In Vitro Assessment of a Doubly Adjuvanted Self-Emulsified Nanoemulsion as a Delivery Vehicle for Antigenic Proteins

<b>Background/Objectives:</b> Leishmaniasis is a prevailing infectious disease transmitted via infected phlebotomine sandflies. The lack of an efficient vaccine with respect to immunogenic antigens and adjuvanted delivery systems impedes its control. Following the induction of immune res...

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Main Authors: Evgenia Tsanaktsidou, Maritsa Margaroni, Evdokia Karagouni, Costas Kiparissides, Olga Kammona
Format: Article
Language:English
Published: MDPI AG 2025-07-01
Series:Pharmaceutics
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Online Access:https://www.mdpi.com/1999-4923/17/7/870
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author Evgenia Tsanaktsidou
Maritsa Margaroni
Evdokia Karagouni
Costas Kiparissides
Olga Kammona
author_facet Evgenia Tsanaktsidou
Maritsa Margaroni
Evdokia Karagouni
Costas Kiparissides
Olga Kammona
author_sort Evgenia Tsanaktsidou
collection DOAJ
description <b>Background/Objectives:</b> Leishmaniasis is a prevailing infectious disease transmitted via infected phlebotomine sandflies. The lack of an efficient vaccine with respect to immunogenic antigens and adjuvanted delivery systems impedes its control. Following the induction of immune responses in mice vaccinated with multi-epitope <i>Leishmania</i> peptides (LeishPts) encapsulated in doubly adjuvanted self-nanoemulsifying drug delivery systems (ST-SNEDDSs), this study aims to assess ST-SNEDDS-based nanoemulsions as vehicles for the delivery of antigenic proteins. <b>Methods:</b> Model antigens (e.g., BSA-FITC, OVA) were encapsulated in ST-SNEDDS after being complexed with the cationic phospholipid dimyristoyl phosphatidylglycerol (DMPG) via hydrophobic ion pairing. The nanoemulsions were characterized with respect to droplet diameter, zeta potential, stability, protein loading, protein release from the nanodroplets in different release media and cell uptake. <b>Results:</b> Both model antigens exhibited high encapsulation efficiency (>95%) and their release from the nanodroplets was shown to be strongly affected by the type of release medium (e.g., PBS, FBS 10% <i>v</i>/<i>v</i>) and the ratio of its volume to that of the oily phase, in agreement with predictions of protein release. Protein-loaded nanoemulsion droplets labeled with Cy-5 were found to be efficiently taken up by macrophages (J774A.1) in vitro. However, no colocalization of the labeled nanodroplets and BSA-FITC could be observed. <b>Conclusions:</b> It was revealed that in contrast with LeishPts, whole protein molecules may not be appropriate antigenic cargo for ST-SNEDDS formulations due to the rapid protein release from the nanodroplets in release media simulating in vitro culture and in vivo conditions such as FBS 10% <i>v</i>/<i>v</i>.
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spelling doaj-art-030efd28fa0247d4adbc7c3c05616cc82025-08-20T03:08:06ZengMDPI AGPharmaceutics1999-49232025-07-0117787010.3390/pharmaceutics17070870In Vitro Assessment of a Doubly Adjuvanted Self-Emulsified Nanoemulsion as a Delivery Vehicle for Antigenic ProteinsEvgenia Tsanaktsidou0Maritsa Margaroni1Evdokia Karagouni2Costas Kiparissides3Olga Kammona4Chemical Process and Energy Resources Institute, Centre for Research and Technology Hellas, P.O. Box 60361, 57001 Thessaloniki, GreeceImmunology of Infection Laboratory, Hellenic Pasteur Institute, 11521 Athens, GreeceImmunology of Infection Laboratory, Hellenic Pasteur Institute, 11521 Athens, GreeceChemical Process and Energy Resources Institute, Centre for Research and Technology Hellas, P.O. Box 60361, 57001 Thessaloniki, GreeceChemical Process and Energy Resources Institute, Centre for Research and Technology Hellas, P.O. Box 60361, 57001 Thessaloniki, Greece<b>Background/Objectives:</b> Leishmaniasis is a prevailing infectious disease transmitted via infected phlebotomine sandflies. The lack of an efficient vaccine with respect to immunogenic antigens and adjuvanted delivery systems impedes its control. Following the induction of immune responses in mice vaccinated with multi-epitope <i>Leishmania</i> peptides (LeishPts) encapsulated in doubly adjuvanted self-nanoemulsifying drug delivery systems (ST-SNEDDSs), this study aims to assess ST-SNEDDS-based nanoemulsions as vehicles for the delivery of antigenic proteins. <b>Methods:</b> Model antigens (e.g., BSA-FITC, OVA) were encapsulated in ST-SNEDDS after being complexed with the cationic phospholipid dimyristoyl phosphatidylglycerol (DMPG) via hydrophobic ion pairing. The nanoemulsions were characterized with respect to droplet diameter, zeta potential, stability, protein loading, protein release from the nanodroplets in different release media and cell uptake. <b>Results:</b> Both model antigens exhibited high encapsulation efficiency (>95%) and their release from the nanodroplets was shown to be strongly affected by the type of release medium (e.g., PBS, FBS 10% <i>v</i>/<i>v</i>) and the ratio of its volume to that of the oily phase, in agreement with predictions of protein release. Protein-loaded nanoemulsion droplets labeled with Cy-5 were found to be efficiently taken up by macrophages (J774A.1) in vitro. However, no colocalization of the labeled nanodroplets and BSA-FITC could be observed. <b>Conclusions:</b> It was revealed that in contrast with LeishPts, whole protein molecules may not be appropriate antigenic cargo for ST-SNEDDS formulations due to the rapid protein release from the nanodroplets in release media simulating in vitro culture and in vivo conditions such as FBS 10% <i>v</i>/<i>v</i>.https://www.mdpi.com/1999-4923/17/7/870self-nanoemulsifying drug delivery systemsantigenic proteinsnanoemulsionprotein releaseadjuvantvaccine carrier
spellingShingle Evgenia Tsanaktsidou
Maritsa Margaroni
Evdokia Karagouni
Costas Kiparissides
Olga Kammona
In Vitro Assessment of a Doubly Adjuvanted Self-Emulsified Nanoemulsion as a Delivery Vehicle for Antigenic Proteins
Pharmaceutics
self-nanoemulsifying drug delivery systems
antigenic proteins
nanoemulsion
protein release
adjuvant
vaccine carrier
title In Vitro Assessment of a Doubly Adjuvanted Self-Emulsified Nanoemulsion as a Delivery Vehicle for Antigenic Proteins
title_full In Vitro Assessment of a Doubly Adjuvanted Self-Emulsified Nanoemulsion as a Delivery Vehicle for Antigenic Proteins
title_fullStr In Vitro Assessment of a Doubly Adjuvanted Self-Emulsified Nanoemulsion as a Delivery Vehicle for Antigenic Proteins
title_full_unstemmed In Vitro Assessment of a Doubly Adjuvanted Self-Emulsified Nanoemulsion as a Delivery Vehicle for Antigenic Proteins
title_short In Vitro Assessment of a Doubly Adjuvanted Self-Emulsified Nanoemulsion as a Delivery Vehicle for Antigenic Proteins
title_sort in vitro assessment of a doubly adjuvanted self emulsified nanoemulsion as a delivery vehicle for antigenic proteins
topic self-nanoemulsifying drug delivery systems
antigenic proteins
nanoemulsion
protein release
adjuvant
vaccine carrier
url https://www.mdpi.com/1999-4923/17/7/870
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