– DECLARATION The work was carried out at the Department of Biomaterials & Bioengineering - Institute of Applied Materials Science (IAMS) - Vietnam Academy of Science and Technology (VAST) in Ho Chi Minh City. I hereby declare that this is my research work under the scientific guidance of Assoc. Nguyen Dai Hai. The research contents and results presented in this thesis are honest and completely based on my research results.
The results of this study have not been published on any thesis of the same level. Ho Chi Minh, December 30th 2022 NGUYEN THI NGOC HOI ii ACKNOWLEDGEMENTS First of all, I would like to express my most profound gratitude to my supervisor Assoc. Nguyen Dai Hai - Vice Director of the Institute of Applied Materials Science, and Head of Department of Biomaterials and Bioengineering. He has given me the delightful lessons, inspiration, constant motivation and enthusiasm that have surely encouraged and helped me to overpass the difficulties encountered, and exerted great aids for my accomplishment of this thesis research.
Secondly, my sincere gratitude also goes to the enthusiastic help and favorable supports during my PhD course from Graduate University of Science and Technology (GUST) and the Institute of Applied Materials Science (IAMS) - Vietnam Academy of Science and Technology (VAST). Furthermore, it is impossible not to mention the valuable support from MSc. Nguyen Dinh Tien Dung and BS. Truong Thi Ngoc Hang.
They contributed great help during the experiments at IAMS. Last but not least, I am grateful to have my family and friends, who always encourage and support all over time that makes my thesis experience more meaningful. Ho Chi Minh, December 30th 2022 NGUYEN THI NGOC HOI iii TABLE OF CONTENTS Page DECLARATION. ii TABLE OF CONTENTS.
iii LIST OF ABBREVIATIONS. vii LIST OF FIGURES. ix LIST OF DIAGRAMS. xiv LIST OF TABLES.
Overview of cancer and cancer treatment. Overview of cancer. Common cancer treatment therapies. Nanomaterials in cancer treatment.
Nanomaterials in anti-cancer drug delivery applications. Silica nanomaterials in anti-cancer drug delivery applications. Recent progress of nano silica particle applications in drug delivery. Hollow mesoporous silica nanoparticles (HMSN).
Structure of HMSN. Synthesis methods of HMSN. Reaction mechanisms in the synthesis of HMSN by silica based hard- template method. Modular factors in HMSN fabrication.
Modifications of HMSN. Multiple-Drug Loading HMSN. MATERIALS AND EXPERIMENTAL METHODS. Synthesis of HMSN.
The effect of PEG on the mesoporous shell thickness of HMSN. The effect of non-ionic surfactants on the mesopore diameter of HMSN. Surface Modification Method of HMSNs with Pluronics. The effect of Pluronics on dual-drugs delivery characteristics of HMSN- Plu.
Physicochemical Analysis Methods. Drug loading and in vitro release study. Cell culture and MTT assay. A MODIFIED HARD-TEMPLATE METHOD FOR HOLLOW MESOPOROUS SILICA NANOPARTICLES SYNTHESIS WITH SUITABLE PARTICLE SIZE AND SHORTENED SYNTHETIC TIME.
Synthesis of silica hard-template. Etching over time of silica hard-template in the synthesis of HMSN. Characterizations of synthesized HMSN. Cytotoxicity of synthesized HMSN.
SIMPLY AND EFFECTIVELY CONTROL THE SHELL THICKNESS OF HOLLOW MESOPOROUS SILICA NANOPARTICLES BY POLYETHYLENE GLYCOL FOR DRUG DELIVERY APPLICATIONS. Effect of PEG molecular weight on the mesoporous shell thickness of dSiO2@MSN. Effect of PEG weight percentage on the mesoporous shell thickness of dSiO2@MSN. Characterizations of the synthesized HMSNs.
Drug loading and in vitro drug release study of the synthesized HMSN. Cytotoxicity of the synthesized HMSN. NON-IONIC SURFACTANTS AS CO-TEMPLATES TO CONTROL THE MESOPORE DIAMETER OF HOLLOW MESOPOROUS SILICA NANOPARTICLES FOR DRUG DELIVERY APPLICATIONS. Preparation of mixed micelles of non-ionic surfactants with CTAB.
Effect of non-ionic surfactants on the mesoporous shell thickness of dSiO2@MSN. Effect of non-ionic surfactants on the mesopore diameter of dSiO2@MSN. Characterizations of the synthesized HMSNs. Drug loading and in vitro drug release study of the synthesized HMSNs.
Cytotoxicity of the synthesized HMSNs. SURFACE MODIFICATION OF HOLLOW MESOPOROUS SILICA NANOPARTICLES WITH PLURONICS FOR DUAL DRUGS DELIVERY. Activation Pluronic with NPC. Amination of HMSNs’ surface.
Modification of HMSNs’ surface with Pluronics via amine intermediate 96 6. Dual-drug loading capacity and in vitro release behavior of HMSN-Plu. In vitro drug release behavior of HMSN-Plu. Cytotoxicity of HMSN-Plu.
Characterizations of HMSN-F127. Cancer cell killing ability of DOX.QUE@HMSN-Plu. 109 CONCLUSIONS AND FUTURE PERSPECTIVES. 111 Novelty of the thesis.
112 LIST OF PUBLICATIONS. 115 vii LIST OF ABBREVIATIONS APTES (3-Aminopropyl)triethoxysilane BET Brunauer-Emmett-Teller BJH Barret Joyner and Halenda BTES Bis (triethoxysilylpropyl) disulfide C18TMS n-octadecyltrimethoxysilan CMC Critical micelle concentration CTAB Cetyltrimethylammonium Bromide CTAC Cetyltrimethylammonium Chloride DI Deioned water DLC Drug loading capacity DLE Drug loading efficiency DLS Dynamic Light Scattering DOX Doxorubicin dSiO2 dense Silicone dioxide EDX Energy Dispersive X-ray EPR Enhanced Permeability and Retention FDA Food and Drug Administration FE-SEM Field Emission Scanning Electron Microscope FT-IR Fourier Transform Infrared GPC Gel Permeation Chromatography HMSN Hollow Mesoporous Silica Nanoparticles HPLC High Performance Liquid Chromatography MCM-41 Mobil Composition of Matter No. 41 MCM-48 Mobil Composition of Matter No. 48 MCM-50 Mobil Composition of Matter No.
50 MDR Multidrug Resistance MON Mesoporous Organosilica Nanoparticle MSN Mesoporous Silica Nanoparticles PAA Poly (Acrylic Acid) PBS Phosphate Buffered Saline PEG Polyethylene Glycol PEO Poly(Ethylene Oxide) PPO Poly(Propylene Oxide) PMMA Polymethylmethacrylate viii PS Polystyrene PVP Polyvinylpyrolidone QUE Quercetin RB Rose Bengal SBA-15 Santa Barbara Amorphous-15 SEM Scanning electron microscope TGA Thermogravimetric analysis TEA Triethanolamine TEM Transmission electron microscopy TEOS Tetraethyl orthosilicate XRD X-ray Diffraction ix LIST OF FIGURES Figure 1. Global cancer data in 2020: a) Female, b) Male [1]. Common treatments for cancers [2]. Popular nanomaterials applied in drug delivery [6].
Members of the M41S family [8]. Structural classification of Mesoporous Silica Nanoparticles [9]. Structure of Hollow Mesoporous Silica Nanoparticle (HMSN): a) 2D radial section; b) 3D model; and c) Mesoporous structure of the shell. Synthesis methods of HMSN.
Hydrolysis and condensation of TEOS precursors in alcohol-water- ammonia medium. Multistage growth diagram of silica particles by hydrolysis of TEOS in alcohol-water-ammonia medium [62]. Illustration of the formation mechanism of the mesoporous shell (MCM-41) [18]. Etching process of hard template dSiO2 by Na2CO3 [64].
Etching mechanism of hard template dSiO2 to form HMSN by Na2CO3: a) Etching process with the presence of CTAB micelles, and b) Etching process without CTAB micelles [64]. Modular factors of the HMSN. Adjustable shell thickness of microporous hollow core@shell silica nanoparticles for controlled release of doxorubicin [70]. The size of biodegradable silica nanoparticles was reduced for efficient curcumin loading [74].
The effect of polyethylene glycol on shape and size of SrTiO3 nanoparticles [83]. Self-assembly of mixed micelle of CTAB and P123 used as mesoporous templates in MSN particle synthesis [96]. Aminated HMSN using 3-Aminopropyl)triethoxysilane for better DOX loading capacity and controlled release [40]. Molecular structure of Pluronics.
Conjugation of polyamidoamine dendrimer and pluronics for hydrophobic drug delivery [108]. Molecular formulas of the used non-ionic surfactants versus CTAB. Characterizations of the synthesized hard-template dSiO2: a) Zeta potential; b) DLS particle size distribution; c) SEM image; d) TEM image. SEM and TEM images of HMSN over etching time.
a) TEM image of dSiO2@MSN; b) TEM image of HMSN, c) N2 adsorption-desorption isotherms of HMSN and d) Pore size distributions of HMSN. Characterizations of the synthesized HMSN: a) FT-IR spectrum; b) EDX parttern; c) Zeta potential; d) DLS particle size distribution; e) XRD pattern; and f) TGA graph. a) Cell viability assay by MTT assay with variable concentrations of HMSN on MCF-7 cells; b) Morphology of MCF-7 cells treated by HMSN at different concentrations. Size dispersion by DLS measurement and field-emission scanning electron microscopy (FE-SEM) images of (a, a’)dSiO2, (b, b’) dSiO2@MSN, (c, c’) dSiO2@MSN-P1k, (d, d’) dSiO2@MSN-P2k, (e, e’) dSiO2@MSN-P4k and (f, f’) dSiO2@MSN-P6k.
Size dispersion by DLS measurement and field-emission scanning electron microscopy (FE-SEM) images of (a, a’) dSiO2@MSN-P1%, (b, b’) xi dSiO2@MSN-P2%, (c, c’) dSiO2@MSN-P3%, (d, d’) dSiO2@MSN-P4% and (e, e’) dSiO2@MSN-P5%. The structure of PEG changes from a) zigzag chains to b) ordered net structure in the solution. Characterizations of the synthesized silica nanoparticles: a) TEM images of dSiO2@MSN-0 and a’) dSiO2@MSN-P; TEM images of b) HMSN-0 and b’) HMSN-P; Size distribution of c) HMSN-0 and c’) HMSN-P; Zeta potential of d) HMSN-0 and d’) HMSN-P. The N2 adsorption-desorption isotherms and pore size distributions of dSiO2@MSN (a and b) and dSiO2@MSN-P (a’ and b’).
Characterizations of the synthesized HMSN-0 (square dot) and HMSN- P (solid): a) EDX patterns; b) FT-IR spectra. DOX loading capacity (DLC - grey) and DOX loading efficiency (DLE - black) of HMSN-0 and HMSN-P (a); In vitro release profile of Dox@HMSN-0 (empty circle) and Dox@HMSN-P (solid circle) (b). The marked points correspond to 0, 1, 3, 6, 9, 12, 24, 36 and 48 h, respectively. Cell viability by MTT assay with variable concentrations of HMSN-0 and HMSN-P on MCF-7 cells (a); MCF-7 cells treated by HMSN-0 and HMSN-P at different concentrations (b).
a) Viscosity of mixed micelles versus molar ratio of non-ionic surfactants and CTAB. Molar concentration of CTAB remained constantly at 0. b) Hydrodynamic diameter of mixed micelles versus molar ratio. Molar concentration of CTAB in each mixture was 50 mM in the presence of 1 mM KBr.
Size distribution by DLS measurement of a) dSiO2@MSN-T20, b) dSiO2@MSN-T80, and c) dSiO2@MSN-BS10. Illustration of the effect of non-ionic surfactants in mixed micelles on the mesoporous shell thickness of dSiO2@MSN. The N2 adsorption-desorption isotherms and pore size distributions of a) dSiO2@MSN, b) dSiO2@MSN-T20, c) dSiO2@MSN-T80 and d) dSiO2@MSN- BS10. SEM images, TEM images, Size distribution and Zeta potential of HMSN, HMSN-T20, HMSN-T80 and HMSN-BS10.
a) XRD patterns and b) FT-IR spectra of HMSN, HMSN-T20, HMSN- T80 and HMSN-BS10. (a) Rose bengal (RB) loading capacity (DLC - grey) and loading efficiency (DLE - black) of HMSN, HMSN-T20, HMSN-T80 and HMSN-BS10; (b) In vitro release profile of RB from HMSN, HMSN-T20, HMSN-T80 and HMSN- BS10. The marked points correspond to 0, 1, 3, 6, 9, 12, 24, 36, 48, 60 and 72 h, respectively. a) Cell viability by MTT assay on MCF-7 cells; and b) MCF-7 cells treated by HMSN, HMSN-T20, HMSN-T80 and HMSN-BS10 at different concentrations.
FT-IR spectra of NPC-Plu-OH. 1H-NMR spectra of NPC-Plu-OH: a) NPC-L64-OH, b) NPC-F68-OH, c) NPC-F127-OH, d) Annotation the molecular structure of NPC-Plu-OH. Characterizations of HMSN and HMSN-NH2: a) Zeta potential; b) Hydrodynamic particle diameter; c) FT-IR spectra; and d) EDX patterns. Characterizations of HMSN-L64, HMSN-F68 and HMSN-F127: a) Zeta potential; b) Hydrodynamic particle diameter; c) FT-IR spectra; and d) TGA graphs.
In vitro release behaviour of free drugs and loaded drugs in different conditions of temperatures and pH values. Illustration of release behavior of HMSN-Plu in different conditions 103 xiii Figure 6. a) Cytotoxicity by MTT assay of HMSN-Plu on Hela cells; b) Morphology of Hela cells treated by HMSN at different concentrations. TEM images and Size distribution of a), a’) HMSN and b), b’) HMSN- F127.
The N2 adsorption-desorption isotherms and pore size distributions of HMSN (a, a’) and HMSN-F127 (b, b’). a) Cell viability and b) Mophorlogy of Hela cells treated by Free DOX, Free QUE and DOX. 108 xiv LIST OF DIAGRAMS Diagram 1. Sol-Gel synthesis of a) hard template dSiO2 and b) mesoporous shell MSN.
The preparation of the hard template dSiO2. The preparation of core@shell structure dSiO2@MSN. The selective etching of dSiO2@MSN to form HMSN. Mesoporous silica layer coating step in HMSN synthesis process with the presence of PEG.
Mesoporous silica layer coating step in HMSN synthesis process with the presence of non-ionic surfactants as co-templates. The surface activation of HMSN with APTES. The activation of Pluronic with NPC. The preparation of HMSN-Plu from HMSN-NH2 and NPC-Plu-OH .