(LUẬN văn THẠC sĩ) nghiên cứu chế tạo và đánh giá băng gạc kháng khuẩn ba lớp định hướng ứng dụng trong điều trị vết thương

MINISTRY OF EDUCATION AND TRAINING VIETNAM ACADEMY OF SCIENCE AND TECHNOLOGY GRADUATE UNIVERSITY OF SCIENCE AND TECHNOLOGY TRẦN MINH CHIẾN Trần Minh Chiến INORGANIC CHEMISTRY INVESTIGATION OF TRILAYER MEMBRANE ORIENTATED FOR ANTIBACTERIAL WOUND DRESSING: FABRICATION, CHARACTERIZATION, AND EVALUATION MASTER THESIS Inorganic Chemistry 2021 Ho Chi Minh city - 2021 download by : skknchat@gmail.com MINISTRY OF EDUCATION VIETNAM ACADEMY OF AND TRAINING SCIENCE AND TECHNOLOGY GRADUATE UNIVERSITY OF SCIENCE AND TECHNOLOGY Tran Minh Chien INVESTIGATION OF TRILAYER MEMBRANE ORIENTATED FOR ANTIBACTERIAL WOUND DRESSING: FABRICATION, CHARACTERIZATION, AND EVALUATION Major: Code: Inorganic Chemistry 8440113 MASTER THESIS SUPERVISOR: Assoc Prof Nguyen Thi Hiep Ho Chi Minh City – 2021 download by : skknchat@gmail.com Tran Minh Chien – Master Thesis DECLARATION OF INTERESTS I hereby declare that this thesis represents my own work which has been done after registration for the Master degree at Graduate University of Science and Technology, and has not been previously included in a thesis or dissertation submitted to this or any other institution for a degree, diploma or other qualifications All the results are correct and impartial, if wrong, I take full responsibility Ho Chi Minh city, September 10th, 2021 Trần Minh Chiến download by : skknchat@gmail.com Tran Minh Chien – Master Thesis ACKNOWLEDGMENTS This thesis has received numerous guidance and assistance from my supervisors and colleagues at Tissue Engineering & Regenerative Medicine laboratory (TERM) It would not have been possible without these individuals who contributed generously to the completion of this thesis First of all, I am especially thankful to my supervisor, Prof Nguyen Thi Hiep for providing me this interesting topic, for the opportunity to work independently, and for valuable discussions She had always trusted and encouraged me during this thesis Her guidance helped me in all the time of research and writing of this thesis I would also thank her for teaching me how to work efficiently, how to solve problems, and how to research independently Secondly, I want to express my gratitude to Prof Nguyen Phuong Tung, who offered me the research position at Nanomaterials and Petroleum additives Lab, Institute of Applied Materials Science Even though the time I worked with her was short, she gave me many precious lessons and experiences If it was not for her encouragement, I could never engage further in science I would like to give my sincere thanks to all the lecturers from Graduate University of Science and Technology for their invaluable guidance throughout my studies They provided me with many in-depth insights into the field of Chemistry that aided my research Next, I would like to thank all the help from my seniors, especially Hieu Minh, Khanh Vinh, and Thao Nhi who welcomed and helped me during my time at TERM I am also grateful to the Department of Biomedical Engineering at the International University for the facility support Last but not least, I would like to express my deepest gratitude to my family They supported and gave me all the best things during my thesis work Their mental and physical supports helped me overcome many difficulties It is my honor to have all of you supporting me Without any of you, I may not make a successful thesis like this Once again, thank you very much Sincerely Yours, Tran Minh Chien download by : skknchat@gmail.com Tran Minh Chien – Master Thesis TABLE OF CONTENT DECLARATION OF INTERESTS ACKNOWLEDGMENTS TABLE OF CONTENT LIST OF FIGURES LIST OF TABLES LIST OF ABBREVIATION ABSTRACT 10 INTRODUCTION 12 CHAPTER 1: LITERATURE REVIEW 15 CHAPTER 2: MATERIALS AND METHODS 19 2.1 MATERIALS 19 2.2 PREPARATION AND CHARACTERIZATION OF PCL-AG SUSPENSIONS 19 2.2.1 Preparation of PCL-Ag suspensions 19 2.2.2 Characterization of PCL-Ag suspensions 19 2.3 FABRICATION AND CHARACTERIZATION OF PCL-AG MEMBRANES 20 2.3.1 Electrospinning 20 2.3.2 Morphological observation of electrospun membranes 20 2.3.3 Nanoparticles analysis 20 2.3.4 Mechanical properties of electrospun membranes 21 2.3.5 Wettability 21 2.3.6 Moisture permeability 21 2.3.7 In vitro Ag release kinetic 21 2.3.8 Antibacterial activity 21 2.3.9 Cytotoxicity assay 22 download by : skknchat@gmail.com Tran Minh Chien – Master Thesis 2.4 PREPARATION OF PCL-AG-COS MEMBRANE 22 2.4.1 Preparation of PCL-Ag/POX membrane 22 2.4.2 Preparation of COS/PVP solution 22 2.4.3 COS/PVP coating on PCL-Ag/POX membrane 23 2.5 CHARACTERIZATION OF PCL-AG-COS MEMBRANE 23 2.5.1 Morphological study 23 2.5.2 Fourier-transform infrared spectroscopy (FTIR) analysis 23 2.5.3 Asymmetric wettability 23 2.5.4 Mechanical properties 24 2.5.5 Water absorbability 24 2.5.6 Moisture permeability 24 2.5.7 In vitro Ag release kinetic 24 2.5.8 Antibacterial activity 25 2.5.9 Cytotoxicity assay 25 2.6 STATISTICAL ANALYSIS 26 CHAPTER 3: RESULTS AND DISCUSSION 27 3.1 CHARACTERIZATION OF PCL-AG SUSPENSIONS 27 3.2 CHARACTERIZATION OF ELECTROSPUN MEMBRANES 29 3.2.1 Morphology of electrospun membranes 29 3.2.2 Nanoparticles analysis 31 3.2.3 Mechanical properties of PCL-Ag membranes 33 3.2.4 Wettability 35 3.2.5 Moisture permeability 36 3.2.6 In vitro Ag release kinetic 37 3.2.7 Antibacterial activity 38 3.2.8 Cytotoxicity assay 39 3.3 CHARACTERIZATION OF PCL-AG-COS MEMBRANE 41 download by : skknchat@gmail.com Tran Minh Chien – Master Thesis 3.3.1 Morphological study 41 3.3.2 FTIR study 42 3.3.3 Asymmetric wettability 43 3.3.4 Mechanical properties 44 3.3.5 Water absorbability and moisture vapor transmission rate 46 3.3.6 In vitro silver release kinetic 48 3.3.7 Antibacterial activity 49 3.3.8 Cytotoxicity assay 51 CHAPTER 4: CONCLUSION AND IMPLICATIONS 53 REFERENCES 54 APPENDIX 64 download by : skknchat@gmail.com Tran Minh Chien – Master Thesis LIST OF FIGURES Figure 2.1 Graphical illustration of the trilayer PCL-Ag-COS membrane fabrication process 26 Figure 3.1 (a) Photographs of PCL-Ag 250 ppm, PCL-Ag 500 ppm, and PCL-Ag 1000 ppm irradiated at gamma dose of 7.5, 15, 25 kGy, respectively Images were taken at 7-day intervals for 28 days Comparison of UV-Vis spectra of PCL-Ag suspensions at day (b) and day 28 (c) 27 Figure 3.2 SEM micrographs of electrospun raw PCL (a1), PCL-Ag 250 ppm (a2), PCL-Ag 500 ppm (a3) and PCL-Ag 1000 ppm (a4) (Scale bar: 50 µm) Histogram of fiber diameter and pore size distribution of raw PCL (b1, c1), PCL-Ag 250 ppm (b2, c2), PCL-Ag 500 ppm (b3, c3) and PCL-Ag 1000 ppm (b4, c4) (n=30) 30 Figure 3.3 X-ray diffraction (XRD) patterns of SNPs incorporated PCL membranes 32 Figure 3.4 Transmission electron microscopy (TEM) micrographs of PCL-Ag 250 ppm (a1), PCL-Ag 500 ppm (b1), PCL-Ag 1000 ppm (c1), and their size distribution histograms (a2, b2, c2) (Scale bars: 200 nm- a1, 500 nm-b1, c1, n=30) 33 Figure 3.5 Tensile strength - strain curves of PCL-Ag compared with raw PCL (n = 3) 34 Figure 3.6 Contact angles of raw PCL, PCL-Ag 250 ppm, PCL-Ag 500 ppm, and PCL-Ag 1000 ppm membranes The photographs above each column illustrate the water droplets on the membrane surface (data = mean ± SD, n=5, *: p0.05) 35 Figure 3.7 Quantification of the in vitro release of silver from the PCL-Ag 250 ppm, PCL-Ag 500 ppm, and PCL-Ag 1000 ppm in PBS solution (pH=5.5) Aliquots were taken after 1, 3, 6, 12, and 24 hours, and quantified by ICP-MS technique (data = mean ± SD, n=3) 37 Figure 3.8 (a) Photographs of the inhibition zones of raw PCL, PCL-Ag 250 ppm, PCL-Ag 500 ppm, and PCL-Ag 1000 ppm against P aeruginosa and S aureus strains and (b) inhibition zone diameters (Scale bar: 10 mm, data = mean ± SD, n=4, *: p0.05) 38 Figure 3.9 Cytotoxicity test of PCL-Ag 250 ppm, PCL-Ag 500 ppm, and PCL-Ag 1000 ppm on L929 murine fibroblast cell (data = mean ± SD, n=3, *: p0.05) 40 Figure 3.10 SEM micrographs of electrospun PCL-Ag 500 ppm, PCL-Ag/POX, and PCL-Ag-COS membranes from (a) top-down view and (b) cross-section view (Scale bar: 10 µm) 41 download by : skknchat@gmail.com Tran Minh Chien – Master Thesis Figure 3.11 FT-IR spectra of PCL-Ag 500 ppm, PCL-Ag/POX and PCL-Ag-COS membranes 42 Figure 3.12 (a) Images of water droplets on PCL-Ag 500 ppm, PCL-Ag/POX, and PCL-Ag-COS over time (b) Dynamic contact angle of PCL-Ag 500 ppm, PCL-Ag/POX, and PCL-Ag-COS (n=3) 44 Figure 3.13 Tensile strength-strain curves of PCL-Ag 500 ppm, PCL-Ag/POX, and PCL-Ag-COS (n = 3) 45 Figure 3.14 Water absorbability of PCL-Ag 500 ppm, PCL-Ag/POX, and PCL-Ag-COS membranes (data = mean ± SD, n = 5, ns: p> 0.05, *: p< 0.05) 47 Figure 3.15 Quantification of the in vitro release of silver from the PCL-Ag 500 ppm, PCL-Ag/POX and PCL-Ag-COS in PBS solution (pH=5.5) Aliquots were taken after 1, 3, 6, 12, and 24 hours, and quantified by ICP-MS technique (data = mean ± SD, n=3) 49 Figure 3.16 Image of (a) the Zones of inhibition formed by the raw PCL, PCL-Ag 500 ppm, and PCL-Ag-COS membranes against P aeruginosa and S aureus and (B) the measured zone diameters (Scale bar: 10 mm, data = mean ± SD, n = 4, ns: p > 0.05, *: p < 0.05) 50 Figure 3.17 Viability (%) of fibroblasts after 24h of incubation in different concentrations of extracted solution of PCL-Ag 500 ppm and PCL-Ag-COS membranes (data = mean ± SD, n = 3, ns: p> 0.05, *: p< 0.05) 51 download by : skknchat@gmail.com Tran Minh Chien – Master Thesis LIST OF TABLES Table 3.1 Viscosity of PCL, PCL-Ag suspensions before and after gamma exposure The deviations between samples were lower than the machine’s error range 29 Table 3.2 Silver content of PCL-Ag 250 ppm, PCL-Ag 500 ppm, and PCL-Ag 1000 ppm membrane 31 Table 3.3 Tensile properties of electrospun PCL membranes incorporated with different concentration of SNPs (data = mean ± SD, n = 3) 34 Table 3.4 Moisture vapor transmission rate of PCL-Ag membranes (data = mean ± SD, n=5) 36 Table 3.5 Average fiber diameter and pore size of PCL-Ag 500 ppm and PCL-Ag/POX membranes (data = mean ± SD, n=30) 41 Table 3.6 Tensile properties of PCL-Ag 500 ppm, PCL-Ag/POX, and PCL-Ag-COS membranes (data = mean ± SD, n = 3) 46 Table 3.7 Moisture vapor transmission rate of PCL-Ag 500 ppm, PCL-Ag/POX and PCL-Ag-COS membranes (data = mean ± SD, n=5) 46 LIST OF ABBREVIATION PCL: Polycaprolactone SNPs: Silver nanoparticles COS: Oligomer Chitosan PVP: Poly (N-vinyl pyrrolidone) POX: Poloxamer 407 DMSO: Dimethyl sulfoxide S aureus: Staphylococcus aureus P aeruginosa: Pseudomonas aeruginosa DMEM: Dulbecco`s Modified Eagle Media PBS: Phosphate-buffered saline UV-Vis: Ultraviolet- Visible spectroscopy SEM: Scanning electron microscopy download by : skknchat@gmail.com Tran Minh Chien – Master Thesis Chapter 3: Results and Discussion Gram-positive and Gram-negative bacteria Several researchers hypothesized that the thicker cell wall of Gram-positive bacteria due to more peptidoglycan composition leads to less susceptibility than Gram-negative bacteria because it can prevent SNPs from anchoring and penetrating the cell wall[95] Likewise, COS was reported to be more effective on Gram-negative bacteria thanks to the more positive charge (NH3+), which promotes higher adsorption through cell walls [96, 97] The inhibition zone formed by PCL-Ag-COS was larger compared to that of PCL-Ag 500 ppm This outcome was more likely due to COS significantly enhancing the antibacterial effect 3.3.8 Cytotoxicity assay Figure 3.17 Viability (%) of fibroblasts after 24h of incubation in different concentrations of extracted solution of PCL-Ag 500 ppm and PCL-Ag-COS membranes (data = mean ± SD, n = 3, ns: p> 0.05, *: p< 0.05) The resazurin reduction assay was carried out to measure cell viability after being cultured in the extract solution for 24 hours, thereby evaluating the cytotoxicity of the materials According to the results shown in Figure 3.17, cell viability was 102% after being cultured in the 50% extract solution from PCL-Ag-COS sample However, being 51 download by : skknchat@gmail.com Tran Minh Chien – Master Thesis Chapter 3: Results and Discussion exposed to 100% concentration reduced survival rate of cell population down to 10%, which was significantly lower compared to that of PCL-Ag 500 ppm (76%) The wound dressing must not cause cytotoxicity or damage to the exposed tissue However, the PCL-Ag-COS membrane seemed to be strongly harmful to L929 cells, which killed mostly 90% of the cell population Aside from SNPs effect, this could also be the results of several factors of COS in the culture extract such as concentration, molecular weight, positive charge density, or deacetylation degree [98, 99] In specific, the high molecular weight polymer with extended conformation and positive charge groups could readily bind to the cell membrane and obstruct their metabolism and respiration Despite the extract from the PCL-Ag-COS membrane annihilated most of the cells, we cannot state that PCL-Ag-COS is inappropriate for wound dressing application Since there are vast differences between in vitro and in vivo experiments For example, in the resazurin assay, the cells were entirely exposed in extract solution containing all the COS and SNPs that were released in 24 hours However, in the in vitro tests, COS and SNPs were slowly diffused into the wound bed for a long period, hence, causing less stress for the damaged tissue Therefore, an in vivo experiment is required to accurately evaluate the potential of PCL-Ag-COS in wound treatment 52 download by : skknchat@gmail.com Tran Minh Chien – Master Thesis Chapter 4: Conclusion and Implications CHAPTER 4: CONCLUSION AND IMPLICATIONS In the first stage, I succeeded in directly synthesizing of SNPs in PCL solution using gamma irradiation for the electrospinning process of PCL-Ag membranes The physicochemical properties of the electrospun membranes were characterized and their in vitro properties were assessed in this study The SNPs produced by gamma irradiation were stable in PCL solution for 28 days, anticipating the potency to synthesize nanoparticles in a non-aqueous solvent Images captured by SEM implied that the rise in silver components slightly increased the fiber diameter XRD spectra confirmed the existence of the SNP crystalline phase, while TEM micrographs demonstrated the uniform distribution of SNPs across the electrospun fibers Further, the electrospun PCL-Ag membrane possessed surface hydrophobic properties and excellent tensile tolerance The PCL-Ag 500 ppm and 1000 ppm samples expressed excellent antimicrobial effects against both S aureus and P aeruginosa strains However, only PCL-Ag 500 ppm membranes showed good biocompatibility and could be promising for wound dressing applications In the second stage, the study has reported the effect of coating the COS/PVP onto PCL-Ag 500 ppm membrane for wound treatment In conclusion, the PCL-Ag-COS membrane was prepared successfully by coating COS/PVP solution onto the PCL-Ag 500 ppm membrane with the help of the PCL-POX layer Morphology observation illustrated an evenly coated surface of the membrane The supplementation of COS/PVP imparted the membrane with asymmetric wettability, water absorbability, while on the other hand, reduced mechanical strength and water vapor transmission rate The gradual release of SNPs after 24 hours was also confirmed Despite having an excellent bacterial inhibitory, the PCL-Ag-COS membrane showed a limitation in the viability of cells, which might be caused by the excessive amount of COS and Ag content in the membrane Future work will be focused on investigating in vivo study of the membrane before being used as a first aid wound treatment dressing, which could provide an optimal environment condition to promote wound healing rate as well as preventing the invasion of bacteria 53 download by : skknchat@gmail.com Tran Minh Chien – Master Thesis References REFERENCES Mukhopadhyay, P.; Maity, S.; Mandal, S.; Chakraborti, A S.; Prajapati, A.; Kundu, P., Preparation, characterization and in vivo evaluation of pH sensitive, safe quercetin-succinylated chitosan-alginate core-shell-corona nanoparticle for diabetes treatment, Carbohydrate Polymers, 2018, 182, 42-51 Yao, Q.; Liu, Y.; Selvaratnam, B.; Koodali, R T.; Sun, H., Mesoporous silicate nanoparticles/3D nanofibrous scaffold-mediated dual-drug delivery for bone tissue engineering, Journal of Controlled Release, 2018, 279, 69-78 Lu, J.; Sun, J.; Li, F.; Wang, J.; Liu, J.; Kim, D.; Fan, C.; Hyeon, T.; Ling, D J., Highly sensitive diagnosis of small hepatocellular carcinoma using pH-responsive iron oxide nanocluster assemblies, Journal of the American Chemical Society, 2018, 140 (32), 10071-10074 Rai, M.; Yadav, A.; Gade, A., Silver nanoparticles as a new generation of antimicrobials, Biotechnology Advances, 2009, 27 (1), 76-83 Tang, S.; Zheng, J., Antibacterial activity of silver nanoparticles: structural effects, Advanced Healthcare Materials, 2018, (13), 1701503 Ivask, A.; Kurvet, I.; Kasemets, K.; Blinova, I.; Aruoja, V.; Suppi, S.; Vija, H.; Käkinen, A.; Titma, T.; Heinlaan, M., Size-dependent toxicity of silver nanoparticles to bacteria, yeast, algae, crustaceans and mammalian cells in vitro, 2014, PloS One, (7), e102108 Pal, S.; Tak, Y K.; Song, J M., Does the antibacterial activity of silver nanoparticles depend on the shape of the nanoparticle? A study of the gram-negative bacterium Escherichia coli, Applied and Environmental Microbiology, 2007, 73 (6), 1712-1720 Shrivastava, S.; Bera, T.; Roy, A.; Singh, G.; Ramachandrarao, P.; Dash, D., Characterization of enhanced antibacterial effects of novel silver nanoparticles, Nanotechnology, 2007, 18 (22), 225103 Sundaramurthi, D.; Krishnan, U M.; Sethuraman, S., Electrospun Nanofibers as Scaffolds for Skin Tissue Engineering, Polymer Reviews, 2014, 54 (2), 348-376 10 Joseph, B.; Augustine, R.; Kalarikkal, N.; Thomas, S.; Seantier, B.; Grohens, Y., Recent advances in electrospun polycaprolactone based scaffolds for wound healing and skin bioengineering applications, Materials Today Communications, 2019, 19, 319335 54 download by : skknchat@gmail.com Tran Minh Chien – Master Thesis 11 References Serrano, M.; Pagani, R.; Vallet-Regı, M.; Pena, J.; Ramila, A.; Izquierdo, I.; Portoles, M., In vitro biocompatibility assessment of poly (ε-caprolactone) films using L929 mouse fibroblasts, Biomaterials, 2004, 25 (25), 5603-5611 12 Ho, M H.; Do, T B.-T.; Dang, N N.-T.; Le, A N.-M.; Ta, H T.-K.; Vo, T V.; Nguyen, H T., Effects of an acetic acid and acetone mixture on the characteristics and scaffold–cell interaction of electrospun polycaprolactone membranes, Applied Sciences, 2019, (20), 4350 13 Lim, M M.; Sultana, N., In vitro cytotoxicity and antibacterial activity of silvercoated electrospun polycaprolactone/gelatine nanofibrous scaffolds, Biotechnology, 2016, (2), 211-211 14 Thanh, N T.; Hieu, M H.; Phuong, N T M.; Thuan, T D B.; Thu, H N T.; Do Minh, T.; Dai, H N.; Thi, H N., Optimization and characterization of electrospun polycaprolactone coated with gelatin-silver nanoparticles for wound healing application, Materials Science and Engineering: C, 2018, 91, 318-329 15 Thomas, R.; Soumya, K.; Mathew, J.; Radhakrishnan, E.; biotechnology, Electrospun polycaprolactone membrane incorporated with biosynthesized silver nanoparticles as effective wound dressing material, Biotechnology and Applied Biochemistry, 2015, 176 (8), 2213-2224 16 Lee, S H.; Jun, B.-H., Silver nanoparticles: synthesis and application for nanomedicine, International Journal of Molecular Sciences, 2019, 20 (4), 865 17 Iravani, S.; Korbekandi, H.; Mirmohammadi, S V.; Zolfaghari, B., Synthesis of silver nanoparticles: chemical, physical and biological methods, Research in Pharmaceutical Sciences, 2014, (6), 385-406 18 Sun, Y.; Chmielewski, A G., Applications of ionizing radiation in materials processing Institute of Nuclear Chemistry and Technology: 2017 19 Uttayarat, P.; Eamsiri, J.; Tangthong, T.; Suwanmala, P., Radiolytic Synthesis of Colloidal Silver Nanoparticles for Antibacterial Wound Dressings Advances in Materials Science and Engineering, 2015, 2015, 376082 20 Grzelczak, M.; Liz-Marzán, L M., The relevance of light in the formation of colloidal metal nanoparticles Chemical Society Reviews, 2014, 43 (7), 2089-2097 21 Juby, K.; Dwivedi, C.; Kumar, M.; Kota, S.; Misra, H.; Bajaj, P., Silver nanoparticle-loaded PVA/gum acacia hydrogel: Synthesis, characterization and antibacterial study, Carbohydrate Polymers, 2012, 89 (3), 906-913 55 download by : skknchat@gmail.com Tran Minh Chien – Master Thesis 22 References Perez, R A.; Won, J.-E.; Knowles, J C.; Kim, H.-W., Naturally and synthetic smart composite biomaterials for tissue regeneration, Advanced drug delivery reviews, 2013, 65 (4), 471-496 23 Huang; Fu, Naturally derived materials-based cell and drug delivery systems in skin regeneration, Journal of Controlled Release, 2010, 142 (2), 149-159 24 Lu, M.; Xing, H.; Jiang, J.; Chen, X.; Yang, T.; Wang, D.; Ding, P., Liquisolid technique and its applications in pharmaceutics, Asian Journal of Pharmaceutical Sciences, 2017, 12 (2), 115-123 25 Abou-Aiad, T.; Abd-El-Nour, K.; Hakim, I.; Elsabee, M., Dielectric and interaction behavior of chitosan/polyvinyl alcohol and chitosan/polyvinyl pyrrolidone blends with some antimicrobial activities, Polymer, 2006, 47 (1), 379-389 26 Fedorowicz, J.; Sączewski, J.; Konopacka, A.; Waleron, K.; Lejnowski, D.; Ciura, K.; Tomašič, T.; Skok, Ž.; Savijoki, K.; Morawska, M., Synthesis and biological evaluation of hybrid quinolone-based quaternary ammonium antibacterial agents, European Journal of Medicinal Chemistry, 2019, 179, 576-590 27 Kang, J.; Dietz, M J.; Li, B J P o., Antimicrobial peptide LL-37 is bactericidal against Staphylococcus aureus biofilms, PLoS One, 2019, 14 (6), e0216676 28 Liao, S.; Zhang, Y.; Pan, X.; Zhu, F.; Jiang, C.; Liu, Q.; Cheng, Z.; Dai, G.; Wu, G.; Wang, L., Antibacterial activity and mechanism of silver nanoparticles against multidrug-resistant Pseudomonas aeruginosa, International Journal of Nanomedicine, 2019, 14, 1469 29 Celebioglu, A.; Topuz, F.; Yildiz, Z I.; Uyar, T J C p., One-step green synthesis of antibacterial silver nanoparticles embedded in electrospun cyclodextrin nanofibers, Carbohydrate Polymers, 2019, 207, 471-479 30 Lee, B.; Lee, D., Synergistic antibacterial activity of gold nanoparticles caused by apoptosis‐like death, Journal of Applied Microbiology, 2019, 127 (3), 701-712 31 Arafa, M G.; El-Kased, R F.; Elmazar, M J S r., Thermoresponsive gels containing gold nanoparticles as smart antibacterial and wound healing agents, Scientific Reports, 2018, (1), 1-16 32 Joe, A.; Park, S.-H.; Shim, K.-D.; Kim, D.-J.; Jhee, K.-H.; Lee, H.-W.; Heo, C.-H.; Kim, H.-M.; Jang, E.-S., Antibacterial mechanism of ZnO nanoparticles under dark conditions, Journal of Industrial and Engineering Chemistry, 2017, 45, 430-439 56 download by : skknchat@gmail.com Tran Minh Chien – Master Thesis 33 References Barbosa, T M.; Levy, S B., The impact of antibiotic use on resistance development and persistence, Antimicrobial and Anticancer Chemotherapy, 2000, (5), 303-311 34 Percival, S L.; Salisbury, A.-M.; Chen, R., Silver, biofilms and wounds: resistance revisited, Critical Reviews in Microbiology, 2019, 45 (2), 223-237 35 Punjataewakupt, A.; Napavichayanun, S.; Aramwit, P.; Diseases, I., The downside of antimicrobial agents for wound healing, European Journal of Clinical Microbiology & Infectious Diseases, 2019, 38 (1), 39-54 36 Le, A N.-M.; Nguyen, T T.; Ly, K L.; Dai Luong, T.; Ho, M H.; Tran, N M.-P.; Dang, N N.-T.; Van Vo, T.; Tran, Q N.; Nguyen, T H, Modulating biodegradation and biocompatibility of in situ crosslinked hydrogel by the integration of alginate into N, O-carboxylmethyl chitosan–aldehyde hyaluronic acid network, Polymer Degradation and Stability, 2020, 180, 109270 37 Nguyen, N T.-P.; Nguyen, L V.-H.; Tran, N M.-P.; Nguyen, D T.; Nguyen, T N.-T.; Tran, H A.; Dang, N N.-T.; Van Vo, T.; Nguyen, T.-H., The effect of oxidation degree and volume ratio of components on properties and applications of in situ cross-linking hydrogels based on chitosan and hyaluronic acid, Materials Science and Engineering: C, 2019, 103, 109670 38 Pham, L.; Truong, M D.; Nguyen, T H.; Le, L.; Nam, N D.; Bach, L G.; Nguyen, V T.; Tran, N Q., A dual synergistic of curcumin and gelatin on thermalresponsive hydrogel based on Chitosan-P123 in wound healing application, Biomedicine & Pharmacotherapy, 2019, 117, 109183 39 Li, S.; Pei, M.; Wan, T.; Yang, H.; Gu, S.; Tao, Y.; Liu, X.; Zhou, Y.; Xu, W.; Xiao, P., Self-healing hyaluronic acid hydrogels based on dynamic Schiff base linkages as biomaterials, Carbohydrate Polymers, 2020, 250, 116922 40 Nguyen, N T.-P.; Nguyen, L V.-H.; Thanh, N T.; Van Toi, V.; Quyen, T N.; Tran, P A.; Wang, H.-M D.; Nguyen, T.-H., Stabilization of silver nanoparticles in chitosan and gelatin hydrogel and its applications, Materials Letters, 2019, 248, 241245 41 Pannerselvam, B.; Jothinathan, M K D.; Rajenderan, M.; Perumal, P.; Thangavelu, K P.; Kim, H J.; Singh, V.; Rangarajulu, S K., An in vitro study on the burn wound healing activity of cotton fabrics incorporated with phytosynthesized silver nanoparticles in male Wistar albino rats, European Journal of Pharmaceutical Sciences, 2017, 100, 187-196 57 download by : skknchat@gmail.com Tran Minh Chien – Master Thesis 42 References Tao, G.; Cai, R.; Wang, Y.; Liu, L.; Zuo, H.; Zhao, P.; Umar, A.; Mao, C.; Xia, Q.; He, H., Bioinspired design of AgNPs embedded silk sericin-based sponges for efficiently combating bacteria and promoting wound healing, Materials & Design, 2019, 180, 107940 43 Sonseca, A.; Madani, S.; Rodríguez, G.; Hevilla, V.; Echeverría, C.; Fernández-García, M.; Moz-Bonilla, A.; Charef, N.; López, D., Multifunctional PLA blends containing chitosan mediated silver nanoparticles: Thermal, mechanical, antibacterial, and degradation properties, Nanomaterials, 2020, 10 (1), 22 44 Nhi, T T.; Khon, H C.; Hoai, N T T.; Bao, B C.; Quyen, T N.; Van Toi, V.; Hiep, N T., Fabrication of electrospun polycaprolactone coated withchitosan-silver nanoparticles membranes for wound dressing applications, Journal of Materials Science: Materials in Medicine, 2016, 27 (10), 156 45 Qian, Y.; Zhang, Z.; Zheng, L.; Song, R.; Zhao, Y., Fabrication and characterization of electrospun polycaprolactone blended with chitosan-gelatin complex nanofibrous mats, Journal of Nanomaterials, 2014 46 Xue, J.; He, M.; Liu, H.; Niu, Y.; Crawford, A.; Coates, P D.; Chen, D.; Shi, R.; Zhang, L., Drug loaded homogeneous electrospun PCL/gelatin hybrid nanofiber structures for anti-infective tissue regeneration membranes, Biomaterials, 2014, 35 (34), 9395-9405 47 Fang, Y.; Zhu, X.; Wang, N.; Zhang, X.; Yang, D.; Nie, J.; Ma, G J E P J., Biodegradable core-shell electrospun nanofibers based on PLA and γ-PGA for wound healing, European Polymer Journal , 2019, 116, 30-37 48 Liu, X.; Nielsen, L H.; Kłodzińska, S N.; Nielsen, H M.; Qu, H.; Christensen, L P.; Rantanen, J.; Yang, M., Ciprofloxacin-loaded sodium alginate/poly (lactic-coglycolic acid) electrospun fibrous mats for wound healing, European Journal of Pharmaceutics and Biopharmaceutics, 2018, 123, 42-49 49 Williamson, M R.; Black, R.; Kielty, C., PCL–PU composite vascular scaffold production for vascular tissue engineering: attachment, proliferation and bioactivity of human vascular endothelial cells, Biomaterials, 2006, 27 (19), 3608-3616 50 Minh, H H.; Hiep, N T.; Hai, N D.; Toi, V V., Fabrication of polycaprolactone/polyurethane loading conjugated linoleic acid and its antiplatelet adhesion, International Journal of Biomaterials, 2017 51 Eskitoros-Togay, Ş M.; Bulbul, Y E.; Tort, S.; Korkmaz, F D.; Acartürk, F.; Dilsiz, N J I j o p., Fabrication of doxycycline-loaded electrospun PCL/PEO 58 download by : skknchat@gmail.com Tran Minh Chien – Master Thesis References membranes for a potential drug delivery system, International Journal of Pharmaceutics, 2019, 565, 83-94 52 Augustine, R.; Kalarikkal, N.; Thomas, S., Electrospun PCL membranes incorporated with biosynthesized silver nanoparticles as antibacterial wound dressings, Applied Biochemistry and Biotechnology, 2016, (3), 337-344 53 Kalantari, K.; Afifi, A M.; Jahangirian, H.; Webster, T., Biomedical applications of chitosan electrospun nanofibers as a green polymer–Review, Carbohydrate Polymers, 2019, 207, 588-600 54 Adnan, S.; Ranjha, N M.; Hanif, M.; Asghar, S., O-Carboxymethylated chitosan; A promising tool with in-vivo anti-inflammatory and analgesic properties in albino rats, International Journal of Biological Macromolecules, 2020, 156, 531-536 55 Torkaman, S.; Rahmani, H.; Ashori, A.; Najafi, S H M J C P., Modification of chitosan using amino acids for wound healing purposes: A review, Carbohydrate Polymers, 2021, 117675 56 Doan, V K.; Ly, K L.; Tran, N M.-P.; Ho, T P.-T.; Ho, M H.; Dang, N T.N.; Chang, C.-C.; Nguyen, H T.-T.; Ha, P T.; Tran, Q N J M., Characterizations and Antibacterial Efficacy of Chitosan Oligomers Synthesized by Microwave-Assisted Hydrogen Peroxide Oxidative Depolymerization Method for Infectious Wound Applications, Materials, 2021, 14 (16), 4475 57 Yeh, J T.; Chen, C L.; Huang, K.; Nien, Y.; Chen, J.; Huang, P., Synthesis, characterization, and application of PVP/chitosan blended polymers Journal of applied polymer science, 2006, 101 (2), 885-891 58 Suknuntha, K.; Tantishaiyakul, V.; Vao‐Soongnern, V.; Espidel, Y.; Cosgrove, T., Molecular modeling simulation and experimental measurements to characterize chitosan and poly (vinyl pyrrolidone) blend interactions Journal of Polymer Science Part B: Polymer Physics, 2008, 46 (12), 1258-1264 59 Afify, T.; Saleh, H.; Ali, Z., Structural and morphological study of gamma‐ irradiation synthesized silver nanoparticles, Polymer Composites, 2017, 38 (12), 26872694 60 Yaqoob, A A.; Umar, K.; Ibrahim, M N M., Silver nanoparticles: various methods of synthesis, size affecting factors and their potential applications–a review Applied Nanoscience, 2020, 10 (5), 1369-1378 61 Nguyen, D T.; Ly, K L.; Tran, N M.-P.; Ho, M H.; Tran, T T.-P.; Nguyen, T.-H.; Nhi, D N T.; Vo, V T., Effect of Microwave Irradiation on Polyvinyl Alcohol 59 download by : skknchat@gmail.com Tran Minh Chien – Master Thesis References as a Carrier of Silver Nanoparticles in Short Exposure Time, International Journal of Polymer Science, 2019, 3623907 62 Darroudi, M.; Ahmad, M B.; Zak, A K.; Zamiri, R.; Hakimi, M., Fabrication and characterization of gelatin stabilized silver nanoparticles under UV-light International journal of molecular sciences, 2011, 12 (9), 6346-56 63 Zhen, J.-B.; Kang, P.-W.; Zhao, M.-H.; Yang, K.-W., Silver nanoparticle conjugated star PCL-b-AMPs copolymer as nanocomposite exhibits efficient antibacterial properties, Bioconjugate Chemistry, 2019, 31 (1), 51-63 64 Wasikiewicz, J M.; Yoshii, F.; Nagasawa, N.; Wach, R A.; Mitomo, H., Degradation of chitosan and sodium alginate by gamma radiation, sonochemical and ultraviolet methods, Radiation Physics and Chemistry, 2005, 73 (5), 287-295 65 de Cassan, D.; Hoheisel, A L.; Glasmacher, B.; Menzel, H., Impact of sterilization by electron beam, gamma radiation and X-rays on electrospun poly-(εcaprolactone) fiber mats, Journal of Materials Science: Materials in Medicine, 2019, 30 (4), 42 66 Li, T.; Park, H G.; Choi, S.-H., γ-Irradiation-induced preparation of Ag and Au nanoparticles and their characterizations, Materials Chemistry and Physics, 2007, 105 (2-3), 325-330 67 Zhou, Y.; Zhao, Y.; Wang, L.; Xu, L.; Zhai, M.; Wei, S.; Chemistry, Radiation synthesis and characterization of nanosilver/gelatin/carboxymethyl chitosan hydrogel, Radiation Physics and Chemistry, 2012, 81 (5), 553-560 68 Tan, S.-H.; Inai, R.; Kotaki, M.; Ramakrishna, S., Systematic parameter study for ultra-fine fiber fabrication via electrospinning process, Polymer, 2005, 46 (16), 6128-6134 69 Zhang, H.; Peng, M.; Cheng, T.; Zhao, P.; Qiu, L.; Zhou, J.; Lu, G.; Chen, J., Silver nanoparticles-doped collagen–alginate antimicrobial biocomposite as potential wound dressing, Journal of Materials Science, 2018, 53 (21), 14944-14952 70 Haseeb, M T.; Hussain, M A.; Abbas, K.; Youssif, B G.; Bashir, S.; Yuk, S H.; Bukhari, S N., Linseed hydrogel-mediated green synthesis of silver nanoparticles for antimicrobial and wound-dressing applications, International Journal of Nanomedicine, 2017, 12, 2845 71 Alhokbany, N.; Ahama, T.; Naushad, M.; Alshehri, S M J C P B E., AgNPs embedded N-doped highly porous carbon derived from chitosan based hydrogel as 60 download by : skknchat@gmail.com Tran Minh Chien – Master Thesis References catalysts for the reduction of 4-nitrophenol, Composites Part B: Engineering, 2019, 173, 106950 72 Chellamani, K.; Vignesh Balaji, R.; Veerasubramanian, D., Quality evaluation methods for textile substrates based wound dressings, Indian Journal of Fibre and Textile Research, 2014, 4, 811-17 73 Cipitria, A.; Skelton, A.; Dargaville, T.; Dalton, P.; Hutmacher, D., Design, fabrication and characterization of PCL electrospun scaffolds—a review, Journal of Materials Chemistry, 2011, 21 (26), 9419-9453 74 Gonzalez, A C d O.; Costa, T F.; Andrade, Z d A.; Medrado, A., Wound healing - A literature review, An Bras Dermatol, 2016, 91 (5), 614-620 75 Dhivya, S.; Padma, V V.; Santhini, E., Wound dressings - a review, Biomedicine, 2015, (4), 22-22 76 Gu, S.-Y.; Wang, Z.-M.; Ren, J.; Zhang, C.-Y., Electrospinning of gelatin and gelatin/poly (l-lactide) blend and its characteristics for wound dressing, Materials Science and Engineering: C, 2009, 29 (6), 1822-1828 77 Miguel, S P.; Ribeiro, M P.; Coutinho, P.; Correia, I., Electrospun polycaprolactone/aloe vera_chitosan nanofibrous asymmetric membranes aimed for wound healing applications, Polymers, 2017, (5), 183 78 Yan, X.; He, B.; Liu, L.; Qu, G.; Shi, J.; Hu, L.; Jiang, G., Antibacterial mechanism of silver nanoparticles in Pseudomonas aeruginosa: proteomics approach, Metallomics, 2018, 10 (4), 557-564 79 Yin, I X.; Zhang, J.; Zhao, I S.; Mei, M L.; Li, Q.; Chu, C H., The Antibacterial Mechanism of Silver Nanoparticles and Its Application in Dentistry, International Journal of Nanomedicine, 2020, 15, 2555 80 Hiep, N T.; Khon, H C.; Niem, V V T.; Toi, V V.; Ngoc Quyen, T.; Hai, N D.; Ngoc Tuan Anh, M., Microwave-Assisted Synthesis of Chitosan/Polyvinyl Alcohol Silver Nanoparticles Gel for Wound Dressing Applications International Journal of Polymer Science, 2016, 1584046 81 Raza, M A.; Kanwal, Z.; Rauf, A.; Sabri, A N.; Riaz, S.; Naseem, S., Sizeand shape-dependent antibacterial studies of silver nanoparticles synthesized by wet chemical routes, Nanomaterials, 2016, (4), 74 82 Kharaghani, D.; Jo, Y K.; Khan, M Q.; Jeong, Y.; Cha, H J.; Kim, I S., Electrospun antibacterial polyacrylonitrile nanofiber membranes functionalized with 61 download by : skknchat@gmail.com Tran Minh Chien – Master Thesis References silver nanoparticles by a facile wetting method, European Polymer Journal, 2018, 108, 69-75 83 Shao, J.; Yu, N.; Kolwijck, E.; Wang, B.; Tan, K W.; Jansen, J A.; Walboomers, X F.; Yang, F., Biological evaluation of silver nanoparticles incorporated into chitosan-based membranes, Nanomedicine, 2017, 12 (22), 2771-2785 84 Ho, T T.-P.; Doan, V K.; Tran, N M.-P.; Nguyen, L K.-K.; Le, A N.-M.; Ho, M H.; Trinh, N.-T.; Van Vo, T.; Dai Tran, L.; Nguyen, T.-H., Fabrication of chitosan oligomer-coated electrospun polycaprolactone membrane for wound dressing application, Materials Science and Engineering: C, 2021, 120, 111724 85 Sarmadi, M., Advantages and disadvantages of plasma treatment of textile materials, 21st International Symposium on Plasma Chemistry (ISPC 21), 2013 86 Bryaskova, R.; Pencheva, D.; Nikolov, S.; Kantardjiev, T., Synthesis and comparative study on the antimicrobial activity of hybrid materials based on silver nanoparticles (AgNps) stabilized by polyvinylpyrrolidone (PVP), Journal of Chemical Biology, 2011, (4), 185-191 87 Zahedi, P.; Rezaeian, I.; Ranaei‐Siadat, S O.; Jafari, S H.; Supaphol, P., A review on wound dressings with an emphasis on electrospun nanofibrous polymeric bandages, Polymers for Advanced Technologies, 2010, 21 (2), 77-95 88 Thomas, S.; Uzun, M., Testing dressings and wound management materials, Advanced Textiles for Wound Care (Second Edition), Rajendran, S., Ed Woodhead Publishing: 2019; pp 23-54 89 Chellamani, K.; Balaji, R V.; Veerasubramanian, D., Quality evaluation methods for textile substrates based wound dressings, IJETAE, 2014, 4, 811-17 90 Thomas, S.; Uzun, M., Testing dressings and wound management materials, Advanced Textiles for Wound Care, Elsevier: 2019; pp 23-54 91 Xu, R.; Xia, H.; He, W.; Li, Z.; Zhao, J.; Liu, B.; Wang, Y.; Lei, Q.; Kong, Y.; Bai, Y., Controlled water vapor transmission rate promotes wound-healing via wound re-epithelialization and contraction enhancement, Scientific reports, 2016, (1), 1-12 92 Mogrovejo-Valdivia, A.; Rahmouni, O.; Tabary, N.; Maton, M.; Neut, C.; Martel, B.; Blanchemain, N., In vitro evaluation of drug release and antibacterial activity of a silver-loaded wound dressing coated with a multilayer system, International Journal of Pharmaceutics, 2019, 556, 301-310 62 download by : skknchat@gmail.com Tran Minh Chien – Master Thesis 93 References Wolcott, R D.; Rhoads, D D.; Dowd, S E., Biofilms and chronic wound inflammation, Journal of wound care, 2008, 17 (8), 333-341 94 Ather, S.; Harding, K.; Tate, S., Wound management and dressings, Advanced textiles for wound care, Elsevier: 2019; pp 1-22 95 Tang, S.; Zheng, J., Antibacterial Activity of Silver Nanoparticles: Structural Effects, Advanced healthcare materials, 2018, (13), e1701503 96 Chung, Y.-C.; Su, Y P.; Chen, C.-C.; Jia, G.; Wang, H L.; Wu, J G.; Lin, J G., Relationship between antibacterial activity of chitosan and surface characteristics of cell wall, Acta pharmacologica sinica, 2004, 25 (7), 932-936 97 Kim, S.-K.; Rajapakse, N., Enzymatic production and biological activities of chitosan oligosaccharides (COS): A review, Carbohydrate Polymers, 2005, 62 (4), 357368 98 Huang, M.; Khor, E.; Lim, L.-Y., Uptake and cytotoxicity of chitosan molecules and nanoparticles: effects of molecular weight and degree of deacetylation, Pharmaceutical Research, 2004, 21 (2), 344-353 99 Fischer, D.; Li, Y.; Ahlemeyer, B.; Krieglstein, J.; Kissel, T., In vitro cytotoxicity testing of polycations: influence of polymer structure on cell viability and hemolysis, Biomaterials, 2003, 24 (7), 1121-1131 63 download by : skknchat@gmail.com Tran Minh Chien – Master Thesis Appendix APPENDIX Figure 1A: EDS mapping of C, O, and Ag elements in meshes (a) PCL-Ag 250 ppm, (b) PCL-Ag 500 ppm and, (c) PCL-Ag 1000 ppm (d) Ag element ratio analysis Scale bar: µm Table 1A: Concentration of residual acetone in raw PCL and PCL-Ag membrane Sample Acetone concentration (ppm) PCL-Ag 250 ppm 1647.5±50.5 PCL-Ag 500 ppm 511.9±20.5 PCL-Ag 1000 ppm 144.5±7.5 Raw PCL (12%PCL-AC) 76.7±4.2 64 download by : skknchat@gmail.com Tran Minh Chien – Master Thesis Appendix Figure 2A: COS release profile from the PCL-Ag-COS membrane in PBS solution (pH=5.5) Aliquots were taken after 5, 10, 15, 20, 40, 60 and 180 minutes, and quantified by Florescence absorbance technique at the wavelength of excitation at 420nm and that of emission at 460nm using microplate reader (Varioskan LUX, ThermoScientific) (data = mean ± SD, n=3) 65 download by : skknchat@gmail.com ... are attractive because of their high effectiveness against bacterial growth with a broad bactericidal spectrum [4] Even though the antibacterial mechanisms of SNPs have yet to be fully elucidated... quantified by ICP-MS technique (data = mean ± SD, n=3) 3.3.7 Antibacterial activity The antibacterial activities of the PCL-Ag-COS against two bacterial strains P aeruginosa and S aureus were investigated... measured zone diameters (Scale bar: 10 mm, data = mean ± SD, n = 4, ns: p > 0.05, *: p < 0.05) 50 Figure 3.17 Viability (%) of fibroblasts after 24h of incubation in different concentrations