A microfluidic approach to fabricate sucrose decorated liposomes with increased uptake in breast cancer cells

Khorshid, Shiva; Montanari, Mariele; Benedetti, Serena; Moroni, Sofia; Aluigi, Annalisa; Canonico, Barbara; Papa, Stefano; Tiboni, Mattia; Casettari, Luca

doi:10.1016/j.ejpb.2022.07.015

Cited by 14 publications

(4 citation statements)

References 69 publications

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“…LNPs were formulated by using a 3D-printed polypropylene microfluidic chip that was manufactured and kindly donated by Prof. Tiboni. The device has a “split and recombine” design with microchannels having a mean internal diameter of 500 µm, 63 , 64 and a syringe pump was used to control the flow of the aqueous and organic solutions (IPS-14, Inovenso, Boston, MA, USA). A total of 5 mg/mL of lipid mixture was solubilized in methanol and injected in the microfluidic chip against TE buffer (Tris.HCl 10 mM, EDTA 1 mM pH 8.0 in DNAse free dH 2 O), using a total flow rate (TFR) of 10 mL/min and a Flow Rate Ratio (FRR) of 1:1.…”

Section: Methodsmentioning

confidence: 99%

Strategies for Improved pDNA Loading and Protection Using Cationic and Neutral LNPs with Industrial Scalability Potential Using Microfluidic Technology

Ottonelli,

Adani,

Bighinati

et al. 2024

IJN

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Purpose In recent years, microfluidic technologies have become mainstream in producing gene therapy nanomedicines (NMeds) following the Covid-19 vaccine; however, extensive optimizations are needed for each NMed type and genetic material. This article strives to improve LNPs for pDNA loading, protection, and delivery, while minimizing toxicity. Methods The microfluidic technique was optimized to form cationic or neutral LNPs to load pDNA. Classical “post-formulation” DNA addition vs “pre” addition in the aqueous phase were compared. All formulations were characterized (size, homogeneity, zeta potential, morphology, weight yield, and stability), then tested for loading efficiency, nuclease protection, toxicity, and cell uptake. Results Optimized LNPs formulated with DPPC: Chol:DOTAP 1:1:0.1 molar ratio and 10 µg of DOPE-Rhod, had a size of 160 nm and good homogeneity. The chemico-physical characteristics of cationic LNPs worsened when adding 15 µg/mL of pDNA with the “post” method, while maintaining their characteristics up to 100 µg/mL of pDNA with the “pre” addition remaining stable for 30 days. Interestingly, neutral LNPs formulated with the same method loaded up to 50% of the DNA. Both particles could protect the DNA from nucleases even after one month of storage, and low cell toxicity was found up to 40 µg/mL LNPs. Cell uptake occurred within 2 hours for both formulations with the DNA intact in the cytoplasm, outside of the lysosomes. Conclusion In this study, the upcoming microfluidic technique was applied to two strategies to generate pDNA-LNPs. Cationic LNPs could load 10x the amount of DNA as the classical approach, while neutral LNPs, which also loaded and protected DNA, showed lower toxicity and good DNA protection. This is a big step forward at minimizing doses and toxicity of LNP-based gene therapy.

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Section: Methodsmentioning

confidence: 99%

Strategies for Improved pDNA Loading and Protection Using Cationic and Neutral LNPs with Industrial Scalability Potential Using Microfluidic Technology

Ottonelli,

Adani,

Bighinati

et al. 2024

IJN

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“…The integration of nanotechnology-based drugs along with 3D printing (“nanoprinting”) brings printed personalized nanomedicines within the most innovative perspectives for the coming years [ 176 , 177 , 178 ], enabling continuous manufacturing through the use of microfluidic chips, facilitating their translation into clinical practice. Finally, the combination of 3D printing with biopharmaceuticals opens multiple windows to improve current therapies and develop further 3D cultures used in tissue engineering.…”

Section: Conclusion and Future Perspectivesmentioning

confidence: 99%

3D Printing Technologies in Personalized Medicine, Nanomedicines, and Biopharmaceuticals

et al. 2023

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3D printing technologies enable medicine customization adapted to patients’ needs. There are several 3D printing techniques available, but majority of dosage forms and medical devices are printed using nozzle-based extrusion, laser-writing systems, and powder binder jetting. 3D printing has been demonstrated for a broad range of applications in development and targeting solid, semi-solid, and locally applied or implanted medicines. 3D-printed solid dosage forms allow the combination of one or more drugs within the same solid dosage form to improve patient compliance, facilitate deglutition, tailor the release profile, or fabricate new medicines for which no dosage form is available. Sustained-release 3D-printed implants, stents, and medical devices have been used mainly for joint replacement therapies, medical prostheses, and cardiovascular applications. Locally applied medicines, such as wound dressing, microneedles, and medicated contact lenses, have also been manufactured using 3D printing techniques. The challenge is to select the 3D printing technique most suitable for each application and the type of pharmaceutical ink that should be developed that possesses the required physicochemical and biological performance. The integration of biopharmaceuticals and nanotechnology-based drugs along with 3D printing (“nanoprinting”) brings printed personalized nanomedicines within the most innovative perspectives for the coming years. Continuous manufacturing through the use of 3D-printed microfluidic chips facilitates their translation into clinical practice.

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“…They displayed proper bioavailability, long half-lives, size-control measures, low risk-to-benefit ratios, and control release features [ 72 ]. Our experience [ 73 ] with these entities pointed out several advantages of intracellular internalization in triple-negative breast cancer cells (MDA-MB-231) for sucrose-decorated liposomes loaded with berberine hydrochloride. Recently, liposomes demonstrated their significant efficacy in SARS-CoV-2 vaccines [ 74 ].…”

Section: Extracellular Vesicles: An Overview Of Their Origin and Comp...mentioning

confidence: 99%

Extracellular Vesicles as New Players in Drug Delivery: A Focus on Red Blood Cells-Derived EVs

et al. 2023

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The article is divided into several sections, focusing on extracellular vesicles’ (EVs) nature, features, commonly employed methodologies and strategies for their isolation/preparation, and their characterization/visualization. This work aims to give an overview of advances in EVs’ extensive nanomedical-drug delivery applications. Furthermore, considerations for EVs translation to clinical application are summarized here, before focusing the review on a special kind of extracellular vesicles, the ones derived from red blood cells (RBCEVs). Generally, employing EVs as drug carriers means managing entities with advantageous properties over synthetic vehicles or nanoparticles. Besides the fact that certain EVs also reveal intrinsic therapeutic characteristics, in regenerative medicine, EVs nanosize, lipidomic and proteomic profiles enable them to pass biologic barriers and display cell/tissue tropisms; indeed, EVs engineering can further optimize their organ targeting. In the second part of the review, we focus our attention on RBCEVs. First, we describe the biogenesis and composition of those naturally produced by red blood cells (RBCs) under physiological and pathological conditions. Afterwards, we discuss the current procedures to isolate and/or produce RBCEVs in the lab and to load a specific cargo for therapeutic exploitation. Finally, we disclose the most recent applications of RBCEVs at the in vitro and preclinical research level and their potential industrial exploitation. In conclusion, RBCEVs can be, in the near future, a very promising and versatile platform for several clinical applications and pharmaceutical exploitations.

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A microfluidic approach to fabricate sucrose decorated liposomes with increased uptake in breast cancer cells

Cited by 14 publications

References 69 publications

Strategies for Improved pDNA Loading and Protection Using Cationic and Neutral LNPs with Industrial Scalability Potential Using Microfluidic Technology

Strategies for Improved pDNA Loading and Protection Using Cationic and Neutral LNPs with Industrial Scalability Potential Using Microfluidic Technology

3D Printing Technologies in Personalized Medicine, Nanomedicines, and Biopharmaceuticals

Extracellular Vesicles as New Players in Drug Delivery: A Focus on Red Blood Cells-Derived EVs

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