Search Results (1,352)

The paradigm of craniomaxillofacial (CMF) reconstruction is shifting from traditional bone grafting and biochemical adjuncts toward a nucleomechanical framework that leverages the cell nucleus as a mechanosensitive organelle. By utilizing computer-aided design and computer-aided manufacturing (CAD/CAM)-derived scaffolds with 10 µm micropillar arrays and specific interfacial stiffness (25–40 kPa), surgeons can physically manipulate the Linker of Nucleoskeleton and Cytoskeleton (LINC) complex to achieve a nuclear aspect ratio above 2.5. This structural deformation mechanically expands nuclear pores to trigger cytoskeletal and molecular responses, such as Yes-associated protein (YAP) and transcriptional coactivator with PDZ-binding motif (TAZ) translocation. Resultantly, this physical tension pulls open chromatin fibers to activate master osteogenic regulators like RUNX2, effectively bypassing the risks and limitations associated with supraphysiologic growth factor delivery (e.g., rhBMP-2). Clinically, translating these principles involves moving away from absolute rigid internal fixation toward advanced resorbable biomaterials that permit controlled micro-motions (100–200 µm) under functional masticatory loads. This review provides a structured synthesis of the field, outlining deterministic topographic criteria, clinical boundary conditions, and the potential strategies needed to overcome age-related mechanosensory blockades. Ultimately, we establish a multidisciplinary framework that bridges precision bioengineering with native oral and maxillofacial surgical realities to drive living, biophysically mediated bone repair. Full article

Biomaterial-associated infection remains a persistent challenge for implantable devices, catheters, wound dressings, and tissue-engineering scaffolds. This structured narrative review critically evaluates biomimetic superwetting polysaccharide-based composite hydrogel interfaces based on chitosan, alginate, hyaluronic acid, cellulose/nanocellulose, bacterial cellulose, and dextran. The analysis links polymer network design and cross-linking strategy to pore architecture, swelling, mechanical properties, hydration-layer stabilization, protein adsorption, bacterial adhesion, biofilm development, and cytocompatibility. Stable interfacial hydration can increase the energetic penalty for protein and bacterial approach, but high water uptake alone is insufficient: excessive swelling, low network density, poorly controlled pore interconnectivity, and weak wet-state fixation can compromise durability or provide protected sites for bacterial retention. Study-level comparisons therefore emphasize reported values for network structure, swelling, mechanics, wettability or hydration, and antibacterial/antibiofilm performance, with unreported parameters identified as such. Notably, interactions among biomaterials, bacteria, and host tissues exhibit synergistic and co-evolutionary characteristics, forming a dynamically evolving microecological balance. This eco-synergistic perspective provides a useful conceptual framework for proposing antifouling strategies that aim to regulate rather than eradicate bacterial colonization. Future work should prioritize eco-synergistic design, durable hydration, mechanically stable and porous-yet-cleanable networks, selective interfacial regulation, dynamic characterization, standardized testing, and manufacturable formulations with the minimum necessary active components. Full article

Bacteriophage technologies have evolved from classical cloning vectors into programmable platforms for genome engineering and molecular selection. Phage display couples a surface-presented binding phenotype to its encoding genotype, enabling iterative selection, sequence recovery, and optimization of peptides, antibody fragments, and other protein binders. This review links phage morphology, genome organization, infection strategy, host defense, and engineering method to practical platform choice. It compares λ, N15, M13, and T7 systems; examines biopanning bias and candidate developability; and evaluates artificial intelligence-assisted, sequencing-guided, and structure-guided workflows. Vaccine applications are considered alongside constraints arising from anti-phage immunity, antigen density, route of administration, and repeat dosing. Progress will depend on experimentally validated closed-loop workflows that integrate library design, selection, high-throughput analytics, structural characterization, and early manufacturability assessment. Full article

Carbon nanotube (CNT)-based biosensors have emerged as promising platforms for non-invasive biofluid analysis because of their high electrical conductivity, large surface area, tunable optical properties, and versatile surface chemistry, enabling miniaturized, flexible sensing devices. Sweat, saliva, tears, and urine are increasingly recognized as attractive alternatives to blood for point-of-care diagnostics because they enable repeated, non-invasive sampling while containing clinically relevant metabolites, electrolytes, proteins, hormones, nucleic acids, pathogens, and other biomarkers. However, the low abundance of many analytes, matrix complexity, biofouling, and biofluid-specific variability present significant analytical challenges. This review critically examines the different CNT-based sensor architectures, and their recent advances in non-invasive analysis of sweat, saliva, tears, and urine. It integrates sensor architecture, biofluid-specific analytical challenges, sample-validation level, and translational readiness within a single comparative framework. Representative applications are discussed for metabolic monitoring, renal health assessment, infectious disease testing, and other clinically relevant uses. Beyond clinical diagnostics, emerging non-clinical applications, including drug-of-abuse detection, forensic body-fluid identification, and occupational or environmental exposure assessment, are also highlighted. Finally, we discuss key barriers limiting real-world translation of CNT biosensors, including material reproducibility issues, biofouling, physiological interpretation of biofluid biomarkers, scalable manufacturing, and long-term operational stability, and outline future strategies to advance these platforms toward robust, reliable, and widely deployable biosensing technologies. Full article

Poultry nutrition simultaneously affects animal health, productive efficiency, product quality and the environmental burdens associated with feed production and manure management. Nutritional interventions may generate synergies among these outcomes, but they can also shift costs or environmental impacts across biological and supply-chain boundaries. This review critically evaluates low-protein diets, functional additives, feed enzymes, alternative protein sources, precision-feeding technologies and nutritional approaches to manure-emission mitigation. The evidence indicates that improved intestinal function and nutrient retention can reduce feed requirements and nutrient excretion, whereas excessive protein restriction, unstable additive responses, limited alternative-ingredient supply and poorly validated digital systems may compromise performance, product quality or economic feasibility. Environmental benefits are also sensitive to ingredient origin, processing, transport, crystalline amino acid and additive manufacture, coproduct allocation and life cycle assessment assumptions. To integrate these considerations, we propose a four-level conceptual decision framework comprising physiological adequacy, precision nutrient supply, constraint-specific functional intervention and whole-chain verification. The framework treats health, productive performance, product quality, economic feasibility and environmental impact as simultaneous decision criteria, while genotype, production stage, disease pressure, climate, ingredient availability and market objectives define the conditions of application. Standardized life cycle assessment and commercial validation are required to distinguish reproducible net benefits from improvements confined to individual indicators. This approach provides a structured basis for selecting poultry nutritional strategies that remain biologically effective, economically accessible and environmentally beneficial under practical production conditions. Full article

D-lactic acid is a key chiral intermediate widely applied in agriculture, pharmaceuticals, and polylactic acid synthesis, with microbial fermentation as its main production method. Commercial D-lactic acid production depends heavily on refined carbohydrates and yeast extract as carbon and nitrogen sources, resulting in prohibitive raw material costs. Abundant in microbially available proteins and carbohydrates, renewable resources can act as carbon and nitrogen substrates for efficient D-lactic acid production. Repurposing these resources reduces production expenses and is essential for the industrial scaling of D-lactic acid manufacturing. This paper reviews research advances in D-lactic acid fermentation using renewable feedstocks including agricultural wastes, sugar industry residues, oil processing wastes, and cheese processing by-products as carbon and nitrogen sources. Full article

Hydrogel-forming microneedles (HFMNs) are minimally invasive interfaces that access interstitial fluid (ISF) through skin penetration, swelling-mediated uptake, analyte diffusion, and hydrated sensor integration. This review examines HFMN architectures, skin–device interfaces, ISF transport, molecular-recognition and signal-transduction strategies, analytical performance, benchmarking, wear-associated failure modes, therapeutic monitoring, and translational priorities. The field has expanded from glucose sensing to metabolites, ions, hormones, proteins, nucleic acids, microbial and wound biomarkers, and therapeutic drugs, enabled by advances in hydrogel chemistry, conductive networks, nanostructured electrodes, catalysis, affinity recognition, molecular imprinting, optical readouts, and multiplexed wearables. Performance remains context dependent and requires physiological range, calibration stability, biofouling resistance, reliable insertion, validated ISF-reference correlations, and interpretable thresholds. Evidence is strongest in artificial matrices, ex vivo tissue, and animals, while human validation remains limited. Translation will require standardized mechanics and transport reporting, longer wear studies, sterilization-compatible chemistries, scalable manufacturing, and clinical validation. HFMNs may complement rather than replace blood-based diagnostics. Full article

A single-chain variable fragment (scFv) is an engineered antibody derivative that retains antigen-binding specificity while having a much smaller size than an antibody, improved tissue penetration, and enhanced versatility for genetic manipulation. When an scFv is fused with diverse protein payloads, multifunctional biologics can be created for targeted delivery and therapy. Over the past decade, scFv fusion proteins have gained significant traction in oncology, where they have been incorporated into immunotoxins, immunocytokines, bispecific antibodies, Chimeric Antigen Receptor (CAR)-T cells constructs, and immune cell engagers. In addition, advances in blood–brain barrier (BBB)-targeting strategies have enabled the exploration of scFv-based therapeutics for neurodegenerative diseases, including Alzheimer’s disease and Parkinson’s disease. Despite promising preclinical and clinical outcomes, challenges such as structural instability, short half-life, immunogenicity, and manufacturing complexity remain. This review provides an in-depth and up-to-date overview of scFv fusion protein engineering and its therapeutic applications in cancer and neurodegenerative disorders. We also highlight the clinical translations and design principles of scFv fusion proteins. Full article

Extracellular vesicles (EVs), including small extracellular vesicles (sEVs) and medium/large extracellular vesicles (MVs), have emerged as promising therapeutic vectors and diagnostic biomarkers across various branches of biomedicine. However, the clinical translation of EV-based technologies remains constrained by persistent challenges in manufacturing: insufficient yield from primary cell sources, limited control over cargo composition, and the absence of scalable, standardized production platforms. Nanosecond pulsed electric fields (nsPEF) represent an emerging biophysical approach that can address several of these limitations. Unlike conventional electroporation, which targets the plasma membrane using microsecond-to-millisecond pulses, nsPEF delivers ultrashort (1–300 ns), high-amplitude (10–300 kV/cm) pulses that penetrate intracellularly to directly perturb endosomal membranes, the endoplasmic reticulum, and the multivesicular body (MVB) compartment, the very organelles where small EVs (exosome) biogenesis and cargo sorting occur. Through coordinated effects on intracellular calcium mobilization, cytoskeletal remodeling, SNARE-mediated membrane fusion, and phospholipid redistribution, nsPEF can stimulate rapid, non-lethal vesicle release, a process labeled as “electro-exocytosis.” Emerging and growing evidence suggests that nsPEF does not merely increase EV yield but actively modulates the proteomic, lipidomic, and nucleic acid composition of released vesicles, offering a potential route to cargo engineering. In addition, the same biophysical principles that drive electro-exocytosis can be exploited in reverse: nsPEF-mediated transient permeabilization of EV membranes allows for post-isolation loading of exogenous therapeutic cargo, small molecules, nucleic acids, or proteins into pre-formed vesicles without destroying their structural integrity. This review discusses current knowledge on EV biogenesis and release mechanisms, introduces the biophysical foundations of nsPEF–cell and nsPEF–membrane interactions, and, by evaluating the experimental evidence supporting nsPEF-driven EV engineering, outlines a translational roadmap for the application and development of this technology toward clinical-grade EV manufacturing. Full article

The therapeutic monoclonal antibody bevacizumab is typically purified using protein A affinity chromatography, a highly effective but costly method. Affinity-based precipitation for antibody purification is a lower-cost approach. In this work, a precipitation protocol was developed for bevacizumab purification using a branched peptide (Ac-PHQGQHIG-Ahx₃)₂-K-Ahx₃-PHQGQHIG-NH₂, which contains the epitope PHQGQHIG that is responsible for interacting with bevacizumab. The peptide was synthesised by a microwave-assisted solid-phase peptide method, employing LiCl as an additive to prevent aggregation and ensure high purity and yield. Three molecules of 6-aminohexanoic acid were introduced between each epitope branch as spacer arms to promote the formation of cyclic complexes. Bevacizumab purification from cell-free culture broth was achieved through a fractional precipitation process. First, a negative precipitation step using (NH₄)₂SO₄ 1.18 M was performed to remove contaminants. Afterwards, 5 moles of peptide per mol of bevacizumab was added to the supernatant, together with additional (NH₄)₂SO₄, to reach a final concentration of 1.20 M. Under these conditions, bevacizumab was recovered in the precipitate with 98% purity and a yield of 73%. In addition to being recyclable, the peptide’s relatively low production cost could enable the development of a single-use purification process, which would be particularly advantageous for biopharmaceutical manufacturing. Full article

Plant-derived exosome-like nanoparticles (PELNs) are lipid bilayer nanostructures containing endogenous lipids, proteins, nucleic acids, and phytochemicals, which have attracted increasing interest as bioinspired carriers for cancer therapy. This review evaluates how plant source, isolation, purification procedures, vesicle composition, cargo-loading strategy, and administration route shape the quality and tumor-delivery performance of PELNs. The available evidence indicates that plant source and processing are major determinants of particle size, purity, surface charge, cargo profile, and biological activity. Ultracentrifugation remains widely used but is limited by contaminant co-isolation and poor scalability, whereas density-gradient purification and size-exclusion chromatography improve purity, and ultrafiltration and tangential flow filtration offer greater potential for large-scale manufacturing. Passive incubation generally preserves vesicle integrity and is most suitable for hydrophobic small molecules, whereas electroporation, sonication, and extrusion can increase cargo loading but may cause aggregation, membrane remodeling, or loss of endogenous components. Preclinical studies suggest that PELNs can exert intrinsic antitumor effects, modulate the tumor microenvironment, improve chemotherapeutic delivery, and help overcome drug resistance. However, evidence for in vivo tumor-targeting remains less robust than evidence for cellular uptake, and direct comparisons with established nanocarriers remain scarce. Clinical translation will require standardized nomenclature and characterization, reproducible manufacturing, quantitative loading and release assays, route-specific biodistribution studies, and repeated-dose safety evaluation. These findings provide a framework for the rational development of PELNs as reproducible tumor-oriented nanocarriers. Full article

Chimeric antigen receptor (CAR) T-cell therapies have demonstrated remarkable clinical efficacy in hematological malignancies, yet their broader application is constrained by manufacturing complexity and variability, particularly in autologous settings. The development of allogeneic CAR T-cell therapies offers a promising alternative by enabling scalable, “off-the-shelf” production; however, these approaches introduce additional challenges related to donor variability, genome editing, and product consistency. Robust analytical strategies are therefore required to ensure safety, efficacy, and batch-to-batch reproducibility. Conventional analytical methods, such as flow cytometry and enzyme-linked immunosorbent assays, provide targeted, high-confidence measurements of predefined cellular and soluble markers but are inherently limited in their ability to capture the full molecular and functional complexity of CAR T-cell products. In current manufacturing paradigms, these assays are typically deployed as isolated quality control readouts rather than as components of an integrated control strategy that links donor variability, gene-editing material quality, in-process metabolic state, final product critical quality attributes, and clinical biomarker responses. In this context, mass spectrometry (MS) has emerged as a powerful platform for high-dimensional molecular characterization, enabling analysis of gene-editing reagents, proteins, metabolites, lipids, and both culture and spent-media composition across the CAR T-cell manufacturing workflow. In this review, we examine the various applications and tools of MS across key stages of the allogeneic CAR T-cell workflow, including donor characterization, analysis of gene-editing materials, in-process culture monitoring, drug product quality assessment, and post-infusion biomarker evaluation. Collectively, these approaches demonstrate the potential of MS-driven analytics to address current limitations in CAR T-cell manufacturing by improving process understanding, enabling comprehensive quality assessment, and supporting regulatory decision-making. The integration of MS into CAR T-cell workflows may ultimately facilitate the development of more consistent, scalable, and effective cell therapies. Full article

Background/Objectives: Glycosylation can affect vaccine antigen structure and function, making site-specific glycan characterization relevant to antigen quality and comparability. However, quantitative approaches for comparing glycan microheterogeneity remain limited. This study evaluated the utility of the glycopeptide abundance distribution spectra framework for measuring similarity among site-specific glycosylation profiles in vaccines and antigen reference reagents across manufacturing conditions. Methods: Intact N-linked glycopeptides were characterized by nanoflow liquid chromatography–tandem mass spectrometry with stepped-energy fragmentation. Products included monovalent and quadrivalent influenza antigens produced in embryonated eggs, Madin–Darby canine kidney cells, or Spodoptera frugiperda cells, together with a SARS-CoV-2 spike vaccine produced in Spodoptera frugiperda cells and a Chinese hamster ovary cell-produced varicella-zoster virus glycoprotein E vaccine. Site-specific glycan distributions were represented as distribution spectra and compared using NIST MS Search software. Dot-product scores ranging from 0 to 999 quantified similarity. Results: Across measured glycosylation sites, distributions clustered into six recurrent classes. Similarity was high for replicate analyses, conserved influenza components across annual formulations, and matched components from different suppliers within the same production platform (similarity scores = 978, 961, and 960, respectively). Similarity was lower between sites within the same protein, between influenza strains, and between production sources (similarity scores = 554, 540, and 209, respectively). Among production-source comparisons, egg- and Madin–Darby canine kidney-derived profiles were most similar, and the overall ordering of glycosylation similarity was consistent with broad phylogenetic relatedness among production hosts. Conclusions: Distribution spectra-based similarity scoring of vaccine glycoproteins provides a quantitative, reusable approach for documenting site-specific glycosylation microheterogeneity. Using this method, we can conclude that production source is the dominant contributor to variation, whereas replicates, annual formulations, and suppliers within the same production platform are highly consistent. Full article

Fourier transform infrared spectroscopy (FTIR) has become an increasingly valuable analytical tool in the dairy industry due to its rapid, non-destructive nature and ability to capture complex chemical information. This study evaluated the feasibility of using FTIR in combination with chemometric modeling to predict the freezing point (FP) of ice cream mix (ICM), an important quality attribute that influences product texture, freezing behavior, and manufacturing efficiency. Traditional methods for FP determination are often labor-intensive and can present challenges when analyzing high-solids dairy systems. In this study, FP values were obtained through a cryoscopic method. FTIR spectra were collected for all samples, and partial least squares (PLS) regression models were developed to relate spectral information to FP values. Model performance metrics indicated that FTIR spectra contained information associated with FP variation; however, overall predictive performance was limited. The relatively weak model accuracy was attributed to the complex nature of FP as a colligative property influenced by multiple compositional factors, including sugars, salts, proteins, and stabilizers, as well as the narrow compositional range and limited sample size of the dataset. Despite these limitations, the results demonstrated the potential of FTIR as a rapid screening tool for FP estimation and provided a foundation for future model development using larger and more compositionally diverse datasets and improved reference methodologies. Full article

Transformative therapeutic innovation should not begin with a molecule—or even a molecular target. It should begin with a clearly defined unmet clinical need. Here, we present a seven-step Translational Therapeutic Development Roadmap that systematically connects an unmet medical need to a developable drug candidate through the disciplined sequence of (i) defining the need, (ii) understanding disease and resistance biology, (iii) building a mechanistic hypothesis, (iv) defining a target product profile (TPP), (v) molecular design and experimental validation, (vi) developability and manufacturability assessment, and (vii) clinical translation. A central conclusion emerging from this review is that resistance biology should be viewed not merely as a cause of therapeutic failure, but as a primary design input for next-generation therapeutic innovation. Our analysis identifies continuous alignment among unmet clinical needs, resistance biology, mechanistic hypothesis, molecular design, developability, and clinical translation as the defining characteristic of successful therapeutic development. We use dual-payload antibody–drug conjugates (ADCs) as a contemporary and highly illustrative case study of this resistance-informed therapeutic development approach. Single-payload ADCs such as trastuzumab deruxtecan and sacituzumab govitecan have transformed treatment across multiple solid tumors, yet most patients ultimately relapse through antigen loss, defective intracellular trafficking, drug efflux, payload-target alterations, and tumor heterogeneity, creating an emerging post-ADC treatment gap. Dual-payload ADCs, which deliver two mechanistically distinct warheads from a single antibody, represent a form of molecular combination therapy designed to increase the barrier to resistance and address multiple escape pathways simultaneously, as well as provide a clinically relevant model for resistance-informed therapeutic design. Using dual-payload ADCs as a worked example, we demonstrate how resistance biology directly informs payload pairing, molecular architecture, conjugation strategy, experimental validation, and developability. Our analysis indicates that successful dual-payload ADC design depends not simply on combining two cytotoxic payloads, but on selecting complementary mechanisms with non-overlapping resistance liabilities while satisfying predefined target product profiles and manufacturability requirements. We further summarize resistance-guided payload pairing strategies, including topoisomerase I plus tubulin inhibitors, topoisomerase I plus DNA-damage-response inhibitors, cytotoxic plus immunomodulatory payloads, and cell-permeable plus non-permeable combinations; the conjugation chemistries that enable defined dual-payload products; the preclinical validation, pharmacological optimization, and developability hurdles that separate promising biology from viable therapeutics; and the rapidly expanding clinical landscape, including the first-in-human program KH815 and emerging bispecific dual-payload constructs. Finally, we demonstrate that the same translational roadmap extends beyond ADCs to radiopharmaceutical conjugates, multispecific antibodies, targeted protein degraders, and cell and gene therapies, indicating that it represents a general framework for therapeutic innovation rather than an ADC-specific strategy. Collectively, this review supports the concept that therapeutic innovation is most successful when unmet clinical needs, resistance biology, molecular design, developability, and clinical translation are considered as an integrated continuum rather than as independent stages of drug discovery. This Translational Therapeutic Development Roadmap provides an organizing framework for guiding the rational development of next-generation targeted therapeutics across diverse therapeutic modalities. Full article