AbstractNanoparticulate delivery systems have been widely used in recent decades, available in a wide variety of structures, for targeted drug delivery. They provide controlled and prolonged release for drugs, peptides and biopharmaceuticals. Ceramic nanoparticles are one of the various nanocarriers, which have been employed in local targeted delivery, most commonly in the area of orthopaedic drug delivery to enhance treatment therapies.
This thesis therefore focused on the development of aquasomes, a ceramic nanoparticulate carrier system, for the delivery of proteins, growth factors and antibiotics for its potential application in bone regeneration in fracture healing. The suitability of non-aqueous silicone elastomer gels (NASEGs) as a topical/transdermal delivery system for proteins as well as protein-loaded aquasomes was also investigated.
Through process optimisation, a suitable lyophilisation method was developed and used for the preparation of bioactive aquasome formulations of growth factors, bone morphogenetic protein (BMP-2), vascular endothelial growth factor (VEGF-121), and antibiotic, gentamicin.
Physical characterisation of aquasomes using zeta potential and optimisation of preliminary aquasome formulations were optimised by utilising smaller nanocore sizes. In addition, scanning electron microscopy (SEM), confocal microscopy analysis and entrapment efficiency studies were performed to ascertain the drug loading efficiency of the different aquasome formulations. BMP-2 loading aquasomes exhibited an entrapment efficiency of 98.9% Protein loading on aquasomes yielded a higher negative zeta potential in comparison to blank nanocores. Confocal microscopy images elucidated the behaviour of nanocore particles showing agglomeration of nanocores and the presence of fluorescent drug adsorbed onto nanocores.
The bioactivity of the aquasome formulations were analysed via in vitro cell culture model assays and microbiological assays. BMP-2-loaded aquasomes were investigated for enhanced osteogenic proliferation and differentiation effects on osteoblast-like cells, MG63 cells. The enhanced osteogenic effect of HUVECs in co-culture with these cells was also examined. In addition, the committed differentiation of ATMSCs into osteoblasts induced by their exposure to BMP-2 -loaded aquasomes was also investigated. Results exhibited the enhanced osteogenic differentiation effect, analysed by alkaline phosphatase (ALP) secretion (a major biochemical marker of osteoblastic differentiation) from MG63 cells was dependent on the protein loading onto the aquasome formulation. However, differentiation of ATMSCs cultured in osteogenic medium was significantly higher than ATMSCs exposed to BMP-2 or VEGF-121 treatments. Gentamicin-loaded aquasomes were investigated for their antimicrobial activity against Staphylococcus aureus, a major pathogen popularly implicated in cases of osteomyelitis. Results showed that gentamicin released from aquasomes exhibited excellent bactericidal activity against bacterial cultures without any reproduction of bacteria in 24 hours. In conclusion, the aquasome formulations were able to offer controlled release of bioactive antimicrobials and growth factors over a prolonged duration. The amount of bio-actives released was dependent on the loading of the bio-actives in the fabrication process of aquasome formulations. However, minute (ng/µg) amounts of adsorbed growth factor/drug were observed in comparison to the loading (high ng/mg) within the duration of study.
It can be inferred these aquasomes can be employed in the sustained local and targeted delivery of antimicrobials and growth factors in orthopaedic treatments for enhanced fracture healing. However, the loading of bio-actives onto aquasome formulations may need to be optimised to increase the amount of bio-actives released to elicit more pronounced pharmacological effects.
|Date of Award||2016|
|Supervisor||Raj Badhan (Supervisor) & Deborah Lowry (Supervisor)|
- ceramic nanoparticles
- VEGF 121
- fracture healing