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20. Development of Focused Ultrasound-Assisted Nanoplexes for RNA Delivery.

Author

Ranjan, Sanjeev, Bosch, Stef, Lukkari, Hannamari, Schirmer, Johanna, Aaltonen, Niina, Nieminen, Heikki J., Lehto, Vesa-Pekka, Urtti, Arto, Lajunen, Tatu, and Rilla, Kirsi

Subjects
HIGH-intensity focused ultrasound, NON-coding RNA, DISTRIBUTION (Probability theory), RNA, ATOMIC force microscopy, DRUG delivery systems
Abstract

RNA-based therapeutics, including siRNA, have obtained recognition in recent years due to their potential to treat various chronic and rare diseases. However, there are still limitations to lipid-based drug delivery systems in the clinical use of RNA therapeutics due to the need for optimization in the design and the preparation process. In this study, we propose adaptive focused ultrasound (AFU) as a drug loading technique to protect RNA from degradation by encapsulating small RNA in nanoliposomes, which we term nanoplexes. The AFU method is non-invasive and isothermal, as nanoplexes are produced without direct contact with any external materials while maintaining precise temperature control according to the desired settings. The controllability of sample treatments can be effectively modulated, allowing for a wide range of ultrasound intensities to be applied. Importantly, the absence of co-solvents in the process eliminates the need for additional substances, thereby minimizing the potential for cross-contaminations. Since AFU is a non-invasive method, the entire process can be conducted under sterile conditions. A minimal volume (300 μL) is required for this process, and the treatment is speedy (10 min in this study). Our in vitro experiments with silencer CD44 siRNA, which performs as a model therapeutic drug in different mammalian cell lines, showed encouraging results (knockdown > 80%). To quantify gene silencing efficacy, we employed quantitative polymerase chain reaction (qPCR). Additionally, cryo-electron microscopy (cryo-EM) and atomic force microscopy (AFM) techniques were employed to capture images of nanoplexes. These images revealed the presence of individual nanoparticles measuring approximately 100–200 nm in contrast with the random distribution of clustered complexes observed in ultrasound-untreated samples of liposome nanoparticles and siRNA. AFU holds great potential as a standardized liposome processing and loading method because its process is fast, sterile, and does not require additional solvents. [ABSTRACT FROM AUTHOR]

Published
2024
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31. The prophylactic value of TNF‐α inhibitors against retinal cell apoptosis and optic nerve axon loss after corneal surgery or trauma.

Author

Paschalis, Eleftherios I., Zhou, Chengxin, Sharma, Jyoti, Dohlman, Thomas H., Kim, Sarah, Lei, Fengyang, Chodosh, James, Vavvas, Demetrios, Urtti, Arto, Papaliodis, George, and Dohlman, Claes H.

Subjects
ADALIMUMAB, OPTIC nerve, AXONS, TRAUMA surgery, CORNEAL transplantation, CELL death, NEURONS, RETINAL surgery
Abstract

Background and Purpose: Late secondary glaucoma is an often‐severe complication after acute events like anterior segment surgery, trauma and infection. TNF‐α is a major mediator that is rapidly upregulated, diffusing also to the retina and causes apoptosis of the ganglion cells and degeneration of their optic nerve axons (mediating steps to glaucomatous damage). Anti‐TNF‐α antibodies are in animals very effective in protecting the retinal cells and the optic nerve—and might therefore be useful prophylactically against secondary glaucoma in future such patients. Here we evaluate (1) toxicity and (2) efficacy of two TNF‐α inhibitors (adalimumab and infliximab), in rabbits by subconjunctival administration. Methods: For drug toxicity, animals with normal, unburned corneas were injected with adalimumab (0.4, 4, or 40 mg), or infliximab (1, 10, or 100 mg). For drug efficacy, other animals were subjected to alkali burn before such injection, or steroids (for control). The rabbits were evaluated clinically with slit lamp and photography, electroretinography, optical coherence tomography, and intraocular pressure manometry. A sub‐set of eyes were stained ex vivo after 3 days for retinal cell apoptosis (TUNEL). In other experiments the optic nerves were evaluated by paraphenylenediamine staining after 50 or 90 days. Loss of retinal cells and optic nerve degeneration were quantified. Results: Subconjunctival administration of 0.4 mg or 4.0 mg adalimumab were well tolerated, whereas 40.0 mg was toxic to the retina. 1, 10, or 100 mg infliximab were also well tolerated. Analysis of the optic nerve axons after 50 days confirmed the safety of 4.0 mg adalimumab and of 100 mg infliximab. For efficacy, 4.0 mg adalimumab subconjunctivally in 0.08 mL provided practically full protection against retinal cell apoptosis 3 days following alkali burn, and infliximab 100 mg only slightly less. At 90 days following burn injury, control optic nerves showed about 50% axon loss as compared to 8% in the adalimumab treatment group. Conclusions: Subconjunctival injection of 4.0 mg adalimumab in rabbits shows no eye toxicity and provides excellent neuroprotection, both short (3 days) and long‐term (90 days). Our total. accumulated data from several of our studies, combined with the present paper, suggest that corneal injuries, including surgery, might benefit from routine administration of anti‐TNF‐α biologics to reduce inflammation and future secondary glaucoma. [ABSTRACT FROM AUTHOR]

Published
2024
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32. The prophylactic value of TNF‐α inhibitors against retinal cell apoptosis and optic nerve axon loss after corneal surgery or trauma

Author

Paschalis, Eleftherios I., primary, Zhou, Chengxin, additional, Sharma, Jyoti, additional, Dohlman, Thomas H., additional, Kim, Sarah, additional, Lei, Fengyang, additional, Chodosh, James, additional, Vavvas, Demetrios, additional, Urtti, Arto, additional, Papaliodis, George, additional, and Dohlman, Claes H., additional

Published
2023
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36. Mass spectrometry in ocular drug research

Author

del Amo, Eva M., primary, Vellonen, Kati‐Sisko, additional, Urtti, Arto, additional, Terasaki, Tetsuya, additional, Hammid, Anam, additional, Honkakoski, Paavo, additional, and Auriola, Seppo, additional

Published
2023
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37. Microemulsions

Author

Pappinen, Sari, Urtti, Arto, Dragicevic, Nina, editor, and Maibach, Howard I., editor

Published
2016
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41. Pharmacokinetic aspects of retinal drug delivery

Author

del Amo, Eva M., Rimpelä, Anna-Kaisa, Heikkinen, Emma, Kari, Otto K., Ramsay, Eva, Lajunen, Tatu, Schmitt, Mechthild, Pelkonen, Laura, Bhattacharya, Madhushree, Richardson, Dominique, Subrizi, Astrid, Turunen, Tiina, Reinisalo, Mika, Itkonen, Jaakko, Toropainen, Elisa, Casteleijn, Marco, Kidron, Heidi, Antopolsky, Maxim, Vellonen, Kati-Sisko, Ruponen, Marika, and Urtti, Arto

Published
2017
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43. Artificially cloaked viral nanovaccine for cancer immunotherapy

Author

Fusciello, Manlio, Fontana, Flavia, Tähtinen, Siri, Capasso, Cristian, Feola, Sara, Martins, Beatriz, Chiaro, Jacopo, Peltonen, Karita, Ylösmäki, Leena, Ylösmäki, Erkko, Hamdan, Firas, Kari, Otto K., Ndika, Joseph, Alenius, Harri, Urtti, Arto, Hirvonen, Jouni T., Santos, Hélder A., and Cerullo, Vincenzo

Published
2019
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45. Tailoring surface properties of liposomes for dexamethasone intraocular administration.

Author

UCL - SSS/LDRI - Louvain Drug Research Institute, Al-Amin, M D, Mastrotto, Francesca, Subrizi, Astrid, Sen, Merve, Turunen, Tiina, Arango-Gonzalez, Blanca, Ueffing, Marius, Malfanti, Alessio, Urtti, Arto, Salmaso, Stefano, Caliceti, Paolo, UCL - SSS/LDRI - Louvain Drug Research Institute, Al-Amin, M D, Mastrotto, Francesca, Subrizi, Astrid, Sen, Merve, Turunen, Tiina, Arango-Gonzalez, Blanca, Ueffing, Marius, Malfanti, Alessio, Urtti, Arto, Salmaso, Stefano, and Caliceti, Paolo

Abstract

Diseases of the posterior eye segment are often characterized by intraocular inflammation, which causes, in the long term, severe impairment of eye functions and, ultimately, vision loss. Aimed at enhancing the delivery of anti-inflammatory drugs to the posterior eye segment upon intravitreal administration, we developed liposomes with an engineered surface to control their diffusivity in the vitreous and retina association. Hydrogenated soybean phosphatidylcholine (HSPC)/cholesterol liposomes were coated with (agmatinyl)-maltotriosyl-acetamido-N-(octadec-9-en-1-yl)hexanamide (Agm-M-Oleate), a synthetic non-peptidic cell penetration enhancer (CPE), and/or 5% of mPEG-DSPE. The zeta potential of liposomes increased, and the mobility in bovine vitreous and colloidal stability decreased with the Agm-M-Oleate coating concentration. Oppositely, mPEG-DSPE decreased the zeta potential of liposomes and restored both the diffusivity and the stability in vitreous. Liposomes with 5 mol% Agm-M-Oleate coating were well tolerated by ARPE-19 retina cells either with or without mPEG-DSPE, while 10 mol% Agm-M-Oleate showed cytotoxicity. Agm-M-Oleate promoted the association of liposomes to ARPE-19 cells with respect to plain liposomes, while mPEG-DSPE slightly reduced the cell interaction. Dexamethasone hemisuccinate (DH) was remotely loaded into liposomes with a loading capacity of ~10 wt/wt%. Interestingly, mPEG-DSPE coating reduced the rate of DH release and enhanced the disposition of Agm-M-Oleate coated liposomes in the ARPE-19 cell cytosol resulting in a more efficient anti-inflammatory effect. Finally, mPEG-DSPE enhanced the association of DH-loaded Agm-M-Oleate coated liposomes to explanted rat retina, which reflected in higher viability of inner and outer nuclear layer cells.

Published
2023

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