Bioengineering & Translational Medicine最新文献

Spinal cord injury (SCI) triggers an immediate influx of immune cells that secrete pro-inflammatory cytokines and reactive oxygen species that cause tissue damage that is secondary to the initial physical trauma. We aim to reprogram these immune cells to promote a less inflammatory and more pro-regenerative environment. Herein, we investigated the window in time during which poly(lactide-co-glycolide) nanoparticles (NPs) administration can successfully modulate the immune response and promote functional sparing. The dynamics of immune cell infiltration and secondary tissue damage were studied following the injection of NPs intravenously every 24 h for 7 days following injury, with the first injection starting at 2, 4, or 24 hours post-injury (hpi). At 7 days post-injury (dpi), early NP intervention decreased the number of infiltrating macrophages and neutrophils, but delaying treatment until 24 hpi increased the number of neutrophils above control. All mice that received NPs had greater neuronal sparing contralateral to the injury, but mice that received NPs at early timepoints had greater neuromuscular junction innervation and motor endplate sparing. The increased sparing of neurons and neural circuits in the 2 hpi NP group corresponded with increased motor function, as measured by a ladder beam test. Collectively, these results suggest that early intervention with NPs can modulate the inflammatory response and preserve motor function and circuits following SCI.

Tissue engineering provides a promising avenue for treating meniscus defects. In this study, a novel polycaprolactone (PCL)/collagen type I (COL I) meniscus scaffold was fabricated using low temperature deposition manufacturing (LDM) 3D printing technology. The scaffold had a ring and radial fiber structure, and its composition and structure were double bionic of the natural meniscus. In vitro experiments showed that the scaffold had good biological properties, which could promote the proliferation of meniscus fibrochondrocytes (MFCs) and increase the secretion of collagen and glycosaminoglycan. Moreover, the scaffold had excellent mechanical properties and could withstand various stress loads from the femur and tibia. The integrity of the scaffold structure was maintained to provide sufficient time and space for tissue regeneration. The PCL/ COL I scaffold has shown good therapeutic effect in a rabbit meniscus defect model and promotes meniscus regeneration. The results of experiments in rabbits suggest that the scaffold may recruit stem cells and differentiate into fibrochondrocytes in the knee joint, which needs to be verified by further experiments. This study introduces a method of fabricating a new structural composition double bionic meniscus scaffold by LDM technology and verifies its ability to promote cell proliferation, increase the secretion of the extracellular matrix of fibrocartilage, and regulate the microenvironment of cell growth. In addition, this scaffold has achieved good results in repairing meniscus defects in small animal models. Our findings strongly indicate that the PCL/COL I biomimetic meniscus scaffold prepared using 3D-LDM technology holds great promise for repairing and regenerating damaged menisci.

Genome sequencing has identified numerous mutations in the DEAD-box RNA helicases, DDX3X and DDX3Y, associated with cancer and other diseases, but monitoring of their functional consequences remains a challenge. Conventional helicase assays are laborious, often technically difficult, and are performed in cell-free systems that do not address biologically relevant questions. Here, we developed an engineered DDX3 reporter cell system capable of interrogating helicase activities of DDX3X and DDX3Y and their mutational variants. For this, we deleted the endogenous DDX3X in human 293T cells using CRISPR/Cas9. DDX3Y is absent in 293T cells being a female-derived line. We transfected cells with firefly luciferase plasmids that provided bioluminescence signals, depending on helicase activities of exogenously expressed wild-type or mutant DDX3X or DDX3Y, and inserted Aequorea coerulescens Green Fluorescent Protein (AcGFP) as an internal control separated by an internal ribosome entry site (IRES). The developed reporter system can be applied to screen compound libraries targeting DDX3X or DDX3Y in living cells and study their functional roles in health and disease.

Diseases and disorders of dental, oral, and craniofacial (DOC) tissues represent a significant global health burden and have been found to have the greatest age-standardized prevalence and incidence of all reported diseases worldwide. While the application of novel therapies has been suggested to address the different types of oral health diseases, only a limited number of interventional regenerative therapies have been reported to improve clinical therapeutic outcomes. The lack of novel therapies in DOC tissue regeneration may be in part attributed to the highly resource-intensive translational path from preclinical models to clinical trials. Recently, stakeholders and regulatory agencies have begun to encourage the use of alternative preclinical models using human tissues for testing therapeutic interventions in place of animal models. This advocacy may provide an opportunity to reduce or eliminate animal testing, ultimately limiting resource expenditure and providing a more efficient regulatory pathway for the approval of novel DOC therapies. While the complexity of DOC physiology, defects, and diseases is not effectively recapitulated in traditional 2D or 3D in vitro culture models, the emergence of more sophisticated in vitro models (or so-called microphysiological systems that include spheroid, organoid and organ on-chip (OoC) systems) has enabled effective modeling of clinically simulated disease states in several DOC tissue and organ systems. Here, we aim to provide an overview and collective comparison of these microphysiological systems, outline their current uses in DOC research, and identify important gaps in both their utilization and abilities to recapitulate essential features of native oral-craniofacial physiology, towards enabling the therapeutic performance of de novo interventions targeted at regeneration outcomes in vivo.

Malignant brain tumors, particularly glioblastoma multiforme (GBM), are aggressive and fatal cancers. The clinical efficacy of current standard-of-care treatments against brain tumors has been minimal, with no significant improvement over the past 30 years. Driven by the success of chimeric antigen receptor (CAR)-T cells in the clinic for treating certain types of cancer, adoptive cell therapies have been of interest as a hopeful therapeutic modality for brain tumors. Clinical trials of GBM-targeting cell therapies, including CAR-T cells, have been initiated; however, none of them have been approved yet, and new challenges have emerged from the completed clinical trials. These issues are being addressed in ongoing clinical trials and recent preclinical research efforts. Herein, we present an overview of the clinical landscape of cell therapies against brain tumors. We analyze past and active 203 clinical trials focusing on cell therapies for brain tumors, discuss limitations for their clinical translation, and highlight emerging approaches to address these challenges. In addition, we review select preclinical studies that show promise to improve the therapeutic efficacy of therapeutic cells on brain tumors and discuss future prospects.

Non-small cell lung cancer (NSCLC) presents significant therapeutic challenges, often characterized by aggressive proliferation and metastasis. This study investigates the role of SLC7A11, a ferroptosis-related gene, in NSCLC progression and the potential of engineered bacterial extracellular vesicles (BEVs) expressing SLC7A11-targeting siRNA as a therapeutic strategy. Using TCGA and GEO databases, we identified that SLC7A11 was significantly upregulated in NSCLC tissues. Functional assays demonstrated that SLC7A11 knockdown in NSCLC cell lines (NCI-H2122 and NCI-H647) via qPCR, Western blot, and immunofluorescence resulted in impaired proliferation, migration, and invasion abilities. In vivo xenograft models further revealed that SLC7A11 knockdown inhibited tumor growth and metastasis, corroborated by histological analyses. To enhance targeted delivery of SLC7A11 siRNA, we engineered BEVs with a lung cell targeting peptide, verifying their structure and function through transmission electron microscopy (TEM) and nanoparticle tracking analysis (NTA). In vivo toxicity assessments indicated safety for these bioengineered vesicles. Importantly, treatment with BEVs-LCTP-siSLC7A11 not only impaired tumorigenesis but also activated ferroptosis pathways, as evidenced by altered expression levels of SLC7A11 and transferrin in tumor and metastatic tissues. Our findings suggest that targeting SLC7A11 through engineered BEVs presents a promising approach to inhibit NSCLC progression while activating ferroptosis, offering insights into novel therapeutic strategies against lung cancer.

Lentiviral vectors (LVVs) are emerging as an enabling tool in gene and cell therapies, yet the toolkit for purifying them at scale is still immature. A pivoting moment in LVV isolation technology was marked by the introduction of affinity ligands for LVVs pseudo-typed with Vesicular Stomatitis Virus G (VSV-G) protein. Camelid antibody ligands were initially discovered and utilized to functionalize a resin with a capacity of 10¹⁴ LVV particles per liter (vp/L). Shortly thereafter, our team introduced VSV-G-targeting peptides and assessed their application as ligands for purifying LVVs from HEK293 cell harvests. In this study, we utilized these peptides to develop novel affinity resins and—first in this field—affinity membranes with optimal binding capacity, productivity, and removal of host cell contaminants. To that end, we evaluated resins of different material, particle and pore size, and functional density, as well as membranes with different fiber morphology, porosity, and ligand distribution. The lead peptide-functionalized resin and membrane featured high capacity (5 × 10⁹ and 1.2 × 10⁹ transducing LVV units per mL of adsorbent, TU/mL) and productivity (2.9 × 10⁹ and 1.7 × 10⁹ TU/mL min) and afforded a substantial enrichment of cell-transducing LVVs and reduction of contaminants (110–170-fold) in the eluates. Finally, we demonstrated an LVV purification process in four steps: clarification and nuclease treatment, affinity capture in bind-and-elute mode, polishing in flow-through mode, and ultra/dia-filtration and sterile filtration. The processes afforded yields of 33%–46%, a residual HCP level below 5 ng/mL, and productivity of 1.25–1.5 × 10¹⁴ active LVV particles per hour and liter of adsorbent.

Nipple reconstruction in patients who undergo total mastectomy or nipple-sparing mastectomy is currently limited by a consistent and significant loss of nipple projection over time, which can negatively affect patient satisfaction and quality of life. To address this issue, we have previously shown that 3D-printed poly-4-hydroxybutyrate (P4HB) nipple-shaped scaffolds promote long-term maintenance of nipple projection in a rat model. Herein, we further optimize the 3D printing parameters (filament diameter and infill density) of absorbable P4HB latticework scaffolds as well as scaffolds fabricated from rolled P4HB knitted mesh to facilitate tissue formation with similar biomechanical properties of the native nipple, while maintaining long-term shape and projection. Over 12 months of in vivo implantation in a dorsal, bilateral CV-flap rat model of nipple reconstruction, 3D-printed P4HB latticework and knitted mesh scaffolded groups demonstrated significantly greater maintenance in projection (80–100% of initial value) when compared to the Cook Biodesign® Nipple Cylinder (~40% of initial projection), resulting from the infiltration of healthy fibrovascular adipose tissue, which demonstrated biomechanical qualities that approached those of the native human nipple. Overall, our results demonstrate that using a 3D-printed P4HB latticework and rolled P4HB knitted mesh scaffolds significantly improved long-term results in our animal model of nipple reconstruction and hold promise for improving nipple reconstruction outcomes in future clinical practice.

Adalimumab (Humira) represents a major advance in rheumatoid arthritis (RA) therapy. However, with long-term administration of Adalimumab, anti-idiotypic antibody (anti-Id Ab) accelerates the Adalimumab clearance rate and reduces the therapeutic effect. To avoid the interference of anti-Id Ab, we used an autologous hinge region as a spatial-hindrance-based Ab lock and connected it to the N-terminal of the light chain and heavy chain via substrate peptides (MMP-2/9) to cover the CDR binding site of Adalimumab to generate pro-Adalimumab. The Ab lock masks the complementarity-determining regions (CDRs) of Adalimumab, thus avoiding interference from anti-Id Ab. Pro-Adalimumab demonstrated a 241.6 times weaker binding ability to TNFɑ than Adalimumab. In addition, pro-Adalimumab showed a 46.6-fold greater blocking of anti-Adalimumab Id Ab in comparison to Adalimumab prior to activation. Similar results were observed with other clinical antibodies, such as pro-Infliximab (anti-TNFɑ Ab) and pro-Nivolumab (anti-PD-1). Furthermore, pro-Adalimumab maintained consistent pharmacokinetics regardless of the presence of anti-Adalimumab Id antibodies, while Adalimumab showed a 49% clearance increase, resulting in a near complete loss of function. Additionally, pro-Adalimumab was able to avoid neutralization and efficiently reduce RA progression in the presence of anti-Adalimumab Id Ab in vivo. In summary, we developed a pro-Adalimumab that avoids interference from anti-Id Abs, thereby addressing the biggest issue limiting clinical efficacy. The findings enclosed herein may have potentially broad application in antibody therapies.