一種以絲膠蛋白與聚乙烯醇為基礎的多孔性泡綿製備與其生物醫學應用

簡易檢索 / 詳目顯示

回結果列表

研究生：	冼祖怡 Joey - Sin
論文名稱：	一種以絲膠蛋白與聚乙烯醇為基礎的多孔性泡綿製備與其生物醫學應用 A porous foam made of silk sericin combining with poly(vinyl alcohol) as a dressing for potential use in biomedical engineering
指導教授：	白孟宜 Meng-Yi Bai
口試委員:	許維君 Wei-Chun Hsu 李嘉甄 Chia-Chen Li
學位類別：	碩士 Master
系所名稱：	應用科技學院 - 醫學工程研究所 Graduate Institute of Biomedical Engineering
論文出版年：	2017
畢業學年度：	105
語文別：	中文
論文頁數：	79
中文關鍵詞：	絲膠蛋白、聚乙烯醇、多孔性泡綿
外文關鍵詞：	Silk sericin, Poly(vinyl alcohol), porous foam
相關次數：	點閱：402 下載：0
分享至:	分享至facebook 分享至twitter

查詢本校圖書館目錄查詢臺灣博碩士論文知識加值系統勘誤回報

在蠶絲純化之過程中，絲膠蛋白通常會當作廢棄物丟棄，然而絲膠蛋白是可回收及利用應用於醫療、化妝品及製藥上。在此研究中，利用冷凍真空乾燥之技術製備一新型的 Sercin/PVA 泡綿。將絲膠絲膠蛋白利用脫膠法將絲膠蛋白從蠶繭中分離，經過計算平均脫膠率為 28.75 2.09%。Sericin/PVA 泡綿之微觀結構透過掃描式電子顯微鏡(SEM)觀察。結果顯示 Sericin/PVA 泡綿具有多孔性結構以及孔隙率約為 50%。材料特性經過傅立葉轉換紅外線光譜進行鑑定。我們發現經過冷凍乾燥程序所製備的 Sericin/PVA 泡綿中的材料特性不會有所改變。 Sericin/PVA 泡綿的透氣率約為 916.109 70.254 g/m2。在生物毒性測試中， Sericin/PVA 泡綿的細胞活性明顯高於控制組，表示 Sericin/PVA 泡綿有良好的生物相容性以及增加細胞增生的能力。因在生物毒性測試中，已證明 Sericin/PVA 泡綿具有細胞增生之能力，故於細胞抗發炎測試中，測試結果相較於控制及 LPS 組別的發炎反應來得強，因此現階段對於 Sericin/PVA 泡綿抗發炎測試結果仍持保留態度，有待進一步釐清。經由以上一系列實驗了解所製備的 Sericin/PVA 泡綿未來具有做為的傷口敷料的潛力應用於皮膚組織再生。

Silk sericin derived form silkworm Bombyx Mori was normally discarded during raw silk processing, sericin can be recovered and reused for other applications, like, medical device, cosmetics product, and pharmaceutical agent. In this study, a novel silk sericin combining with poly(vinyl alcohol) (PVA) was developed by freeze drying to generate a foam. Sericin were harvested from degumming processes of cocoon. The average degummed rate was 28.75 2.09%. The morphology of sericin/PVA foam was investigate by using scanning electron microscope (SEM). The result showed that sericin/PVA foam had porous structure and its porosity was about 52.793 8.347%. The surface characteristic was detected by Fourier transform infrared spectroscopy (FT-IR). We found that after freeze drying processes the material surface properties revealed hydrophilic amide I and amide II functional groups. The moisture vapor transmission rate (MVTR) of the sericin/PVA foam was determined to be 916.109 70.254 g/m2. In the cytotoxicity test, the cell viability of sericin/PVA foam was approximately 80~100% than control group. Indicate that sericin/PVA foam had good biocompatibility. As the sericin has been shown to have the ability of promoting cell proliferation, the relevant anti-inflammatory test is still needed to make clear at this stage. In conclusion, sericin/PVA foam will to be a potential material for wound management.

摘要 .............................................................................................................................. I 
Abstract ....................................................................................................................... II 
全名與縮寫對照表 ......................................................................................................... III 
致謝 ............................................................................................................................. IV 
圖目錄 ......................................................................................................................... VIII 
表目錄 .......................................................................................................................... IX
第一章 緒論 .................................................................................................................... 1
1 前言 .......................................................................................................................... 1 
2 實驗動機與目的 ........................................................................................................ 2
3 實驗設計與規劃 ........................................................................................................ 2
第二章 文獻回顧 ............................................................................................................ 4
1 皮膚 ......................................................................................................................... 4
1.1 皮膚構造與功能...................................................................................................... 4
1.2 傷口癒合機制.......................................................................................................... 7
2 敷料 .......................................................................................................................... 8
2.1 敷料種料 ................................................................................................................. 8 
2.2 敷料結構 ................................................................................................................ 10
2.3 敷料功能性 ............................................................................................................. 11
3 蠶繭 .......................................................................................................................... 12
3.1 蠶繭結構 ................................................................................................................. 12
3.2 絲膠蛋白 ................................................................................................................. 13
3.3 絲膠生醫上的應用.................................................................................................... 15
4 冷凍真空乾燥原理 .......................................................................................................16
5 聚乙烯醇(Poly(vinyl alcohol), PVA) .............................................................................17
第三章 材料與方法 ........................................................................................................... 18 
1 實驗材料 .....................................................................................................................18
2 實驗器材與儀器 ..........................................................................................................20
3 蠶繭脫膠萃取絲膠蛋白方法 ........................................................................................22
3.1 絲膠蛋白脫膠方式.................................................................................................... 22 
4 冷凍乾燥技術製備 Sericin/PVA 泡綿 ...........................................................................23 
4.1 製備絲膠蛋白泡綿.................................................................................................... 23 
4.2 製備 Sericin/PVA 泡綿............................................................................................. 24
5 透氣率測試 .................................................................................................................25 
6 體外細胞實驗...............................................................................................................25 
6.1 細胞種類 ................................................................................................................. 25
6.2 細胞培養基配置....................................................................................................... 26 
6.3 細胞培養 ................................................................................................................ 28
6.4 細胞毒性測試(Cytotoxicity model based on fibroblast cell).................................... 32
6.5 細胞抗發炎測試(LPS-induced macrophage model) ................................................ 34 
7 統計方式..................................................................................................................... 36
第四章 結果與討論 ........................................................................................................... 37 
1 絲膠蛋白脫膠率 ...........................................................................................................37
2 Sericin/PVA 泡綿表面型態...........................................................................................38
2.1 孔隙率分析 .............................................................................................................. 39
3 傅立葉轉換紅外線光譜(FT-IR)分析泡綿表面化學組成 ..................................................40
4 透氣率測試結果 ...........................................................................................................41
5 纖維母細胞之毒性測試 ................................................................................................42
6 抗發炎及誘導發炎之能力測試 ......................................................................................43
第五章 結論 ...................................................................................................................... 44 
第六章 未來展望................................................................................................................ 45
第七章 參考文獻................................................................................................................ 47
                                

1. 吳芳禎, 毛文霖, and 施英隆, 蠶絲絲膠蛋白之純化與應用研究. 科學與工程技術期刊, 2016.12(1): p. 21-30.
2. Zhang, Y.-Q., Applications of natural silk protein sericin in biomaterials. Biotechnology Advances, 2002. 20: p. 91-100.
3. Teramoto, H., T. Kameda, and Y. Tamada, Preparation of gel film from Bombyx mori silk sericin and its characterization as a wound dressing. Biosci Biotechnol Biochem, 2008.72(12): p. 3189-96.
4. Honari, G. and H. Maibach, Skin Structure and Function. 2014: p. 1-10.
5. Maryam K. Mamou, B., RN, CWOCN; Michael Jay Katz, MD, PhD, Wound Care. 2016.
6. Barbieri, J.S., K. Wanat, and J. Seykora, Skin: Basic Structure and Function.2014: p. 1134-1144.
7. Weinstein, G.D. and E.J. van Scott, Autoradiographic Analysis of Turnover Times of Normal and Psoriatic Epidermis. Journal of Investigative Dermatology, 1965. 45(4): p. 257-262.
8. BERGSTRESSER, P.R., and J. RICHARD TAYLOR., Epidermal ‘turnover time’—a new examination. British Journal of Dermatology, 1977. 96(5): p.503-506.
9. Ather, S. and K. Hargding, Wound management and dressings. Advanced Textiles for Wound Care, 2009: p. 3-19.
10. 靳燕芬, 敷料與傷口換藥疼痛. 護理雜誌, 2006. 53(6): p. 73-77.
11. Boateng, J.S., et al., Wound healing dressings and drug delivery systems: a review. J Pharm Sci, 2008. 97(8): p. 2892-923.
12. Qin, Y., Functional wound dressings. Medical Textile Materials, 2016: p.89-107.
13. Dhivya, S., V.V. Padma, and E. Santhini, Wound dressings - a review. Biomedicine (Taipei), 2015. 5(4): p. 22.
14. Jones, V., J.E. Grey, and K.G. Harding, Wound dressings. British Medical Journal, 2006. 7544: p. 777-780.
15. Lou, C.W., et al., Properties Evaluation of Tencel/Cotton Nonwoven Fabric Coated with Chitosan for Wound Dressing. Textile Research Journal, 2008. 78(3): p. 248-253.
16. Woo, H.-D., et al., Preparation of UV-curable gelatin derivatives for drug immobilization on polyurethane foam: Development of wound dressing foam. Macromolecular Research, 2015. 23(11): p. 994-1003.
17. Zehrer, C.L., et al., A comparison of the in vitro moisture vapour transmission rate and in vivo fluid-handling capacity of six adhesive foam dressings to a newly reformulated adhesive foam dressing. Int Wound J, 2014. 11(6): p. 681-90.
18. Mandal, A. and D. Chakrabarty, Characterization of nanocellulose reinforced semi-interpenetrating polymer network of poly(vinyl alcohol) & polyacrylamide composite films. Carbohydr Polym, 2015. 134: p. 240-50.
19. Singh, D. and R. Singh, Papain incorporated chitin dressings for wound debridement sterilized by gamma radiation. Radiation Physics and Chemistry, 2012. 81(11): p. 1781-1785.
20. K.P. Chellamani, R.S.V.B., D. Veerasubramanian and J. Sudharsan, Wound Healing Ability of Herbal Drug Incorporated PCL(Poly(ε-caprolactone)) Wound Dressing. Journal of Academia and Industrial Research, 2014. 2(11): p. 622-626.
21. Padamwar, M.N. and A.P. Pawar, Silk sericin and its applications: A review. Journal of Scientific & Industrial Research, 2004. 63: p. 323-329.
22. Omenetto, F.G. and D.L. Kaplan, New opportunities for an ancient material. Science, 2010. 329(5991): p. 528-31.
23. Altman, G.H., et al., Silk-based biomaterials. Biomaterials, 2003. 24(3): p. 401-416.
24. Gulrajani, M.L., Degumming of silk. Review of Progress in Coloration and Related Topics, 1992. 22(1): p. 79-89.
25. Chopra, S., and M. L. Gulrajani, Comarative evaluation of the various methods of degumming silk. Indian Journal of Fibre and Textile Research, 1994. 19: p. 76-83.
26. Jiang, P., et al., Tensile behavior and morphology of differently degummed silkworm (Bombyx mori) cocoon silk fibres. Materials Letters, 2006. 60(7): p. 919-925.
27. Aramwit, P., et al., The effect of sericin from various extraction methods on cell viability and collagen production. Int J Mol Sci, 2010. 11(5): p. 2200-11.
28. Gulrajani, M.L., Agarwal, R., Grover, A., & Suri, M., Degumming of silk with lipase and protease. Journal of Fibre and Textile Research, 2000. 25(1): p. 69-74.
29. Khan, M.R., et al., Physical properties and dyeability of silk fibers degummed with citric acid. Bioresour Technol, 2010. 101(21): p. 8439-45.
30. Freddi, G., R. Mossotti, and R. Innocenti, Degumming of silk fabric with several proteases. Journal of Biotechnology, 2003. 106(1): p. 101-112.
31. Rockwood, D.N., et al., Materials fabrication from Bombyx mori silk fibroin. Nat Protoc, 2011. 6(10): p. 1612-31.
32. Nagura, M., et al., Structures and physical properties of cross-linked sericin membranes. Journal of Insect Biotechnology and Sericology, 2001. 70(2): p. 149-153.
33. Kato, N., et al., Silk Protein, Sericin, Inhibits Lipid Peroxidation and Tyrosinase Activity. Bioscience, Biotechnology, and Biochemistry, 2014. 62(1): p. 145-147.
34. Zhaorigetu, S., et al., Inhibitory effects of silk protein, sericin on UVB-induced acute damage and tumor promotion by reducing oxidative stress in the skin of hairless mouse. Journal of Photochemistry and Photobiology B: Biology, 2003. 71(1-3): p. 11-17.
35. Aramwit, P. and A. Sangcakul, The Effects of Sericin Cream on Wound Healing in Rats. Bioscience, Biotechnology, and Biochemistry, 2014. 71(10): p. 2473-2477.
36. Kweon, H.-Y. and C.-S. Cho, Biomedical applications of silk protein. International Journal of Industrial Entomology, 2001. 3(1): p. 1-6.
37. Tao, W., M. Li, and R. Xie, Preparation and Structure of Porous Silk Sericin Materials. Macromolecular Materials and Engineering, 2005. 290(3): p.188-194.
38. Joshi, M., et al., Ecofriendly antimicrobial finishing of textiles using bioactive agents based on natural products. Indian journal of fibre and textile research, 2009. 34(3): p. 295-304.
39. Takahashi, M., et al., A sericin-derived peptide protects sf9 insect cells from death caused by acute serum deprivation. Biotechnol Lett, 2005. 27(13): p. 893-7.
40. Terada, S., et al., Preparation of silk protein sericin as mitogenic factor for better mammalian cell culture. J Biosci Bioeng, 2005. 100(6): p. 667-71.
41. KozoTsubouchi, Wound covering material. U.S. Patent No. 5,951,506., 1999.
42. Sarovart, S., et al., The use of sericin as an antioxidant and antimicrobial for polluted air treatment. Rev Adv Mater Sci, 2003. 5(3): p. 193-198.
43. 徐崇榮, et al., 冷凍真空乾燥. 科學發展, 2012. 473: p. 70-75.
44. Franks, F., Freeze-drying of bioproducts: putting principles into practice. European Journal of Pharmaceutics and Biopharmaceutics, 1998. 45(3): p. 221-229.
45. USP Pharmacists’ Pharmacopeia. 2009.
46. Liang, J., et al., Molecular-Level Dispersion of Graphene into Poly(vinyl alcohol) and Effective Reinforcement of their Nanocomposites. Advanced Functional Materials, 2009. 19(14): p. 2297-2302.
47. Ino, J.M., et al., Plasma functionalization of poly(vinyl alcohol) hydrogel for
cell adhesion enhancement. Biomatter, 2013. 3(4).
48. Cutiongco, M.F., et al., In vitro and ex vivo hemocompatibility of off-the-shelf modified poly(vinyl alcohol) vascular grafts. Acta Biomater, 2015. 25: p. 97-108.
49. van Meerloo, J., G.J. Kaspers, and J. Cloos, Cell sensitivity assays: the MTT assay. Methods Mol Biol, 2011. 731: p. 237-45.
50. Sargent, J.M. and C.G. Taylor, Appraisal of the MTT assay as a rapid test of chemosensitivity in acute myeloid leukaemia. British journal of cancer, 1989. 60(2): p. 206-210.
51. Green, S.J., Mellouk, S., Hoffman, S. L., Meltzer, M. S., & Nacy, C. A., Cellular mechanisms of nonspecific immunity to intracellular infection- Cytokine-induced synthesis of toxic nitrogen oxides from l-arginine by macrophages and hepatocytes. Immunology letters, 1990. 25(1): p. 15-19.
52. 邱清裕, 內毒素特性與檢測方法之簡介. 腎臟與透析, 2008. 20(3).
53. Todar, K., Mechanisms of bacterial pathogenicity: endotoxins. . Todar's Online Textbook of Bacteriology. Madison: University of Wisconsin,Department of Bacteriology, 2008. 8.

全文公開日期 2022/02/14 (校內網路)
全文公開日期本全文未授權公開 (校外網路)
全文公開日期本全文未授權公開 (國家圖書館：臺灣博碩士論文系統)

簡易檢索 / 詳目顯示

相關論文