Production and purification of outer membrane vesicles encapsulating green fluorescent protein from Escherichia coli: a step towards scalable OMV technologies DOI Creative Commons
Julian Daniel Torres-Vanegas,

Nicolas Rincon-Tellez,

Paula Guzmán-Sastoque

et al.

Frontiers in Bioengineering and Biotechnology, Journal Year: 2024, Volume and Issue: 12

Published: Nov. 14, 2024

Outer membrane vesicles (OMVs) are spherical structures that contain a small fraction of the periplasm Gram-negative bacteria, surrounded by its outer membrane. They naturally produced and detached from bacterial surface, participate in diverse biological processes, their diameter size is range 10–300 nm. OMVs have gained interest different applications, such as development biosensors, vaccines, protein chips, encapsulation heterologous proteins peptides expressed these microorganisms. However, use applications limited due to low yields high purification costs. In this study, we green fluorescent (GFP) encapsulated into using Escherichia coli JC8031 transformed with pTRC99A-ssTorA-GFP establish production route. Results showed motility strain prevents immobilization alginate, which hampers OMVs. To address issue, zeolite-based column was used chromatographically separate smaller particles. Further experiments will be focused on standardizing at scalable level.

Language: Английский

Classical and emerging approximations for the screening of antimicrobial peptide libraries DOI
Cristian F. Rodríguez, Valentina Quezada,

Valentina Andrade-Pérez

et al.

Elsevier eBooks, Journal Year: 2025, Volume and Issue: unknown, P. 195 - 232

Published: Jan. 1, 2025

Language: Английский

Citations

2

Nanotheranostics Revolutionizing Gene Therapy: Emerging Applications in Gene Delivery Enhancement DOI Creative Commons
Paula Guzmán-Sastoque, Cristian F. Rodríguez,

María Camila Monsalve

et al.

Journal of Nanotheranostics, Journal Year: 2025, Volume and Issue: 6(2), P. 10 - 10

Published: April 9, 2025

Nanotheranostics—where nanoscale materials serve both diagnostic and therapeutic functions—are rapidly transforming gene therapy by tackling critical delivery challenges. This review explores the design engineering of various nanoparticle systems (lipid-based, polymeric, inorganic, hybrid) to enhance stability, targeting, endosomal escape genetic payloads. We discuss how real-time imaging capabilities integrated into these platforms enable precise localization controlled release genes, improving treatment efficacy while reducing off-target effects. Key strategies overcome barriers (such as proton sponge effect photothermal disruption) achieve nuclear are highlighted, along with recent advances in stimuli-responsive that facilitate spatiotemporal control expression. Clinical trials preclinical studies demonstrate expanding role nanotheranostics managing cancer, inherited disorders, cardiovascular neurological diseases. further address regulatory manufacturing hurdles must be for widespread clinical adoption nanoparticle-based therapies. By synthesizing progress ongoing challenges, this underscores transformative potential effective, targeted, image-guided delivery.

Language: Английский

Citations

1

Enhancing Magnetic Micro- and Nanoparticle Separation with a Cost-Effective Microfluidic Device Fabricated by Laser Ablation of PMMA DOI Creative Commons
Cristian F. Rodríguez, Paula Guzmán-Sastoque,

Carolina Muñoz-Camargo

et al.

Micromachines, Journal Year: 2024, Volume and Issue: 15(8), P. 1057 - 1057

Published: Aug. 22, 2024

Superparamagnetic iron oxide micro- and nanoparticles have significant applications in biomedical chemical engineering. This study presents the development evaluation of a novel low-cost microfluidic device for purification hyperconcentration these magnetic particles. The device, fabricated using laser ablation polymethyl methacrylate (PMMA), leverages precise control over fluid dynamics to efficiently separate particles from non-magnetic ones. We assessed device’s performance through Multiphysics simulations empirical tests, focusing on separation magnetite blue carbon dots microparticles polystyrene at various total flow rates (TFRs). For nanoparticle separation, achieved recall up 93.3 ± 4% precision 95.9 1.2% an optimal TFR 2 mL/h, significantly outperforming previous models, which only 50% recall. Microparticle demonstrated accuracy 98.1 1% mL/h both experimental conditions. Lagrangian model effectively captured microparticle microparticles, with close agreement between simulated results. Our findings underscore robust capability distinguishing nanoscales. highlights potential low-cost, non-cleanroom manufacturing techniques produce high-performance devices, thereby expanding their accessibility applicability industrial research settings. integration continuous magnet, as opposed segmented magnets designs, was identified key factor enhancing efficiency.

Language: Английский

Citations

5

A Novel Method for In Vivo Gene Editing in the Brain of Guppies Using Unique Nanoparticles as Delivery Vehicles DOI Creative Commons

María Camila Monsalve,

Miguel Asmad, M.A. Subieta Vásquez

et al.

Research Square (Research Square), Journal Year: 2025, Volume and Issue: unknown

Published: April 1, 2025

Abstract Background Uncovering the genomic basis of traits has advanced rapidly in evolutionary biology and neuroscience, largely through phenotypic with adaptive value been advancing biology, behavior, due to research using non-traditional model systems. Direct gene editing adult brain represents a crucial next step linking genotype phenotype, avoiding confounding effects that arise from modifications during development. However, implementing these technologies beyond traditional laboratory models remains challenging delivery limitations. Methods We developed an intracranial microinjection protocol for guppies (Poecilia reticulata) deliver gene-editing elements cells. designed magnetic nanoparticles functionalized novel translocating agent, non-viral carrier capable transporting linearized nucleic acids across cellular nuclear membranes. comprehensively assessed nanoparticle uptake, colocalization, potential health impacts histological analysis, liver enzyme activity assays, behavioral assessments. Results Our successfully entered cells colocalized nuclei at rates exceeding 50% after two weeks, demonstrating their efficient in vivo editing. Health assessments showed no significant cell death (> 80% viability), toxicity (normal ALT, AST, ALP levels), alterations individual social behaviors, confirming nanoparticles’ biocompatibility systemic safety. Conclusions results, combined previous vitro work our are effective system editing, show they can be used safe interventions P. reticulata. This overcomes major technical barrier acid-carrying vehicle brain. approach provides versatile platform studying genetic mechanisms underlying behavior small freshwater fish while helping overcome limitations conducting functional studies on non-model organisms.

Language: Английский

Citations

0

Magnetoliposomes for nanomedicine: synthesis, characterization, and applications in drug, gene, and peptide delivery DOI
Cristian F. Rodríguez, Paula Guzmán-Sastoque,

Alan Santacruz-Belalcazar

et al.

Expert Opinion on Drug Delivery, Journal Year: 2025, Volume and Issue: unknown

Published: May 15, 2025

Magnetoliposomes represent a transformative advancement in nanomedicine by integrating magnetic nanoparticles with liposomal structures, creating multifunctional delivery platforms that overcome key limitations of conventional drug carriers. These hybrid systems enable precision targeting through external fields, controlled release via hyperthermia, and real-time theranostic capabilities, offering unprecedented spatiotemporal control over therapeutic administration. This manuscript focused primarily on studies from 2023-2025 however, few select older references were included to provide background context.This review examines the fundamental design principles Magnetoliposomes, including bilayer composition, nanoparticle integration strategies, physicochemical properties governing their biological performance. We comprehensively assess synthesis methodologies - traditional thin-film hydration advanced microfluidic approaches highlighting impact colloidal stability, encapsulation, scaling potential. Characterization techniques essential for quality regulatory approval are systematically reviewed, followed applications across oncology, gene delivery, neurology, infectious disease treatment, supported recent experimental evidence. While magnetoliposomes show remarkable versatility, clinical translation requires addressing biocompatibility concerns, manufacturing scalability, hurdles. Integration artificial intelligence, organ-on-chip technologies, personalized medicine will likely accelerate development toward reality, potentially revolutionizing treatment paradigms complex diseases tailored interventions.

Language: Английский

Citations

0

Production and purification of outer membrane vesicles encapsulating green fluorescent protein from Escherichia coli: a step towards scalable OMV technologies DOI Creative Commons
Julian Daniel Torres-Vanegas,

Nicolas Rincon-Tellez,

Paula Guzmán-Sastoque

et al.

Frontiers in Bioengineering and Biotechnology, Journal Year: 2024, Volume and Issue: 12

Published: Nov. 14, 2024

Outer membrane vesicles (OMVs) are spherical structures that contain a small fraction of the periplasm Gram-negative bacteria, surrounded by its outer membrane. They naturally produced and detached from bacterial surface, participate in diverse biological processes, their diameter size is range 10–300 nm. OMVs have gained interest different applications, such as development biosensors, vaccines, protein chips, encapsulation heterologous proteins peptides expressed these microorganisms. However, use applications limited due to low yields high purification costs. In this study, we green fluorescent (GFP) encapsulated into using Escherichia coli JC8031 transformed with pTRC99A-ssTorA-GFP establish production route. Results showed motility strain prevents immobilization alginate, which hampers OMVs. To address issue, zeolite-based column was used chromatographically separate smaller particles. Further experiments will be focused on standardizing at scalable level.

Language: Английский

Citations

1