نوع مقاله : مقاله کامل پژوهشی

نویسندگان

1 دانشجوی کارشناسی ارشد مهندسی پزشکی، دانشکده مهندسی پزشکی، واحد علوم و تحقیقات، دانشگاه آزاد اسلامی، تهران

2 دانشیار، گروه بیومواد، پژوهشکده فناوری نانو و مواد پیش‌رفته، پژوهشگاه مواد و انرژی، کرج

3 استادیار، گروه مواد دندانی، دانشکده دندان‌پزشکی، دانشگاه علوم پزشکی شهید بهشتی، تهران

10.22041/ijbme.2014.14709

چکیده

داربست­های سه­بعدی که شباهت ریزساختاری بسیاری به ماتریکس خارج سلولی (ECM)داشته و از جنس کوپلیمر لاکتیک گلیکولیک­اسید (PLGA)/ژلاتین هستند به روش ریخته­گری انجمادی تهیه شدند. با این روش، امکان انجماد جهت­دار محلول پلیمری مهیّا شده و اثرهای مطلوب آن بر خواص فیزیکی/مکانیکی داربست­ها مورد ارزیابی قرار گرفت. برای انحلال دو پلیمر PLGA و ژلاتین از استیک­اسید به عنوان حلال مشترک استفاده شد. تصاویر به دست آمده از میکروسکوپ الکترونی روبشی (SEM) نشان­گر دست­یابی به داربست­هایی با تخلخل­های باز، درصد تخلخل بیش­از 95 درصد و با توزیع ابعادی حدود 400-50 میکرومتر در مقطع عمود بر جهت انجماد و 300-50 میکرومتر در مقطع موازی با جهت انجماد هستند. نتایج تخلخل­سنجی جیوه­ای، توزیع تخلخل 200-100 میکرومتر را نشان داد. نتایج طیف­سنجی فروسرخ (FTIR) حاکی­از عدم تغییر ساختاری مواد پس­از ساخت داربست­ها است. آزمون استحکام فشاری (MPa 2/3) نشان داد که داربست­های ساخته شده از استحکام مناسبی برخوردار هستند. نتایج آزمون جذب آب (950%) و زیست تخریب پذیری داربست­ها، گویای حفظ پایداری ساختار و تبادل­های سلولی درطی دوره­ی تخریب است. نتایج نشان می­دهد که داربست­های مذکور ویژگی­های اولیه و خواص مطلوبی برای استفاده در مهندسی بافت دارند و گزینه­ی مناسبی برای حمایت از چسبندگی سلولی و حفظ پایداری ساختاری در بازه­ی زمانی مورد نظر هستند.

کلیدواژه‌ها

موضوعات

عنوان مقاله [English]

Morphological and Physiochemical Characteristics of biodegradable PLGA-Gelatin Scaffolds by Unidirectional Freezing Technique

نویسندگان [English]

  • Farnaz Ghorbani 1
  • Ali Zamanian 2
  • Hanie Noje Dehian 3

1 M. Sc Student, Department of Biomedical Engineering, Tehran Science and Research Branch, Islamic Azad University, Tehran, Iran

2 Associate Professor, Department of Nanotechnology and Advanced Materials, Materials and Energy Research Center, Karaj, Iran

3 Assistant Professor, Department of Dental Materials, Dental School, Shahid Beheshti University of Medical Science, Tehran, Iran

چکیده [English]

In this study, we fabricated 3-dimentional PLGA-gelatin scaffolds with aligned-oriented pores by freeze casting technique which is similar to Extra Cellular Matrix (ECM), and evaluated its effect on both physical and mechanical features. Dissolving synthetic (PLGA) and natural (Gelatin) polymers in common solvent was one of the strengths of this investigation. Scanning electron microscopy (SEM) micrographs indicated that scaffolds contained 95% interconnected pores with diameter about 50-400 µm in horizontal direction and 50-200 µm in vertical direction. Moreover, the results of mercury intrusion porosimetry represented diameter of pores in range of 100–300 µm. According to fourieres transform infrared (FTIR) spectrum there was no inappropriate interactions during processing. Additionally, mechanical analysis (3.2 MPa) of PLGA-gelatin constructs illustrated that polymeric scaffolds can withstand mechanical loads in freezing direction. Based on the water absorption (950%) and biodegradation results, samples can support cellular interactions and prevent their integrity during tissue regeneration. In brief, freeze casted PLGA-gelatin scaffolds can provide unidirectional matrix with desired physical and mechanical characters to regenerate lesions.

کلیدواژه‌ها [English]

  • Lactic-co-Glycolic acid
  • Gelatin
  • freeze casting
  • polymeric scaffolds
  • Acetic acid
[1]   Y. Hagiwara, A. Nakashima, S. Itoh, T. Sakakura, C. Otsuji, E. Yamagishi, H. Shimizu, "Clinical application of PGA-tube for regeneration of intrapelvic nerves during extended surgery for intrapelvic recurrent rectal cancer, Gan to Kagaku Ryoho" Cancer & Chemotherapy 29, pp 2202–2204, 2002.
[2]   W. Noordenbos, P. D. Wall, "Implications of the failure of nerve resection and graft to cure chronic pain produced by nerve lesions" Journal of Neurology, Neurosurgery & Psychiatry 44, pp 1068–1073, 1981.
[3]   T. Nakamura, Y. Inada, S. Fukuda, M. Yoshitani, A. Nakada, S. Itoi, et al. "Experimental study on the regeneration of peripheral nerve gaps through a polyglycolic acid-collagen (PGA-collagen) tube" Brain Research 1027, pp 18–29, 2004.
[4]   K. Mizuseki, T. Sakamoto, K. Watanabe, K. Muguruma, M. Ikeya, A. Nishiyama, et al. "Generation of neural crest-derived peripheral neurons and floor plate cells from mouse and primate embryonic stem cells" Proceedings of the National Academy of Sciences of the United States of America 100, pp 5828–33, 2003.
[5]   D. Y. Lee, B. H. Choi, J. H. Park, S. J. Zhu, B. Y. Kim, J. Y. Huh, et al. "Nerve regeneration with the use of a poly (l-lactide-co-glycolic acid)-coated collagen tube filled with collagen gel" Journal of Cranio-Maxillo-Facial Surgery: Official Publication of the European Association for Cranio-Maxillo-Facial Surgery 34, pp 50–6, 2006.
[6]   B. Liu, S. X. Cai, K. W. Ma, Z. L. Xu, X. Z. Dai, L. Yang, et al. "Fabrication of a PLGA-collagen peripheral nerve scaffold and investigation of its sustained release property in vitro" Journal of Materials Science. Materials in Medicine 19, pp 1127–32, 2008.
[7]   L. Song, D. Baksh, R. S. Tuan, "Mesenchymal stem cell-based cartilage tissue engineering: cells, scaffold and biology" Cytotherapy 6, pp 596–601, 2004.
[8]   J. L. Drury, D. J. Mooney, "Hydrogels for tissue engineering: scaffold design variables and applications" Biomaterials 24, pp 4337–4351, 2003.
[9]   Z. X. Meng, Y. S. Wang, C. Ma, W. Zheng, L. Li, Y. F. Zheng, "Electrospinning of PLGA/gelatin randomly-oriented and aligned nanofibers as potential scaffold in tissue engineering" Materials Science and Engineering: C. 30, pp 1204–1210, 2010.
[10] X. Y. Zhao, J. Zhao, Y. P. Zhang, W. Yuan, "Electrospinning of PLGA/Gt blend system" Chemical Journal of Chinese Universities 30, pp 391–395, 2009.
[11] G. Wang, X. Hu, W. Lin, C. Dong, H. Wu, "Electrospun PLGA-silk fibroin-collagen nanofibrous scaffolds for nerve tissue engineering" In Vitro Cellular & Developmental Biology Animal 47, pp 234–40, 2011 .
[12] H. Ghaleh, F. Abbasi, M. Alizadeh, A. B. Khoshfetrat, "Mimicking the quasi-random assembly of protein fibers in the dermis by freeze-drying method" Materials Science & Engineering C Materials for Biological Applications 49, pp 807–15, 2015.
[13] N. Arabi, A. Zamanian, "Effect of cooling rate and gelatin concentration on the microstructural and mechanical properties of ice template gelatin scaffolds" Biotechnology and Applied Biochemistry 60, pp 573–9, 2013.
[14] A. Zamanian, S. Farhangdoust, M. Yasaei, M. Khorami, "The Effect of Particle Size on the Mechanical and Microstructural Properties of Freeze-Casted Macroporous Hydroxyapatite Scaffolds" International Journal of Applied Ceramic Technology 10, pp 1–10, 2013.
[15] Y. Zhou, S. Fu, Y. Pu, S. Pan, A. J. Ragauskas, "Preparation of aligned porous chitin nanowhisker foams by directional freeze-casting technique" Carbohydrate Polymers 112, pp 277–83, 2014.
[16] Y. Tang, Q. Miao, S. Qiu, K. Zhao, L. Hu, "Novel freeze-casting fabrication of aligned lamellar porous alumina with a centrosymmetric structure" Journal of the European Ceramic Society 34, pp 4077–4082, 2014.
[17] Y. Tang, K. Zhao, L. Hu, Z. Wu, "Two-step freeze casting fabrication of hydroxyapatite porous scaffolds with bionic bone graded structure" Ceramics International 39, pp 9703–9707, 2013.
[18] T. H. Qazi, R. Rai, A. R. Boccaccini, "Tissue engineering of electrically responsive tissues using polyaniline based polymers: A review" Biomaterials pp 1–19, 2014.
[19] P. van de Witte, P. J. Dijkstra, J. W. a. van den Berg, J. Feijen, "Phase separation processes in polymer solutions in relation to membrane formation" Journal of Membrane Science 117, pp 1–31, 1996.
[20] S. C. Subia, B. Kundu, J. Kundu, "Biomaterial scaffold fabrication techniques for potential tissue engineering applications" in: Tissue Engineering: pp 141–159, 2010.
[21] M. Alizadeh, F. Abbasi, A. B. Khoshfetrat, H. Ghaleh, "Microstructure and characteristic properties of gelatin/chitosan scaffold prepared by a combined freeze-drying/leaching method" Materials Science & Engineering C, Materials for Biological Applications 33, pp 3958–67, 2013.
[22]   S. Deville, "Freeze Casting of Porous Ceramics: A Review of Current Achievements" Advanced Engineering Materials pp 115–169, 2008.
[23]   M. Ho, P. Kuo, H. Hsieh, T. Hsien, L. Hou, "Preparation of porous scaffolds by using freeze-extraction and freeze-gelation methods" Biomaterials 25, pp 129–138, 2004.
[24]   X. K. Li, S. X. Cai, B. Liu, Z. L. Xu, X. Z. Dai, K. W. Ma, et al. "Characteristics of PLGA-gelatin complex as potential artificial nerve scaffold" Colloids and Surfaces B, Biointerfaces 57, pp 198–203, 2007.
[25]   X. Dai, L. Wang, K. Ma, K. Pan, "Characterization of a Hybridization Scaffold Based on PLGA/Acellular Pigskin for Nerve Regeneration" Journal of Medical and Biological Engineering 33, 2012.
[26]   F. Zamani, M. Latifi, M. Amani-Tehran, M. A. Shokrgozar, "Effects of PLGA nanofibrous scaffolds structure on nerve cell directional proliferation and morphology" Fibers and Polymers 14, pp 698–702, 2013.
[27]   Q. Cai, G. Shi, J. Bei, S. Wang, "Enzymatic degradation behavior and mechanism of Poly (lactide-co-glycolide) foams by trypsin" Biomaterial 24, pp 629–638, 2003.
[28]   V. K. Nandagiri, P. Gentile, V. Chiono, C. Tonda-Turo, A. Matsiko, Z. Ramtoola, et al. "Incorporation of PLGA nanoparticles into porous chitosan-gelatin scaffolds: influence on the physical properties and cell behavior" Journal of the Mechanical Behavior of Biomedical Materials 4, pp 1318–27, 2011.
[29]   Z. X. Meng, X. X. Xu, W. Zheng, H. M. Zhou, L. Li, Y. F. Zheng, et al. "Preparation and characterization of electrospun PLGA/gelatin nanofibers as a potential drug delivery system" Colloids and Surfaces B, Biointerfaces 84, pp 97–102, 2011.
[30]   C. Ye, P. Hu, M. X. Ma, Y. Xiang, R. G. Liu, X. W. Shang, "PHB/PHBHHx scaffolds and human adipose-derived stem cells for cartilage tissue engineering" Biomaterials 30, pp 4401–6, 2009.
[31]   A. L. Luís, J. M. Rodrigues, S. Geuna, S. Amado, Y. Shirosaki, J. M. Lee, et al. "Use of PLGA 90:10 scaffolds enriched with in vitro-differentiated neural cells for repairing rat sciatic nerve defects" Tissue Engineering Part A 14, pp 979–93, 2008.
[32]   H. Nojehdehian, F. Moztarzadeh, H. Baharvand, H. Nazarian, M. Tahriri, "Preparation and surface characterization of poly-L-lysine-coated PLGA microsphere scaffolds containing retinoic acid for nerve tissue engineering: in vitro study" Colloids and Surfaces B, Biointerfaces 73, pp 23–29, 2009.
[33]   G. R. D. Evans, K. Brandt, S. Katz, P. Chauvin, L. Otto, M. Bogle, et al. "Bioactive poly (L-lactic acid) conduits seeded with Schwann cells for peripheral nerve regeneration" Biomaterials 23, pp 841–8, 2002.
[34]   P. A. Webb, "Volume and Density Determinations for Particle Technologists" Micromeritics Instrument Corp 2001.
[35]   J. Khazaei, "Water Absorption Characteristics" Cercetari Agronomice in Moldova XLI, pp 5–16, 2008.
[36]   S. G. Lévesque, R. M. Lim, M. S. Shoichet, "Macroporous interconnected dextran scaffolds of controlled porosity for tissue-engineering applications" Biomaterials 26, pp 7436–46, 2005.