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Research output: Thesis › Master's Thesis
Research output: Thesis › Master's Thesis
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TY - THES
T1 - Development of peptide inhibitor nanoparticles (PINPs) for treatment of Alzheimer’s Disease
AU - Michail, Christos
PY - 2015
Y1 - 2015
N2 - Purpose: To investigate the best carrier technology for our β-amyloid (Aβ)aggregation inhibitors by developing three types of liposomes (a) plain liposomes, (b)MAL-PEG liposomes, and finally the combination of retro-inverted peptide RI-OR2-TAT (Ac-rGffvlkGrrrrqrrkkrGyc-NH2) attached onto the surface of MAL-PEGliposomes, creating Peptide Inhibitor Nanoparticles (PINPs) of three different sizes(50, 100 and 200 nm). In addition, these nanoliposomes (NLPs) (with particular focuson PINPs) were examined for their ability to affect Aβ aggregation, and to protectagainst Aβ cytotoxicity.Methods: The creation of NLPs was carried out by the use of a mini extruder, whilethe elution of PINPs from a size exclusion column was assessed by Dynamic LightScattering (DLS). The quantification of peptide bound to liposomes was determinedby bicinchoninic acid (BCA) assay, while phospholipid content was quantified byWako phospholipid assay. The effects of the different types of liposomes on Aβtoxicity and viability of SHSY-5Y neuronal cells were examined by MTS assay, whereaseffects on Aβ aggregation were determined by Thioflavin-T (Th-T) assay. In addition,a cell penetration assay was carried out in order to examine the ability of liposomesto penetrate into neuroblastoma SHSY-5Y cells.Results: Low concentrations of PINPs 0.1 μM inhibited Aβ aggregation and toxicity invitro. MAL-PEG liposomes and PINPs were able to penetrate into neuroblastomaSHSY-5Y cells and were also more stable than simple liposomes. Stability means theability of liposomes to keep their size and their shape stable for long time. Inaddition, the three types of liposomes were not toxic towards SHSY-5Yneuroblastoma cells. Cytotoxicity is the quality of being toxic to cells. So, none of thethree types of our liposomes showed any negative effect on the viability towardsSHSY-5Y neuroblastoma cells.Conclusion: NLPs are an ideal carrier for our aggregation inhibitors because theyaffect Aβ aggregation and toxicity at low doses, and according to other datagenerated by our group, can cross the Blood Brain Barrier (BBB).
AB - Purpose: To investigate the best carrier technology for our β-amyloid (Aβ)aggregation inhibitors by developing three types of liposomes (a) plain liposomes, (b)MAL-PEG liposomes, and finally the combination of retro-inverted peptide RI-OR2-TAT (Ac-rGffvlkGrrrrqrrkkrGyc-NH2) attached onto the surface of MAL-PEGliposomes, creating Peptide Inhibitor Nanoparticles (PINPs) of three different sizes(50, 100 and 200 nm). In addition, these nanoliposomes (NLPs) (with particular focuson PINPs) were examined for their ability to affect Aβ aggregation, and to protectagainst Aβ cytotoxicity.Methods: The creation of NLPs was carried out by the use of a mini extruder, whilethe elution of PINPs from a size exclusion column was assessed by Dynamic LightScattering (DLS). The quantification of peptide bound to liposomes was determinedby bicinchoninic acid (BCA) assay, while phospholipid content was quantified byWako phospholipid assay. The effects of the different types of liposomes on Aβtoxicity and viability of SHSY-5Y neuronal cells were examined by MTS assay, whereaseffects on Aβ aggregation were determined by Thioflavin-T (Th-T) assay. In addition,a cell penetration assay was carried out in order to examine the ability of liposomesto penetrate into neuroblastoma SHSY-5Y cells.Results: Low concentrations of PINPs 0.1 μM inhibited Aβ aggregation and toxicity invitro. MAL-PEG liposomes and PINPs were able to penetrate into neuroblastomaSHSY-5Y cells and were also more stable than simple liposomes. Stability means theability of liposomes to keep their size and their shape stable for long time. Inaddition, the three types of liposomes were not toxic towards SHSY-5Yneuroblastoma cells. Cytotoxicity is the quality of being toxic to cells. So, none of thethree types of our liposomes showed any negative effect on the viability towardsSHSY-5Y neuroblastoma cells.Conclusion: NLPs are an ideal carrier for our aggregation inhibitors because theyaffect Aβ aggregation and toxicity at low doses, and according to other datagenerated by our group, can cross the Blood Brain Barrier (BBB).
M3 - Master's Thesis
PB - Lancaster University
ER -