Luận án tiến sĩ Vật lý: Synthesis of biodegradable porous silica nanoparticles and the use of optical techniques for characterizing the material properties and its drug delivery capabilities

VIETNAM NATIONAL UNIVERSITY HCMC UNIVERSITY OF SCIENCE MAI NGOC XUAN DAT SYNTHESIS OF BIODEGRADABLE POROUS SILICA NANOPARTICLES AND THE USE OF OPTICAL TECHNIQUES FOR CHARACTERIZING THE MATERIALS PROPERTIES AND ITS DRUG DELIVERY CAPABILITY PhD THESIS OF PHYSICS HOCHIMINH CITY - 2022 VIETNAM NATIONAL UNIVERSITY HCMC UNIVERSITY OF SCIENCE MAI NGOC XUAN DAT SYNTHESIS OF BIODEGRADABLE POROUS SILICA NANOPARTICLES AND THE USE OF OPTICAL TECHNIQUES FOR CHARACTERIZING THE MATERIALS PROPERTIES AND ITS DRUG DELIVERY CAPABILITY Speciality: Optics Code: 62440109 Reviewer 1: Assoc. Huynh Dai Phu Reviewer 2: Assoc. Nguyen Dai Hai Reviewer 3: Assoc. Nguyen Manh Tuan Independent reviewer 1: Assoc.

Tran Ngoc Quyen Independent reviewer 2: Assoc. Huynh Dai Phu Supervisor: Prof. Phan Bach Thang HOCHIMINH CITY - 2022 ACKNOWLEDGEMENTS First of all, I would like to thank Faculty of Physics and Engineering Physics, University of Science, VNU-HCM, for support of my PhD course. I would like to thank my supervisor, Professor Phan Bach Thang, for his support of my doctoral program with his supervision and guidance during the whole period.

He always supports the best conditions for me to finish this thesis and improve my research. I would like to especially thank my group leader, Dr. Doan Le Hoang Tan. He helped me build up strong foundations and develop ideas when I started this new field.

In addition, he always was willing to give me profound comments, insightful discussions, and encouragement to complete all projects. I greatly appreciate all your help. Also, I want to thank Professor Tamanoi, Dr. Kotaro Matsumoto, and all members of his group in iCeMS, Kyoto University.

It was my great honor to have a chance to attend his group. I would like to thank Dr. Uyen-Chi Nguyen Le, Dr. Lien-Thuong Thi Nguyen, Dr.

Long Binh Vong, Dr. Thi-Hiep Nguyen, Dr. Tri Minh Le, Mr. Ha Van Nguyen for their collaboration in my research.

I also thank Dr. Kim Ngoc Pham, Ms. Hanh Kieu Thi Ta, and Dr. Tran Thi Nhu Hoa for their encouragement and support whenever I feel depressed.

I also thank all members of INOMAR for their friendship and help during my research. Special thanks to Nhoi Nhoi group, Thieu, Tay Ninh for their kind inspiration to help me relieve stress. Finally, I would like to thank my family and my parents for their encouragement in my life. I greatly acknowledge my parents for giving me invaluable things and always believing in me.

This dissertation was supported by VietNam National University Ho Chi Minh City (NCM2019-50-01). TABLE OF CONTENTS LIST OF 0902. vi LIST OF FIGURES 115. Nanoparticle-based drug delivery in cancer treafmenf.

Application of biodegradable periodic mesoporous organosilicas as nanocarriers and optical methods in anticancer drug delivery. Biodegradable periodic mesoporous organosilica nanoparticles as 40091900 58i1/101966 51T. Effective anticancer drugs in cancer freatmehI. Efficient optical methods in the study of nanocarriers in drug delivery.

Fourier transform infrared spectrOSCOpy. X-ray photoelectron spectrOSCOpy. Dynamic light MicrOSCOPY. Confocal laser scanning MICTOSCOPE.

Biological models for evaluation of the cytotoxicity of nanoparticles. Spheroids - Jn vitro three-dimensional (3D) cell model. Tumor-bearing chicken embryo model. - G0 ng nh HH Hàng 34 ii PIN.

Synthesis of biodegradable periodic mesoporous organosilica nanoparticles 35 2. Synthesis of ethane-containing tetrasulfide-based biodegradable periodic mesoporous organosilica nanoparticles (E4S). Synthesis of fluorescent ethane-containing tetrasulfide-based biodegradable periodic mesoporous organosilica nanoparticles (RTC-E4S). Synthesis of phenylene-containing tetrasulfide-based biodegradable periodic mesoporous organosilica nanoparticles (P4S).

Synthesis of inorganic mesoporous silica nanopartIcÌe. Physicochemical characterization by optical techniques. Degradation and drug loading behavior experiments.scscscssssssseseees 42 J SP ái o9on. Anticancer drug ÏOaHIng.

In Vitro TÏ©fAS€. SH HT HH TH HT HT HT HH HH 45 2. Cytotoxicity of materials in biological models. Cell viability analysis.

Uptake of nanoparticles by 3D tumor spheroid. Evaluation of 3D tumor spheroid ØørOW(H. Ovarian cancer tumor formation on chorioallantoic membrane. Investigation of nanoparticles biodistribution on chicken egg model.

Evaluation of tumor elimination effect of DNR-loaded nanoparticles. SYNTHESIS OF TETRASULFIDE-BASED BIODEGRADABLE PERIODIC MESOPOROUS ORGANOSILICA NANOPARTICLES AND THEIR CHARACTERIZATION USING OPTICAL METHODS.1 Synthesis, physicochemical characterization of tetrasulfide-based biodegradable periodic mesoporous organosilica nanoparticles by optical 3. Evaluation of biodegradability of synthesized tetrasulfide-based BPMO. Biodistribution and cytotoxicity of tetrasulfide-based BPMOs in tumor spheroid using confocal mÏCTOSCOJDV.

Biodistribution and cytotoxicity of tetrasulfide-based BPMOs in chicken egg Model using confocal MICTOSCOPY. TAILORING PARTICLE SIZE TAILORING AND IN VITRO CYTOTOXICITY EVALUATION OF TETRASULFIDE-BASED PERIODIC MESOPOROUS ORGANOSILICA NANOPARTICLES. Synthesize and compare physicochemical properties of tailored particle size of tetrasulfide-based periodic mesoporous organosilica nanoparticles using Optical Methods. Comparative in vitro biodegradability of tetrasulfide-based BPMOs via transmission electron IÏCTOSCOJDS.

Cordycepin loading capacity and release behavior of tetrasulfide-based IV 4. Cytotoxicity of tetrasulfide-based biodegradable mesoporous organosilica nanoparticles in 2D mO(€ÌL. CONCLUSIONS AND FUTURE PERSPECTTIVES. Novelty of the dÏssS€rfAafÏOT.- << 5 TH TH TH 000000009005, 104 LIST OF PUBLICA TIONS.

LH HH HT HH Hiệp 106 REFERENCES ậ. 108 LIST OF TABLES Table 1. Differences between two cell culture models !Ở,. IR band assignments of E4S nanoparticles .- -- 5-5 -< «<< £++xc<seexse 56 Table 3.

Average of tumor weight 3 days after InJeCfION. Reacting conditions for synthesis and properties of obtained E4S particles79 Table 4. Properties of various E4S BPMOS. Porosity, surface area, pore size and loading capacity of silica materials.

Influence of solvent loading in capacity of E4S NPs.c--S2 93 vi LIST OF FIGURES Figure 1. Nanovectors as an effective alternative in cancer therapies!?. Overview of outstanding properties and biomedical applications of Mesoporous Silica and organosilical? NA. Modern drug delivery systems response to internal stimuli conditions”’.

The formation of silica and silsesquioxane materials through the hydrolysis and condensation of silanes a) and organosilanes b), respectively. The creation of MSNs, MONs, and PMO NPs by templated sol—gel processes €)!9. Obtained morphology and size of the BPMOs by adjusting the ratio between two organosilica precursors. TEM images show various sizes and shapes of synthesized manoparticles?.

Degradable nanoparticles are synthesized by Moghaddam et al. Transmission electron microscope images of obtained nanoparticles synthesized by various combinations of inorganosilica precursor and organosilica precursors". TEM images showed the biodegradation of HMONs in PBS with 10 mM GSH for various times: (a) 0, (b) 3, (c) 7 days, and (d) 14 days. (e) DLS results showed the degradation and (f) the mass of Si in HMONs during the degradation in PBS with different GSH concentrations.

(g) A scheme of the reactions occurred during the degradation and biodegradation process Of HMONS. Structure of daunorubicin anticancer drug .-- --- -s«++s£+ss+sexssss 18 Figure 1. Cordyceps fungi and cordycepin structure ”Ì.---‹s-+ss+s+x+x+xsxerzesesree 19 Figure 1. Efficient optical methods applied in the study of nanocarriers in drug 6i 1à.

Huang’s group performed FT-IR to confirm the presence of vinyl in the mesoporous silica nanoparticles and DOX-conjugated polymer grafted MSNsŸ. Mekaru et al. reported the reduction of the disulfide bonds to thiol groups of biodegradability of disulfide-organosilica nanoparticles by means of XPS including (a) S 2p, (b) C 1s, (c) O 1s, and (d) Si 2p?. Morphology of disulfide-doped silica nanoparticles was reported by I P1331 1801 8n a1Ầ-”'^”-'-:-:-3.

SEM images of submicrometric capsules synthesized by Zyuzin et al., including SiO2 capsules with (A) low and (B) high amounts of tetraethyl orthosilicate, (C) (DEXS/PARG}x and (PSS/PAH)4 capsules”. Croissant et al. applied TEM images to evaluate the degradation of oxamide-phenylene-based mesoporous organosilica nanoparticles in PBS with trypsin at various times ((B) 24 h and (C) 48 by. TEM images of (a) pristine, (b) FITC-doped silica nanoparticles, and (c-f) individual anticancer drugs silica nanoparticles were reported by Sully et al.

Zyuzin et al. performed confocal laser scanning microscopy to study the transfection of SiOz capsules in HeLa cells. (A) Various exposure conditions were applied to evaluate the transfection of SiOz. Scale bar: 100 um.

(B) Transfected cells with high magnification using SiOz. Scale bar: 20 IIrm”,. 5+5: ss+x+c+zvzvzesesxssee 29 Figure 1. Confocal microscopy images of MCF-7 cells (a) as a control, (b) incubated with chitosan-modified APTES-FITC-containing mesoporous silica nanoparticles, and (c) incubated with MTX-loaded chitosan-modified APTES-FITC-containing MSNs in 0110100000208] {010g nà.

Spheroids formation on human breast cancer cell (MCF-7)!°3. The experiment timeline of the CAM model using chicken embryoŸ”. Procedure for synthesis of ethane-containing tetrasulfide-based biodegradable periodic mesoporous organosilica nanoparfICÌes. Experimental procedure for the synthesis of fluorescent ethane-containing tetrasulfide-based biodegradable periodic mesoporous organosilica nanoparticles.

Experimental procedure for the synthesis of phenylene-containing tetrasulfide-based biodegradable periodic mesoporous organosilica nanoparticles. Experimental procedure for the synthesis of inorganic mesoporous silica I0 1091614110118. Experimental procedure for evaluating the degradation of nanoparticles in VATIOUS CONIIONS .-- 2G 1 1 910191 9119 1 301901 nh ni HH TH nhờ 42 Figure 2. Experimental procedure for daunorubicin loading.

Experimental procedure for cordycepin loading. Experimental procedure for evaluating the in vitro release profile of cordycepin from nanoparticles dd. Experimental procedure for evaluating cytotoxicity of nanoparticles. Experimental procedure for evaluating the uptake of E4S NPs by 3D tumor SPherOIids.

Experimental procedure for evaluating the biodistribution of E4S on Chicken egg TmO(GÌ.- - 5s 1 1991193191130 1 91 hi HH HH nước 51 Figure 2. Experimental procedure for evaluating the tumor elimination of daunorubicin-loaded E4S on chicken egg models. Schematic illustration of a synthesis process of ethane-containing tetrasulfide-based BPMOs (E⁄4S). 00101111 111g v11 ng vn ng ky 53 Figure 3.

SEM images Of E⁄4S.-- óc HH HH TH TH HH HH ng rệt 54 Figure 3. TEM images Of E⁄4S.- 5 càng HH HH Hà HH 55 Figure 3. Elemental mapping of E4S: Si and S. FT-IR spectra Of E4S.- c9 TH HH HH ng giết 56 Figure 3.

XPS survey Of EAS oo. XPS spectra of C 1s in E⁄4S. XPS spectra of S 2p in E4S .-- -- Hn HH HnHHHkHkt 59 Figure 3. XPS spectra of Si 2p in E4S oo.

eeeeceseenececeeeeceeeeseceeeeseceeceaeeseeeseeneenee 60 ix Figure 3. XPS spectra of O 1s In E4S. -- -- cv SH ng, 61 Figure 3. Thermogravimetric analysis Of E4S.

Na adsorption-desorption isotherm of E4S.- --- --«csss+sscssssreeses 63 Figure 3. In vitro biodegradation of E4S in the reducing environment (10 mM glutathione in phosphate-buffered saline). Scale bar: 200 nm. TEM images and respective DLS results exhibited the degradation of E4S after 3 days of incubation in the reducing environment (10 mM glutathione in phosphate- buffered saline or in simulated body fÏU1(]).

In vitro biodegradation of MSN in the reducing environment (10 mM glutathione in phosphate-buffered saline). Uptake of Rhodamine B-labeled E4S BPMO into 3D ovarian tumor spheroids after 18 h incubation. Green fluorescent protein (GFP) expressed human ovarian cancer cells (OVCAR-8). RFP showed the fluorescence of daunorubicin.

Scale bar equals 100 UM. Optical images of ovarian tumor spheroid after treatment with no injection (Control), free E4S (BPMO), free DNR, or DNR-loaded E4S (DNR-BPMO) over a period of 7 days. The scale bar equals 100 Hm. 25 55553 £++++vE+eeeeEeseeeerees 69 Figure 3.

Calculated spheroid volume after treatment with no injection (Control), free EAS (BPMO), free DNR, or DNR-loaded E4S (DNR-BPMO) over 7 days. Error bars SHOW standard errr. The spheroid volume was calculated after treatment with no injection (Control) and free E4S (BPMO) (2.5, 5, and 10 ug) for 7 days. Error bars show standard Figure 3.

The spheroid volume was calculated after treatment with no injection (Control) and free DNR (0.2 ug) for 7 days. Error bars show standard error Figure 3. The spheroid volume was calculated after treatment with no injection (Control) and DNR-loaded E4S (DNR-BPMO) (2.5, 5, and 10 ug) for 7 days. Error bars SHOW Standard errr nntatR.

Chicken egg tumor model. OVCAR-8 tumor produced by transplanting ovarian cancer cells expressing GFP. Rhodamine B-labeled E4S BPMO (0.100 pL’ ') was intravenously injected into the chicken egg blood vessel. Bright-field and fluorescence images indicate the preferential accumulation in the tumor of E4S BPMO.

Rhodamine B-labeled E4S BPMO (0.100 pL!) was intravenously injected into the chicken egg blood vesSelÌ. - --‹-+-s<+<<<x<+ex+sess 73 Figure 3. Confocal images of the accumulation of E4S BPMO in the chicken egg embryo.