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Nature Communications Aug 2023Adenosylcobalamin (AdoCbl), a biologically active form of vitamin B (coenzyme B), is one of the most complex metal-containing natural compounds and an essential vitamin...
Adenosylcobalamin (AdoCbl), a biologically active form of vitamin B (coenzyme B), is one of the most complex metal-containing natural compounds and an essential vitamin for animals. However, AdoCbl can only be de novo synthesized by prokaryotes, and its industrial manufacturing to date was limited to bacterial fermentation. Here, we report a method for the synthesis of AdoCbl based on a cell-free reaction system performing a cascade of catalytic reactions from 5-aminolevulinic acid (5-ALA), an inexpensive compound. More than 30 biocatalytic reactions are integrated and optimized to achieve the complete cell-free synthesis of AdoCbl, after overcoming feedback inhibition, the complicated detection, instability of intermediate products, as well as imbalance and competition of cofactors. In the end, this cell-free system produces 417.41 μg/L and 5.78 mg/L of AdoCbl using 5-ALA and the purified intermediate product hydrogenobyrate as substrates, respectively. The strategies of coordinating synthetic modules of complex cell-free system describe here will be generally useful for developing cell-free platforms to produce complex natural compounds with long and complicated biosynthetic pathways.
Topics: Animals; Vitamin B 12; Cell-Free System; Vitamins; Aminolevulinic Acid; Biocatalysis
PubMed: 37620358
DOI: 10.1038/s41467-023-40932-4 -
ArXiv Aug 2023Our ability to produce human-scale bio-manufactured organs is critically limited by the need for vascularization and perfusion. For tissues of variable size and shape,...
Our ability to produce human-scale bio-manufactured organs is critically limited by the need for vascularization and perfusion. For tissues of variable size and shape, including arbitrarily complex geometries, designing and printing vasculature capable of adequate perfusion has posed a major hurdle. Here, we introduce a model-driven design pipeline combining accelerated optimization methods for fast synthetic vascular tree generation and computational hemodynamics models. We demonstrate rapid generation, simulation, and 3D printing of synthetic vasculature in complex geometries, from small tissue constructs to organ scale networks. We introduce key algorithmic advances that all together accelerate synthetic vascular generation by more than 230 -fold compared to standard methods and enable their use in arbitrarily complex shapes through localized implicit functions. Furthermore, we provide techniques for joining vascular trees into watertight networks suitable for hemodynamic CFD and 3D fabrication. We demonstrate that organ-scale vascular network models can be generated in silico within minutes and can be used to perfuse engineered and anatomic models including a bioreactor, annulus, bi-ventricular heart, and gyrus. We further show that this flexible pipeline can be applied to two common modes of bioprinting with free-form reversible embedding of suspended hydrogels and writing into soft matter. Our synthetic vascular tree generation pipeline enables rapid, scalable vascular model generation and fluid analysis for bio-manufactured tissues necessary for future scale up and production.
PubMed: 37645046
DOI: No ID Found -
Food Science & Nutrition Jul 2023Functional stirred yogurt samples were manufactured with combinations of grape pomace (GP) and flaxseed oil (FO) in microencapsulated or free forms (2% w/w) and quality...
Functional stirred yogurt manufactured using co-microencapsulated or free forms of grape pomace and flaxseed oil as bioactive ingredients: Physicochemical, antioxidant, rheological, microstructural, and sensory properties.
Functional stirred yogurt samples were manufactured with combinations of grape pomace (GP) and flaxseed oil (FO) in microencapsulated or free forms (2% w/w) and quality characteristics of yogurts were investigated during 21 days of storage. The incorporation of GP and FO in microencapsulated or free forms caused a significant decrease in pH, syneresis, and a significant increase in acidity, water holding capacity, and viscosity of stirred yogurt ( < .05). While stirred yogurt containing GP and FO in free form had the highest loss modulus (G″), all yogurt samples represented solid-like behavior. Stirred yogurts containing the microencapsulated form of GP and FO showed the highest amount of phenolics and antioxidant activity compared with the two other yogurt samples ( < .05). More compact structure and higher gel strength were observed in stirred yogurts formulated with the microencapsulated or free form of GP and FO, compared to the control yogurt sample. The overall sensory acceptability of stirred yogurt manufactured using the encapsulated form of GP and FO was not significantly different from the control yogurt sample ( > .05). In conclusion of this competitive study, GP and FO as bioactive compounds could be used in the microencapsulated form in order to develop functional stirred yogurt with specific quality characteristics.
PubMed: 37457195
DOI: 10.1002/fsn3.3385 -
Pharmaceutics Jul 2023Currently, the field of medicine is drastically advancing, mainly due to the progress in emerging areas such as nanomedicine, regenerative medicine, and personalized...
Currently, the field of medicine is drastically advancing, mainly due to the progress in emerging areas such as nanomedicine, regenerative medicine, and personalized medicine. For example, the development of novel drug delivery systems in the form of nanoparticles is improving the liberation, absorption, distribution, metabolism, and excretion (LADME) properties of the derived formulations, with a consequent enhancement in the treatment efficacy, a reduction in the secondary effects, and an increase in compliance with the dosage guidelines. Additionally, the use of biocompatible scaffolds is translating into the possibility of regenerating biological tissues. Personalized medicine is also benefiting from the advantages offered by additive manufacturing. However, all these areas have in common the need to develop novel materials or composites that fulfill the requirements of each application. Therefore, the aim of this Special Issue was to identify novel materials/composites that have been developed with specific characteristics for the designed biomedical application.
PubMed: 37514123
DOI: 10.3390/pharmaceutics15071938 -
Signal Transduction and Targeted Therapy Mar 2024The Orthopoxvirus genus, especially variola virus (VARV), monkeypox virus (MPXV), remains a significant public health threat worldwide. The development of therapeutic...
The Orthopoxvirus genus, especially variola virus (VARV), monkeypox virus (MPXV), remains a significant public health threat worldwide. The development of therapeutic antibodies against orthopoxviruses is largely hampered by the high cost of antibody engineering and manufacturing processes. mRNA-encoded antibodies have emerged as a powerful and universal platform for rapid antibody production. Herein, by using the established lipid nanoparticle (LNP)-encapsulated mRNA platform, we constructed four mRNA combinations that encode monoclonal antibodies with broad neutralization activities against orthopoxviruses. In vivo characterization demonstrated that a single intravenous injection of each LNP-encapsulated mRNA antibody in mice resulted in the rapid production of neutralizing antibodies. More importantly, mRNA antibody treatments showed significant protection from weight loss and mortality in the vaccinia virus (VACV) lethal challenge mouse model, and a unique mRNA antibody cocktail, Mix2a, exhibited superior in vivo protection by targeting both intracellular mature virus (IMV)-form and extracellular enveloped virus (EEV)-form viruses. In summary, our results demonstrate the proof-of-concept production of orthopoxvirus antibodies via the LNP-mRNA platform, highlighting the great potential of tailored mRNA antibody combinations as a universal strategy to combat orthopoxvirus as well as other emerging viruses.
Topics: Animals; Mice; Orthopoxvirus; Combined Antibody Therapeutics; Vaccinia; Antibodies, Viral; Vaccinia virus
PubMed: 38531869
DOI: 10.1038/s41392-024-01766-8 -
Frontiers in Immunology 2024The clinical success of chimeric antigen receptor-modified T cells (CAR-T cells) for hematological malignancies has not been reproduced for solid tumors, partly due to...
INTRODUCTION
The clinical success of chimeric antigen receptor-modified T cells (CAR-T cells) for hematological malignancies has not been reproduced for solid tumors, partly due to the lack of cancer-type specific antigens. In this work, we used a novel combinatorial approach consisting of a versatile anti-FITC CAR-T effector cells plus an FITC-conjugated neuroblastoma (NB)-targeting linker, an FITC-conjugated monoclonal antibody (Dinutuximab) that recognizes GD2.
METHODS
We compared cord blood (CB), and CD45RA-enriched peripheral blood leukapheresis product (45RA) as allogeneic sources of T cells, using peripheral blood (PB) as a control to choose the best condition for anti-FITC CAR-T production. Cells were manufactured under two cytokine conditions (IL-2 IL-7+IL-15+IL-21) with or without CD3/CD28 stimulation. Immune phenotype, vector copy number, and genomic integrity of the final products were determined for cell characterization and quality control assessment. Functionality and antitumor capacity of CB/45RA-derived anti-FITC CAR-T cells were analyzed in co-culture with different anti-GD2-FITC labeled NB cell lines.
RESULTS
The IL-7+IL-15+IL-21 cocktail, in addition to co-stimulation signals, resulted in a favorable cell proliferation rate and maintained less differentiated immune phenotypes in both CB and 45RA T cells. Therefore, it was used for CAR-T cell manufacturing and further characterization. CB and CD45RA-derived anti-FITC CAR-T cells cultured with IL-7+IL-15+IL-21 retained a predominantly naïve phenotype compared with controls. In the presence of the NB-FITC targeting, CD4+ CB-derived anti-FITC CAR-T cells showed the highest values of co-stimulatory receptors OX40 and 4-1BB, and CD8+ CAR-T cells exhibited high levels of PD-1 and 4-1BB and low levels of TIM3 and OX40, compared with CAR-T cells form the other sources studied. CB-derived anti-FITC CAR-T cells released the highest amounts of cytokines (IFN-γ and TNF-α) into co-culture supernatants. The viability of NB target cells decreased to 30% when co-cultured with CB-derived CAR-T cells during 48h.
CONCLUSION
CB and 45RA-derived T cells may be used as allogeneic sources of T cells to produce CAR-T cells. Moreover, ex vivo culture with IL-7+IL-15+IL-21 could favor CAR-T products with a longer persistence in the host. Our strategy may complement the current use of Dinutuximab in treating NB through its combination with a targeted CAR-T cell approach.
Topics: Humans; T-Lymphocytes; Receptors, Chimeric Antigen; Interleukin-15; Interleukin-7; Fluorescein-5-isothiocyanate; Cytokines; Neuroblastoma
PubMed: 38601159
DOI: 10.3389/fimmu.2024.1375833 -
Advanced Science (Weinheim,... Aug 2023Aerogel-based biomaterials are increasingly being considered for biomedical applications due to their unique properties such as high porosity, hierarchical porous... (Review)
Review
Aerogel-based biomaterials are increasingly being considered for biomedical applications due to their unique properties such as high porosity, hierarchical porous network, and large specific pore surface area. Depending on the pore size of the aerogel, biological effects such as cell adhesion, fluid absorption, oxygen permeability, and metabolite exchange can be altered. Based on the diverse potential of aerogels in biomedical applications, this paper provides a comprehensive review of fabrication processes including sol-gel, aging, drying, and self-assembly along with the materials that can be used to form aerogels. In addition to the technology utilizing aerogel itself, it also provides insight into the applicability of aerogel based on additive manufacturing technology. To this end, how microfluidic-based technologies and 3D printing can be combined with aerogel-based materials for biomedical applications is discussed. Furthermore, previously reported examples of aerogels for regenerative medicine and biomedical applications are thoroughly reviewed. A wide range of applications with aerogels including wound healing, drug delivery, tissue engineering, and diagnostics are demonstrated. Finally, the prospects for aerogel-based biomedical applications are presented. The understanding of the fabrication, modification, and applicability of aerogels through this study is expected to shed light on the biomedical utilization of aerogels.
Topics: Biocompatible Materials; Tissue Engineering; Desiccation; Wound Healing
PubMed: 37217831
DOI: 10.1002/advs.202204681 -
Photodiagnosis and Photodynamic Therapy Apr 2024Unlike surgical instruments and endoscopic equipment, Photodynamic Therapy (PDT) devices are not readily available or accessible to the clinicians who may like to add... (Review)
Review
Unlike surgical instruments and endoscopic equipment, Photodynamic Therapy (PDT) devices are not readily available or accessible to the clinicians who may like to add this form of treatment modality for selected patients and on an ad hock basis. There is in fact a vacuum in finding the "tools" of PDT for those clinicians who are not part of a "Centre" with a built-in knowledge base and contacts for manufacturers. In this compendium the Yorkshire Laser Centre /YLC in the UK, (the Project of the Moghissi Laser Trust - (Charity number 326689) requested three experienced clinicians (RA, ZH, KM) to produce essential information on the use of and equipment for PDT in the clinic. The YLC also sponsored a researcher (ID) to search and compile a detailed but non-exhaustive list of approved photosensitizing agents, pharmaceutical companies, light sources and laser manufacturers with appropriate delivery devices for PDT. Thus, this Mini -Compendium is the end result of what is hoped to be a useful adjunct for practitioners, scientists and students of PDT.
Topics: Photochemotherapy; Photosensitizing Agents; Humans
PubMed: 38513811
DOI: 10.1016/j.pdpdt.2024.104058 -
Journal of Functional Biomaterials Oct 2023Additively manufactured synthetic bone scaffolds have emerged as promising candidates for the replacement and regeneration of damaged and diseased bones. By employing...
Additively manufactured synthetic bone scaffolds have emerged as promising candidates for the replacement and regeneration of damaged and diseased bones. By employing optimal pore architecture, including pore morphology, sizes, and porosities, 3D-printed scaffolds can closely mimic the mechanical properties of natural bone and withstand external loads. This study aims to investigate the deformation pattern exhibited by polymeric bone scaffolds fabricated using the PolyJet (PJ) 3D printing technique. Cubic and hexagonal closed-packed uniform scaffolds with porosities of 30%, 50%, and 70% are utilized in finite element (FE) models. The crushable foam plasticity model is employed to analyze the scaffolds' mechanical response under quasi-static compression. Experimental validation of the FE results demonstrates a favorable agreement, with an average percentage error of 12.27% ± 7.1%. Moreover, the yield strength and elastic modulus of the scaffolds are evaluated and compared, revealing notable differences between cubic and hexagonal closed-packed designs. The 30%, 50%, and 70% porous cubic pore-shaped bone scaffolds exhibit significantly higher yield strengths of 46.89%, 58.29%, and 66.09%, respectively, compared to the hexagonal closed-packed bone scaffolds at percentage strains of 5%, 6%, and 7%. Similarly, the elastic modulus of the 30%, 50%, and 70% porous cubic pore-shaped bone scaffolds is 42.68%, 59.70%, and 58.18% higher, respectively, than the hexagonal closed-packed bone scaffolds at the same percentage strain levels. Furthermore, it is observed in comparison with our previous study the μSLA-printed bone scaffolds demonstrate 1.5 times higher elastic moduli and yield strengths compared to the PJ-printed bone scaffolds.
PubMed: 37888161
DOI: 10.3390/jfb14100496 -
ACS Omega Feb 2024Metallic foam is a popular topic due to its diverse industrial applications and unique combination of properties. Metallic foam is significantly lighter than nonfoam... (Review)
Review
Metallic foam is a popular topic due to its diverse industrial applications and unique combination of properties. Metallic foam is significantly lighter than nonfoam metal materials due to its porous structure, which incorporates a substantial amount of air or voids. This lower density makes metallic foam advantageous in applications in which weight reduction is critical. This makes it ideal for the aerospace, automotive, and construction industries; also, its versatile nature continues to make it an attractive material for various industrial applications such as impact absorbers, heat exchangers, and biomedical and marine engineering. However, the choice between metallic foam and nonfoam metal also depends on other factors like mechanical properties, cost, and specific application requirements. This review describes various fabrication methods of metallic foam that include the liquid metallurgy route which uses liquid or semiliquid metal, the powder metallurgy route uses metal in powder form, metal ion, and the metal vapor route which uses electrolytic deposition method to produce metallic foam. These methods include direct gas injection, adding blowing agents in solid or liquid metals, investment casting, the addition of a space holder in the precursor, metallic ion, vapor deposition on a polymer sponge, and many more. The morphology of metallic foam depends upon the method that is chosen for fabrication, and up to 98% porosity can be achieved by these methods. Additive manufacturing for metallic foam fabrication is an emerging field based on selective laser melting and electron beam melting principles. It has exceptional possibilities for generating complicated 3D shapes and customizing the material characteristics. The main purpose of this review article is to give significant insights into the various production procedures for metallic foams to researchers, engineers, and industry experts, assisting in the selection of acceptable methods depending on individual application needs. This review investigates the manufacturing conditions for metallic foams and finally discusses their advantages, drawbacks, and obstacles in mass production. The findings add to current efforts to expand metallic foam technology and encourage its wider application across diverse sectors, opening the path for future research and development.
PubMed: 38371845
DOI: 10.1021/acsomega.3c08613