Sageretia thea (Osbeck.) mediated synthesis of multifunctional nanoparticles and their possible applications By Ali Talha Khalil Department of Biotechnology Faculty of Biological Sciences Quaid-i-Azam University Islamabad, Pakistan 2017 i Sageretia thea (Osbeck.) mediated synthesis of multifunctional nanoparticles and their possible applications A thesis Submitted to the Department of Biotechnology in the partial fulfillment of the requirement for the degree of Doctor of Philosophy In Biotechnology By Ali Talha Khalil Department of Biotechnology Faculty of Biological Sciences Quaid-i-Azam University Islamabad, Pakistan 2017 ii iii iv v PRAYER In the name of ALLAH, The most beneficent, The most merciful. Praise to Allah, Lord of worlds The beneficent, The merciful Master of the Day of Judgment Thee we worship, Thee we ask for help Show us the straight path The path of those whom thou worth favored Neither of who earn Thins anger Nor of those who go astray (Ameen) Al-Quran Surrah Al-Fatiha vi Dedicated to My parents, Siblings, And Family. vii Table of contents Contents Chapter 1: Introduction .................................................................................................................... 1 st 1.1 Nanoscience in 21 century ................................................................................................... 2 1.2 Metal oxide nanoparticles ...................................................................................................... 2 1.2.1 Zinc oxide (ZnO) ............................................................................................................ 3 1.2.2 Iron oxide (γ-Fe2O3) ........................................................................................................ 4 1.2.3 Nickel oxide (NiO) ......................................................................................................... 5 1.2.4 Cobalt oxide (Co3O4) ...................................................................................................... 5 1.2.5 Lead oxide (PbO) ............................................................................................................. 5 1.3 Biosynthesis of nanoparticles ................................................................................................ 6 1.4 Sageretia thea (Osbeck.) ........................................................................................................ 8 1.5 Importance of study ............................................................................................................... 8 1.6 Specific objectives ............................................................................................................... 10 Chapter 2: Sageretia thea (Osbeck.) mediated synthesis of ZnO nanoparticles and its biological applications .................................................................................................................................... 11 2.1 Introduction .......................................................................................................................... 14 2.2 Materialand methods ............................................................................................................ 15 2.2.1 Plant material processing .............................................................................................. 15 2.2.2 Biosynthesis of ZnO nanoparticles ............................................................................... 17 2.2.3 Characterizations ........................................................................................................... 17 2.2.4 Test sample preparation ................................................................................................ 18 2.2.5 Antibacterial activity ..................................................................................................... 18 2.2.6 Antifungal activity ........................................................................................................ 19 2.2.7 Protein kinase inhibition ............................................................................................... 19 2.2.8 Alpha amylase inhibition .............................................................................................. 20 2.2.9 Antileishmanial activity (amastigotes and promastigotes) ........................................... 20 2.2.10 Brine shrimp cytotoxicity............................................................................................ 21 2.2.11 Antioxidant activities .................................................................................................. 21 2.2.12 Biocompatibility of ZnO nanoparticles with RBC’s ................................................... 22 2.2.13 Biocompatibility of ZnO nanoparticles with human macrophages ............................. 22 2.3 Results and discussion ......................................................................................................... 23 2.3.1 Biosynthesis and characterization of ZnO nanoparticles .............................................. 23 viii 2.3.2 Physical characterization............................................................................................... 25 2.3.3 Antibacterial activity ..................................................................................................... 33 2.3.4 Antifungal activity ........................................................................................................ 38 2.3.5 Antileishmanial potential (promastigotes and amastigotes) ......................................... 40 2.3.6 Brine shrimp cytotoxicity.............................................................................................. 40 2.3.7 Biocompatibility assessment using Human RBC’s and macrophages .......................... 40 2.3.8 Enzyme inhibition activity (protein kinase inhibition and alpha amylase inhibition) .. 42 2.3.9 Antioxidant activities .................................................................................................... 44 Chapter 3: Biosynthesis of iron oxide (Fe2O3) nanoparticles via aqueous extracts of Sageretia thea (Osbeck.) and their pharmacognostic properties .................................................................... 46 3.1 Introduction .......................................................................................................................... 49 3.2 Material and Methods .......................................................................................................... 50 3.2.1 Plant material processing .............................................................................................. 50 3.2.2 Biosynthesis of IONP’s ................................................................................................. 51 3.2.3 Physical characterizations ............................................................................................. 53 3.4 Antimicrobial activities ........................................................................................................ 53 3.4.1 Antibacterial activity ..................................................................................................... 53 3.4.2 Antifungal activity ........................................................................................................ 54 3.5 Cytotoxic activities .............................................................................................................. 54 3.5.1 Antileishmanial activity (Promastigotes and Amastigotes) .......................................... 54 3.5.2 Brine shrimp cytotoxicity.............................................................................................. 55 3.6 Biocompatibility assays ....................................................................................................... 55 3.6.1 Human RBC’s ............................................................................................................... 55 3.6.2 Human Macrophages .................................................................................................... 55 3.7 Enzyme inhibition assays ..................................................................................................... 56 3.7.1 Protein kinase inhibition ............................................................................................... 56 3.7.2 Alpha amylase inhibition .............................................................................................. 56 3.8 Antioxidant activities ........................................................................................................... 56 3.8.1 DPPH free radical scavenging ...................................................................................... 56 3.8.2 Total Antioxidant Capacity ........................................................................................... 57 3.8.3 Total Reducing Power ................................................................................................... 57 3.9 Results and Discussion ........................................................................................................ 57 3.9.1 Biosynthesis of IONPs .................................................................................................. 57 3.9.2 Room temperature physical characterizations .............................................................. 58 ix 3.9.3 Antimicrobial properties of bioinspired IONPs ............................................................ 63 3.9.4 Cytotoxic activities ......................................................................................................