The aims of this study were to prepare poly (ethylcyanoacrylate) (PECA) nanoparticles on the basis of different types of microemulsions, to investigate the entrapment within and release of a bioactive from these particles and to establish a set of delivery systems with varying entrapment and release characteristics, thereby giving the formulator the opportunity of a more tailor-made approach in the development of a delivery system. Furthermore the scale up of particle preparation and the possible enhancement of the immunogenic properties of PECA particles by incorporation of the adjuvant Quil A was investigated.
Methods: Four phase triangles were established and microemulsion samples, used as a template to prepare nanoparticles, were characterised by viscosity and conductivity measurements, polarising light microscopy, freeze fracture transmission electron microscopy (TEM), cryo field emission electron microscopy (cryo FESEM) and self-diffusion NMR to determine their microemulsion type (droplet, bicontinuous, solution type). PECA nanoparticles were prepared from different types of microemulsions by interfacial polymerisation. Particle size, polydispersity index (PI) and [zeta]-potential were measured by photon correlation spectroscopy and electrophoretic mobility respectively. Normal scanning electron microscopy (SEM) and cryo FESEM were used to visualise particles. Fluorescently labelled ovalbumin (FITC-OVA) was used as a model protein/antigen and entrapment within and release from nanoparticles was investigated. To scale up nanoparticle preparation an instrumental set-up with reactor, peristaltic pump and stirrer was used. A 2⁷ fractional factorial study was designed to observe possible factors or their interactions that could influence particle formation under scale up conditions. For an immunological study freeze dried formulations of PECA nanoparticles, having FITC-OVA and Quil A entrapped, were prepared, and activation and uptake of formulations by murine bone marrow derived dendritic cells (DCs) and T cells in vitro were monitored.
Results: Results obtained from the measurements described above, for formulations from the four different phase triangles, indicated that microemulsions of w/o droplet, bicontinuous or solution type could be formed. It was possible to prepare PECA nanoparticles from all of the different types of microemulsions. Particles had an average size of 265 nm � 24, with an average PI of 0.18 � 0.05 and an average negative [zeta]-potential of -17 mV � -5. Particle size, PI and [zeta]-potential were not influenced by the type of microemulsion that was used as a polymerisation template. Entrapment and release were however influenced by the type of microemulsion and although entrapment of FITC-OVA was generally high for PECA particles, it was highest for particles prepared from a droplet type microemulsion. Entrapment could also be increased by increasing amounts of monomer. The rate of release was dependent on the amount of monomer used for polymerisation and the type of microemulsion used for particle preparation, with nanoparticles prepared from a w/o droplet type microemulsion showing the slowest release. Furthermore it was shown that particle preparation could be scaled-up with the instrumental set-up used in this study, but conditions need to be refined as the average particle size and polydispersity index were considerably larger (441 nm � 101, 0.68 � 0.14) when compared to particles prepared by the beaker-pipette method (see above). The adjuvant Quil A could efficiently be entrapped into PECA nanoparticles together with FITC-OVA. Incubation of DCs and T cells with the various formulations did, however, not result in increased uptake or activation.
Conclusions: PECA nanoparticles with high entrapment efficiency of antigen and adjuvant can be prepared from different types of microemulsions. Particles show different rates of entrapment and release depending on the type of microemulsion used as a polymerisation template, possibly because two different types of nanoparticles form. Nanocapsules are believed to form on the basis of droplet type microemulsions and nanospheres form on the basis of bicontinuous and solution type microemulsions. Freeze dried formulations of PECA nanoparticles, containing Quil A and FITC-OVA, were not able to induce an immune response, which might be due to charge repulsion effects between the negatively charged PECA nanoparticles and the negatively charged surface of dendritic cells. Moreover, no adjuvant effect of Quil A was apparent, perhaps caused by total encapsulation of the compound into the particle matrix with no active groups extending out displaying adjuvanticity.
| Identifer | oai:union.ndltd.org:ADTP/217567 |
| Date | January 2005 |
| Creators | Krauel, Karen, n/a |
| Publisher | University of Otago. School of Pharmacy |
| Source Sets | Australiasian Digital Theses Program |
| Language | English |
| Detected Language | English |
| Rights | http://policy01.otago.ac.nz/policies/FMPro?-db=policies.fm&-format=viewpolicy.html&-lay=viewpolicy&-sortfield=Title&Type=Academic&-recid=33025&-find), Copyright Karen Krauel |
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