remission and relapse, and underscored the amelioration of proinflammatory immune profiles. NMU-expressing IGHV1-69+ plasma cells represent a pathogenic effector population driving IgG4-RD recurrence. These findings establish a mechanistic framework for B-cell depletion and identify specific cellular and molecular biomarkers to improve disease monitoring and relapse prediction in clinical practice.
The chromatin organizer SATB1 is indispensable for thymic regulatory T cell (Treg cell) development and T helper cell induction. Several gene loci have been described to be SATB1-controlled, including the transcription factor GATA3 and the cytokine loci IL-4 and IL-17. However, the global effects of SATB1 on fully differentiated human CD4 conventional T cells (Tconv cells) and Treg cells, and thus the potential of SATB1 as a target for T-cell engineering, are poorly understood. Here, we describe SATB1-regulated gene signatures as largely subset-specific, with broader effects on Treg cells. Despite distinct gene-regulatory patterns, we observe overarching dysregulated cytokine and JAK-STAT signaling after SATB1 ablation. Functionally, SATB1 KO reduces suppressive capacities of human Treg cells but boosts tumor clearance via CD4 CAR T cells in a preclinical, humanized mouse model. Taken together, Treg destabilization and simultaneous increased activation of CD4 CAR T cells by SATB1 modulation may be a strategy to boost the efficiency of CAR T cell therapies.
Natural killer (NK) cells are key for tumor immune defense. Tissue-resident NK subsets often differ from classical CD16+ NKs and can be immunosuppressive. The exact traits and mechanisms of these tissue-resident NKs in the tumor microenvironment (TME) of non-small cell lung cancer (NSCLC) are still unclear. This study aimed to comprehensively analyze NK cells in the NSCLC tumor microenvironment to identify distinct tissue-resident subsets, assess their clinical relevance, and investigate their functional properties and underlying mechanisms in shaping immunosuppression. We identified CD49a+ NK cells as a distinct tissue-resident subset in non-small cell lung cancer surgical specimens. Single-cell RNA sequencing revealed that this subset is associated with poor pathological response to neoadjuvant therapy. The abundance of CD49a+ NK cells negatively correlates with the formation of tertiary lymphoid structures (TLS). These cells exhibit an altered phenotype, characterized by upregulated immune checkpoint genes, downregulated cytotoxicity-related genes, and uniquely elevated expression of CSF-1. This CD49a+ NK cell subset is functionally linked to driving M2 macrophage polarization, and M2 polarization tends to inversely correlate with TLS density. Our findings indicate that CD49a+ NK cells contribute to the induction of M2 macrophage differentiation within the tumor stroma and are unfavorable for TLS formation. Our findings identify a specific CD49a+ tissue-resident NK cell subset that fosters an immunosuppressive microenvironment in NSCLC by impairing TLS formation and driving M2 macrophage polarization, a mechanism that likely contributes to adverse responses to neoadjuvant therapy.
Chimeric antigen receptor T-cell (CAR-T) therapy was initially used to treat B-cell malignancies, and it is now considered an effective treatment option for multiple sclerosis (MS). CAR-T therapy selectively targets and depletes pathogenic B cells within lymphoid tissue and the central nervous system (CNS), showing promise for achieving deep, sustained remission and long-term treatment-free disease control in patients with refractory MS. A comprehensive analysis was carried out by searching multiple keywords with combinations such as "CAR-T", "MS", "Demyelination", "Autoimmunity", "CD19", "Inflammation", "B cells", T cells", "Neurodegeneration ", "Neurological Disorders", "Immunity", etc. The review included preclinical and clinical research articles publicly available till March 2026. This study was conducted to explore the mechanisms of action, clinical effectiveness, safety profile, and prospects for CAR-T treatment for MS. From the beginning clinical testing indicates that CD19 targeted CAR-T cells can efficiently and permanently destroy through B-cells, leading to a significant decrease in disease progression, a recovery of impairment, and an immense reduction in inflammatory markers in individuals who have progressive MS. New techniques for engineering such as allogeneic CAR-T cells and enhanced CRISPR-based safety switches, are being investigated for making things safer and easier for individuals. CAR-T treatment represents a revolutionary approach for individuals with refractory MS. With ongoing improvements in safety and specificity, it has the potential to transform the therapeutic paradigm toward a sustainable immunological reset and prolonged remission in clinical neuroimmunology.
Progress in immuno-oncology has been stymied by poorly immunogenic 'cold' tumours and the focus on T cells at the cost of other immune cells. Here we report that AT-1965, a small molecule in a lipid nanoparticle, induces rapid regression of poorly immunogenic tumours with the formation of immune memory through interaction with Cap-specific RNA (nucleoside-2'-O-)-methyltransferase 2 (CMTR2) in cancer cells, triggering an innate inflammatory viral defence response. AT-1965-treated tumours were found to be highly infiltrated with B cells, which are known to act as early responders to viral signatures. Genetic knockout of functional B cells abrogated the anti-tumour efficacy of AT-1965, directly implicating B cells in the anti-tumour outcome. Our results rationalize clinical data showing that patients with high CMTR2 expression in tumours have a poor prognosis and that B cell infiltration is associated with long-term survival in multiple tumour types. The AT-1965 nanomedicine-inspired discovery of CMTR2 as a potential cancer target and B cell recruitment opens a new frontier for immuno-oncology.
Patients with lipopolysaccharide-responsive beige-like anchor protein (LRBA) deficiency typically suffer from severe B cell dysfunction. However, the underlying mechanisms remain incompletely understood. In this study, we identify non-muscle myosin IIA (NMIIA) as an interaction partner of LRBA in B cells, and uncover a role for LRBA in regulating actin cytoskeleton dynamics during B cell activation. LRBA-deficient B cells exhibit abnormal migration, impaired F-actin polymerization, and reduced B cell receptor signalling and polarization upon activation. In addition, LRBA deficiency severely disrupts immune synapse formation as evidenced by diminished central SMAC formation, reduced microtubule organizing center translocation and disrupted BCR and lysosome polarization. Consistent with these defects, internalization of the BCR-antigen complex is also impaired. Mechanistically, NMIIA activation, assessed by myosin light chain (MLC) phosphorylation, is reduced in LRBA-deficient cells. In addition, LRBA co-localizes with active NMIIA during both migration and immune synapse formation. Collectively, our findings establish LRBA as an important regulator of cytoskeleton dynamics during B cell activation, which may contribute to the defective humoral immunity observed in LRBA-deficient patients.
Fibroadipogenic progenitors (FAPs) play a key role in skeletal muscle homeostasis and regeneration. They produce extracellular matrix components and secrete cytokines that regulate muscle stem cell function. The number of FAPs and their activity need to be dynamically regulated to avoid their chronic accumulation and overproduction of fibrosis. However, the intrinsic factors by which FAPs control their cell fate decisions remain elusive. Here, we show that FAPs-secreted prostaglandin-E2 (PGE2) functions as a key autoregulatory factor. Using lipidomics and single cell transcriptomics, we show that FAPs are a main cellular source of PGE2 in resting muscle and during regeneration. FAP-secreted PGE2 exerts paracrine effects that maintain the muscle stem cell pool at steady state and stimulate their proliferation post-injury. Moreover, it functions as an autocrine regulator of FAP fate decisions (apoptosis) and subpopulation dynamics. Acute or chronic administration of non-steroidal anti-inflammatory drugs that inhibit the prostaglandin-synthesizing enzyme COX2 increases FAPs content post-injury. Using Pdgfrα-CreERT2_Cox2flox mice, we showed that conditional ablation of COX2 specifically in FAPs increases FAPs content, fibrosis, and impairs muscle regeneration, which can be rescued by PGE2 administration. Furthermore, we show that PGE2 production in FAPs is impaired in mouse models of Duchenne muscular dystrophy, and we provide a proof-of-concept that PGE2 administration can reduce FAPs numbers, fibrosis accumulation, and increase muscle strength. Overall, we uncover a novel role for PGE2 beyond its function in inflammation and identify a mechanism by which FAPs intrinsically regulate their own cell fate, revealing therapeutic potential for muscle injury and dystrophic conditions.
Immunosuppression greatly drives oral squamous cell carcinoma (OSCC) progression and impairs patient prognosis, so exploring microbial regulators of the tumor immune microenvironment carries essential research value. While oral bacteria are tightly associated with worsening OSCC, it remains poorly understood how critical periodontal bacteria reshape immune cell composition to support tumor expansion. Here, we demonstrate that the common periodontal pathogen Porphyromonas gingivalis (P. gingivalis, P. g) stimulates cancer cells to secrete C-C motif chemokine ligand 20 (CCL20). This key signaling protein specifically recruits immunosuppressive regulatory T cells (Tregs). This bacterium elevates immune regulatory molecules, strengthens local immune suppression, and accelerates tumor growth in experimental OSCC models. Clinically, abundant P. gingivalis relates to advanced disease and poorly prognosis, and blocking C-C chemokine receptor type 6 (CCR6)-the immune cell receptor for CCL20-reverses the tumor-promoting effect. This study establishes a novel immune-mediated connection between periodontal infection and OSCC progression, and provides promising molecular targets to improve future prevention and targeted treatment for OSCC.
The spatial interactions between malignant and immune cells in the tumor microenvironment are important for tumor immunobiology and patient outcomes. However, analytical tools that can extract rigorous yet interpretable spatial features and link them to patient outcomes remain limited. We propose a framework integrating TDA with statistical approaches to extract interpretable spatial features characterizing malignant-immune interactions. We introduce Topological Malignant Region (TopMR), which uses topological persistence to automatically define regions of malignant cells, providing an objective reference for spatial analysis even when tumor boundaries are ambiguous. Global-scale infiltration is quantified using signed distances from immune cells to the TopMR boundary and local-scale interactions are captured via malignant cell density around individual immune cells. These global and local features are integrated into a unified signed distance-density (sDD) space, enabling comprehensive characterization of spatial patterns. We apply this framework to high-resolution multiplex immunofluorescence images of diffuse large B-cell lymphoma, analyzing both malignant-enriched and tumor border regions. Two-stage hierarchical clustering stratifies patients based on spatial interaction patterns, revealing associations with survival outcomes. This framework provides an end-to-end pipeline from spatial feature extraction to clinical interpretation, suggesting how region-aware spatial analysis can capture biologically meaningful patterns linked to patient survival.
Regeneration of the dentin-pulp complex is essential for tooth integrity and function. However, the inherent cell heterogeneity limits our understanding of lineage-specific subsets critical for efficient odontogenesis and regenerative outcomes. Here, we demonstrated that CD24+ human dental papilla cells (hDPCs) exhibit robust odontogenic differentiation capacity and drive coordinated regeneration of well-vascularized pulp and structurally integrated dentin tissues in both ectopic murine and preclinical in situ minipig models, significantly outperforming conventional dental pulp stem cells. Mechanistically, we delineate a BMP2/SIRT1 axis where elevated BMP signaling sustains SIRT1 expression and promotes mitochondrial metabolism and odontogenic capacity. Furthermore, BMP signaling induces VEGF expression, enhancing neovascularization via paracrine effects. CD24 is also a downstream marker of BMP signaling, though it does not directly mediate differentiation. Together, CD24+ hDPCs represent a regeneration-competent subpopulation that integrates mitochondrial metabolism and signaling crosstalk to enable coordinated dentin-pulp regeneration, representing a translationally relevant cell source for dental tissue engineering.
The evolutionary transition from unicellular to multicellular organisms exhibited a sharp increase in metabolic rates, yet the physical conditions supporting this transition remain poorly understood. Here we propose that wide-spread hydrodynamic enhancement of nutrient delivery to the microorganisms in the Neoproterozoic Era constituted a necessary enabling condition for early multicellularity, complementing the effect of rising oxygen levels occurring at the same time. Along with conceptual arguments in support of this hypothesis, we define three testable hydrodynamic mechanisms (viscous-diffusive, viscous-convective, and inertial-convective) controlling nutrient uptake and largely applicable for both bed-attached and free-floating microorganisms. We also highlight a flow-bed interface in surface streams as potentially important hotbed for multicellularity breeding, complementing the mainstream perception on the defining role of early oceans. Overall, our results emphasize hydrodynamics as under-represented physical component in existing evolutionary frameworks and generate testable predictions linking organism size, flow conditions, and habitat type.
Idiopathic inflammatory myopathies (IIMs) are autoimmune disorders defined by persistent muscle inflammation, fibrosis, and frequent resistance to current therapies. However, the mechanisms perpetuating disease activity despite immunosuppressive treatment remain elusive. Here, we describe a novel role for tissue-resident stromal cells, specifically fibro-adipogenic progenitors (FAPs), in sustaining skeletal muscle inflammation. Utilizing single-nucleus and spatial transcriptomics in 24 IIM patients and six non-diseased controls, we describe how FAPs adapt to their tissue context, favoring T-cell-centric programs in T-cell environments and myeloid programs in macrophage environments. At the spatial level, FAPs form inflammatory niches by co-localizing with muscle stem cells and activated macrophages, positioning them to participate in cell-to-cell communication with both immune and muscle cells. Trajectory and ligand-receptor analyses suggest a dual-input mechanism whereby infiltrating immune cells (via TGF-β) and myofibers (via epidermal growth factor (EGF)) converge on the AP-1 transcription factor to drive FAP differentiation toward a pro-inflammatory and pro-fibrotic phenotype. Mechanistically, exposure of primary human FAPs to TGF-β and EGF induces a primed state by altering the accessibility to AP-1 regulatory elements. Together, our findings reveal a previously unrecognized role of tissue-resident stromal cells in IIM, highlighting microenvironmental cross-talk centered on FAPs as a promising and actionable therapeutic target.
Stretch-mediated tissue expansion is commonly used to grow extra skin for reconstructive surgeries. To ensure harmonious growth, the two main skin compartments, the epidermis and the dermis, must both expand in a coordinated manner. How fibroblasts respond to stretch-mediated tissue expansion in supporting keratinocyte proliferation remains unclear. Here we map the fibroblast transcriptional response to stretching in vivo and demonstrate that stretching forces fibroblasts to exit their quiescent state and restart proliferation. Concurrently, fibroblasts reduce collagen production and upregulate extracellular matrix remodelling factors, adopting a more embryonic-like program. Because embryonic fibroblasts are widely used as feeder layers to support the expansion of epidermal stem cells for clinical application, we leveraged this model to show that a low collagen state enhances epidermal stem cell self-renewal, thereby coordinating epidermal and dermal responses during skin expansion. These findings provide valuable insights to guide the design of in vivo stretch-mediated tissue expansion protocols and the production of in vitro skin grafts for clinical application.
Multiple myeloma (MM) is a common hematological malignancy, while the prognostic value of tumor-infiltrating immune cells in MM remains elusive. This study aimed to construct an immune-related prognostic model and identify potential therapeutic targets for MM. RNA-seq and clinical data of 751 newly diagnosed MM patients were analyzed. LASSO regression was applied to establish an immune pathway-based prognostic model for patient risk stratification. WGCNA was used to screen hub genes, and in vitro and in vivo experiments validated gene functions and therapeutic effects. Five immune cell pathways were significantly correlated with MM prognosis. MCM2 was identified as the key hub gene associated with risk scores. MCM2 knockdown induced G2-phase cell cycle arrest and suppressed MM proliferation both in vitro and in vivo. Moreover, MCM2 inhibition enhanced the antitumor efficacy of PD1/PDL1 inhibitor BMS1, and CDK inhibitor PHA767491 sensitized MM to immunotherapy. The immune-based model reliably predicts MM prognosis. MCM2 serves as a vital prognostic biomarker. Targeting MCM2 combined with PD1/PDL1 and CDK inhibitors represents a promising therapeutic strategy for MM.
Alternative splicing is essential for the production of diverse messenger RNAs from a single gene. However our understanding of alternative splicing and its regulation during processes such as osteoblast differentiation remains incomplete. Using differentiating hMSC-TERT4 cells as a model, we performed deep short- and long-read RNA sequencing, revealing a highly integrated multiphasic differentiation program. Analysis for splicing revealed extensive changes during lineage commitment, in genes associated with known transcriptional regulators including RUNX2, TEAD1, CTNNB1 and NFATC4. During late-stage matrix osteoblast differentiation (maturation), splicing changes occurred in genes encoding cytoskeletal and extracellular matrix proteins, and Golgi and vesicle trafficking proteins. Analysis of proteomic and phosphoproteomic profiles confirmed the presence of alternative splicing in proteins participating in these biological processes as well as in RNA splicing and autophagy, with splicing involving a variety of protein domains, regions and PTM sites. Together, the transcriptomic and protein splicing landscapes provide a comprehensive description of how mesenchymal stromal cells progressively acquire an osteoblastic phenotype.
Many antiphage defence systems directly bind a specific phage-encoded protein that acts similar to a pathogen-associated molecular pattern to activate an immune response. Such activation is often assumed to occur independent of host factors. Here we demonstrate that the antiphage defence protein CapRelEbc, a fused toxin-antitoxin system from Enterobacter chengduensis, senses the T7 phage-encoded protein Gp0.4 in complex with the host bacterial factor FtsZ, an essential cell division protein. During T7 infection, Gp0.4 sequesters monomeric FtsZ to block its polymerization and thereby inhibit bacterial cell division. Only the complex of Gp0.4-FtsZ, but neither protein alone, triggers CapRelEbc activity. Structural modelling and hydrogen-deuterium exchange mass spectrometry indicate that Gp0.4, FtsZ and CapRelEbc form a ternary complex that activates phage defence. Our work suggests that activation of bacterial immune systems does not always depend exclusively on phage-encoded triggers. Instead, activation can involve host factors targeted by phages, analogous to how eukaryotic innate immune systems detect pathogen-induced perturbations of host cells through effector-triggered immunity.
In this study, we report the synthesis and comprehensive characterization of heparin-capped carbon dots (Hep-C-dots) prepared using D-glucose as a carbon precursor and heparin, a negatively charged polysaccharide belonging to the glycosaminoglycan family, as a capping and stabilizing agent. The obtained Hep-C-dots exhibited a uniform nanoscale size distribution with an average diameter of 2.5 ± 0.5 nm and a high negative surface charge (- 36.8 mV). Full characterization of as-synthesized Hep-C-dots has been done by several state-of-the-art analytical techniques, such as transmission electron microscopy (TEM), X-ray diffraction (XRD), UV-visible spectroscopy, Fourier-transform infrared (FTIR) spectroscopy, and zeta potential analysis. The interactions between Hep-C-dots and key human proteins, namely human methemoglobin (HB) and human serum albumin (HSA), were investigated via fluorescence spectroscopy, demonstrating significant binding affinities with Ksv values of 2.61 ± 0.5 × 107 M- 1 for HSA and 1.83 ± 0.4 × 107 M- 1 for HB. Cytotoxicity assays performed on A549 lung cancer cells revealed that Hep-C-dots exhibit slightly higher toxicity compared to bare carbon dots (C-dots), with IC50 values of 176.21 µg/mL and 200.4 µg/mL, respectively. Moreover, hemolysis assessment using bovine red blood cells (RBCs) showed that Hep-C-dots induce negligible hemolysis (0.002% at 200 µg/mL), confirming their excellent hemocompatibility. These findings suggest that Hep-C-dots hold promise as biocompatible nanomaterials for biomedical applications.