Search Results (149)

Adult hematopoietic stem cells (HSCs) and bone marrow (BM) mesenchymal stem/stromal cells (MSCs) are essential for lifelong hematopoiesis, skeletal homeostasis, immune competence, and tissue regeneration. The use of electronic cigarettes (E-cigs) among women of reproductive age continues to rise, raising concerns about potential adverse developmental effects; however, the long-term consequences of maternal E-cig vaping on offspring BM stem cell function and hematopoietic homeostasis remain incompletely understood. Here, using a rat model of maternal E-cig exposure (containing nicotine) during gestation, combined with longitudinal in vivo analyses and complementary ex vivo studies of human cells, we show that prenatal E-cig exposure is associated with persistent alterations in offspring BM stem cell function and lineage commitment. Gestational E-cig exposure was associated with expansion of the CD11b/c⁺ myeloid-enriched compartment, increased CD90⁺ stromal cells, and impaired osteogenic differentiation in rat offspring. Complementary experiments using primary human cells showed that nicotine exposure was associated with reduced T-cell proliferation and impaired cytotoxic activity in a proof-of-principle co-culture assay. Mechanistically, transcriptomic profiling followed by Gene Ontology and pathway enrichment analyses identified alterations in molecular programs associated with KLF4–Notch1 signaling, mitochondrial biogenesis, inflammation, and stem cell regulation in the BM of E-cig-exposed rat offspring. Changes in CCL11, FTO, and RUNX2 were additionally associated with an inflammatory and aging-related molecular phenotype that persisted from early life into adulthood, although these findings do not establish a causal CCL11–FTO–RUNX2 signaling axis or direct cellular senescence. Collectively, our study provides a phenotypic and mechanistic framework for understanding how maternal E-cig exposure may influence long-term offspring hematopoietic, skeletal, and immune health while highlighting the need for further studies to establish causal molecular mechanisms and determine their relevance to maternal E-cig use in humans. Full article

Advancing age is often accompanied by a gradual decline in musculoskeletal (MSK) health and integrity, making the skeleton not just a structural framework; but a dynamic indicator of biological ageing. Bone tissue undergoes continuous remodelling through tightly regulated interactions among osteocytes, osteoblasts, osteoclasts, progenitor cells or mesenchymal stem/stromal cells (MSCs) and growth factors. With advancing age, this balance shifts toward increased resorption, increased adipose tissue formation, reduced bone formation, and deterioration of bone microarchitecture. This eventually can lead to frailty, increased fracture risk and other age-related diseases (ARDs) due to ageing, cellular senescence and inflammaging. Interestingly, bone-derived changes often precede clinical manifestations of these ARDs, suggesting its potential use as an early biomarker for biological ageing and longevity risks. In our review, we explore the possibility of using the bone as a biomarker for ageing, followed by anti-ageing strategies to encourage bone longevity. We begin by discussing our current knowledge of the bone, the bone marrow (BM), changes in the BM with ageing and current methods of tracking bone ageing; both clinically and experimentally. We then explore studies that investigated the anti-ageing strategies, specifically focused on diet, exercises and lifestyle to shift towards enhanced bone health and longevity. Finally, we discuss the limitations of achieving this, what the future of this field may look like and what methods may be used to translate the idea of bone as a biomarker of ageing, from bench to bed. Full article

Cardiovascular diseases (CVDs) remain the foremost cause of death globally, responsible for 19.2 million deaths and 437 million disability-adjusted life years in 2023, with prevalent cases having more than doubled since 1990. No approved therapy restores myocardium lost to infarction. The adult heart replaces cardiomyocytes at approximately 1% per year in young adults, declining to about 0.45% per year with ageing, far below what is needed to recover the more than one billion cells destroyed by a large myocardial infarction. Cell-based regenerative strategies have been investigated for more than two decades, encompassing bone marrow mononuclear cells (BM-MNCs), mesenchymal stromal cells (MSCs), cardiac progenitor cells, cardiosphere-derived cells (CDCs), skeletal myoblasts, and induced pluripotent stem cell-derived cardiomyocytes (iPSC-CMs). Safety has been consistent. Efficacy has been modest and variable: the CADUCEUS trial demonstrated scar mass reduction with CDCs without proportionate ejection fraction improvement; the Phase 1/2 POSEIDON trial confirmed MSC safety in 30 patients; and the Phase 3 DREAM-HF trial, enrolling 537 patients, failed its primary endpoint (HR 1.2, p = 0.406). Mechanistic work has established that transplanted cells engraft poorly and exert their benefit principally through paracrine signalling mediated by secreted extracellular vesicles and exosomes carrying microRNAs, trophic factors, and immunomodulatory proteins. For iPSC-CMs, electrophysiological immaturity and arrhythmogenic risk in primate models remain unresolved barriers. Emerging strategies include CRISPR-engineered hypoimmune iPSC lines, bioengineered cardiac patches, injectable hydrogel scaffolds, and engineered exosome platforms. This review provides a comprehensive synthesis of preclinical and clinical evidence, examines translational barriers, and identifies the scientific and regulatory priorities required before these therapies can enter routine clinical practice. Full article

This in vitro study investigates the effects of therapeutic magnetic resonance (TMR^®), a novel biophysical stimulation technology, on mesenchymal stromal cells (MSCs) to support its potential application in regenerative medicine. The role of pulsed electromagnetic fields in bone healing is already established. We evaluated whether TMR^® influences MSC proliferation, differentiation, and immunomodulatory properties in vitro. Bone marrow-derived MSCs (BM-MSCs) and adipose-derived MSCs (AD-MSCs) were cultured with or without TMR^® exposure and assessed through flow cytometry, karyotype analysis, senescence assays, proliferation tests, gene expression analysis, and osteogenic differentiation assays. TMR^® did not alter MSC phenotype, proliferation, senescence, or genomic stability, confirming its safety profile. Notably, treated MSCs showed enhanced osteogenic differentiation, with increased early expression of key osteogenic markers (RUNX2, ALP, and COL1A1) and greater collagen deposition compared to untreated controls. TMR^® reduced peripheral blood mononuclear cell proliferation and MSC ROS production, suggesting anti-inflammatory and antioxidative effects. Overall, TMR^® appears to be a safe, non-invasive stimulus able to promote osteogenic differentiation, supporting its potential clinical application in bone regeneration. Further in vitro studies and clinical trials are needed to confirm these findings. Full article

Stage-specific embryonic antigen-3-positive pluripotent-like/macrophage-like multilineage-differentiating stress-enduring (Muse) cells are a distinct subpopulation of mesenchymal stromal cells (MSCs), accounting for 1% to several percent of MSCs. Although stem cell therapy for spinal cord injury (SCI) typically targets the subacute phase to avoid the hostile acute environment, the therapeutic window for Muse cells remains unclear. C57BL/6J mice with severe T9 contusion SCI received a single tail vein injection of human bone marrow-derived (BM) Muse cells, BM-MSCs (both 5 × 10⁴ cells), or vehicle at 2, 8, 14, or 28 days post-injury (DPI) without immunosuppressants. Among the different administration time points, the 2-DPI Muse cell group exhibited significantly higher Basso Mouse Scale scores than the BM-MSC and vehicle groups from 14 days after injection, while no significant differences were observed at the other administration time points. The 2-DPI Muse cell group showed significantly greater homing to the injured spinal cord than the BM-MSC group, with persistent engraftment and neural-lineage marker expression at day 42. Ablation of engrafted Muse cells at day 42 partially reversed locomotor recovery, suggesting that engrafted Muse cells contributed to functional recovery. These findings suggest that intravenous Muse cell therapy exerts timing-dependent therapeutic effects after SCI, with greater efficacy during the early post-injury phase. Full article

The development of reusable and biocompatible biomaterial-based culture substrates is increasingly relevant for improving sustainability in biomedical research workflows. In this study, polyethylene terephthalate glycol-modified (PETG) was evaluated as a potential alternative to conventional polystyrene (PS) for in vitro cell culture applications. PETG substrates were fabricated through laser cutting and tested for their ability to support cell adhesion, viability, proliferation, and lineage-specific differentiation across multiple human cell models, including calvarial mesenchymal stromal cells (CMSCs), bone marrow-derived mesenchymal stromal cells (hBM-MSCs), dermal fibroblasts, LHCN-M2 myoblasts, and SH-SY5Y neuroblastoma cells. Morphological and immunofluorescence analyses demonstrated that PETG supported cell attachment and focal adhesion formation, comparable to standard PS surfaces. Cell viability and proliferation assays confirmed metabolic activity and growth over time. Furthermore, PETG substrates supported osteogenic, adipogenic, myogenic, and neuronal differentiation, as demonstrated by histological staining, myotube formation, neurite outgrowth, and lineage-specific gene expression analyses. Finally, PETG maintained CMSC morphology and metabolic activity after repeated recovery, ethanol/UV treatment, and gelatin re-coating, with comparable results between new substrates and those reused for up to three cycles. These findings support PETG as a biocompatible culture substrate with preliminary short-term reuse potential and possible sustainability benefits for laboratory workflows. Full article

Background: Osteochondral lesions of the talus (OLT) remain a challenging condition due to the limited regenerative potential of articular cartilage. Conventional bone marrow stimulation (BMS) techniques often result in fibrocartilage formation with inferior biomechanical properties. This study aimed to evaluate the safety and preliminary clinical efficacy of an arthroscopically assisted, single-stage injection of a heparin-conjugated fibrin hydrogel (HCFH) for OLT treatment. Methods: Twelve patients with symptomatic OLT underwent arthroscopic debridement, microfracturing, and HCFH injection containing autologous mesenchymal stromal cells (MSCs) and growth factors. Safety was assessed through systematic monitoring of adverse events (graded according to Common Terminology Criteria for Adverse Events criteria), wound healing, and serial laboratory inflammatory markers (leukocytes, erythrocyte sedimentation rate, C-reactive protein) during early and late follow-up. Clinical outcomes were evaluated using the Visual Analog Scale (VAS) and American Orthopedic Foot and Ankle Society score (AOFAS) preoperatively and at 6 and 12 months. Morphological assessment was performed using magnetic resonance imaging (MRI) with the modified Magnetic Resonance Observation of Cartilage Repair Tissue (MOCART) scoring system, evaluated independently by two blinded musculoskeletal radiologists. Results: No serious adverse events (Grade III–IV) were observed during the 12-month follow-up. All adverse events were mild (Grade I) and self-limited. A transient postoperative elevation in inflammatory markers was observed, returning to clinically acceptable levels by day 14. Significant improvements were noted in pain (VAS decreased from 6.0 to 2.0) and ankle function (AOFAS increased from 70.0 to 90.6) (p < 0.001). MRI demonstrated progressive morphological improvement, with the MOCART score increasing from 34.16 ± 17.1 at 6 months to 75 ± 5.43 at 12 months (p < 0.001). This increase corresponded with imaging features consistent with tissue maturation over time. The favorable MOCART outcomes observed in this study may be explained by the regenerative properties of heparin-conjugated fibrin hydrogels; however, larger randomized controlled trials with longer follow-up are needed to confirm the durability of the regenerated tissue. Interobserver agreement was substantial to almost perfect for MOCART scoring (κ = 0.68–0.84), with perfect agreement observed for surface assessment, bony defect/overgrowth, and cysts. Conclusions: Within the limitations of this study, single-stage HCFH injection demonstrated an acceptable safety profile and favorable preliminary clinical and radiological outcomes at 12 months. These findings suggest potential regenerative capability; however, controlled studies with larger cohorts and longer follow-up are required to determine comparative efficacy and long-term durability. Full article

Mesenchymal stromal cell (MSC) adhesion and retention at sites of cartilage degeneration are critical for improving cartilage repair. This study investigated whether platelet-rich plasma (PRP) enhances the adhesion and short-term retention of bone marrow-derived MSCs (BM-MSCs) and chondrocytes under in vitro and ex vivo conditions. BM-MSCs and chondrocytes were treated with PRP or pretreated with PRP for 10 or 30 min, and cell adhesion to collagen-coated surfaces was evaluated using a cell viability assay. Ex vivo adhesion and short-term retention of BM-MSCs on osteochondral discs with varying lesion severity were assessed by fluorescence imaging analysis. PRP significantly enhanced the adhesion of both BM-MSCs and chondrocytes in a time-dependent manner, with the 30 min PRP pretreatment group showing the greatest effect. BM-MSC attachment in the 30 min PRP pretreatment group was significantly higher than that in the untreated control group after 30 min of incubation (p < 0.001), whereas chondrocyte attachment was also significantly increased following PRP pretreatment. In addition, PRP pretreatment significantly enhanced BM-MSC attachment compared with PRP treatment alone at 20 and 30 min of incubation (both p < 0.001). In ex vivo experiments, adhesion and short-term retention increased significantly with increasing lesion severity from G1 to G3 (p < 0.05 and p < 0.01, respectively). In G2 and G3 lesions, PRP pretreatment for 30 min significantly enhanced BM-MSC adhesion and short-term retention compared with the control group (both p < 0.01). These findings suggest that PRP may improve the early adhesion and retention of MSCs on damaged cartilage and support the potential use of PRP as a biological adjunct for MSC-based cartilage repair strategies. Full article

(1) Background: Cerium silicate (CeSi) nanoparticles (NPs) have potential as a restorative filler particle with multifunctional properties to improve longevity. To increase the biological activity, these nanoparticles can be fabricated with ultrasmall pores (mesoporous) (MPCeSi-NP) that can be loaded with a polyphosphate inhibitor, such as gallein. (2) Methods: MPCeSi-NPs were custom-synthesized with a microemulsion method, using cetyltrimethylammonium bromide (CTAB) as a template for self-assembly. Biocompatibility with oral keratinocytes/fibroblasts was tested, with the addition of examining the biomineralization potential with human bone-marrow-derived mesenchymal stromal cells (BM-MSCs). MPCeSi-NP, loaded with gallein, was tested against Rothia dentocariosa (Rd). MPCeSi-NP was added to a resin matrix of triethylene glycol dimethacrylate (TEGDMA) and Bisphenol A-glycidyl methacrylate (BisGMA) with subsequent mechanical properties evaluation. (3) Results: MPCeSi-NPs had high biocompatibility with oral keratinocytes and fibroblasts, especially at concentrations below 300 µg/mL. MPCeSi-NPs induced the biomineralization of BM-MSCs. Higher cerium levels increased mineralization. MPCeSi-NP had weak antimicrobial activity against Rd. At 1% wt, MPCeSi-NPs did not reduce the polymerization potential and mechanical properties of a TEGDMA:BisGMA polymer material, with controlled release of gallein in a simulated degradation model. (4) Conclusions: MPCeSi-NPs are highly biocompatible and bioinductive and have the potential to improve the biological response of current restorative materials. Full article

Background: Advanced glycation end products (AGEs) and oxidative stress (OS) have been linked to bone complications related to type 2 diabetes mellitus (T2DM). However, the effects of AGEs and OS on bone marrow mesenchymal stromal cells (BMMSCs), which play a key role in bone homeostasis and repair, remain unclear. Objectives: This study aimed to investigate the effects of AGEs on BMMSCs function and the ability of N-acetylcysteine (NAC) to alleviate AGE-induced OS in a T2DM context. Methods: Bone marrow (BM) and BMMSCs were isolated from Zucker diabetic fatty (ZDF) rats, which serve as a T2DM model, and their lean littermates (ZL, controls) at 24 weeks of age. Results: The results show that long-standing T2DM leads to changes in the BM’s cellular composition and BMMSCs function that are distinct from age-related changes. In vitro, AGEs decreased BMMSCs viability, proliferation, and migration. The effects of AGEs were stronger in BMMSCs derived from a T2DM microenvironment. In both T2DM- and ZL-BMMSCs, AGEs induced cytoplasmic ROS, which was differentially reduced by NAC. The effect of NAC on T2DM-BMMSCs was greater when the cells were pre-treated with NAC 24 h before exposure to AGEs, whereas simultaneous exposure to both resulted in a smaller effect. Conclusions: These results show that AGEs impair BMMSCs expansion and functionality. AGE-induced ROS generation may be a critical factor in this impairment, while NAC was able to reduce OS in BMMSCs from a T2DM context. These findings highlight the vicious negative effects of the T2DM microenvironment on BMMSCs and underscore the need for further studies to better understand the underlying mechanisms and to explore strategies aimed at mitigating OS in the T2DM context. Full article

Mesenchymal stromal cells (MSCs) have shown immunomodulatory effects and great promise in many inflammatory diseases such as acute respiratory distress syndrome (ARDS). However, several barriers to translation remain such as cell availability and potency. This study evaluates the therapeutic potentials of three types of MSCs, bone marrow-derived MSCs (BM-MSC), the human induced pluripotent stem cell-derived MSC wild type (iMSC WT) and β2 microglobulin-knockout iMSCs (iMSC B2M KO) with or without proinflammatory cytokine preconditioning. BM-MSC, iMSC WT and iMSC B2M KO were preconditioned with a proinflammatory cytokine cocktail (Cytomix: IL-1β, IFN-γ and TNF-α). Immunoregulatory biomarkers were analysed by flow cytometry and cytokines released by ELISA. MSC antimicrobial properties were analysed via CFU assays while the MSCs’ immunomodulatory effects were evaluated using macrophage activation and T cell proliferation assays. Proinflammatory cytokine preconditioning enhanced the therapeutic potency of all three types of MSCs by increasing immunomodulatory marker expression, enhancing the antimicrobial effects and improving MSC-mediated inhibition of T cell proliferation. These findings provided new insights into the therapeutic potencies of MSCs in inflammation. Further studies are required for in vitro characterisation of the MSCs and in vivo efficacy verification of these MSCs prior to their clinical application. Full article

Articular cartilage, a highly specialised and avascular tissue, exhibits limited regenerative potential following trauma or degenerative conditions such as osteoarthritis (OA). Conventional surgical interventions, including microfracture and autologous chondrocyte implantation (ACI), have shown limited long-term efficacy due to donor site morbidity and restricted cell proliferation. In this context, mesenchymal stromal cells (MSCs) have emerged as a promising alternative owing to their multipotency, self-renewal capacity, and low immunogenicity. While bone marrow (BM) remains the traditional source of MSCs, recent studies have reported that peripheral blood-derived mesenchymal stromal cells (PB-MSCs) may possess chondrogenic, osteogenic, and adipogenic potential comparable to that of BM-derived MSCs. PB-MSCs can be harvested through minimally invasive methods, thereby avoiding the complications associated with BM aspiration. Experimental evidence indicates that PB-MSCs exhibit strong cell viability, proliferative potential, and the ability to synthesise cartilage-specific extracellular matrix proteins, such as type II collagen and sulphated glycosaminoglycans, within three-dimensional scaffolds. Immunophenotypically, PB-MSCs express mesenchymal markers including CD29, CD44, CD90, and CD105 while lacking hematopoietic markers CD34 and CD45. Flow cytometry analyses reveal that CD105⁺ populations increase following cryopreservation, highlighting their clinical utility. In contrast to these experimentally defined PB-MSCs, the term peripheral blood stem cells (PBSCs) is used in clinical studies to describe heterogeneous, non-cultured peripheral blood-derived cell preparations, typically enriched in hematopoietic stem and progenitor cells following granulocyte colony-stimulating factor (G-CSF) mobilisation, without full mesenchymal characterisation. In vitro studies confirm successful tri-lineage differentiation, whereas in vivo investigations have demonstrated effective cartilage regeneration using PB-based clinical approaches, including postoperative intra-articular administration of hyaluronic acid (HA) combined with PBSCs, as well as implantation of PBSCs covered with a collagen membrane. Furthermore, advancements in biomaterial engineering, such as poly(ethylene glycol)–cysteine–arginine–glycine–aspartic acid (PEG-CRGD) hydrogels, have enhanced PB-MSC adhesion, proliferation, and chondrogenic differentiation while promoting immunomodulation through M2 macrophage polarisation. Despite these promising outcomes, the available evidence remains limited and heterogeneous, with substantial variability in cell definitions, experimental models, and clinical study designs, which currently constrains definitive conclusions regarding clinical efficacy. Future research should focus on optimising isolation protocols, understanding molecular pathways governing PB-MSC chondrogenesis, and standardising clinical applications. Overall, PB-MSCs represent a viable, less invasive, and translationally relevant cell source for cartilage regeneration and regenerative orthopaedic therapies Full article

Background/Objectives: B-cell precursor acute lymphoblastic leukemia (B-ALL), the most common pediatric acute leukemia (AL), is frequently characterized by aberrant antigen expression, which aids diagnosis and prognosis. The myeloid antigen CD66c is notably frequent in B-ALL and has been proposed as a marker of disease aggressiveness and treatment response. Evaluating CD66c in Mexican pediatric patients may provide insights into disease biology. Methods: A cohort of 128 pediatric patients was referred to the Laboratory of Oncoimmunology and Cytomics of Childhood Cancer (OCL) at Instituto Mexicano del Seguro Social (IMSS) for immunophenotyping tests between March 2022 and November 2023. Additionally, control bone marrow (BM) samples were assessed. Aberrant antigen expression in hematopoietic populations and BM microenvironment stroma phenotyping were performed. Results: In total, 84.38% of B-ALL patients exhibited aberrant expression of ≥1 myeloid antigen. Among CD66c-positive patients, 13.79% had detectable measurable residual disease (MRD) during follow-up and 20.69% died. Mesenchymal stromal cells (MSCs) from patients with positive or low CD66c expression displayed inflammatory profiles. ProB leukemias with low CD66c expression were more likely to exhibit detectable MRD, increased mortality, and reduced survival. Conclusions: Low CD66c expression induces molecular stealth that could favor immune evasion and niche persistence, thereby increasing the risk of relapse and therapeutic failure. Full article

Bone marrow mesenchymal stem/stromal cells (BM-MSCs) are important components of bone marrow, possessing multipotent differentiation potential and the ability to support hematopoiesis. Exposure to ionizing radiation (IR) induces cellular damage in BM-MSCs, such as DNA lesions and mitochondrial dysfunction. Despite their relative radioresistance, most surviving BM-MSCs enter senescence post-irradiation. This senescent state disrupts the bone marrow niche, impairs stem cell proliferation and differentiation, and contributes to acute radiation syndrome (ARS) and myelosuppression. To clarify the impact of IR on BM-MSCs, this review systematically summarizes the general mechanisms of radiation-induced cellular senescence, examines the effects of different radiation types (e.g., gamma rays, X-rays, and heavy-ion radiation) and doses on BM-MSCs senescence, and outlines senotherapeutic strategies targeting BM-MSCs senescence. The analysis indicates that the senescence of BM-MSCs caused by IR is type- and dose-dependent. The review identifies key factors in IR-induced BM-MSCs senescence to guide targeted interventions, highlighting the need for future studies to elucidate the underlying mechanisms of IR-induced BM-MSCs senescence. Full article

Biomorphic hydroxyapatite scaffolds derived from rattan wood (GreenBone) show significant promise in bone tissue engineering due to their inherent structural similarity to natural bone. Laser-drilled GreenBone scaffolds were studied for enhanced porosity, nutrient diffusion, cellular infiltration, and vascularisation. Patient-derived bone marrow mesenchymal stromal/stem cells (BMMSCs) and culture-expanded mesenchymal stem cells (cMSCs) demonstrated high cell viability (>90%), considerable adhesion, and extensive cytoskeletal organisation. Trilineage differentiation confirmed the multipotency of BMMSCs, with osteogenic, adipogenic, and chondrogenic markers being successfully expressed. BMMSCs and cMSCs exhibited enhanced differentiation and gene expression profiles. At week 4, key osteogenic and angiogenic genes such as BMP2, VEGFC, RUNX2, and COL1A1 showed elevated expression, indicating improved bone formation and vascularisation activity. Markers associated with extracellular matrix (ECM) remodelling, including MMP9 and TIMP1, were also upregulated, suggesting active tissue remodelling. ELISA analysis for VEGF further demonstrated increased VEGF secretion, highlighting the scaffold’s angiogenic potential. The improved cellular response and vascular signalling emphasise the translational relevance of laser-modified GreenBone scaffolds for bone tissue engineering, particularly for critical-sized defect repair requiring rapid vascularised bone regeneration. Full article