The success of hematopoietic stem cell transplantation (HSCT) relies significantly on the effective mobilization and collection of hematopoietic stem cells. However, various factors can lead to failure in hematopoietic stem cell mobilization in some patients, including age, type of malignancy, chemotherapy regimens (such as fludarabine, Melphalan, and Lenalidomide), radiotherapy, bone marrow cellularity, and comorbidities. This study investigates the impact of Diabetes Mellitus type 2 on hematopoietic stem cell mobilization outcomes in patients undergoing autologous transplantation. We conducted a retrospective analysis of 43 patients, comparing mobilization efficiency, engraftment times, and blood product transfusion requirements between diabetic and non-diabetic cohorts. Mobilization was achieved using granulocyte colony-stimulating factor (G-CSF), with the specific dosing regimen and timing of apheresis detailed in the methods section. Our findings indicate that diabetic patients exhibited significantly lower CD34 + cell counts and required more engraftment time than their non-diabetic counterparts (P < 0.05). Additionally, diabetic patients had higher rates of blood product transfusions post-transplantation. Our findings have shed light on the impact of Diabetes Mellitus type 2 on Hematopoietic Stem Cells mobilization and autologous transplantation, providing crucial insights for the hematopoietic stem cell transplantation field. It is crucial to recognize that diabetes can significantly affect the outcomes of hematopoietic stem cell transplantation. Factors such as poor stem cell mobilization, prolonged engraftment times, and increased need for blood product transfusions underscore the importance of considering diabetes in HSCT candidates.
Intestinal diseases are imposing a prevalent socioeconomic burden worldwide, which is largely attributed to the disrupted homeostasis of gut-immune crosstalk including gut microbiota dysbiosis and immunomodulatory imbalance. Stem cells are unique cell populations with self-renewal and multi-lineage differentiation properties, and current literature has indicated the remarkable applications in early embryogenesis, disease remodeling and novel cytotherapy via diverse modes of action (e.g., direct- or trans-differentiation, cytokine paracrine, and bidirectional immunomodulation). Of them, human pluripotent stem cells (hPSCs) and the derivatives, including the self-organizing organoids, have been vastly employed for intestinal disease remodeling, while mesenchymal stem/stromal cells (MSCs) and the microvesicles are adopted for multifarious intestinal disorder intervention. In this review article, we mainly focus on the progressions in stem cell-based disease remodeling (intestinal lineage and organoid induction from hPSCs) and cellular therapy for intestinal diseases (e.g., inflammatory bowel disease, infectious intestinal diseases, and colorectal cancer), together with the underlying repair mechanisms (e.g., target genes and signaling pathways). Furthermore, the promising prospective and the concomitant challenges of stem cell-based preclinical and clinical investigations upon intestinal disorders are also outlined, including ethical considerations and supervision, heterogeneity and standardization, integration with novel biotechnologies (e.g., single-cell RNA sequencing, intracellular tracing, gene-editing and three-dimensional bioprinting). Collectively, our findings will supply profitable references and facilitate the development of stem cell-based mechanistic investigation and regenerative medicine for intestinal diseases.
Meristems are the growth centers of plants and fundamental in understanding plant development, morphogenesis, and vegetative propagation. Across all plant groups, the phytohormone auxin controls meristem maintenance, represses the emergence of new meristems (apical dominance), and mediates cellular reprogramming when new meristems regenerate following removal of existing meristems. The liverwort Marchantia produces clonal propagules (gemmae) featuring two apical notches that develop into functional meristems. This presents a tractable experimental system to study meristem developmental biology. I used laser ablation microscopy to precisely disrupt cells in and around the developing premeristem in the apical notches of germinating gemma, finding that the first cell row is indispensable. Within this layer, a contiguous quorum of stem cells is required for activity. Apical notches reorientate in response to damage, demonstrating that the apical notch stem cells act as a communicating population. Feedback from the stem cell population is necessary to maintain notch activity and generate the notch apex. These experiments show communication between notches and regenerating meristems. The apical dominance signal represses cell division and requires both sources and sinks, features of auxin-mediated communication. Central regions of the gemma could transmit these apical dominance signals, but the tissues of the gemma periphery could not. I present a model of Marchantia gemma and apical notch organization, involving intra-, inter-, and extranotch communication. This provides a framework for further study of meristem formation, communication, and maintenance in Marchantia and improving knowledge of plant meristems more generally.
Advancements in stem-cell biology and tissue engineering (TE) are transforming the field of oculoplastic surgery by offering innovative solutions for eyelid, orbital, and periocular reconstruction. Stem cells, including mesenchymal stem cells (MSCs), induced pluripotent stem cells (iPSCs), and adipose-derived stem cells (ASCs), provide regenerative potential through their ability to differentiate into ocular adnexal tissues and modulate wound healing via paracrine effects. Tissue-engineered constructs, combining biocompatible scaffolds, cellular components, and growth factors, enable the development of functional replacements for eyelid skin, tarsus, conjunctiva, and lacrimal structures. These technologies enhance esthetic and functional outcomes, reduce donor-site morbidity, and may overcome limitations of traditional grafts and flaps. Current applications include periocular volume augmentation, eyelid reconstruction, and management of anophthalmic sockets, with ongoing research directed toward bioengineered lacrimal glands and dynamic eyelid substitutes. Although challenges remain, such as optimizing scaffold integration, vascularization, and long-term safety, stem-cell and TE approaches hold significant promise for personalized, restorative, and regenerative strategies in oculoplastic surgery.
The routine mass applications of human pluripotent stem (hPS) cell-derived progenies for regenerative medicine or high-throughput drug screenings will depend on the standardized supply of high-quality cells via controlled, efficient bioprocesses. Recent suspension culture (three-dimensional, 3D) strategies for hPS cell production in stirred-tank bioreactors (STBR) support this development. However, bioprocess inoculation still depends on adherent (two-dimensional, 2D) preculture, which is labor intensive, resource demanding and poorly controlled and limits process automation. Here we describe the controlled in-process production and dissociation of 3D cultured hPS cell aggregates directly in STBRs, tackling these challenges. The resulting cells can be used for the generation of high-density cryostocks, subsequently enabling the direct inoculation of 3D cultures, thereby entirely omitting the need for 2D preculture and the associated limitations. A key feature of this Protocol is the nonenzymatic, EDTA-based hPS cell aggregate dissociation approach in STBRs, enabling the impeller-based mechanical control of the dissociation process. The resulting cell suspension can be used for process reinoculation and seed train-based upscaling, as well as for the cryopreservation of produced hPS cells, ideally via controlled-rate freezing. Together, the protocol enables the efficient and flexible hPS cell suspension culture over multiple passages, maintaining karyotype stability and pluripotency. This Protocol can be easily implemented by any cell culture-educated scientist without extensive bioprocess training. Cell thawing requires 1 h, the 2D preculture requires 9 days, 2D cell passaging requires 1 h, bioreactor preparation requires 2 days, (direct) bioreactor inoculation requires 1.5 h, 3D STBR cultivation requires 3-4 days and STBR-based aggregate dissociation requires 2 h.
Limbal stem cell deficiency (LSCD) is a severe ocular condition that impairs vision and causes chronic pain. Current therapeutic approaches, such as stem cell transplantation and biomaterial scaffolds, are under continuous refinement. This study evaluated a novel treatment strategy combining a biocompatible and biodegradable chitosan-gelatin (CS-Gel) electrospun scaffold with a topical limbal stem cell (LSC) spray for treating alkaline-induced LSCD in a rabbit model. The scaffold was fabricated via electrospinning, stabilized with water vapor, and characterized using SEM, ATR-FTIR, contact angle, weight loss, and swelling analyses. Limbal stem cells were isolated from cadaveric corneas, cultured, and prepared as a spray suspension (104 cells/mL). Nine rabbits with induced alkaline burns were divided into three groups: an untreated control, a group treated with the CS-Gel scaffold alone, and a group receiving the CS-Gel scaffold followed by the LSC spray one week later. Clinical and histopathological evaluations assessed epithelialization, stromal integrity, neovascularization, and inflammation. The results demonstrated that the combined CS-Gel + cell spray treatment significantly enhanced corneal regeneration, reduced inflammation, and improved healing outcomes compared to the other groups. This study confirms the potential of the CS-Gel + cell spray as a promising platform for LSCD therapy, warranting further investigation. Clinical Registration: This study is an animal-based research and does not involve human clinical trials.
Three-dimensional (3D) dynamic conditions are necessary for physiologically relevant mesenchymal stem cell (MSC) culture. Furthermore, bioreactor-based processes and hydrogel-encapsulation substantially improve niche standardization, expansion control, and process scalability. The integration of these technologies can overcome some challenges that prevent the clinical translation of MSCs. However, few studies have investigated the synergistic use of bioreactors and hydrogels, and none have explored the combination of high-throughput encapsulation platforms, such as microfluidics or millifluidics-generated microgels, with standard stirred-tank systems. In this study, we characterized the continuous culture of MSCs in a stirred-tank reactor, encapsulating the cells in gelatin methacryloyl (GelMA) microgels using a low-cost, user-friendly approach. The effects of seeding density, GelMA's degree of functionalization (DoF), bioreactor sampling protocol, and donor screening were assessed using cell metabolic activity, differentiation, proliferation, and extracellular vesicle (EV) secretion. Gentle dynamic culture enhanced MSCs' metabolic activity. However, cell proliferation was inhibited within the microgels, and no cell migration was observed on the hydrogel's surface. GelMA with a low DoF and high cell seeding density favored cell survival during culture, whereas pronounced donor-dependent differences were observed in cell proliferation and metabolism. The yield, size distribution, and protein content of MSC-EVs were affected by seeding density under dynamic 3D conditions. Furthermore, MSC differentiation led to readily measurable changes in the microgels. Our findings highlight the platform's potential for high throughput microtissue generation and efficient assessment of bioactive compounds.
Biodiversity loss in the present era requires new tools for studying nonmodel organisms. Elephants are both an endangered species and excellent models for studying complex phenotypes including size, social behavior and longevity. Here we report the first derivation of elephant (Elephas maximus) induced pluripotent stem (emiPS) cells. We achieved emiPS cells using two approaches: (1) a two-step process of chemical media induction and colony selection followed by over-expression of elephant transcription factors; and (2) a one-step process with transcription factors and HRAS mutant, HRASG12V. For both protocols, we inhibited TP53 retrogenes, which are hypothesized to confer unique cancer resistance in elephants. To confirm their reprogrammed state, we generated a functional omics catalog of emiPS cells. While these emiPS cells remain transgene-dependent, we inactivated the transgenes and differentiated emiPS cells into all three germ layers via tri-lineage differentiation, embryoid body generation and direct differentiation into putative cell types from all three layers. These methods will open new frontiers for cellular models of nonmodel organisms, including for genetic rescue and conservation.
Bone disorders and skeletal defects represent a significant clinical challenge, often requiring transplantation techniques limited by donor site morbidity and insufficient regenerative potential. Tissue engineering and regenerative medicine (TERM) strategies using 3D bioprinting have emerged as promising alternatives, but their efficacy is limited by the difficulty of directing stem cell differentiation in a controlled and reproducible manner. To address this limitation, we are proposing 3D bone printing via ultrasound-mediated osteogenic differentiation of stem cells (referred to here as '3DBonUS'). This biofabrication approach integrates low-intensity pulsed ultrasound (LIPUS) with a microfluidic-assisted 3D bioprinting system. This unprecedented approach enables biophysical stimulation of human bone marrow stromal cells (HBMSCs) during the fabrication of scaffolds, promoting osteogenic differentiation without the need for extensive post-fabrication treatments. In addition, the incorporation of microbubbles (MBs) enhanced the effects of LIPUS by amplifying mechanical signals at the cellular level. Our results revealed that the 3DBonUS system significantly upregulated key osteogenic markers (RUNX-2, ALP, COL1A1, BMP-2, OCN and OPN) as confirmed by immunofluorescence and RT-qPCR analysis. Moreover, the LIPUS-treated constructs showed a significant (p<0.05) increase in alkaline phosphatase (ALP) activity and calcium deposition, indicating enhanced mineralisation. The biofabricated constructs maintained high cell viability while exhibiting improved osteogenic differentiation, surpassing traditional 3D bioprinting approaches in both efficiency and efficacy. The 3DBonUS strategy represents a new modality in skeletal TERM, combining biofabrication with targeted mechanical stimulation, with potential for scalability of scaffold manufacturing and clinical application. Future studies will aim to validate the functional skeletal scaffolds in vivo to assess their regenerative potential, with the goal of advancing patient-specific bone implants with enhanced osteogenic properties.
Extracellular matrix (ECM) stiffness critically regulates stem cell behavior. Previously, we demonstrated that pathological increases in matrix stiffness during aging disrupt stem Leydig cell (SLC) homeostasis, leading to a decline in testosterone. Building on this discovery, we here present a detailed protocol-originally developed in our laboratory-for fabricating polyacrylamide (PA) hydrogels with tunable stiffness to model the testicular microenvironment in vitro. This method enables reproducible casting of gels across a stiffness range of 1-100 kPa, covering physiological to pathological conditions. Key steps include precise mixing of acrylamide/bis-acrylamide, gel swelling equilibration, surface activation with Sulfo-SANPAH, and collagen coating to support SLC adhesion and culture. We provide optimized formulations for target stiffnesses and troubleshooting guidance for common issues such as incomplete polymerization and poor cell attachment. This system allows systematic investigation of how substrate stiffness modulates SLC proliferation, differentiation, and steroidogenic function under defined 2D conditions. Beyond reproductive biology, it also serves as a valuable platform for mechanobiological studies in other cell types and for screening therapeutics targeting stiffness-related dysfunction.
The survival rate of patients diagnosed with non-small cell lung cancer (NSCLC) are significantly influenced by drug resistance and metastasis with cancer stem cells (CSCs). MicroRNAs (miRNAs) impact on the progression of various human diseases, including different types of cancers. This research seeks to clarify how miR-34a-5p influences CSC characteristics and resistance to cisplatin (DDP) in NSCLC. Initially, the levels of expression for miR-34a-5p, KRT5, and DSP were analyzed in both NSCLC tissue samples and cell lines through RT-qPCR and western blot techniques. Subsequently, sphere formation assays, CCK-8 assays, and flow cytometry tests were carried out to evaluate CSCs and their resistance to DDP in NSCLC. The expression of genes related to stemness, such as SOX2, CD133, and ALDH, was assessed using western blotting methods. Functional experiments were also conducted to investigate the regulatory mechanisms linked to miR-34a-5p, KRT5, and DSP. Finally, the effect of miR-34a-5p on the CSC phenotype and resistance to DDP was validated using a xenograft mouse model. Results showed that, in NSCLC patients, miR-34a-5p was found to be decreased, while KRT5 and DSP expressions were elevated. By targeting the KRT5/DSP axis, miR-34a-5p was shown to suppress CSC properties and enhance DDP sensitivity in NSCLC cells. miR-34a-5p directly bound to and inhibited KRT5, resulting in the downregulation of DSP expression. Furthermore, the overexpression of miR-34a-5p in mice resulted in reduced tumor growth. Collectively, this study elucidates the regulatory role of miR-34a-5p on CSC properties, providing insights that may aid in overcoming drug resistance in NSCLC.
Osteoporosis (OP) is a gradual metabolic bone disease characterized by decreased bone mass and degradation of bone microarchitecture. It affects hundreds of millions of people globally and places considerable pressure on healthcare systems. Current pharmacological treatments, such as bisphosphonates, selective estrogen receptor modulators, and anabolic agents, can reduce fracture risk; however, their prolonged use is limited by significant adverse effects, elevated treatment costs, and a lack of sustained disease remission. Their constraints have intensified interest in restorative approaches utilizing mesenchymal stem cells (MSCs). In the past 20 years, MSCs have emerged as attractive treatment options for OP due to their capacity to differentiate into osteoblasts, modulate immune responses, and exert paracrine effects. Bone marrow-MSCs are the best characterized; nevertheless, MSCs obtained from adipose tissue, umbilical cord, and dental pulp have distinct benefits. Preclinical data demonstrate that direct MSC transplantation enhances bone mineral density, promotes osteoblast production, and reestablishes the equilibrium of bone remodeling in many OP models, including Ovariectomy, glucocorticoid-induced OP, and diabetic OP. Nonetheless, significant obstacles persist: insufficient targeting of osteoporotic bone surfaces, suboptimal cell viability and integration, donor heterogeneity, and unresolved safety concerns. The discovery that the secretome and exosomes (EXOs) produced from MSCs recapitulate several therapeutic advantages of the original cells has initiated a transition toward cell-free methodologies. EXOs produced from MSCs include osteogenic microRNAs (including miR-150-3p and miR-21), inhibit NLRP3 inflammasome activation in osteoclasts, promote macrophage polarization toward an M2 phenotype via TRIM25/TREM1 signaling, and facilitate angiogenesis through the activation of the PI3K/Akt pathway. Furthermore, nanoparticle engineering and combinatorial medicines are advancing to enhance targeting and therapeutic efficacy.
Alzheimer's disease (AD) is characterized not only by gray matter lesions but also by significant white matter damage and oligodendrocyte dysfunction. This study elucidates the pyroptosis-suppressive potential of Scutellaria baicalensis Georgi stem-leaf flavonoids (SSFs) in oligodendroglial lineage cells. We co-treated rat oligodendrocytes (OLN-93 cell line) with 7.5 μmol/L Aβ₁₋₄₂ to simultaneously induce pyroptosis and SSFs. We observed cell morphology via microscopy, measured cell viability with the MTT assay, and assessed membrane damage using the LDH release assay. We then used qPCR to measure the mRNA levels of myelin-related genes (MBP, MAG, MOG) and pyroptosis-related genes (NLRP3, Caspase-1, GSDMD). We employed Western blotting to quantify the expression of pyroptosis-related proteins.Immunofluorescence was. used to localize myelin proteins whereas pyroptosis indicators and applied PI/Hoechst staining was used to evaluate cell membrane permeability within cells. SSFs at concentrations of 15-60 mg/L significantly improved cell morphology, increased cell survival, reduced LDH release, modulated the expression of relevant genes and proteins, decreased the proportion of cells with impaired membrane integrity, and reduced the fluorescence signal of the pyroptotic execution protein GSDMD. These effects were comparable to those of the commonly used positive control, 60 mg/L Ginkgo biloba extract (GBE). This study found that SSFs can alleviate Aβ-induced oligodendrocyte pyroptosis by inhibiting the NLRP3 inflammasome pathway, thereby preserving myelin function. The limitations of this study include the exclusive use of cell line models, the unclear identity of the specific active ingredients in SSFs, and the lack of verification of their blood-brain barrier penetration. Further studies, including in vivo animal experiments, active component isolation, and pharmacokinetic research, are required to evaluate its therapeutic potential for AD fully. SSFs protect oligodendrocytes against Aβ-induced injury by inhibiting the NLRP3 inflammasome pathway.
As a prevalent chronic illness across the globe, diabetes mellitus (DM) is marked by disrupted glucose balance in the body. In type 1 diabetes mellitus (T1DM), autoimmune responses destroy pancreatic β-cells entirely, resulting in total insulin insufficiency. Patients with this condition need lifelong supplemental insulin treatment. The present study intends to create a reliable and economical culture platform to generate pancreatic islet organoid-like structures from bone marrow mesenchymal stem cells (BMSCs). This platform can not only ease the shortage of donor islets for transplantation, but also serve as an in vitro model for diabetes-related research. By optimizing the preparation process of porcine pancreatic tissue lysate (centrifugation at 12,000 rpm for 30 min at 4 °C, followed by 0.22 µm filtration for sterilization), BMSCs were successfully induced to differentiate into pancreatic islet organoid-like structures. Morphological validation confirmed that only induced BMSCs formed compact, plump, and highly transparent islet-like aggregates, whereas noninduced BMSCs exhibited significant vacuolization and reduced transparency. Dynamic monitoring showed that cell aggregates (50-100 µm in diameter) formed on day 16 of culture and developed into capsule- like structures (200-300 µm in diameter) by day 22. This method is simple to operate, cost- effective, and does not require complex equipment. The resulting organoids are morphologically similar to pancreatic islets, providing a tool for in vitro studies and related drug screening.
Intraoperative blood salvage (IBS) reduces exposure to allogeneic blood but is accompanied by concerns in oncologic surgery regarding the potential reinfusion of residual tumor cells and subsequent metastatic dissemination. The CATUVAB® procedure was developed to eliminate EpCAM-positive tumor cells from autologous erythrocyte concentrates (EC). As CATUVAB® requires integration of a leukocyte depletion filter (LDF), the present study aimed to evaluate whether three different commercially available LDF meet predefined safety and efficacy criteria within this procedure. In this prospective ex vivo study, intraoperatively collected blood from patients undergoing major oncologic surgery was processed using the CATUVAB® procedure in combination with one of three LDF (Fresenius BioR Flex AT, Haemonetics US RS1, Puriblood LRW-50-04-PS). The primary endpoint was depletion of EpCAM-positive tumor cells. Secondary endpoints included residual catumaxomab levels in the final EC and changes in proinflammatory cytokines (IL-6, IL-8, TNF-α, IFN-γ). Removal of EpCAM/CD133 double-positive cancer stem cells was explored in a subgroup. Thirty-one patients were analyzed. EpCAM-positive tumor cells were detected in intraoperative blood in a substantial proportion of samples, with marked interindividual variability in tumor cell burden. After CATUVAB® processing and final filtration, no EpCAM-positive tumor cells were detectable in any EC, irrespective of the LDF used. EpCAM/CD133 double-positive cancer stem cells were also eliminated in the subgroup. Residual catumaxomab was detectable in a proportion of ECs but consistently remained below the predefined safety threshold of 70 ng per EC. IL-6 and IL-8 levels were markedly reduced during processing across all filter types, whereas tumor necrosis factor-α (TNF-α) and interferon-gamma (IFN-γ) were largely below detection limits. The present study demonstrates that the CATUVAB® procedure, in combination with different leukocyte depletion filters, achieved effective removal of EpCAM-positive tumor cells, accompanied by low residual catumaxomab levels and reduced pro-inflammatory cytokine concentrations. These findings support the feasibility of this approach; however, given the limited sample size and exploratory design, they should be considered preliminary and require confirmation in larger studies.
Autologous stem cell transplantation (ASCT) remains a cornerstone therapy for eligible multiple myeloma (MM) patients, but its use in older adults is debated due to concerns about toxicity and cost-effectiveness (CE). This study aimed to evaluate the CE of upfront ASCT compared with non-transplant therapy in newly diagnosed MM patients aged 65-69 years within a single-payer healthcare system. Using nationwide claims data from the Korean Health Insurance Review and Assessment Service (HIRA database, 2009-2024; study enrollment period, 2017-2024), we conducted a CE analysis of 735 patients, of whom 458 received bortezomib-thalidomide-dexamethasone induction followed by ASCT and 277 received non-transplant therapy. A state-transition semi-Markov model estimated life-years (LYs), quality-adjusted life-years (QALYs), and direct medical costs from a healthcare system perspective. Inverse probability of treatment weighting reduced confounding bias, and deterministic and probabilistic sensitivity analyses assessed parameter uncertainty. ASCT yielded superior overall survival (4-year: 67.8% vs. 55.0%; HR 0.62, P = 0.001) and longer time-to-next-treatment (29.8 vs. 19.7 months, P < 0.001), with 0.71 additional QALYs at an incremental cost of $21,378. The incremental CE ratio was $30,251 per QALY, remaining below Korea's willingness-to-pay threshold of $37,453. Probabilistic sensitivity analysis demonstrated a 70.3% likelihood of CE at this threshold. Upfront ASCT is a cost-effective consolidation strategy for appropriately selected MM patients aged 65-69 years within Korea's single-payer healthcare system, supporting fitness-based rather than age-based transplant eligibility criteria.
The regeneration of vascularized bone tissue requires biomaterials that deliver coordinated osteogenic and angiogenic signals within mechanically robust three-dimensional architectures. Here, we present a decellularized osteo-angiogenic scaffold generated by integrating mineral-coated nanofiber-incorporated human adipose-derived stem cell spheroids into a 3D-printed polymer scaffold. The mineral-coated spheroids enhanced extracellular matrix (ECM) deposition and osteogenic priming during preculture, and subsequent decellularization efficiently removed cellular components while preserving osteoinductive matrix proteins and pro-angiogenic growth factors. The resulting cell-free scaffold established a homogeneous, multifunctional signaling microenvironment that supported host cell infiltration and potently induced coupled osteogenic and angiogenic responses in vitro without exogenous growth factor supplementation. In a murine critical-sized calvarial defect model, the mineralized scaffold achieved significantly enhanced neovascularization (16 ± 1 α-SMA+ arterioles per mm2) and mature lamellar bone formation (66.2 ± 5.9% BV/TV) compared with the non-mineralized controls. This work introduces a stem cell-derived, ECM-enriched 3D scaffold platform that couples osteogenesis and angiogenesis through endogenous bioactive cues, providing a clinical translation strategy for vascularized bone regeneration.
The role of autologous stem cell transplantation (ASCT) is central in AL amyloidosis and continues to be defined in the era of newer therapies. To evaluate contemporary patient selection, practice patterns, and outcomes, we conducted a retrospective, multicenter study across nine tertiary referral centers, including 1047 patients with AL amyloidosis who underwent ASCT between 2010 and 2020. Most patients (67.9%) received full-dose melphalan conditioning, and 66.5% received pre-ASCT induction therapy, predominantly proteasome inhibitor-based regimens. Day-100 all-cause mortality was 3.0%, representing a substantial improvement compared with historical cohorts. Post-ASCT hematological responses were satisfactory in 75.4% of patients. Among patients with paired assessments, ASCT deepened responses in 56.3% who had not achieved a complete response with induction therapy. With a median follow-up of 5.1 years, the median overall survival was 6.3 years. Independent predictors of inferior survival included age ≥60 years, bone marrow plasma cells ≥20%, pre-ASCT dFLC ≥40 mg/L, elevated cardiac biomarkers, lambda-restricted disease, reduced eGFR and reduced-dose conditioning. In this contemporary cohort, ASCT for AL amyloidosis demonstrated low early mortality, high rates of deep hematologic response, and durable survival in carefully selected patients. These findings support its role as an effective consolidative strategy, while its optimal positioning alongside emerging therapies requires further study.
Gastrointestinal graft-versus-host disease (GI-GVHD) is a major complication after allogeneic hematopoietic stem cell transplantation (allo-HSCT), often requiring invasive endoscopy for diagnosis. Fecal calprotectin (FC) offers a non-invasive marker of intestinal inflammation, but its utility in GI-GVHD needs clarification. In this prospective observational study of 165 adult allo-HSCT recipients, FC was measured on days +7, +14, and +21 post-transplant, at GI-GVHD onset, and 7 days post-treatment, alongside clinical, endoscopic, and histological data. GI-GVHD developed in 52.7% of patients, with histological confirmation in 90.3% of cases. FC lacked predictive value on day +7 (AUC = 0.50) but showed moderate-to-high accuracy on days +14 (AUC = 0.69) and +21 (AUC = 0.77), with an optimal day +21 cutoff of 52.5 µg/g (sensitivity 75%, specificity 87%). At diagnosis, median FC was 120 µg/g, correlating with endoscopic severity (r = 0.31; p = 0.02) but not clinical or histological grades. FC declined significantly post-treatment (120 to 51.5 µg/g; p = 0.04), though concurrent infections elevated levels without compromising discriminative ability. FC serves as a dynamic biomarker for predicting, diagnosing, and monitoring GI-GVHD, but requires integrated clinical interpretation due to limited specificity amid other inflammations.
Premature ovarian insufficiency (POI) is a complex endocrine disorder characterized by the loss of ovarian function before age 40, leading to infertility, hypoestrogenism, and systemic complications. Conventional hormone replacement therapies (HRTs) offer only partial relief, failing to replicate the dynamic feedback of endogenous hormone regulation. Advances in tissue engineering, stem cell technology, and microphysiological systems have catalyzed the development of bioengineered platforms that mimic native ovarian endocrine function. This review outlines progress from three-dimensional (3D) follicle-mimetic constructs and scaffold-free organoids to dynamic ovary-on-a-chip systems, emphasizing advances in steroidogenesis reconstitution, innovations in granulosa-theca co-cultures, and the design of cell-based hormone replacement therapies (cHRTs) capable of re-engaging the hypothalamic-pituitary-ovarian (HPO) axis. We also address critical translational challenges including immunoisolation, vascularization, and regulatory pathway. By bridging bioengineering and reproductive medicine, cHRTs represent a paradigm shift, offering the potential to restore endocrine health, fertility, and systemic homeostasis in a personalized and physiologically relevant manner.