Journal of Tissue Engineering最新文献

Pancreatic islet transplantation offers great promise for the treatment of type 1 diabetes, yet the functional decline of islets after isolation remains a major obstacle. Increasing evidence highlights the endoplasmic reticulum (ER) as a critical regulator of islet cell survival under stress. We explored how ex vivo culture conditions affect encapsulated islet resilience under ER-stress. Two conditions were assessed: (i) incorporation of decellularized porcine pancreatic extracellular matrix (ECM) into alginate microcapsules, and (ii) free-fall dynamic pre-conditioning culture. Human islets were encapsulated in alginate with or without ECM, cultured under static or dynamic conditions, and exposed to acute ER-stress followed or not by a recovery period. Dynamic culture preserved viability and enhanced glucose responsiveness. ECM-containing capsules showed reduced inflammatory marker expression, while encapsulation in alginate-only capsules led to more pronounced changes associated with ECM remodeling. Under ER-stress, the dynamic culture, especially combined with ECM, maintained cell function and reduced cell death. Gene profiles indicated improved stress adaptation and ECM remodeling. These results highlight ECM enrichment and dynamic culture as good strategies to maintain islet survival and functionality.

Intervertebral disc degeneration (IDD) is a common condition and a leading cause of chronic low back pain, affecting millions of individuals worldwide. Human Umbilical Cord Mesenchymal Stromal Cell (hUCMSC)-derived extracellular vesicles (EVs) are emerging as a promising therapeutic strategy for IDD. However, the limited production yield and unclear mechanisms by which EV contents mediate their therapeutic effects have hindered the clinical application of EVs. In this study, using transcriptomic data and single-cell RNA sequencing, we identify PANoptosis as a key mechanism driving the progression of IDD. Furthermore, parathyroid hormone (PTH) enhances the secretion of hUCMSC-derived EVs and alters their cargo composition, which may contribute to their improved therapeutic effects. Mechanistically, PTH-preconditioned EVs, enriched with ZDHHC5, ameliorate PANoptosis by modulating ZBP1 transcription through competitive inhibition of YBX1 phosphorylation via palmitoylation. Our findings provide strong support for a cell-free therapeutic strategy utilizing EVs from PTH-preconditioned MSCs for IDD treatment and propose the ZDHHC5/YBX1/ZBP1 axis as a novel molecular target for inhibiting PANoptosis, thus paving the way for clinical translation and broader healthcare applications.

Bisphenol A (BPA), a widely used industrial chemical with endocrine-disrupting properties, raises developmental and cardiotoxicity concerns. We established a stage-specific cardiotoxicity platform using human pluripotent stem cell (hPSC)-derived cardiomyocytes in two-dimensional and three-dimensional (3D) cultures. BPA exposure at ⩾10 µM significantly reduced cell viability and downregulated pluripotency and cardiac lineage markers such as OCT4, NKX2-5, and cTnT in a stage-dependent manner. Electrophysiological analysis revealed that acute exposure to 10 µM BPA disrupted action potentials in hPSC-derived cardiomyocytes, inducing membrane depolarization and rhythm disturbances. Furthermore, 3D cardiac tissues treated with 10 or 50 µM BPA exhibited severe mitochondrial deformation and impaired contractile function, as observed by TEM and beating analysis. Reproducing these effects in a personalized hPSC line validated the platform's applicability for patient-specific toxicity assessment. These findings highlight the importance of integrating developmental stage-specific and 3D human-relevant models for comprehensive cardiotoxicity evaluation of environmental chemicals such as BPA.

Three-dimensional engineered muscle tissues (EMTs) are transformative tools for modeling skeletal muscle physiology and pathology in vitro. Here, we perform a comprehensive comparison of EMTs derived from primary human myoblasts (hP-Myo) and hiPS-derived myoblasts (hiPS-Myo) to evaluate their structural, functional, and transcriptional characteristics. Contractile performance was quantified using a magnetic force-sensing platform, revealing that hP-Myo EMTs generate ~2 fold higher twitch forces and enhanced tetanic responses compared to hiPS-Myo EMTs. Tissue architecture and maturation were assessed and demonstrated significant larger myofiber diameters in hP-Myo EMTs. Transcriptomic profiling highlighted that hP-Myo EMTs maintain a mature skeletal muscle-like signature, marked by enriched pathways linked to sarcomere organization and fast-/slow-twitch fiber specification. To model metabolic dysfunction, hiPS-Myo EMTs were subjected to lipid overload, recapitulating hallmarks of intracellular lipid (IMCL) accumulation, including impaired contractility, blunted force-frequency responses, and dysregulated lipid metabolism genes.

The plasticity of blood mononuclear cells (MCs) and their role in vascular remodeling have been the focus of many studies; however, their in vitro differentiation efficiency remains poorly understood. Herein, we demonstrate that the inflammatory response accelerates the efficiency of MCs differentiation into endothelial-like cells through chemical cues in vitro. RT-PCR and RNA sequencing revealed that the differentiated cells exhibited upregulated pathways associated with vascular remodeling and regeneration. In contrast, MCs collected from normal blood showed a differentiation bias toward macrophages. Notably, under inflammatory conditions, primarily monocytes transitioned into the CD14++/CD16+/CD163+ subset, which contributed significantly to vascular remodeling. This transition was triggered by inflammation, as confirmed by in vitro cytokine treatment.

Angiogenesis is essential for successful tissue regeneration, particularly in clinical contexts such as ischemic injury, wound healing, and reconstructive therapies. However, the establishment of functional vasculature remains a major limitation in organoid-based systems. In this study, we developed vascularized organoid tissue modules (Angio-TMs) by incorporating human umbilical vein endothelial cells (HUVECs) into scaffold-free, self-organized constructs. Remarkably, the inclusion of HUVECs at 1% of the total cell population was sufficient to generate highly reproducible and structurally stable Angio-TMs, which exhibited clear endothelial differentiation and vascular functionality both in vitro and in vivo. Furthermore, inhibition of transforming growth factor (TGF)-β signaling in Angio-TMs led to a 2.5-fold increase in vessel length density, demonstrating a substantial enhancement in angiogenic potential. These findings highlight Angio-TMs as a robust and modular platform for engineering vascularized tissues and underscore their translational relevance in regenerative medicine and tissue transplantation.

NK cell mimics are assemblies of a cell membrane and a template that replicate biomimetic features and physicochemical properties, respectively. To develop this targeted drug delivery system, gelatin microspheres (cG) were fabricated using a water-in-oil emulsion and reinforced via DMTMM cross-linking to exhibit tunable Young's modulus, a critical parameter for cell-material interactions. These microspheres were subsequently coated with membranes derived from the human NK cell line KHYG-1 to form biomimetic NK cell mimics (cGCM), combining physicochemical control with bioinspired functionality. These engineered cGCM were non-toxic, non-inflammatory, and capable of reducing macrophage uptake by ~10% when incubated with differentiated THP-1 cells. In vitro studies demonstrated significant interaction/ proximity of the cGCM with cancer cells in 2D cultures of breast cancer cells (MDA-MB-231), 3D spheroids of liver (HepG2), and colon (HT-29) cancer cell models, and a zebrafish breast cancer xenograft (MDA-MB-231) model. The cGCM also evaded macrophage detection in a Kdrl:EGFP Spil:Ds Red zebrafish model. Furthermore, in a pilot assessment, loading and release of the sialyltransferase inhibitor (STI, 3Fax-Peracetyl Neu5Ac) using cGCM significantly reduced α-2,6 sialylation in 2D cultures of MDA-MB-231 cells, demonstrating the STI's intact functionality in inhibiting sialylation. By integrating bioinspired membranes with mechanically tunable gelatin-based carriers, our system demonstrates a multifunctional immune-mimicking platform with relevance to tissue engineering, tumour modelling, immune modulation, and drug delivery. These findings offer a promising foundation for future therapeutic strategies in cancer research and immuno-engineering.

Hair regrowth through mechano-stimulation and other therapeutic approaches has emerged as a significant area of research in regenerative medicine. This review examines recent advances in hair regeneration strategies, with a particular focus on mechanical stimulation and complementary treatments. Studies have demonstrated that skin stretching can activate hair follicle stem cells and promote hair growth under specific conditions and durations. This process involves intricate signaling interactions, particularly through the WNT and BMP pathways, and follows a two-stage mechanism that recruits and modulates the function of macrophages. Mechanical stimulation induces the release of growth factors such as HGF and IGF-1, which activate stem cells and support hair follicle regeneration. Beyond mechanical activation, emerging hair restoration therapies, including MSC transplantation, MSC secretome therapy, and platelet-rich plasma treatments, have shown promising results. These innovative strategies overcome the limitations of conventional therapies, offering effective solutions for various types of hair loss. Additionally, here we discuss the molecular mechanisms underlying hair follicle growth and repair, the influence of external factors, and novel hair follicle formation processes, such as chimeric follicle development and follicular neogenesis. Special attention is given to the roles of dermal papilla cells and their interactions with mesenchymal cells in promoting hair regrowth. The key strategies and underlying mechanisms discussed in this review will drive future research and potential clinical applications.

[This corrects the article DOI: 10.1177/20417314231196275.].

Pathogenic mutations in Bcl2-associated athanogene 3 (BAG3) cause genetic dilated cardiomyopathy (DCM), a disease characterized by ventricular dilation, systolic dysfunction, and fibrosis. Previous studies have demonstrated that BAG3 mediates sarcomeric protein turnover through chaperone-assisted selective autophagy to maintain sarcomere integrity in the human heart. Although mouse models provide valuable insights into whole-organism effects of BAG3 knockout or pathogenic variants, their utility is limited by species-specific differences in pathophysiology, which often do not translate to humans and contribute to the failure of clinical trials. As a result, the development of induced pluripotent stem cell-derived cardiomyocytes (iCM) and engineered heart tissues presents a promising alternative for studying adult-onset cardiac diseases. Here, we used genome engineering to generate an isogenic pseudo-wild-type control cell line from a patient-derived iPSC line carrying a pathogenic BAG3 variant, clinically presenting with DCM. While monolayer iCMs recapitulated some features of BAG3-mediated DCM, such as reduced autophagy, mitochondrial membrane potential, and decreased HSPB8 stability, they failed to develop the age-associated impairment in sarcomere integrity. Therefore, we developed a mature, patient-specific, human engineered heart tissue model of BAG3-mediated DCM and compared it to its isogenic healthy control. We demonstrated successful recapitulation of adult-like features of the clinically observed disorganized sarcomeres and impaired tissue contractility, thereby providing a platform to investigate BAG3-related pathophysiology and therapeutic interventions.