研究生: |
郭建宏 Chien-Hung Kuo |
---|---|
論文名稱: |
利用電漿沉積微結構圖形並探討對細胞生長之影響 Preparation of Micro-patterns by Plasma Polymerization and the Impacts on Cell Behaviors |
指導教授: |
王孟菊
Meng-Jiy Wang |
口試委員: |
陳克紹
Ko-Shao Chen 陳賜原 Szu-yuan Chen 何郡軒 Jinn-Hsuan Ho 林文賓 Wen-Pin Lin |
學位類別: |
碩士 Master |
系所名稱: |
工程學院 - 化學工程系 Department of Chemical Engineering |
論文出版年: | 2020 |
畢業學年度: | 108 |
語文別: | 中文 |
論文頁數: | 86 |
中文關鍵詞: | 真空電漿 、電漿聚合薄膜 、抗蛋白質貼附 、微米等級圖案 |
外文關鍵詞: | Low pressure plasma, Plasma polymerized thin films, Anti-fouling, Micro-scale pattern |
相關次數: | 點閱:248 下載:0 |
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近年來利用微米級圖案化建構精緻的智能生物材料,以改變微米級圖案,產生不同的生物信號及控制細胞行為,常應用於組織工程。因此本論文選用兩性雙離子型分子SBMA (sulfobetaine methacrylate) 及吡咯 (pyrrole, Py) 進行基材表面修飾。由於SBMA、及pyrrole分別具有良好的抗沾黏特性及生物相容性,可藉由微米級圖案之遮罩進行電漿沉積得到具有圖案且具選擇性抗沾黏特性之薄膜。
本論文第一部分,目標為在材料表面製備具有微米等級之細胞圖案,做法為選用光固化樹脂 (BV-007) 進行3D列印,製備多孔型遮罩,藉由遮罩掩蓋法以真空電漿進行沉積,得到具有微米等級圖案之材料表面。實驗結果顯示,遮罩之設計尺寸與實際沉積圖案之尺寸差異呈現規則的趨勢,圖案實際長度約為所設計之83%,實際寬度約為所設計之75%,實際厚度約為所設計之40%,而實際孔洞邊界約為所設計之150%,藉由此線性關係製作微米等級孔洞之遮罩。
本論文第二部分,為探討SBMA分別與丙烯酸 (acrylic acid, AAc) 及丙烯胺 (allylamine, AAm) 是否成功藉由自由基聚合法分別得到SBMA與AAc之聚合物 (poly(SBMA-co-AAc)) 及SBMA與AAm之聚合物 (poly(SBMA-co-AAm)),因此利用傅立葉轉換紅外線光譜儀 (FTIR)、液態超導核磁共振儀 (NMR)、及凝膠滲透層析儀 (GPC) 進行poly(SBMA-co-AAc)及poly(SBMA-co-AAm)之分析測定。FTIR結果顯示,poly(SBMA-co-AAc)具有SBMA、及AA之特徵峰,因此判斷成功聚合。由NMR結果顯示,poly(SBMA-co-AAc) 組成含有SBMA主鏈之亞甲基原子及AAc結構式中之質子,且分別之積分面積比例為61:39,因此判斷SBMA與AAc成功進行聚合反應。且由GPC分析結果顯示,poly(SBMA-co-AAc) 平均分子量為150 kDa,證明SBMA與AAc成功聚合poly(SBMA-co-AAc)。與 poly(SBMA-co-AAm) 之分析結果相比,SBMA與AAm藉由聚合法得到之聚合物為均聚物,而非共聚產物,因此判斷SBMA與AAm不適用於自由基聚合法進行共聚反應。
最後,探討L-929老鼠纖維母細胞及間充質幹細胞 (mesenchymal stem cell, MSC),在微米等級的生長環境之最適化參數。利用(3-aminopropyl) triethoxysilane (APTES) 將製備之高分子聚合物修飾於聚苯乙烯培養皿 (TCPS) 表面,再以pyrrole作為前驅物進行電漿沉積,當沉積時間為5至30分鐘,薄膜厚度分別從35.2上升至161.7 nm,且薄膜厚度有效地改變圖案範圍內之細胞型態。在施加功率20 W、壓力120 mTorr之電漿參數設計下,由L-929於圖案上之細胞型態結果顯示,當圖案尺寸為600 μm x 600 μm,不論圖案之間距為100 μm或150 μm,皆呈現明顯之細胞圖案,推測圖案間距並非影響 L-929 細胞圖案之參數;另一方面,當圖案之間距為100 μm,不論圖案尺寸為600 μm x 600 μm或是1200 μm x 600 μm,皆呈現明顯之細胞圖案,推測圖案尺寸亦非影響L-929細胞圖案之參數。由MSC於圖案上之細胞型態結果顯示,當圖案尺寸為600 μm x 600 μm,不論圖案之間距為100 μm或150 μm,於各沉積時間下皆沒有明顯之細胞圖案產生。另一方面,圖案尺寸為1200 μm x 600 μm、圖案之間距為100 μm,於各沉積時間皆呈現細胞圖案,因此證明影響MSC細胞圖案最重要之參數為圖案之尺寸。由水接觸角量測結果顯示,當沉積時間5至30分鐘,水接觸角分別從11.2o上升至36.9o,隨著沉積時間增加,水接觸角呈現上升的趨勢。最後MSC之貼附面積相較於L-929之貼附面積較大,故MSC需要較大的圖案面積方能呈現細胞圖案。藉由遮罩技術結合電漿沉積製造微米級圖案,於抗沾黏之材料表面,產生具有良好生物相容性之微米級圖案,具有應用於生醫材料之潛力。
In recent years, micro-scale patterning has been used to construct exquisite intelligent biomaterials. Micro-scale patterning can be applied in tissue engineering to generate different biological signals and control cell behavior. Therefore, the materials selected in this study are zwitterionic molecules of sulfobetaine methacrylate (SBMA) and pyrrole (Py). SBMA and pyrrole show excellent anti-fouling properties and good biocompatibility, therefore the thin films with the aforementioned selective anti-fouling properties will be acquired by plasma deposition through a mask with micro-scale patterns.
At the first part of the thesis, in order to produce micro-scale cell patterns on the surface of the material, UV initiated-monomers and oligomers (BV-007) was selected for 3D printing to prepare a porous mask. The deposition with a vacuum plasma was achieved by using a mask, finally to obtain a micro-scale patterned material. By observing the mask with optical microscope, the difference between the designed size of the mask and the actual size of the mask show a regular trend, in which the actual pattern length is about 83% of the designed pattern length. The actual pattern width is approximately 75% of the designed pattern width. The actual thickness is approximately 40% of the designed thickness. The actual bridge width is about 150% of the designed bridge width. Based on this linear relationship, a mask of micro-scale pores is made.
At the second part of the thesis, in order to explore whether the polymer can be successfully copolymerized by free radical polymerization method, poly(SBMA-co-AAc) and poly(SBMA-co-AAm) were analyzed by FTIR, NMR, and GPC analyses. FTIR analysis showed that poly(SBMA-co-AAc) only showed the characteristic peaks of SBMA, so it can only be inferred that SBMA successfully polymerized. NMR analysis showed that poly(SBMA-co-AAc) consists of the methylene atom of the SBMA main chain and the protons in the AAc structural formula. Therefore, it is inferred that SBMA and AAc successfully polymerized, and the integral area ratios were 61: 39. And GPC analysis showed that the average molecular weight of poly(SBMA-co-AAc) was 150 kDa, which proved that SBMA and AAc successfully polymerized poly(SBMA-co-AAc). Compared with the analysis results of poly(SBMA-co-AAm), the polymer obtained by the free radical polymerization method of SBMA and AAm was the homopolymer, rather than the copolymerized product. Therefore, it is inferred that SBMA and AAm are not suitable for copolymerization by free radical polymerization reaction.
For the third part of the thesis, in order to infer the optimal parameters of L-929 mouse fibroblast and mesenchymal stem cell in micro-scale growth environment. When the pyrrole deposition times increased from 5 to 30 minute, the film thicknesses increased from 35.2 to 161.7 nm. The results show that the thickness of the film effectively changes the cell type in the range of pattern. In this case, the parameter of plasma treatment applied in the experiment with the applied power of 20 W and the pressure of 120 mTorr. The results of the L-929 cell morphology on the pattern show that when the pattern size was 600 μm x 600 μm, no matter the distance between the patterns was 100 μm or 150 μm, clear cell patterns appeared. It is speculated that the distance between each pattern didn’t affect the L-929 cell pattern; On the other hand, when the distance between the patterns was 100 μm, no matter the pattern size was 600 μm x 600 μm or 1200 μm x 600 μm, the cell patterns appeared clearly. It is suggested that the pattern size didn’t affect the formation of the L-929 cell pattern. The results of cell morphology on the pattern by MSC showed that when the pattern size was 600 μm x 600 μm, there were no significant cell patterns were generated at each deposition time regardless of the pattern spacing between 100 μm or 150 μm. On the other hand, when changing the size of the pattern into 1200 μm x 600 μm, and the distance between the patterns is 100 μm, the cell patterns appeared at each deposition time. Therefore, it is proved that the most important parameter affecting the MSC cell pattern was the size of the pattern. The water contact angle results show that when the deposition time increased from 5 to 30 minute, the water contact angles increased from 11.2o to 36.9o. The increasing trend proves that the pyrrole film was affected by the
hydrophobicity of pyrrole. Finally, the attachment area of the MSC is larger than the area of the L-929, so the MSC need a larger pattern area to present the cell pattern. By using the masking technology combined with plasma deposition to create micro-scale patterns, micro-scale patterns with good biocompatibility can be generated on the surface of anti-fouling materials. The research prove that the masking technique combined with plasma deposition on the antifouling material with the potential to be applied in biomedical materials.
[1] A. Ohl, W. Schleinitz, A. Meyer-Sievers, A. Becker, D. Keller, K. Schröder, and J. Conrads, “Design of an UHV reactor system for plasma surface treatment of polymer materials,” Surface and Coatings Technology, vol. 116, pp. 1006-1010, 1999.
[2] P. K. Chu, J. Chen, L. Wang, and N. Huang, “Plasma-surface modification of biomaterials,” Materials Science and Engineering: R: Reports, vol. 36, no. 5-6, pp. 143-206, 2002.
[3] N. Brinkmann, D. Sommer, G. Micard, G. Hahn, and B. Terheiden, “Electrical, optical and structural investigation of plasma-enhanced chemical-vapor-deposited amorphous silicon oxynitride films for solar cell applications,” Solar Energy Materials and Solar Cells, vol. 108, pp. 180-188, 2013.
[4] X. Wang, and G. Grundmeier, “Surface analytical studies of Ar-plasma etching of thin heptadecafluoro-1-decene plasma polymer films,” Applied surface science, vol. 252, no. 23, pp. 8331-8336, 2006.
[5] S. Liu, M. M. Vareiro, S. Fraser, and A. T. A. Jenkins, “Control of attachment of bovine serum albumin to pulse plasma-polymerized maleic anhydride by variation of pulse conditions,” Langmuir, vol. 21, no. 19, pp. 8572-8575, 2005.
[6] S. Fleire, J. Goldman, Y. Carrasco, M. Weber, D. Bray, and F. Batista, “B cell ligand discrimination through a spreading and contraction response,” Science, vol. 312, no. 5774, pp. 738-741, 2006.
[7] B. Geiger, A. Bershadsky, R. Pankov, and K. M. Yamada, “Extracellular matrix–cytoskeleton crosstalk,” Nat Rev Mol Cell Biol, vol. 2, pp. 793-805, 2001.
[8] N. Wang, J. P. Butler, and D. E. Ingber, “Mechanotransduction across the cell surface and through the cytoskeleton,” Science, vol. 260, no. 5111, pp. 1124-1127, 1993.
[9] S. Bhatia, U. Balis, M. Yarmush, and M. Toner, “Microfabrication of hepatocyte/fibroblast co‐cultures: Role of homotypic cell interactions,” Biotechnology progress, vol. 14, no. 3, pp. 378-387, 1998.
[10] J. El-Ali, P. K. Sorger, and K. F. Jensen, “Cells on chips,” Nature, vol. 442, no. 7101, pp. 403, 2006.
[11] A. Folch, A. Ayon, O. Hurtado, M. Schmidt, and M. Toner, “Molding of deep polydimethylsiloxane microstructures for microfluidics and biological applications,” Journal of biomechanical engineering, vol. 121, no. 1, pp. 28-34, 1999.
[12] R. S. Kane, S. Takayama, E. Ostuni, D. E. Ingber, and G. M. Whitesides, "Patterning proteins and cells using soft lithography," The Biomaterials: Silver Jubilee Compendium, pp. 161-174: Elsevier, 1999.
[13] E. A. Roth, T. Xu, M. Das, C. Gregory, J. J. Hickman, and T. Boland, “Inkjet printing for high-throughput cell patterning,” Biomaterials, vol. 25, no. 17, pp. 3707-3715, 2004.
[14] R. J. Goldston, and P. H. Rutherford, Introduction to plasma physics: CRC Press, 1995.
[15] J. A. Bittencourt, Fundamentals of plasma physics: Springer Science & Business Media, 2013.
[16] R. Fitzpatrick, “Introduction to plasma physics,” The University of Texas at Austin: sn, pp. 242, 2008.
[17] P. Bourke, D. Ziuzina, D. Boehm, P. J. Cullen, and K. Keener, “The potential of cold plasma for safe and sustainable food production,” Trends in biotechnology, vol. 36, no. 6, pp. 615-626, 2018.
[18] J. W. Coburn, and H. F. Winters, “Plasma etching—A discussion of mechanisms,” Journal of vacuum Science and Technology, vol. 16, no. 2, pp. 391-403, 1979.
[19] T. S. Williams, H. Yu, and R. F. Hicks, “Atmospheric pressure plasma activation as a surface pre-treatment for the adhesive bonding of aluminum 2024,” Journal of Adhesion Science and Technology, vol. 28, no. 7, pp. 653-674, 2014.
[20] D. Ben Salem, O. Carton, H. Fakhouri, J. Pulpytel, and F. Arefi‐Khonsari, “Deposition of water stable plasma polymerized acrylic acid/MBA organic coatings by atmospheric pressure air plasma jet,” Plasma Processes and Polymers, vol. 11, no. 3, pp. 269-278, 2014.
[21] V. M. Donnelly, and A. Kornblit, “Plasma etching: Yesterday, today, and tomorrow,” Journal of Vacuum Science & Technology A: Vacuum, Surfaces, and Films, vol. 31, no. 5, pp. 050825, 2013.
[22] K. J. Kanarik, T. Lill, E. A. Hudson, S. Sriraman, S. Tan, J. Marks, V. Vahedi, and R. A. Gottscho, “Overview of atomic layer etching in the semiconductor industry,” Journal of Vacuum Science & Technology A: Vacuum, Surfaces, and Films, vol. 33, no. 2, pp. 020802, 2015.
[23] M. Noeske, J. Degenhardt, S. Strudthoff, and U. Lommatzsch, “Plasma jet treatment of five polymers at atmospheric pressure: surface modifications and the relevance for adhesion,” International journal of adhesion and adhesives, vol. 24, no. 2, pp. 171-177, 2004.
[24] C.-M. Chan, T.-M. Ko, and H. Hiraoka, “Polymer surface modification by plasmas and photons,” Surface science reports, vol. 24, no. 1-2, pp. 1-54, 1996.
[25] D. Caschera, A. Mezzi, L. Cerri, T. de Caro, C. Riccucci, G. M. Ingo, G. Padeletti, M. Biasiucci, G. Gigli, and B. Cortese, “Effects of plasma treatments for improving extreme wettability behavior of cotton fabrics,” Cellulose, vol. 21, no. 1, pp. 741-756, 2014.
[26] A. Waterhouse, S. G. Wise, Y. Yin, B. Wu, B. James, H. Zreiqat, D. R. McKenzie, S. Bao, A. S. Weiss, and M. K. Ng, “In vivo biocompatibility of a plasma-activated, coronary stent coating,” Biomaterials, vol. 33, no. 32, pp. 7984-7992, 2012.
[27] H. Chen, Q. Lin, Q. Xu, Y. Yang, Z. Shao, and Y. Wang, “Plasma activation and atomic layer deposition of TiO2 on polypropylene membranes for improved performances of lithium-ion batteries,” Journal of membrane science, vol. 458, pp. 217-224, 2014.
[28] A. Zille, M. M. Fernandes, A. Francesko, T. Tzanov, M. Fernandes, F. R. Oliveira, L. Almeida, T. Amorim, N. Carneiro, and M. F. Esteves, “Size and aging effects on antimicrobial efficiency of silver nanoparticles coated on polyamide fabrics activated by atmospheric DBD plasma,” ACS applied materials & interfaces, vol. 7, no. 25, pp. 13731-13744, 2015.
[29] A. Javid, M. Kumar, L. Wen, S. Yoon, S. B. Jin, J. H. Lee, and J. G. Han, “Surface energy and wettability control in bio-inspired PEG like thin films,” Materials & Design, vol. 92, pp. 405-413, 2016.
[30] M. Kumar, J. X. Piao, S. B. Jin, J. H. Lee, S. Tajima, M. Hori, and J. G. Han, “Low temperature plasma processing for cell growth inspired carbon thin films fabrication,” Archives of biochemistry and biophysics, vol. 605, pp. 41-48, 2016.
[31] J. X. Piao, M. Kumar, A. Javid, S. Yoon, J. H. Lee, and J. G. Han, “Pulsed DC-plasma sputtering induced synthesis of hydrogenated carbon thin films for L-929 cell cultivation,” Surface and Coatings Technology, vol. 307, pp. 1119-1123, 2016.
[32] S. David, C. Polonschii, C. Luculescu, M. Gheorghiu, S. Gáspár, and E. Gheorghiu, “Magneto-plasmonic biosensor with enhanced analytical response and stability,” Biosensors and Bioelectronics, vol. 63, pp. 525-532, 2015.
[33] M. Heuberger, and T. E. Balmer, “The transmission interferometric adsorption sensor,” Journal of Physics D: Applied Physics, vol. 40, no. 23, pp. 7245, 2007.
[34] M. Vandenbossche, L. Bernard, P. Rupper, K. Maniura-Weber, M. Heuberger, G. Faccio, and D. Hegemann, “Micro-patterned plasma polymer films for bio-sensing,” Materials & Design, vol. 114, pp. 123-128, 2017.
[35] D. J. Menzies, T. Gengenbach, J. S. Forsythe, N. Birbilis, G. Johnson, C. Charles, G. McFarland, R. J. Williams, C. Fong, and P. Leech, “One step multifunctional micropatterning of surfaces using asymmetric glow discharge plasma polymerization,” Chemical Communications, vol. 48, no. 13, pp. 1907-1909, 2012.
[36] V. Mishra, and R. Kumar, “Living radical polymerization: A review,” J. Sci. Res, vol. 56, pp. 141-176, 2012.
[37] J. Costerton, L. Montanaro, and C. R. Arciola, “Biofilm in implant infections: its production and regulation,” The International journal of artificial organs, vol. 28, no. 11, pp. 1062-1068, 2005.
[38] L. Tang, P. Thevenot, and W. Hu, “Surface chemistry influences implant biocompatibility,” Current topics in medicinal chemistry, vol. 8, no. 4, pp. 270-280, 2008.
[39] T. Ingverud, "Polymerization of Zwitterionic Sulphobetaine Methacrylate and Modification of Contact Lens Substrate," 2015.
[40] V. B. Damodaran, and N. S. Murthy, “Bio-inspired strategies for designing antifouling biomaterials,” Biomaterials research, vol. 20, no. 1, pp. 18, 2016.
[41] R. F. Zwaal, and A. J. Schroit, “Pathophysiologic implications of membrane phospholipid asymmetry in blood cells,” Blood, vol. 89, no. 4, pp. 1121-1132, 1997.
[42] R. Zwaal, P. Comfurius, and L. Van Deenen, “Membrane asymmetry and blood coagulation,” Nature, vol. 268, no. 5618, pp. 358, 1977.
[43] A. L. Lewis, “Phosphorylcholine-based polymers and their use in the prevention of biofouling,” Colloids and Surfaces B: Biointerfaces, vol. 18, no. 3-4, pp. 261-275, 2000.
[44] K. Ishihara, H. Oshida, Y. Endo, T. Ueda, A. Watanabe, and N. Nakabayashi, “Hemocompatibility of human whole blood on polymers with a phospholipid polar group and its mechanism,” Journal of biomedical materials research, vol. 26, no. 12, pp. 1543-1552, 1992.
[45] G.-H. Wu, and S.-h. Hsu, “polymeric-based 3D printing for tissue engineering,” Journal of medical and biological engineering, vol. 35, no. 3, pp. 285-292, 2015.
[46] W.-H. Kuo, M.-J. Wang, H.-W. Chien, T.-C. Wei, C. Lee, and W.-B. Tsai, “Surface modification with poly (sulfobetaine methacrylate-co-acrylic acid) to reduce fibrinogen adsorption, platelet adhesion, and plasma coagulation,” Biomacromolecules, vol. 12, no. 12, pp. 4348-4356, 2011.
[47] A. Striegel, W. W. Yau, J. J. Kirkland, and D. D. Bly, Modern size-exclusion liquid chromatography: practice of gel permeation and gel filtration chromatography: John Wiley & Sons, 2009.
[48] A. Della-Bona, “Characterizing ceramics and the interfacial adhesion to resin: II-the relationship of surface treatment, bond strength, interfacial toughness and fractography,” Journal of Applied Oral Science, vol. 13, no. 2, pp. 101-109, 2005.
[49] A. Larrañaga, S. Petisco, R. Villanueva, J. J. Iturri, S. Moya, E. Meaurio, and J.-R. Sarasua, "Physicochemical properties of plasma polymerized acrylic acid, ε-caprolactone and lactic acid films."
[50] M. Bashir, and S. Bashir, "Polymerization of acrylic acid using atmospheric pressure DBD plasma jet." p. 012036.
[51] Y. Fan, N. Migliore, P. Raffa, R. K. Bose, and F. Picchioni, “Synthesis of Zwitterionic Copolymers via Copper-Mediated Aqueous Living Radical Grafting Polymerization on Starch,” Polymers, vol. 11, no. 2, pp. 192, 2019.
[52] D.-J. Liaw, and C.-C. Huang, “Dilute solution properties of poly (3-dimethyl acryloyloxyethyl ammonium propiolactone),” Polymer, vol. 38, no. 26, pp. 6355-6362, 1997.
[53] V. Krishnamurthy, I. L. Kamel, and Y. Wei, “Analysis of plasma polymerization of allylamine by FTIR,” Journal of Polymer Science Part A: Polymer Chemistry, vol. 27, no. 4, pp. 1211-1224, 1989.
[54] Y.-F. Yang, Y. Li, Q.-L. Li, L.-S. Wan, and Z.-K. Xu, “Surface hydrophilization of microporous polypropylene membrane by grafting zwitterionic polymer for anti-biofouling,” Journal of Membrane Science, vol. 362, no. 1-2, pp. 255-264, 2010.
[55] I. Ahmed, H. Y. Liu, P. C. Mamiya, A. S. Ponery, A. N. Babu, T. Weik, M. Schindler, and S. Meiners, “Three‐dimensional nanofibrillar surfaces covalently modified with tenascin‐C‐derived peptides enhance neuronal growth in vitro,” Journal of Biomedical Materials Research Part A: An Official Journal of The Society for Biomaterials, The Japanese Society for Biomaterials, and The Australian Society for Biomaterials and the Korean Society for Biomaterials, vol. 76, no. 4, pp. 851-860, 2006.
[56] J. G. Steele, G. Johnson, C. McFarland, B. Dalton, T. Gengenbach, R. Chatelier, P. A. Underwood, and H. Griesser, “Roles of serum vitronectin and fibronectin in initial attachment of human vein endothelial cells and dermal fibroblasts on oxygen-and nitrogen-containing surfaces made by radiofrequency plasmas,” Journal of Biomaterials Science, Polymer Edition, vol. 6, no. 6, pp. 511-532, 1995.
[57] G. Kaklamani, N. Mehrban, J. Bowen, H. Dong, L. Grover, and A. Stamboulis, “Nitrogen plasma surface modification enhances cellular compatibility of aluminosilicate glass,” Materials Letters, vol. 111, pp. 225-229, 2013.
[58] S. Chen, L. Li, C. Zhao, and J. Zheng, “Surface hydration: Principles and applications toward low-fouling/nonfouling biomaterials,” Polymer, vol. 51, no. 23, pp. 5283-5293, 2010.
[59] Z. Zhang, T. Chao, S. Chen, and S. Jiang, “Superlow fouling sulfobetaine and carboxybetaine polymers on glass slides,” Langmuir, vol. 22, no. 24, pp. 10072-10077, 2006.
[60] A. Curtis, J. Forrester, C. McInnes, and F. Lawrie, “Adhesion of cells to polystyrene surfaces,” The Journal of cell biology, vol. 97, no. 5, pp. 1500-1506, 1983.
[61] C. F. Amstein, and P. A. Hartman, “Adaptation of plastic surfaces for tissue culture by glow discharge,” Journal of clinical microbiology, vol. 2, no. 1, pp. 46-54, 1975.