preparation and characterization of cationic nanoemulsion&#45;encapsulated retinoic acid

Evaluering av immunogenisitet av nanoemulsjonsinnkapslet retinsyre hos mus

Adjuvants are often used to enhance the efficacy of a vaccine by stimulating the immune system to respond more strongly to the vaccine, thereby increasing immunity to a particular pathogen1. Nanoemulsion (NE) adjuvant refers to a colloidal dispersion system with thermodynamic stability by emulsifying a certain proportion of oil phase and aqueous phase to produce an emulsion in the form of water-in-oil (W/O) or oil-in-water (O/W)2. O/W nanoemulsion adjuvant can produce cytokines and chemokines at the injection site, induce the rapid aggregation and proliferation of important immune cells such as monocytes, neutrophils, and eosinophils, and enhance the immune response, and improve the immunogenicity of antigens3. Currently, three nanoemulsion adjuvants (MF59, AS03, and AF03) have been licensed for use in vaccines and have shown good safety and efficacy4.
However, mucosal immunity has been poorly addressed by the currently licensed adjuvant formulations in conventional parenteral vaccination5. Retinoic acid (RA) has been reported to induce intestinal homing of immune cells, but its low polarity, poor solubility in water, and poor light and thermal stability limit its use for robust enteric vaccination. Nanoemulsions offer opportunities to increase the bioavailability of highly lipophilic drugs, and the oil core of O/W emulsion adjuvants is suitable for dissolving non-polar substances such as RA6. Therefore, nanoemulsions can be used as carriers for RA in order to achieve the dual response effect of systemic immunity and mucosal immunity.
Compared to neutral or anionic delivery systems, cationic delivery systems have relatively efficient antigen encapsulation and delivery capabilities, which can enhance the immunogenicity of antigens7,8,9. The cationic surface charge of a variety of adjuvant systems is important for their adjuvant effects10,11,12. The cationic charge is an important factor in prolonging vaccine retention at the injection site, increasing antigen presentation and prolonging the stimulation of cellular immunity in the body12.
Based on the above considerations, we developed a cationic nanoemulsion to effectively co-deliver RA and antigens. The particle size and zeta potential of the nanoemulsion were determined using dynamic light scattering (DLS), and the systemic and mucosal immune responses of the nanoemulsion combined with OVA were evaluated by intramuscular injection13.

Forfatter Spotlight: Utvikling og evaluering av et kationisk nanoemulsjonsinnkapslet retinsyresystem for slimhinnevaksinasjon

Kationisk nanoemulsjon-innkapslet retinsyre som en adjuvans for å fremme OVA-spesifikke systemiske og slimhinneresponser

We are working on a mucosal vaccine that produces an immune response as the set of infection, which has the potential to prevent infection and interrupt transmission. We have developed a catatonic nanoemulsion-encapsulated retinoic acid delivery system immunization with ovalbumin by intramuscular injection, which triggers both systemic and mucosal immune responses. In future studies, animal models of intestinal-related diseases can be used to further evaluate the effect and the mechanism of a cationic nanoemulsion-encapsulated retinoic acid in enhancing vaccine protection.

Fremstilling og karakterisering av kationisk nanoemulsjon-innkapslet retinsyre

Begynn oppkjøpspreparatet ved å oppløse 0,15 gram polysorbat 80 i 28,2 ml PBS under omrøring ved 40 grader Celsius. For å forberede oljefaseformuleringen, oppløs Sorbitan Trioleate 85 DOTAP og retinsyre i Squalene under omrøring ved 40 grader Celsius. Deretter legger du til acquiesce-fasen ved en milliliter per minutt til oljefasen under omrøring ved 1400 RPM for primær olje- og vannemulsjonsforberedelse.Pre emulgerer blandingen ved hjelp av en høy skjæremulgator ved 30 000 RPM i seks minutter. Behandle den primære olje- og vannemulsjonen med en høytrykkshomogenisator i 12 sykluser ved 900 bar for å oppnå en homogen nanoemulsjon. Samle emulsjonen i en 50 milliliter kolbe og filtrer den gjennom en sprøyte med et 0,2 mikron PES-filter.Koble kolben til schlink-ledningen gjennom kateteret, drei treveis stoppekranen for å koble systemet til vakuumrøret. Slå på vakuumpumpen for å skape et vakuum i kolben. Drei deretter treveis stoppekran for å koble systemet til argonrøret og fyll det med argon.Bytt kolbekorken og forsegl den med en parafinfilm. Pakk kolben i tinfoil og oppbevar den ved 40 grader Celsius. Slå på det dynamiske lysspredningsinstrumentet og vent i 30 minutter til laserlyskilden stabiliserer seg.Fortynn nanoemulsjonen 50 ganger med ultrarent vann og tilsett en milliliter prøve til den tilsvarende prøvecellen. Sett temperaturen på instrumentet til 25 grader Celsius. Velg vann som dispergeringsmedium og engangs plastfat som målecelle.Legg i prøvene, og mål partikkelstørrelsen. Rapporter middelverdien av tre gjentatte målinger for hver prøve som resultat.

Research

JoVE Journal

Immunology and Infection

Preparation and Characterization of Cationic Nanoemulsion&#45;Encapsulated Retinoic Acid

Cationic Nanoemulsion-Encapsulated Retinoic Acid as an Adjuvant to Promote OVA&#45;Specific Systemic and Mucosal Responses

Cationic nanostructures have emerged as an adjuvant and antigen delivery system that enhances dendritic cell maturation, ROS generation, and antigen ...

Evaluation of Ovalbumin-Specific Antibodies in Mice After Intramuscular Administration of Cationic Nanoemulsion&#45;Encapsulated Retinoic Acid

Evaluering av ovalbuminspesifikke antistoffer hos mus etter intramuskulær administrering av kationisk nanoemulsjon-innkapslet retinsyre