Search Results (6,160)

Background: Konjac (Amorphophallus spp.) is an economically important crop valued for the glucomannan content in its corms. Currently, the konjac industry faces germplasm degeneration due to long-term asexual propagation. Developing tissue culture and genetic transformation techniques is essential for its genetic improvement. The WUSCHEL-related homeobox (WOX) transcription factors are critical regulators of somatic embryogenesis and stem cell maintenance in plants. Methods: In this study, we performed genome-wide identification and characterization of WOX genes in the A. konjac reference genome. Furthermore, comparative transcriptomic analyses and functional verification were conducted in A. bulbifer. Results: A total of 12 AkWOX genes were identified in A. konjac, and their structural features were documented. Comparative transcriptomic analysis of A. bulbifer revealed that AbWOX genes were differentially expressed between embryogenic calli (EC) and non-embryogenic calli (nEC). Notably, AbWOX2 was significantly upregulated in EC. Overexpression of AbWOX2 significantly promoted callus proliferation and shoot regeneration in A. bulbifer. Furthermore, AbWOX2-overexpressing lines exhibited a 5.3-fold increase in genetic transformation efficiency (from 5.12% to 27.31%) compared to the control. Conclusions: We characterized the diverse expression patterns of the WOX gene family in Amorphophallus. Crucially, we identified specific individual members—most notably the markedly upregulated AbWOX2—that function as pivotal drivers of somatic embryogenesis and serve as promising candidates for enhancing regeneration and genetic engineering efficiency in Amorphophallus species. Full article

Human liver stem cells (HLSCs) are a mesenchymal stromal cell (MSC)-like population isolated from adult liver biopsies. HLSCs share key characteristics with MSCs, including phenotype and differentiation capabilities. Previous studies have demonstrated that HLSCs promote regeneration in different experimental models of acute and chronic tissue injury and that HLSC-derived extracellular vesicles (HLSC-EVs) recapitulate the therapeutic effects of the cells of origin. This study aimed to determine whether HLSC-EVs, obtained and characterized under good manufacturing practice (GMP) conditions, can influence the progression of liver fibrosis in vivo. The EV production process was carried out under GMP conditions to generate batches of HLSC-EVs by tangential flow filtration. To assess their therapeutic potential, an in vivo model of hepatic fibrosis was established through administration of thioacetamide (TAA). In TAA-treated mice, EV administrations attenuated fibrosis progression. Molecular analyses showed a significant reduction in the expression levels of key pro-fibrotic genes. At the functional level, EV administration resulted in a significant reduction in plasma alanine aminotransferase levels and an increase in albumin levels, indicating improved liver function. These data indicate that HLSC-EVs, produced under GMP conditions, display antifibrotic effects in a chronic liver disease model, leading to improved liver function and histology. Full article

Background: Malignant salivary gland tumors represent a highly diverse group of neoplasms, their heterogeneity likely arising due to variable origin in different tissue components. Emerging evidence suggests that SOX-2 and EZH-2 play critical roles in salivary gland carcinogenesis, being related to tumor cell stemness potential, along with accelerated tumor progression and unfavorable clinical outcomes. The aim of this study was to assess the association between SOX-2 and EZH-2 expression, survival parameters, and tumors’ pathological characteristics in a group of patients with primary epithelial malignant salivary gland tumors (MSGTs) and to evaluate their value as diagnostic and prognostic markers. Methods: Our study group comprised 104 patients with primary epithelial MSGTs diagnosed in “Sf. Spiridon” County Hospital, Iasi, over a period of fifteen years. Pathological parameters and survival evaluation, along with SOX-2 and EZH-2 immunohistochemistry assessment and scoring, were conducted, and the associations between different parameters were analyzed. Results: High SOX-2 immunoexpression was significantly associated with lymphatic invasion (LY) (p = 0.003), pT stage (p = 0.010), histological tumor type (p = 0.003), and tumor grading (p = 0.037), while high EZH-2 immunoexpression was significantly associated with perineural invasion (PnI) (p < 0.001), vascular invasion (p = 0.038), LY (p = 0.001), tumor grading (p = 0.002), and pathological extranodal extension (pENE) (p = 0.018). The tumors with high SOX-2 and EZH-2 expressions were associated with a reduced overall survival (OS) (p = 0.013 and p = 0.011). Cox regression analysis revealed that pT (HR = 1.826, p = 0.019), LY (HR = 0.318, p = 0.007), and tumor grade (HR = 0.505, p = 0.021) added to high SOX-2 and EZH-2 immunoexpression independently predicted a poor survival outcome (HR = 2.373, p = 0.016 and HR = 2.746, p = 0.015). Conclusions: Our findings suggest that SOX-2 and EZH-2 may serve as biomarkers of aggressive behavior and a poor prognosis in primary epithelial MSGTs, providing potential opportunities for precision-targeted therapies. Full article

Cardiac fibrosis is a pathology induced by various conditions, such as myocardial infarction, or certain cardiomyopathies, and represents one of the most prevalent cardiac abnormalities. This process, defined as the excessive accumulation of extracellular matrix within damaged cardiac tissue, leads to significant complications, including impaired systolic and diastolic function as well as arrhythmias. Conventional therapies focus primarily on slowing down the progression of fibrosis. Recently, there has been a growing research interest in therapies based on stem cells and their derivatives, which hold the potential to greater decrease formation and area of fibrosis. In this review, we aim to systematise the most recent data regarding the application of these approaches. We focus on describing the types of cells employed, methods of their implementation, and strategies for optimising these processes. Particular attention is given to exosomes due to the reports highlighting their use as innovative and potentially effective tools in the treatment of cardiac diseases. Full article

Repairing maxillofacial bone defects remains a major clinical challenge due to inadequate vascularisation and poor integration with host tissue. While bioactive scaffolds have shown promise in supporting osteogenesis and angiogenesis, achieving robust and synchronised dual regenerative outcomes is still elusive. This study presents a multifunctional, cell-free magnetic hydrogel platform designed to biomimetically coordinate osteogenic and angiogenic processes for effective maxillofacial bone regeneration. The composite poly(vinyl alcohol)-vaterite (PVA-Vat) hydrogel scaffold incorporates tuneable magnetic nanoparticles (MNPs) composed of single-domain superparamagnetic iron oxide (Fe₃O₄). By harnessing magneto-mechanical cues to orchestrate bilateral communication between human bone mesenchymal stem cells and endothelial cells, this platform provides a deeper mechanistic understanding of coupled tissue regeneration and delivers superior dual-regenerative performance for maxillofacial bone repair. Under magnetic stimulation, a coculture system demonstrated strong osteogenesis-angiogenesis coupling mediated by reciprocal VEGFA-BMP2 signalling. This reciprocal crosstalk was evidenced by a synergistic amplification of VEGFA and BMP2 expression in coculture compared to monocultures, where MNP-stimulated osteoprogenitors secreted VEGFA to drive endothelial capillary-like network formation, while endothelial cells reciprocally enhanced endogenous BMP2 levels to accelerate osteoblastic mineralisation. These findings establish MNP-integrated hydrogels as a cell-free, multifunctional platform capable of synchronising dual regenerative pathways, offering a biomimetic strategy to overcome vascularisation and integration barriers in maxillofacial bone repair. Full article

Animal studies have shown that early life exposure to general anesthetics may impair brain development. However, the implications of this phenomenon in human patients remain unclear. In this study, we use an induced pluripotent stem cell (iPSC)-derived human brain microphysiological system (bMPS) to investigate the effects of early sevoflurane (SEV) exposure on human brain development. Human iPSCs were cultured and differentiated into neural progenitor cells (NPCs) and then into bMPS. At week 8, bMPSs were exposed to 2.4% SEV for 4 h. Four weeks after exposure, immunofluorescence (IF), Western blotting (WB), and quantitative real-time polymerase chain reaction (qPCR) were conducted to evaluate the alteration of nerve cells in bMPS. After SEV exposure, the number of apoptotic cells increases, and the level of neural differentiation markers decreases. The ratios of mature neurons over NPCs and mature oligodendrocytes over oligodendrocyte progenitor cells (OPCs) are reduced, which leads to a reduction in myelination. SEV also impedes the development of astrocytes and synaptogenesis, especially the formation of excitatory synapses. Meanwhile, SEV increases the expression of molecules in the mammalian target of rapamycin (mTOR) signal pathway. In conclusion, early SEV exposure substantially disrupts the development of human brain tissue, and the mTOR signal pathway is likely to be involved in this alteration. Full article

Titanium implants are widely used in orthopedic and dental fields but often face challenges such as insufficient osseointegration and peri-implant inflammation. While Strontium (Sr) possesses potent bioactive properties, achieving its controlled delivery at the implant-tissue interface remains technically challenging. To address this, we engineered a multidimensional composite coating by constructing a micro/nano-porous TiO₂ substrate via micro-arc oxidation (MAO), followed by polydopamine (PDA)-assisted Sr immobilization. This integrated architecture significantly enhanced surface hydrophilicity and facilitated high-content Sr loading with sustained release kinetics. Biological evaluations demonstrated that the PDA-mediated interface promoted superior initial adhesion and spreading of bone marrow mesenchymal stem cells (BMSCs), synergizing with released Sr²⁺ to markedly upregulate core osteogenic markers (Runx2, ALP). Crucially, the functionalized surface actively optimized the immune microenvironment by inducing M1-to-M2 macrophage polarization and comprehensively suppressing RANKL-induced osteoclastogenesis via the downregulation of TRAP and DC-STAMP. By integrating these pro-osteogenic, anti-inflammatory, and anti-resorptive capabilities, this tri-functional system effectively rebalances the bone remodeling microenvironment. Consequently, it provides a robust, universally applicable strategy for enhancing the therapeutic efficacy of next-generation orthopedic and dental implants. Full article

Sechium edule (Cucurbitaceae), commonly known as chayote, which is a cucurbit of economic relevance, has experienced higher incidence of wilting from Phytophthora capsici in Mexican commercial fields during heavy rainfall. The infection process of this oomycete on chayote stems at the anatomical level had not been documented. This study characterized histological changes in chayote stems infected with P. capsici. Plants were inoculated at the stem base with P. capsici mycelial plugs, while controls received sterile plugs. Stem samples collected at 8, 12, 16, 22, and 30 days post-inoculation were processed and stained using safranin O–fast green. Microscopic observations showed progressive anatomical alterations. At 8 dpi, hyphae appeared in cortical parenchyma and epidermis, with phenolic compound accumulation. By 12 dpi, stromata and sporangia were visible in vascular and cortical tissues, with tyloses formation. At 16 dpi, cell wall collapse and xylem colonization became evident. These effects intensified at 22 and 30 dpi, with tissue degradation and an abundance of hyphae. Control stems maintained intact structures. Macroscopically, plants remained asymptomatic until 12 dpi, when brown lesions appeared. By 22 dpi, leaf yellowing and stem necrosis were observed, leading to plant death by 30 dpi. The results demonstrate the rapid colonization of chayote tissues by P. capsici, and its impact on vascular integrity. This study provides knowledge for future research on host resistance and disease management in chayote crops. Full article

Small extracellular vesicles (sEVs) derived from mesenchymal stem cells (MSCs) are emerging as potent acellular therapeutics; however, their rapid clearance hinders their clinical translation. To address this issue, 3D-bioprinted genipin-crosslinked gelatin (GECL) was engineered for human health. GECL hydrogels were functionalised with human umbilical cord MSC-derived sEVs (hUCMSC-sEVs) to create a bioactive wound-healing platform. These hydrogels demonstrated favourable physicochemical, mechanical, and biodegradable properties while providing an extracellular matrix (ECM)-mimetic environment conducive to tissue regeneration. MSCs were isolated from the umbilical cords, and their small extracellular vesicles (sEVs) were extracted and incorporated into gelatin-based hydrogels via 3D bioprinting. These sEV-loaded scaffolds were embedded in full-thickness wounds in mice, and healing was evaluated through macroscopic observation, histological analysis, collagen deposition, and angiogenesis assessment. Compared with the untreated controls, both the hydrogel-only (B) and sEV-loaded hydrogel (BE) groups significantly accelerated in vivo wound healing. Notably, the BE group achieved complete wound closure within 14 days, restoring the skin architecture, which closely resembled the native tissue with well-organised epidermal and dermal layers, optimal thickness, and skin appendages. Histological and ultrastructural assessments revealed an increased collagen type I deposition, a reduced α-smooth muscle actin (α-SMA) expression, and a robust neovascularisation. The TEM revealed tight junctions and active cellular infiltration, indicating scaffold integration and functional remodelling. Immunohistochemistry further revealed an upregulated CD31 expression with a balanced α-smooth muscle actin (α-SMA) expression, reflecting coordinated angiogenesis and myofibroblast regulation. These results highlight sEV-functionalised GECL hydrogels as robust and clinically translatable acellular therapeutic green products for accelerated wound closure and functional skin regeneration, advancing the fields of regenerative medicine and life expectancy. Full article

Adipose mesenchymal stem cell-derived exosomes (ADSC-Exo) have demonstrated therapeutic effects in liver diseases and injuries. The Augmenter of Liver Regeneration (ALR), a novel hepatic trophic growth factor, promotes hepatic structural and functional recovery. In this study, we constructed ALR-overexpressing ADSC-Exo (ADSC-ALR-Exo) by harnessing the messaging capacity of ADSC-Exo, and analyzed the effects of ADSC-ALR-Exo on hepatic ischemia–reperfusion injury (IRI) combined with partial hepatectomy in a minipig model. Our results indicated that, compared to the ADSC-Exo group, the ADSC-ALR-Exo group exhibited a significant reduction in reactive oxygen species (ROS) levels, alongside a notable increase in the activity of antioxidant enzymes superoxide dismutase (SOD) and catalase (CAT). Furthermore, there was a marked decrease in malondialdehyde (MDA) content. Concurrently, the concentrations of pro-inflammatory factors in the blood (IL-1β, IL-18, and TNF-α) and liver tissue (IL-1β, IL-18, IL-6, and TNF-α) were significantly lower in the ADSC-ALR-Exo group, while the level of the anti-inflammatory factor IL-10 in the blood was significantly elevated. Additionally, ALR enrichment enhanced the inhibitory effect of ADSC-ALR-Exo on endoplasmic reticulum stress-related pathways, specifically ATF6, IRE1α, and PERK. Compared to ADSC-Exo, the ADSC-ALR-Exo intervention was also more effective in reducing the expression levels of NLRP3, caspase-1, and GSDMD, thereby decreasing the incidence of pyroptosis. In conclusion, ADSC-ALR-Exo mitigated liver injury by inhibiting endoplasmic reticulum stress and cellular pyroptosis induced by liver injury. Full article

Craniosynostosis is the premature fusion of skull bones due to loss of stem/progenitor cells located in non-mineralized tissue between growing cranial bones of infants. We generated scaffolds from a biodegradable biomaterial with small interconnected pores (125–250 μm diameter), previously shown to maintain stemness of a mesenchymal cell population, to further develop a method for the creation of an artificial cranial bone stem cell niche. Polymer scaffolds of consistent pore size were fabricated using a molecular-sieved sugar sphere casting technique with poly-l-lactic acid. A rectangular surgical defect within the parietal bone of juvenile mice was created. The three groups included sham animals with surgery but no scaffold, experimental animals with surgery plus an implanted cell-free scaffold, and experimental animals with surgery plus an implanted bone mesenchymal cell-seeded scaffold. Healing at the surgical site was evaluated at 4 and 12 weeks after surgery by micro-CT and histology. Surgical site bone volume fraction and bone mineral density were significantly greater at twelve than four weeks in the sham group but not in either of the scaffold groups. At twelve weeks, the surgical site bone volume fraction and bone mineral density were significantly lower in the cell-seeded scaffold as compared to the sham animal group. At twelve weeks, the anterior and middle cranial vault widths were significantly greater in the cell-seeded scaffold as compared to the sham animal group on the surgery side of the skulls. Less mineralization was evident within the cell-seeded than the cell-free scaffolds by histology. Based on these findings, scaffolds of sufficiently small pore size seeded with autologous bone mesenchymal stem cells could function as an artificial cranial stem cell niche to inhibit surgical-site mineralization and promote cranial growth. Full article

Background: Cutaneous radiation injury arises when ionizing radiation disrupts epidermal barrier integrity, triggering persistent DNA damage, oxidative stress, and senescence-associated inflammatory signaling that drive extracellular matrix degradation and impaired regeneration. Clinical burden is rising due to dose-intensified radiotherapy, but also due to an increased use of energy-based aesthetic procedures that elicit radiation-like dermal injury. Dermal fibroblasts exhibit marked sensitivity to ionizing radiation and rapidly acquire senescence-associated secretory phenotypes that suppress collagen biosynthesis and promote chronic inflammation, underpinning the need for regenerative treatments that restore tissue homeostasis and regenerative competence. Mesenchymal stem cell–derived exosomes have emerged as a promising therapeutic strategy in this setting, with increasing preclinical evidence demonstrating their capacity to attenuate oxidative stress, enhance DNA damage-repair pathways, and normalize fibroblast metabolic function. Methods: In this study, we examine the expression profiles for 14 radiation response–associated genes of irradiated human dermal fibroblasts that were treated with bone marrow and umbilical cord MSC-derived exosomes at different timepoints using quantitative RT-PCR analysis. We also explore functional relationships among these genes through interaction network analysis, and outline a framework to organize pathway-level transcriptional responses to irradiation and exosome treatment. Results: MSC-derived exosome treatment was associated with attenuated early damage response signaling at 24 h, followed by increased expression of genes associated with DNA repair and oxidative stress recovery at intermediate timepoints. Exosome-treated cells also exhibited transcriptional changes consistent with modulation of cell-cycle regulatory pathways and reduced expression of pro-inflammatory markers by 5 d. These findings suggest that MSC-derived exosomes influence the temporal organization of the fibroblast transcriptional response to ionizing radiation and may contribute to molecular programs associated with tissue recovery following ionizing radiation exposure. Full article

Aging is a complex biological process characterized by progressive loss of cellular homeostasis, impaired regenerative capacity, and accumulation of senescent cells that collectively predispose tissues to disease. Traditional two-dimensional culture systems and animal models have provided valuable insights but fail to fully recapitulate the spatial organization, cellular heterogeneity, and microenvironmental cues of aging human tissues. Organoid technology—three-dimensional self-organizing structures derived from adult stem cells or pluripotent stem cells has emerged as a transformative platform to model aging in vitro. These mini-tissues retain the architecture, signaling dynamics, and lineage hierarchy of native organs, making them powerful systems to interrogate age-associated cellular phenotypes, DNA damage responses, and senescence programs. This review discusses how organoid models are advancing our understanding of aging biology across multiple organ systems, from the intestines and liver to the brain and lung. We highlighted key molecular pathways driving cellular senescence within organoids—including p16INK4a/p21CIP1 signaling, SASP activation, mitochondrial dysfunction, and epigenetic drift—and how these can be targeted to restore tissue homeostasis. We further discussed how organoids derived from aged tissues, induced pluripotent stem cells, and engineered oncogene systems reveal new therapeutic opportunities to modulate senescence in age-related disorders, cancer, and regenerative medicine. Finally, we discussed emerging integrative tools such as organoid co-cultures, single-cell omics, and senolytics drug screening that are expanding the potential of organoids as translational platforms for anti-aging and disease intervention. Full article

A simple method for the efficient high-throughput transformation of hybrid poplar (Populus tremula x alba clone 717 1B) is described. Factors considered in developing the method included the ease and efficiency of preparing large numbers of explants for transformation, and selection of culture media that enhanced cell and tissue growth while promoting shoot regeneration competence. We found that petiole explants from in vitro-cultured plantlets can be easily collected and prepared for transformation and regenerate shoots comparable to stem or leaf explants. Culturing petiole explants on Driver Kuniyuki Walnut (DKW) medium resulted in significantly greater tissue growth compared to Murashige Skoog (MS) medium. Moreover, the inclusion of low concentrations of thidiazuron (TDZ) in callus-inducing media (CIM) significantly enhanced shoot regeneration competence of cultured petiole explants. As a consequence, the combination of petiole explants cultured on DKW medium along with 2.2 ug/L TDZ during the callus induction phase resulted in rapid and efficient transformation of this hybrid poplar genotype. When applied to genomic approaches such as activation tagging or CRISPR-Cas9 and Cas12a gene editing, we obtained transformation efficiencies ranging between 70% and 90%. The described protocol provides a simple and efficient method that is easily scalable for high-throughput approaches, which could facilitate genome-wide methods for the rapid and efficient production of transformed hybrid poplars. Full article

Chronic wounds represent a significant global healthcare challenge, affecting millions of patients and imposing substantial economic burdens on healthcare systems. Traditional wound management approaches often fail to address the complex pathophysiology underlying chronic wounds, including persistent inflammation, impaired angiogenesis, and disrupted extracellular matrix remodeling. Three-dimensional (3D) bioprinting has emerged as a transformative technology that enables the fabrication of patient-specific, biomimetic tissue constructs capable of addressing these intricate challenges. This comprehensive review synthesizes recent advances in 3D bioprinting for chronic wound treatment, examining bioprinting technologies, biomaterial innovations, mechanisms of wound healing, and clinical applications. Recent studies demonstrate that bioprinted constructs incorporating living cells, growth factors, and bioactive molecules can significantly accelerate wound closure, enhance vascularization, and restore functional skin architecture. Notable innovations include in situ bioprinting systems, photosynthetic scaffolds for oxygen delivery, and immunomodulatory bioinks. While significant technical challenges remain—including vascularization, scalability, and regulatory approval—the integration of advanced bioprinting techniques with regenerative medicine principles offers unprecedented opportunities for personalized chronic wound care and improved patient outcomes. Full article