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論文中文名稱:電紡 collagen/chitosan/PVA 奈米纖維支架為傷口敷料之研究 [以論文名稱查詢館藏系統]
論文英文名稱:Electrospun Collagen/Chitosan/PVA Nanofibers Scaffolds for Wound dressing [以論文名稱查詢館藏系統]
院校名稱:臺北科技大學
學院名稱:工程學院
系所名稱:化學工程研究所
畢業學年度:99
出版年度:100
中文姓名:郭育仁
英文姓名:Yu-Jen Kuo
研究生學號:98738036
學位類別:碩士
語文別:中文
口試日期:2011-07-25
論文頁數:56
指導教授中文名:林忻怡
口試委員中文名:黃聲東;王大銘
中文關鍵詞:電紡奈米纖維幾丁聚醣膠原蛋白戊二醛
英文關鍵詞:electrospunnanofiberschitosancollagenglutaraldehyde
論文中文摘要:組織工程的重點在於細胞、支架、生長訊息。篇著重於支架的製造,支架要可以模仿天然細胞外間質,且要具有大量的孔隙,主要利用天然高分子及混合人工高分子,並利用電紡絲的方法,來製造出具有大量表面積及孔隙的結構,以模仿天然的細胞外間質。材料我們選擇膠原蛋白、幾丁聚醣及聚乙烯醇,皆是生物可分解材料,具有很好的生物相容性,並利用濃醋酸(90vol%)當成溶劑,因為可以有效降低表面張力,也可以增加電荷密度,以電紡絲的方式成奈米纖維,再藉由戊二醛蒸氣交聯,使機械性質提升,並用掃描式電子顯微鏡觀察表
面型態、傅立葉轉換紅外線光譜得知交聯後材料特徵吸收峰都還在,且三材料間可能會產生複雜的分子間吸以力、流變儀觀察流體的黏度、膨潤率、拉伸側詴測量其機械性質。且使用 MTT 測量細胞活性、並定量 DNA,測詴細胞與支架的生物相容性;細胞的結果顯示支架可能具有細胞毒性,此部分的清洗步驟可能還需更改。
論文英文摘要:Tissue engineering includes three major elements: cells, scaffolds, and signal molecules. We are focuses on how to prepare the mimic extracellular matrices whose components and porous structures are analogous to those of the native extracellular matrices (ECMs). Electrospun nanofibers have amazing characteristics such as very large surface area-to-volume ratio and high porosity with very small pore size. The process was an efficient technique for the fabricationof both natural and synthetic polymers nanofibers and various polymer have been electrospun. Collagen, chitosan, polyvinyl alcohol(PVA) are biodegradable, biocompatible materials. We using
concentrated acetic acid solution(90%) as a solvent. Because more concentrated acetic acid in water progressively decreased surface tension of the chitosan solution and concomitantly increased charge density of jet. Electrospun collagen/chitosan/PVA nanofibers were crosslinked by glutaraldehyde(GTA) vapor to enhanced stability. The morphology of the electrospun collagen/chitosan/PVA nanofiber scaffolds was observed by scanning electron microscopy (SEM) and stabilized by glutaraldehyde (GTA) vapor via crosslinking. Fourier transform infrared spectra analysis showed that the collagen/chitosan/PVA nanofibers scaffold do not change significantly, except for enhanced stability after crosslinking by GTA vapor. The thermal behavior was studied by thermal gravimetric analysis and the mechanical properties were Studied by tensile testing. To assay the biocompatibility of electrospun scaffolds, cellular behavior on the nanofibrous scaffolds was also investigated by SEM, methylthiazol tetrazolium testing(MTT) and dsDNA content. The cyototoxic effecy of GA-vapor crosslinked scaffolds was observed. The scaffolds show almosted no cellular activity.
論文目次:摘要 .................................................... I
ABSTRACT ............................................... II
誌謝 ...................................................III
圖目錄 ..................................................VI
表目錄................................................. VII
1.1 前言 .............................................- 1 -
1.2 人工敷料介紹 .....................................- 2 -
1.2.1 皮膚簡介 ........................................- 2 -
1.2.2 人工敷料 ........................................- 2 -
1.3 靜電紡絲原理(electrospun) .........................................- 3 -
1.3.1 奈米纖維製備方法 ................................- 3 -
1.3.2 電紡絲(Electrospinning)介紹 .....................- 3 -
1.3.3 影響電紡絲結果因素 ..............................- 5 -
1.4 幾丁聚醣(chitosan) ...............................- 7 -
1.5 第一型膠原蛋白(type Ⅰ collagen) ...............- 8 -
1.6 聚乙烯醇(Polyvinyl Alcohol, PVA)介紹 ..............- 9 -
1.7 交聯處理(cross-linking) ..........................- 9 -
1.8 研究動機與目的 ..................................- 10 -
第二章 實驗藥品、儀器及方法 .........................- 11 -
2.1 實驗藥品 ...................................................- 11 -
2.1.1 細胞來源 ......................................- 11 -
v

2.1.2 細胞培養所用藥品及配置 ........................- 11 -
2.1.3 電紡溶液與實驗所需藥品 ........................- 12 -
2.2 實驗儀器 .........................................- 14 -
2.3 實驗方法 ........................................- 16 -
2.3.1 實驗架構 .......................................- 16 -
2.3.2 電紡溶液製備 ..................................- 18 -
2.3.3 電紡架構及參數設定 ............................- 18 -
2.3.4 交聯處理(crosslinking)........................................- 19 -
2.3.5 物理性質測詴 ...................................- 19 -
2.3.6 細胞測詴 ......................................- 21 -
2.3.7 統計分析 ...................................................- 23 -
第三章 實驗結果與討論 ...............................- 24 -
3.1 奈米纖維製備 .....................................- 24 -
3.2 物理性質 ...................................................- 25 -
3.2.1 奈米纖維直徑分佈 ..........................- 25 -
3.2.2 PBS 中降解 SEM 圖 ...................................................- 26 -
3.2.3 傅立葉轉換紅外線光譜 ..........................- 29 -
3.2.4 黏度............................................- 35 -
3.2.5 膨潤率 ........................................- 37 -
3.2.6 拉伸測詴 ......................................- 38 -
3.3 細胞測詴 .........................................- 40 -
3.3.1 細胞活性(MTT assay) ...............................................- 40 -
3.3.2 DNA 含量 ...................................................- 41 -
3.3.3 掃描式電子顯微鏡(SEM) .........................- 43 -
3.4 旋轉收集器 ......................................- 44 -
第四章 結論 .........................................- 49 -
參考文獻 .............................................- 50 -
附錄 .................................................- 56 -
論文參考文獻:A., Formhals (1934). Process and apparatus for preparing artificial threads. U. Patent: 1975504.
Asran, Ashraf Sh, Henning, S. and Michler, Goerg H. (2010). "Polyvinyl alcohol–collagen–hydroxyapatite biocomposite nanofibrous scaffold: Mimicking the key features of natural bone at the nanoscale level." Polymer 51(4): 868-876.
Balasubramani, Manimalha, Kumar, T. Ravi and Babu, Mary (2001). "Skin substitutes: a review." Burns 27: 534-544.
Beele, H. (2002). "Artificial skin: past, present and future." Int J Artif Organs 25(3): 163-173.
Bhattarai, S. R., Bhattarai, N., Yi, H. K., Hwang, P. H., Cha, D. I. and Kim, H. Y. (2004). "Novel biodegradable electrospun membrane: scaffold for tissue engineering." Biomaterials 25(13): 2595-2602.
Buchko, C.J., Chen, L.C., Shen, Y. and Martin, D.C. (1999). "Processing and microstructural characterization of porous biocompatible protein polymer thin films." Polymer 40(26): 7397-7407.
Chen, J. and Chang, G. (2008). "Electrospun collagen/chitosan nanofibrous membrane as wound dressing." Colloids and Surfaces A: Physicochemical and Engineering Aspects 313-314: 183-188.
Chen, Sung-Ching, Wu, Yung-Chih, Mi, Fwu-Long, Lin, Yu-Hsin, Yu, Lin-Chien and Sung, Hsing-Wen (2004). "A novel pH-sensitive hydrogel composed of N,O-carboxymethyl chitosan and alginate cross-linked by genipin for protein drug delivery." Journal of Controlled Release 96(2): 285-300.
Deitzel, J.M., Kleinmeyer, J., D. Harris, N.C. and Tan, Beck (2001). "The effect of processing variables on the morphology of electrospun
nanofibers and textiles." Polymer 42(1): 261-272.
Dong, Bin, Arnoult, Olivier, Smith, Meghan E. and Wnek, Gary E. (2009). "Electrospinning of Collagen Nanofiber Scaffolds from Benign Solvents." Macromolecular Rapid Communications 30(7): 539-542.
Duan, B., Dong, C., Yuan, X. and Yao, K. (2004). "Electrospinning of chitosan solutions in acetic acid with poly(ethylene oxide)." J Biomater Sci Polym Ed 15(6): 797-811.
Feng, Lin, Li, Shuhong, Li, Huanjun, Zhai, Jin, Song, Yanlin, Jiang, Lei and Zhu, Daoben (2002). "Super-Hydrophobic Surface of Aligned Polyacrylonitrile Nanofibers " Angewandte Chemie 114(7): 1269-1271.
Fong, H., Chun, I. and Reneker, D.H. (1999). "Beaded nanofibers formed during electrospinning." Polymer 40: 4585-4592.
Geng, X., Kwon, O. and Jang, J. (2005). "Electrospinning of chitosan dissolved in concentrated acetic acid solution." Biomaterials 26(27): 5427-5432.
Hartwell, R., Leung, V., Chavez-Munoz, C., Nabai, L., Yang, H., Ko, F. and Ghahary, A. (2011). "A novel hydrogel-collagen composite improves functionality of an injectable extracellular matrix." Acta Biomaterialia 7(8): 3060-3069.
Howling, G. I., Dettmar, P. W., Goddard, P. A., Hampson, F. C., Dornish, M. and Wood, E. J. (2001). "The effect of chitin and chitosan on the proliferation of human skin fibroblasts and keratinocytes in vitro." Biomaterials 22(22): 2959-2966.
Hsieh, Anne, Zahir, Tasneem, Lapitsky, Yakov, Amsden, Brian, Wan, Wankei and Shoichet, Molly S. (2010). "Hydrogel/electrospun fiber composites influence neural stem/progenitor cell fate." Soft Matter 6(10): 2227.
Huang, Chen, Chen, Rui, Ke, Qinfei, Morsi, Yosry, Zhang, Kuihua and Mo, Xiumei (2011). "Electrospun collagen–chitosan–TPU nanofibrous scaffolds for tissue engineered tubular grafts." Colloids and Surfaces B: Biointerfaces 82(2): 307-315.
Hutmacher, D. W., Goh, J. C. and Teoh, S. H. (2001). "An introduction to biodegradable materials for tissue engineering applications." Ann Acad Med Singapore 30(2): 183-191.
Itoh, Soichiro, Takakuda, Kazuo, Kawabata, Sigenori, Aso, Yu, Kasai, Kanae, Itoh, Hiroshi and Shinomiya, Kenichi (2001). "Evaluation of cross-linking procedures of collagen tubes used in peripheral nerve repair." Biomaterials 23: 4475-4481.
Jiao, Yanpeng, Liu, Zonghua and Zhou, Changren (2007). "Fabrication and characterization of PLLA–chitosan hybrid scaffolds with improved cell compatibility." Journal of Biomedical Materials Research Part A 80A(4): 820-825.
Khajavi, R. and Damerchely, R. (2007). "Effect of polyvinyl alcohol concentration in spinning dope on diameter, beads and HHS of produced nanofibers." Pak J Biol Sci 10(2): 314-317.
Khil, M. S., Cha, D. I., Kim, H. Y., Kim, I. S. and Bhattarai, N. (2003). "Electrospun nanofibrous polyurethane membrane as wound dressing." J Biomed Mater Res B Appl Biomater 67(2): 675-679.
Kim, Kwan Woo, Lee, Keun Hyung, Khil, Myung Seob, Ho, Yo Seung and Kim, Hak Yong (2004). "The effect of molecular weight and the linear velocity of drum surface on the properties of electrospun poly(ethylene terephthalate) nonwovens." Fibers and Polymers 5: 122-127.
Klossner, Rebecca R., Queen, Hailey A., Coughlin, Andrew J. and Krause, Wendy E. (2008). "Correlation of Chitosan’s Rheological Properties and Its Ability to Electrospin." Biomacromolecules 9: 2947-2953.
Koski, A. (2004). "Effect of molecular weight on fibrous PVA produced by electrospinning." Materials Letters 58(3-4): 493-497.
Kriegel, C., Kit, K. M., McClements, D. J. and Weiss, J. (2009). "Nanofibers as carrier systems for antimicrobial microemulsions. Part I: fabrication and characterization." Langmuir 25(2): 1154-1161.
Lee, Joon Seok, Choi, Kyu Ha, Ghim, Han Do, Kim, Sam Soo, Chun, Du Hwan, Kim, Hak Yong and Lyoo, Won Seok (2004). "Role of molecular weight of atactic poly(vinyl alcohol) (PVA) in the structure and properties of PVA nanofabric prepared by electrospinning." Journal of Applied Polymer Science 93(4): 1638-1646.
Lin, S. Y., Chen, K. S. and Run-Chu, L. (2001). "Design and evaluation of drug-loaded wound dressing having thermoresponsive, adhesive, absorptive and easy peeling properties." Biomaterials 22(22): 2999-3004.
Liu, Shih-Jung, Kau, Yi-Chuan, Chou, Chi-Yin, Chen, Jan-Kan, Wu, Ren-Chin and Yeh, Wen-Ling (2010). "Electrospun PLGA collagen nanofibrous membrane as early-stage wound dressing." Journal of Membrane Science 355: 53-59.
Liu, Ting, Teng, Wai Keng, Chan, Barbara P. and Chew, Sing Yian (2010). "Photochemical crosslinked electrospun collagen nanofibers: Synthesis, characterization and neural stem cell interactions." Journal of Biomedical Materials Research Part A 95A(1): 276-282.
Liu, Xiao Fei, Guan, Yun Lin, Yang, Dong Zhi, Li, Zhi and Yao, Kang De (2000). "Antibacterial action of chitosan and carboxymethylated chitosan." Journal of Applied Polymer Science 79: 1324-1335.
Ma, L. (2003). "Collagen/chitosan porous scaffolds with improved biostability for skin tissue engineering." Biomaterials 24(26): 4833-4841.
Ma, P. X. and Zhang, R. (1999). "Synthetic nano-scale fibrous extracellular matrix." J Biomed Mater Res 46(1): 60-72.
Matthews, Jamil A., Wnek, Gary E., Simpson, David G. and Bowlin, Gary L. (2002). "Electrospinning of Collagen Nanofibers." Biomacromolecules 3(2): 232-238.
Megelski, Silke, Stephens, Jean S., Chase, D. Bruce and Rabolt, John F. (2002). "Micro- and Nanostructured Surface Morphology on Electrospun Polymer Fibers." Macromolecules 35: 8456-8466.
Moel, K. de, Ekenstein, G. O. R. Alberda van, Nijland, H., Polushkin, E. and Brinke, G. ten (2001). "Polymeric Nanofibers Prepared from Self-Organized Supramolecules." Chem. Mater. 13(12): 4580-4583.
Ondarcuhu, T. and Joachim, C. (1998). "Drawing a single nanofibre over hundreds of microns." Europhysics Letters 42(2): 215-220.
Ramakrishna, Seeram, Fujihara, Kazutoshi, Teo, Wee-Eong, Lim, Teik-Cheng and Ma, Zuwei, Eds. (2005). An introduction to Electrospinning and Nanofibers. Singapore, World Scientific.
Ramakrishna, Seeram, Fujihara, Kazutoshi, Wee-Eong, Lom, Teik-Cheng and Ma, Zuwei (2005). An introduction to Electrospinning and Nanofibers. Singapor, World Scientific.
Reneker, Darrell H and Chun, Iksoo (1996). "Nanometre diameter fibres of polymer, produced by electrospinning." Nanotechnology 7(3): 216-223.
Rho, K. S., Jeong, L., Lee, G., Seo, B. M., Park, Y. J., Hong, S. D., Roh, S., Cho, J. J., Park, W. H. and Min, B. M. (2006). "Electrospinning of collagen nanofibers: effects on the behavior of normal human keratinocytes and early-stage wound healing." Biomaterials 27(8): 1452-1461.
Sangsanoh, Pakakrong, Suwantong, Orawan, Neamnark, Artphop, Cheepsunthorn, Poonlarp, Pavasant, Prasit and Supaphol, Pitt (2010). "In vitro biocompatibility of electrospun and solvent-cast chitosan substrata towards Schwann, osteoblast, keratinocyte and fibroblast cells." European Polymer Journal 46(3): 428-440.
Sarti, B. and Scandola, M. (1995). "Viscoelastic and thermal properties of collagen/poly(vinyl alcohol) blends." Biomaterials 16(10): 785-792.
Shin, M., Yoshimoto, H. and Vacanti, J. P. (2004). "In vivo bone tissue engineering using mesenchymal stem cells on a novel electrospun nanofibrous scaffold." Tissue Eng 10(1-2): 33-41.
Sionkowska, Alina, Skopińska, Joanna and Wisniewski, Marcin (2004). "Photochemical stability of collagen/poly (vinyl alcohol) blends." Polymer Degradation and Stability 83(1): 117-125.
Sun, L. P., Wang, S., Zhang, Z. W., Wang, X. Y. and Zhang, Q. Q. (2009). "Biological evaluation of collagen–chitosan scaffolds for dermis tissue engineering." Biomedical Materials 4(5): 055008.
Sung, Hsing-Wen, Liang, I-Lin, Chen, Chiun-Nan, Huang, Rong-Nan and Liang, Hsiang-Fa (2001). "Stability of a biological tissue fixed with a naturally occurring crosslinking agent (genipin)." Journal of Biomedical Materials Research 55: 538-546.
Teo, W. E. and Ramakrishna, S. (2006). "A review on electrospinning design and nanofibre assemblies." Nanotechnology 17(14): R89-R106.
W., Graessley William (1980). "Polymer chain dimensions and the dependence of viscoelastic properties on concentration, molecular weight and solvent power." Polymer 21: 258.
Wang, T. W., Sun, J. S., Wu, H. C., Tsuang, Y. H., Wang, W. H. and Lin, F. H. (2006). "The effect of gelatin-chondroitin sulfate-hyaluronic acid skin substitute on wound healing in SCID mice." Biomaterials 27(33): 5689-5697.
Widmer, M. S., Gupta, P. K., Lu, L., Meszlenyi, R. K., Evans, G. R., Brandt, K., Savel, T., Gurlek, A., Patrick, C. W., Jr. and Mikos, A. G. (1998). "Manufacture of porous biodegradable polymer conduits by an extrusion process for guided tissue regeneration." Biomaterials 19(21): 1945-1955.
Yang, Ye, Zhu, Xinli, Cui, Wenguo, Li, Xiaohong and Jin, Yan (2009). "Electrospun Composite Mats of Poly[(D,L-lactide)-co-glycolide] and Collagen with High Porosity as Potential Scaffolds for Skin Tissue Engineering." Macromolecular Materials and Engineering 294(9): 611-619.
Zeugolis, Dimitrios I., Khew, Shih T., Yew, Elijah S. Y., Ekaputra, Andrew K., Tong, Yen W., Yung, Lin-Yue L., Hutmacher, Dietmar W., Sheppard, Colin and Raghunath, Michael (2008). "Electro-spinning of pure collagen nano-fibres – Just an expensive way to make gelatin?" Biomaterials 29(15): 2293-2305.
Zhang, C., Yuan, X., Wu, L., Han, Y. and Sheng, J. (2005). "Study on morphology of electrospun poly(vinyl alcohol) mats." European Polymer Journal 41(3): 423-432.
Zhang, Yuanyuan, Huang, Xiaobo, Duan, Bin, Wu, Lili, Li, Shuo and Yuan, Xiaoyan (2007). "Preparation of electrospun chitosan/poly(vinyl alcohol) membranes." Colloid and Polymer Science 285(8): 855-863.
Zhou, Yingshan, Yang, Dongzhi, Chen, Xiangmei, Xu, Qiang, Lu, Fengmin and Nie, Jun (2008). "Electrospun Water-Soluble Carboxyethyl Chitosan/Poly(vinyl alcohol) Nanofibrous Membrane as Potential Wound Dressing or Skin Regeneration." Biomacromolecules 9: 349-354.
Zong, Xinhua, Kim, Kwangsok, Fang, Dufei, Ran, Shaofeng, Hsiao, Benjamin S. and Chu, Benjamin (2002). "Structure and process relationship of electrospun bioabsorbable nanofiber membranes." Polymer 43(16): 4403-4412.
洪, 琇議 (2009). "靜電紡絲技術製備水溶性幾丁聚醣/聚乙烯醇奈米纖維薄膜於抗菌之應用." 國立台北科技大學 有機高分子研究所 碩士學位論文.
葉, 志宗 (2009). "以幾丁聚醣為基質製備應用於藥物釋放之組織工程多孔性支架." 國立台北科技大學 化學工程研究所 碩士學位論文.
劉士榮 (2005). 高分子流變學.
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