Frontiers in Bioengineering and Biotechnology最新文献

Intervertebral disc degeneration (IDD) is a primary cause of chronic low back pain, severely impacting patients' quality of life. Conventional treatments focus on symptom relief but fail to restore disc structure and function. Recent bioengineering advances offer regenerative solutions, integrating cell therapy, tissue-engineered scaffolds, gene therapy, and mechanobiology. Cell therapy leverages mesenchymal stem cells (MSCs) from bone marrow, adipose tissue, or umbilical cord blood, with biomaterial carriers enhancing survival in the harsh disc microenvironment. Scaffolds-natural (collagen, chitosan) or synthetic (PLGA, PCL)-mimic native extracellular matrix (ECM) and provide mechanical support, often combined with growth factors for controlled release. Gene therapy targets ECM synthesis, inflammation, and degradation pathways via viral or non-viral vectors, while mechanobiology reveals how mechanical forces regulate disc cell behavior, guiding scaffold design. Animal models validate these therapies, and early clinical trials show promise in pain reduction and disc height restoration. However, challenges remain, including low cell survival, scaffold mechanical adaptation, and gene delivery safety. Multidisciplinary collaboration is key to translating preclinical progress into effective clinical interventions, addressing the unmet medical need for IDD treatment.

Introduction: Acute-on-chronic liver failure (ACLF) is a highly lethal clinical syndrome with limited effective therapeutic options. Urine-derived stem cells (USCs) represent a non-invasive and readily accessible cell source, but whether USCs obtained from patients with severe liver dysfunction retain therapeutic and immunomodulatory potential remains unclear.

Methods: To address this question, USCs derived from ACLF patients (LF-USCs) were evaluated in a Concanavalin A (Con A)-induced immune-mediated acute liver injury mouse model. Hydrogel-encapsulated LF-USCs were transplanted, and therapeutic efficacy was assessed by survival analysis, serum biochemical parameters, histological examination, and inflammatory cytokine profiling.

Results: Transplantation of hydrogel-encapsulated LF-USCs significantly improved mouse survival, reduced serum transaminase levels, and alleviated hepatocellular necrosis (p < 0.05). At the mechanistic level, LF-USC treatment was associated with decreased systemic inflammatory cytokine levels, attenuation of intrahepatic inflammatory injury, and dynamic modulation of macrophage-associated inflammatory signatures.

Discussion: These findings demonstrate that functionally competent USCs can be successfully obtained from ACLF patients and highlight their potential as a readily accessible autologous cell source for immune modulation and liver tissue repair in immune-mediated acute liver injury.

Genetic manipulation technologies have revolutionized our ability to control cellular activities with high precision. Five major approaches-chemogenetics, optogenetics, odorgenetics, magnetogenetics, and sonogenetics-offer distinct advantages for different research and therapeutic scenarios. However, a unified framework for systematic comparison across these technologies has been lacking.This review provides a comprehensive analysis of these five genetic manipulation technologies from three perspectives: molecular mechanisms, quantitative performance metrics, and application scenarios. We first dissect the signaling pathways and key molecular components underlying each technology. We then establish a seven-dimensional evaluation framework encompassing spatiotemporal resolution, tissue penetration, cell-type specificity, reversibility, multiplexing capability, biosafety, and technical accessibility. Using this framework, we systematically score and compare the five technologies, revealing that optogenetics excels in spatiotemporal precision (millisecond/micrometer scale), chemogenetics offers superior clinical translatability, while sonogenetics and magnetogenetics provide advantages for non-invasive deep tissue applications. We further analyze optimal application scenarios for each technology, including neural circuit dissection, chronic disease management, and deep tissue intervention.This comparative analysis provides researchers with an evidence-based guide for technology selection. We propose that future developments should focus on hybrid approaches combining the strengths of multiple technologies, and on addressing current limitations in delivery efficiency and long-term biosafety for clinical translation.

Introduction: The leading cause of preventable traumatic death is hemorrhage. Early detection of hemorrhagic shock remains a critical challenge. For the early prediction of hemorrhagic shock-related cardiovascular decompensation, our team has developed the compensatory reserve measurement (CRM) algorithm. CRM uses a photoplethysmography waveform to quantify the body's capacity to compensate during hypovolemia. This study focuses on the development and use of an application that can predict CRM in real-time (CRM_RT) during simulated hypovolemia experiments.

Methods: The CRM_RT application was developed in Python to generate CRM predictions and highlight trend trajectories in real-time (RT). Data were collected during a human research protocol that was reviewed and approved by the Institutional Review Board. Participants (n = 20) meeting the inclusion criteria underwent a simulated hypovolemia procedure in a lower-body negative pressure chamber while wearing a Masimo® MightySat® Rx pulse oximeter. Data were streamed in RT via a Bluetooth® connection to a computer running the CRM_RT application.

Results: CRM was successfully implemented for RT data capture during the research study. The CRM_RT application achieved a median performance error of -0.95%, while the median absolute performance error was higher at 19.00%. CRM_RT resulted in an average early prediction time of 18.3 min by tracking the slope trend changes in RT.

Discussion: The CRM_RT application effectively tracked CRM during simulated hypovolemia using a wearable non-invasive sensor. Predictions served as an earlier indicator of hemorrhage compared to traditional vital signs, addressing a limitation of current triage practices. Overall, the CRM_RT application represents a promising advancement toward RT prediction of hypovolemic decompensation.

Burn injuries remain a significant global health challenge, often leading to prolonged healing, infection, and scarring. Addressing these complications requires innovative therapeutic strategies that can accelerate tissue repair and minimize adverse outcomes. Quercetin, a widely distributed plant-derived flavonoid, has emerged as a promising multi-functional agent in burn wound management. This review presents a comprehensive overview of quercetin's therapeutic potential, emphasizing its pharmacological versatility and the need for advanced topical delivery systems such as nanostructured lipid carriers and hydrogels to overcome its limited bioavailability. Quercetin's efficacy is rooted in its multi-targeted mechanisms of action. It exhibits potent antioxidant activity by directly scavenging reactive oxygen and nitrogen species and activating the Nrf2-ARE signaling pathway, thereby enhancing endogenous antioxidant defenses. Its anti-inflammatory effects are mediated through the inhibition of NF-κB and MAPK pathways, leading to the suppression of key pro-inflammatory cytokines. Quercetin also promotes angiogenesis via upregulation of vascular endothelial growth factor (VEGF), supports fibroblast proliferation and extracellular matrix remodeling, and facilitates re-epithelialization. This review consolidates dispersed experimental findings into a unified mechanistic and translational framework, highlighting why quercetin is uniquely positioned among phytochemicals for burn therapy. By integrating pharmacodynamics, formulation science, and clinical feasibility, the article clarifies current evidence gaps and outlines practical directions for therapeutic development. Such synthesis is timely because research on flavonoid-based wound therapeutics is expanding rapidly but remains fragmented across disciplines.

Introduction: The synthesis of nanomaterials aims to integrate scientific innovation with a commitment to address the UN's Sustainable Development Goals (SDGs). Our study reports two biotechnological protocols for the synthesis of colloidal Ag@ AgCl nanoparticles (Ag@AgCl-NPs) using the endemic Mauritian herb Psiadia terebinthina via microwave extraction (ME) and heat extraction (HE) methods.

Methods: The Ag-based nanoparticles (NPs) containing colloids and extracts were tested for antimicrobial, antioxidant, and anti-inflammatory activities. The successful bottom-up phytosynthesis of Ag@AgCl-NPs was characterized by transmission electron microscopy (TEM) with chemical verification using energy-dispersive X-ray spectroscopy (EDS), crystallinity assessment using X-ray diffraction (XRD), UV-VIS and Fourier transform-infrared (FT-IR) spectroscopy, and investigation of colloidal properties. Green synthesis approaches favored NP formation and colloidal stability. Antimicrobial activity was tested against Gram-positive (G+) and Gram-negative (G-) bacteria and fungi using the broth microdilution assay. Antioxidant activity was determined by the 2,2-diphenyl-1-picrylhydrazyl (DPPH) method, while anti-inflammatory activity was examined using the bovine serum albumin (BSA) test.

Results: Our results confirmed the successful green bionanotechnological synthesis of Ag@AgCl-NPs with varying relative mass ratios, exhibiting spherical morphology and good crystallinity. Additionally, both colloids with Ag@AgCl-NPs exhibited exceptional synergistic antimicrobial effects against highly resistant G+ and G- bacterial pathogens and the yeast Candida albicans. They also demonstrated significant anti-inflammatory activity compared to ME or HE extracts, which, surprisingly, showed greater antioxidant activity.

Discussion: Thus, eco-friendly phytosynthesis from Psiadia terebinthina extracts yielded multifunctional hybrid products with potential utility in pharmacology, medicine, and toxicology.

Introduction: Mammalian cell cultures are widely used for producing complex biopharmaceuticals that require human-like post-translational modifications, such as antibody-based therapeutics. Traditionally, serum-supplemented media support high cell viability and productivity; however, regulatory and scientific requirements demand serum-free conditions for clinical-grade manufacture. Recently, a novel fusion protein, the anti-huCD20(hγ1)-IL2no-alpha immunocytokine (IC), was presented as a promising therapeutic alternative, mostly for relapsed or refractory (r/r) B-cell non-Hodgkin lymphoma (B-NHL) patients, considering currently approved therapies.

Methods: Three Chinese hamster ovary clones (K1 strain) producing the anti-huCD20(hγ1)-IL2no-alpha IC were generated and adapted to serum-free suspension culture. We performed a kinetic characterization of one clone in two culture media with different nutritional compositions, evaluating cell growth, productivity, cell cycle progression and mTOR signaling. The IC was purified by Protein A, then evaluated for identity, aggregation profile, CD20 recognition, CTLL-2 cytokine activity, ex vivo B-cell depletion in PBMC from r/r B-NHL patients and antitumor efficacy in immunocompetent C57BL/6 mice bearing EL4-hCD20- cells.

Results: The results demonstrated noticeable differences in cell growth and productivity in both batch and pseudo-perfusion performance, likely due to an influence on cell-cycle progression and mTOR signaling. The purified IC maintained its structural integrity while exhibiting an improved aggregation profile compared to serum-containing cultures. Furthermore, key biological activities, including B-cell depletion and antitumoral effects, remained intact.

Discussion: This research highlights the successful serum-free production of a functional anti-huCD20(hγ1)-IL2no-alpha IC, reinforcing its potential for biopharmaceutical development.

Background: Human gingival fibroblasts (HGFs) are stromal cells that maintain periodontal tissue structure and extracellular matrix (ECM) dynamics. ECM stiffness serves as a physical cue that regulates HGF behavior and secretory profiles. This study investigated how substrate stiffness modulates the secretome of HGFs and observed the subsequent effects of this secretome on the osteogenic differentiation of human periodontal ligament cells (HPDLCs).

Methods: HGFs isolated from healthy donors were cultured on polydimethylsiloxane substrates, representing soft or hard periodontal tissue under normal and lipopolysaccharide (LPS)-induced inflammatory conditions. The expression of cytokines and chemokines was analyzed using qRT-PCR and ELISA, with p38 MAPK inhibitors used to identify stiffness-associated signaling involvement. HPDLCs were treated with conditioned medium from HGFs (HGF-CM) under osteogenic induction, osteogenic marker expression was examined using qRT-PCR and immunofluorescence, with mineralization assessed by Alizarin red S staining. To establish mechanistic causality, functional blocking was conducted using a C-X-C motif chemokine receptor 4 (CXCR4) inhibitor.

Results: Hard substrates significantly increased the expression of anti-inflammatory cytokines and the C-X-C motif chemokine ligand 12 (CXCL12) in HGFs, whereas inhibition of p38 mitogen-activated protein kinase (MAPK) activity attenuated stiffness-associated CXCL12 expression. Under LPS-induced inflammatory conditions, hard substrates-maintained matrix metalloproteinase-9 suppression and tissue inhibitor of metalloproteinases 1 upregulation, although CXCL12 protein levels were decreased. Furthermore, HPDLCs treated with HGF-CM derived from hard substrates in osteogenic induction media exhibited elevated CXCR4 expression, increased osteogenic marker levels at days 14 and 21, and enhanced mineral deposition compared to those treated with HGF-CM from soft substrates. In addition, functional blocking with a CXCR4 inhibitor significantly reduced the expression of osteogenic markers (ALP, RUNX2, COL1A1, and OSX) and confirmed a subsequent decrease in matrix mineralization.

Conclusion: Substrate stiffness modulated the paracrine behavior of HGFs, with CXCL12 serving as a representative example of a stiffness-responsive factor. These alterations in the HGF-derived secretome were associated with altered osteogenic and inflammatory responses in HPDLCs. These findings support the influence of the physical microenvironment on fibroblast-periodontal ligament cell interactions on anti-inflammatory response and periodontal tissue stabilization.

Chimeric antigen receptor T (CAR-T) cell therapy has achieved transformative success in hematological malignancies; however, its translation to solid tumors remains severely limited by tumor heterogeneity, immunosuppressive microenvironments, and safety concerns such as on-target/off-tumor toxicity. A major contributor to these challenges is the lack of preclinical models capable of faithfully recapitulating human tumor architecture and tumor-immune interactions. Conventional two-dimensional cell cultures and animal models frequently fail to predict CAR-T efficacy, resistance, and toxicity observed in patients. Organoid technology, particularly patient-derived organoids (PDOs) and immune-integrated organoid systems, has emerged as a next-generation platform that bridges this translational gap. By preserving patient-specific genetic, phenotypic, and spatial heterogeneity, organoids provide a physiologically relevant and scalable system for interrogating CAR-T cell behavior in human-like tumor contexts. Recent advances in tumor-immune co-culture, vascularized organoids, and microfluidic organoid-on-a-chip platforms have further expanded their utility for dynamic assessment of CAR-T infiltration, cytotoxicity, cytokine release, and adaptive resistance mechanisms. In this review, we comprehensively examine how organoid-based models are reshaping the CAR-T development pipeline, spanning target discovery and validation, functional efficacy assessment, safety profiling, and optimization of combination therapies. We further discuss emerging applications of organoids as patient-specific "avatars" for personalized CAR-T selection and response prediction. Finally, we highlight current technical limitations and future bioengineering directions required to enable clinical translation. Collectively, organoid platforms represent a transformative tool for accelerating precision development of next-generation CAR-T cell therapies and advancing human-relevant immuno-oncology research.