Yang Cao

Mesenchymal stem cells (MSCs) exhibit dynamic immunomodulatory capacity, transitioning between immunosuppressive and immunosupportive states in response to microenvironmental cues. However, the molecular signals that regulate this functional plasticity across diverse MSC populations remain poorly defined. Here, we report that monocyte chemoattractant protein-1 (MCP-1) promotes a T-cell-supportive phenotype in selected MSC populations (such as MSCs from bone marrow, adipose tissue, pancreas, and gingiva). In CTX-induced immunosuppressed mice, MCP-1-pretreated selected MSCs protected T cells from apoptosis and partially restored lymphoid organ atrophy. In vitro, MCP-1 inhibited IFN-γ binding to TLR2 and promoted CXCL10 expression in LNMSCs, supporting T-cell survival and proliferation. SPR confirmed dose-dependent MCP-1–TLR2 binding, and molecular dynamics simulations provide a computational structural rationale for this interference. These findings identify MCP-1 as a promoter of T-cell-supportive function in selected MSC populations, with responses dependent on tissue source and cellular context.

Background: Osteoporosis is a systemic skeletal disorder characterized by reduced bone strength and increased fracture risk. Conventional evaluation relies mainly on bone mineral density and microarchitecture, but these measures do not fully capture the tissue-level material properties that contribute to fragility. Here, we integrated Raman-derived compositional information with microarchitectural, local mechanical, and single-cell transcriptomic data to map a multi-scale coupling landscape and identify a conserved compositional fingerprint of osteoporotic trabecular bone. Methods: Trabecular bone alterations were profiled by Raman spectroscopy in murine models of natural aging and ovariectomy (OVX)-induced osteoporosis. A linear support vector machine (LSVM) classifier was trained for automated phenotyping of Raman spectra. Raman-defined spectral features were then integrated with micro-CT-based microarchitectural measurements, nanoindentation-derived local mechanical properties, and single-cell RNA sequencing (scRNA-seq) of bone marrow mesenchymal stem cells (BMMSCs) to contextualize compositional changes across structure, mechanics, and remodeling programs. Results: We identified a conserved osteoporotic Raman fingerprint characterized by reduced phosphate and collagen signals and increased lipid-associated bands. These compositional signatures were strongly associated with micro-CT-defined structural deterioration and nanoindentation-derived local mechanical alterations, specifically reduced hardness and increased elastic modulus. Furthermore, scRNA-seq revealed shifts in BMMSC transcriptomic programs related to mineral, extracellular matrix, and lipid metabolism that paralleled the Raman-defined changes. The OVX model further confirmed the etiological robustness of this Raman fingerprint in capturing multi-scale alterations in bone quality. Conclusions: Raman-based compositional fingerprinting provides a multidimensional readout that can be integrated with structural imaging, mechanical testing, and transcriptomic profiling. This cross-scale framework refines osteoporosis evaluation, supports the development of advanced diagnostic strategies, and offers mechanistic insight into bone fragility beyond conventional structural metrics.

Aim: Regeneration of tubular dentine structure is key to its biological function, and the polarity of odontoblasts is crucial for this, but the mechanism is unclear. On the basis of differential gene expression data comparing odontoblasts and donor‐matched osteoblasts, we hypothesized that forkhead box Q1 (Foxq1), a key regulator in embryonic development, plays a significant role in the differentiation of polarized odontoblasts. This study aimed to investigate the role of Foxq1 in odontoblast polarization and tubular‐dentine formation, and to explore its relationship with Wingless‐type family member 5A (Wnt5a) signalling. Methodology: We examined Foxq1 's spatiotemporal expression in tooth germs from embryonic to postnatal stages and studied its effect on tooth development by local Foxq1 level manipulation in mesenchymes. Dental papilla stem cells (DPSCs) were isolated from Embryonic 14.5 (E14.5) mouse embryos to evaluate osteogenic, odontogenic, and polarization marker expression and further in vitro studies. The interaction between Foxq1 and Wnt5a was assessed by co‐immunoprecipitation and surface plasmon resonance for protein–protein interaction, and dual‐luciferase reporter assays and chromatin immunoprecipitation assay for protein‐gene regulation. These findings were subsequently validated using an in vivo cell‐inoculated scaffolds subcutaneous implantation animal model. Results: Foxq1 was expressed in dental mesenchyme, and its overexpression promoted dentine thickness and odontoblastic polarization, whereas suppression reduced tooth germ size and disrupted polarity and dentine formation. WNT5A protein reversed these changes caused by Foxq1 suppression. We proved Foxq1 could bind to the promoter region of Wnt5a and dentine sialophosphoprotein ( Dspp ). In vivo, both Foxq1 + DPSCs and Wnt5a + DPSCs promoted regeneration of de novo tubular‐dentine‐like structures on the basis of original tubular dentine. However, where there is no access to tubular dentine opening, only irregular “osteodentin” was formed. Conclusions: The activation of the Foxq1‐Wnt5a signalling axis synergizes with acellular dentine scaffolds to promote tubular dentine regeneration.

Temporomandibular joint osteoarthritis (TMJ‐OA) is a progressive degenerative disorder, for which therapeutic interventions remain limited. The disruption of metabolic homeostasis plays a critical role in the pathogenesis and advancement of TMJ‐OA. However, it remains unclear whether extracellular vesicles (EVs) as cellular metabolites are correlated with the pathogenesis, treatment and diagnosis of TMJ‐OA. In this study, we demonstrated that autologous circulating extracellular vesicles (C‐EVs) possessed significant therapeutic potential for TMJ‐OA through the targeted removal of senescent chondrocytes. In a randomized clinical trial (ChiCTR2200063153), C‐EV administration was found to significantly enhance condylar bone regeneration and alleviate symptoms relative to hyaluronic acid controls, without eliciting any adverse effects. Comparative analysis revealed that joint cavity‐derived EVs from TMJ‐OA patients (OA‐EVs) exhibited structural abnormalities, diminished expression of canonical EV markers, and pro‐inflammatory characteristics. In contrast, C‐EVs were significantly enriched with functional proteins C1q binding protein (C1QBP). And the level of C1QBP‐positive EVs was positively correlated with therapeutic outcomes, thereby establishing C1QBP as a potential predictive biomarker for TMJ‐OA. Furthermore, C‐EVs reestablished joint homeostasis by regulating the immune microenvironment and tissue regeneration capacity. Mechanistically, C1QBP ^high C‐EVs upregulated the expression of membrane C1q on senescent chondrocytes, thereby initiating C1q–C1QBP binding, p14ARF translocation to mitochondria, and subsequent cytochrome C/caspase‐3‐dependent apoptosis. Our findings demonstrate that C‐EVs serve a dual therapeutic role by facilitating the clearance of senescent cells via the C1QBP/C1q/p14ARF axis, while promoting tissue regeneration and regulating metabolites homeostasis, offering a novel biological strategy for TMJ‐OA treatment.

Aim: To identify novel non‐syndromic hereditary gingival fibromatosis (nsHGF)–associated pathogenic variants and discover therapeutic targets for innovative, minimally invasive therapies. Materials and Methods: Whole‐genome sequencing was performed to identify the pathogenic variant in a family with nsHGF. Levels of fibrosis markers and the yes‐associated protein/transcriptional coactivator with PDZ‐binding motif (YAP/TAZ) in gingival fibroblasts were measured by qPCR, western blot and immunofluorescence. Conditioned medium from gingival fibroblasts was used to treat THP‐1‐derived macrophages. In vivo pro‐fibrotic behaviour of CHYMASE ‐silenced gingival fibroblasts and verteporfin‐loaded exosome efficacy were evaluated in NOD/SCID mice. Results: We identified a novel CHYMASE ( CMA1 ) nonsense mutation (c.114C>A, p.Tyr38*) in the nsHGF family. This mutation caused chymase deficiency in the patient's gingival fibroblasts, directly leading to extracellular matrix (ECM) overproduction through YAP/TAZ activation. Moreover, CHYMASE ‐silenced gingival fibroblasts promoted interleukin‐6 (IL‐6) secretion by macrophages, thereby amplifying pro‐fibrotic responses in gingival fibroblasts. The YAP inhibitor verteporfin suppressed ECM overproduction in CHYMASE ‐silenced gingival fibroblasts. In vivo, topical administration of verteporfin‐loaded exosomes significantly attenuated chymase deficiency–induced fibrosis. Conclusion: Our findings support the pathogenic role of the CHYMASE mutation in nsHGF, establish chymase deficiency and consequent YAP/TAZ activation as the underlying mechanism and propose verteporfin‐loaded exosomes as a promising therapeutic strategy for nsHGF‐associated gingival overgrowth.

Objectives Balancing the aesthetic and mechanical effects remains challenging in terms of attachments. This study proposed a strategy by introducing laser surface texturing in clear aligners fabrication to enhance the retention and grip of clear aligners and minimize the size of attachments. Materials and methods A grid pattern with a spacing of 1 mm on the surface of thermoplastic polyurethane substrate was textured using a CO₂ laser. We used LST-modified TPU to fabricate aligners and evaluated its mechanical and surface properties. Results TPU after laser surface texturing exhibited increased roughness, and therefore enhanced the retention and gripping force of LST-modified clear aligners. Although the average transmittance of the LST-modified TPU was 2.32% lower, LST had little negative effect on aesthetic of aligners. Conclusions Laser surface texturing, as a novel auxiliary approach in aligners fabrication, has the potential to enhance mechanical properties such as roughness, retention and force delivery of TPU-fabricated clear aligners. Clinical relevance The mechanical enhancement of LST-modified clear aligners could have a better control on teeth movements and reduce the sizes of attachments without sacrificing their mechanical effect, helping to balance the aesthetics and mechanic effects of attachments.

Dental caries is a prevalent biofilm-related oral infectious disease, in which Streptococcus mutans plays an important role in the pathological process. The discovery of novel agents with inhibitory effects on S. mutans is essential to develop effective strategies against dental caries. Plant extracts are a rich source of novel chemicals for the discovery of more effective antimicrobial agents. Coptis chinensis Franch. is a traditional Chinese herbal medicine with various pharmacological properties. This study aimed to evaluate the antimicrobial activity of berberine (BBR), a bioactive alkaloid isolated from Coptis chinensis Franch., on the biofilm formation of S. mutans and to investigate the relevant mechanism of its action. The study found that BBR inhibited the growth of planktonic S. mutans , as demonstrated by Minimum Inhibitory Concentration, Minimum Bactericidal Concentration, and growth curve assay. BBR was found to inhibit biofilm formation and exhibit antimicrobial effects on S. mutans , as demonstrated by crystal violet staining assay, confocal laser scanning microscopy, and CFU counting assay. Moreover, the inhibitory effect of BBR on the virulence of S. mutans was evaluated by the anthrone-sulfuric method and lactic acid assay. Bacterial aggregation assay, membrane potential assay, and transmission electron microscopy were used to preliminarily elucidate the membrane-destructive bactericidal mechanism of BBR. The expression of virulence genes gtfB , gtfC , gtfD , ldh , vicR , liaR , and comD was measured using quantitative real-time PCR (qRT-PCR). The cell viability assay on human oral cells and macrophages demonstrated that BBR exhibited no cytotoxicity to the tested host cells. In conclusion, the study demonstrates that BBR has significant anticaries activity against S. mutans and has the potential to be further developed as a novel anticaries agent for clinical use. IMPORTANCE: Dental caries is a chronic disease affecting people worldwide, which could cause a series of oral problems. Plant-derived compounds have shown antimicrobial properties against a wide range of pathogens. This study investigated the antimicrobial effect of BBR on Streptococcus mutans , including the inhibitory effect on the planktonic bacteria growth, biofilm formation, virulence factors, and explored the mechanism and biocompatibility of BBR. This study demonstrated the significant anticaries activity of BBR against S. mutans and its potential to be further developed as a novel anticaries agent for clinical prevention and treatment of dental caries.

Craniomaxillofacial sutures play a critical role in craniomaxillofacial development through continuous bone reconstruction and regeneration, processes modulated by mechanical tension. Bone suture stem cells (SuSCs) are central to these functions. Distraction osteogenesis, which promotes craniomaxillofacial suture growth, is a common therapeutic approach for craniofacial deformities. However, the underlying mechanisms by which mechanical forces drive suture and bone remodeling remain poorly understood, posing significant clinical challenges. To investigate these mechanisms, we established a rapid maxillary expansion (RME) model in mice to widen the midpalatal suture. Single-cell RNA sequencing (scRNA-seq) was employed to identify subsets of SuSCs responsive to mechanical tension and analyze their differentiation potential under varying conditions. Further functional studies were conducted to explore the role of DALR anticodon binding domain containing 3 (Dalrd3) and its associated tRNA 3-methylcytosine (m3C) modification in SuSCs under mechanical tension. Our study identified a subset of SuSCs with multidirectional differentiation potential that shifted from a chondrogenic to an osteogenic trajectory in response to mechanical tension. Mechanical tension also upregulated Dalrd3 expression and its associated tRNA m3C modification in activated SuSCs. Knockdown of Dalrd3 in SuSCs significantly impaired osteogenic differentiation, proliferation, migratory capacity, and translational activity within the bone morphogenetic protein (BMP) signaling pathway. Furthermore, Dalrd3 knockdown suppressed the translational activity of inhibitor of DNA binding 3 (Id3), a key BMP-induced mediator of osteoblastogenesis. Restoring Id3 expression in Dalrd3-deficient SuSCs rescued their osteogenic, proliferative, and migratory functions. These findings reveal a translational regulatory mechanism in SuSCs activated by mechanical tension and underscore the pivotal role of Dalrd3 in suture remodeling and bone formation. The insights provided by this study have the potential to guide targeted therapeutic strategies for optimizing distraction osteogenesis and other treatments for craniofacial deformities.