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Original Article

Vol. 3 No. 1 (2020): March-September

Synthesis and Characterization of S-IPN Hydrogels of Chitosan/PVA/PNIPAm to be Used in the Design of Nucleus Pulposus Prosthesis

DOI
https://doi.org/10.25061/ijamb.v3i1.30
Published
2020-01-03

Abstract

Hydrogels (HG) have been widely used in biomedical applications due to their high-water content which improves their biocompatibility with living tissue. In this study, Chitosan (CS) hydrogels cross-linked with Genipin and semi interpenetrated network (S-IPN) with PVA/PNIPAm were prepared to be used in the design of nucleus pulposus (NP) prosthesis. Chemical structure, morphology, swelling ratio (SR), mechanical properties and cytotoxicity were evaluated through a variation of the Genipin percentage and CS/PVA/PNIPAm proportions. Those experiments were carried out through Fourier-Transform Infrared Spectroscopy, Scanning Electron Microscopy, swelling studies, dynamic rheology, and hemocompatibility tests. The results showed that regardless of the Genipin percentage or polymers proportions, all HGs had interconnective porous structure. What did change microstructurally, was the pore size, its size distribution and the wall thickness. Firstly, an increment in the Genipin percentage and in the CS proportion concluded in an augmentation of the pore size. Secondly, an augmentation in the PVA proportion ended up producing smaller pores, with larger wall thickness and more homogeneous pore size distribution. The variation in PNIPAm proportion didn´t influence the morphology, but did have an impact on the SR and storage modulus (G´) augmenting in both cases as the PNIPAm proportion. The swelling ratio turned out to be related to the pore morphology; as smaller the pore size, smaller the SR. Likewise, the storage modulus rose insofar the SR diminished. In these S-IPN HGs, G´ varied between 77 Pa and 27000 Pa, values below and above G´ reported for human NP. Also, ? varied between 1.4° and 13.17° while the ? reported for NP is 23°-31°. Finally, the hemocompatibility tests did not show cellular lysis for any formulation. These outcomes demonstrated that from the rheological and hemocompatibility point of view, this kind of as semi interpenetrated networks (S-IPNs) HGs can be tailored to attain the NP´s properties

References

  1. Qi-Bin Bao, Geoffrey M. Mccullen, Paul A. Higham, John H. Dumbleton and Hansen A.Yuan "Review: The artificial disc: theory, design and materials" Biomaterids , Vol. 17, pp. 1157-1167, 1996.
  2. C. Jongeneelen, Biomechanics in the intervertebral disc, A literature review, Eindhoven: University of Technology, pp 1-15, 2006.
  3. Chan, S. C. y B., Gantenbein Ritter "Intervertebral disc regeneration or repair with biomaterials and stem cell therapy – feasible or fiction?" Swiss Med Weekly, vol. 142, pp. 1-12, 2012.
  4. Iatridis JC1, Nicoll SB, Michalek AJ, Walter BA, Gupta MS. "Role of biomechanics on intervertebral disc degeneration and regenerative therapies: What needs repairing in the disc and what are promising biomaterials for its repair?" Spine Journal, vol. 13, nº 3, p. 243–262, 2013.
  5. Iatridis, James C.; Weidenbaum, Mark; Setton, Lori A.; Mow, Van C., "Is the nucleus pulposus a solid or a fluid? Mechanical behaviors of the nucleus pulposus of the human intervertebral disc" Spine, vol. 21, nº 10, pp. 1174-1184, 1996.
  6. Nikolai Bogduk, "Clinical Anatomy of the Lumbar Spine and Sacrum", London/New York/ Oxford/Philadelphia/St Louis/Sydney/Toronto: Elsevier, pp. 1-77, 2005.
  7. Zhiyu Zhou, Manman Gao, Fuxin Wei, Jiabi Liang, Wenbin Deng, Xuejun Dai, Guangqian Zhou and Xuenong Zou, "Shock Absorbing Function Study on Denucleated Intervertebral Disc with or without Hydrogel Injection through Static and Dynamic Biomechanical Tests In Vitro" BioMed Research International, pp. 1-7, 2014.
  8. Irving M. Shapiro and Makarand V., "The Intervertebral Disc", Philadelphia: Springer, pp.3-52, 109-124 and 139-170, 2014.
  9. Peter J. Roughley, "Biology of intervertebral disc aging and degeneration: Involvement of the extracellular matrix" Spine, vol. 29, nº 23, pp. 2691-2699, 2004.
  10. James N. Parker, and Philip M. Parker, "Degenerative Disc Disease", San Diego: ICON Health Publications, 2004.
  11. D.W., Meakin J.R. and Hukins, "Effect of removing the nucleus pulposus on the deformation of the annulus fibrosus during compression of the intervertebral dis" Biomechanics, vol. 33, nº 5, pp. 575-580, 2000.
  12. Manohar Panjabi, Mark Brown, Sven Lindahl, Lars Irstam, and Martin Hermens, "Intrinsic disc pressure as a measure of integrity of the lumbar spine" JournalSpine, vol. 13, nº 8, pp. 913-917, 1988.
  13. Richard E. Seroussi, Martin H. Krag, David L. Muller and Malcolm H. Pope, "Internal deformations of intact and denucleated human lumbar discs subjected to compression, flexion, and extension loads" Orthopaedic Research, vol. 7, nº 1, p. 122–131, 1989.
  14. Iatridis JC, Setton L.A., Weidenbaum M., Mow V.C., "Alterations in the mechanical behavior of the human lumbar nucleus pulposus with degeneration and aging" J Orthop Res., vol. 15, nº 2, pp. 318-322, 1997.
  15. Brinkman P, Grootenbore H. ,"Change of disc height, radial disc bulge and intradiscal pressure from discectomy" Spine, vol. 16, pp. 641-646, 1991.
  16. Nishiyama K, Nye T., "Kinematics of whole lumbar spine, effect of discectomy" Spine, vol. 10, pp. 543-554, 1985.
  17. Michael A. Adams, "Biomechanics of back pain" Acupuncture in Medicine, vol. 22, nº 4, pp. 178-188, 2004.
  18. Simon Dagenais, Jaime Caro, and Scott Haldeman "A systematic review of low back pain cost of illness studies in the United States and internationally" The Spine Journal, vol. 8, pp. 8-20, 2008.
  19. Surgeons, American Academy of Orthopaedic "The Burden of Musculoskeletal Diseases in the United States," Rosemont, IL: Bone and Joint Decade, 2008. Available: http:// www.boneandjointburden.org/.
  20. Jensen M.C., Kelly A.P., Brant-Zawadzki M.N., "MRI of degenerative disease of the lumbar spine" Magn Reson Q., vol. 10, nº 3, p. 173–190, 1994.
  21. Shono Y., Kaneda K,Abumi K., "Stability of posterior spinal instrumentation and its effects on adjacent motion segments in the lumbosacral spine" Spine, vol. 23, pp. 1550-1558, 1998.
  22. Lehman T. R., Spratt K. F., "Long-term follow-up of lower lumbar fusion patients" Spine, vol. 12, pp. 97-104, 1987.
  23. Lee C. K. and Langrana N. A., " Lumbosacral spinal fusion: a biomechanical study" Spine, vol. 9, pp. 574-581, 1984.
  24. O'Connell G.D., Malhotra N.R., Vresilovic E.J., Elliott D.M., "The Effect of Discectomy and the Dependence on Degeneration of Human Intervertebral Disc Strain in Axial Compression" Spine, vol. 36, nº 21, p. 1765–1771, 2011.
  25. Judith R. Meakin, Thomas W. Redpath, David W.L. Hukins, "The Effect of Partial Removal of the Nucleus Pulposus from the intervertebral Disc on the Responsse of the Human Annulus Fibrosus to Compression" Clinical Biomechanics, vol. 16, pp. 121-128, 2001.
  26. Neil R. Malhotra, Woojin M. Han, Jesse Beckstein, Jordan Cloyd, "An Injectable Nucleus Pulposus Implant Restores Compressive Range of Motion in the Ovine Disc" Spine, vol. 37, nº 18, p. 1099–1105, 2013.
  27. Gillet P., "The fate of the adjacent motion segments after lumbar fusion" J Spinal Disord Tech, vol. 16, nº 4, pp. 338-45, 2003.
  28. Rahm M.D., Hall B.B., "Adjacent segment degeneration after lumbar fusion with instrumentation: a retrospective study" Spinal Disord., vol. 9, nº 5, pp. 392-400, 1996.
  29. Chunpeng Ren, Yueming Song, Limin Liu, Youdi Xue, "Adjacent segment degeneration and disease after lumbar fusion compared with motion-preserving procedures: a meta-analysis,» Orthop Surg Traumatol, vol. 24, nº 1, p. 245–253, 2014.
  30. Casey K. Lee, Vijay K. Goel, "Artificial disc prosthesis: design concepts and criteria" The Spine Journal, vol. 4, pp. 209-218, 2004.
  31. Aetna, "Intervertebral Disc Prostheses" 17/01/2018. Available: http://www.aetna.com/cpb/medical/data/500_599/0591.html.
  32. Angelo Carpy, "Progress in Molecular and Environmental Bioengineering - From Analysis and Modeling to Technology Applications", Rijeka, Croatia, InTech, 2011, pp. 117-150.
  33. Hassan Serhan, Devdatt Mhatre, Henri Defossez, "Motion-preserving technologies for degenerative lumbar spine: The past, present, and future horizons" SAS Journal , vol. 5, pp. 75-89, 2011.
  34. Raphael M. Otternbrite, Kinam Park, Teruo Okano "Biomedical Applications of Hidrogels Handbook", Ner York-London: Springer, pp 227-242.
  35. Shoufeng Yang, Kah-Ah-Fai Leong, Zhaohui Du y Chee-Kai Chua, "The Design of Scaffolds for Use in Tissue Engineering. Part I. Traditional Factors" Tissue Engineering, vol. 7, nº 6, pp. 679-689, 2001.
  36. Cheng Y.H., Yang S.H., Su W.Y., Chen Y.C., Yang K.C., Cheng W.T., Wu S.C., Lin F.H., "Thermosensitive chitosan-gelatin- glycerol phosphate hydrogels as a cell carrier for nucleus pulposus re- generation: an in vitro study" Tissue Eng Part A. , vol. 16, nº 2, pp. 695-703, 2010.
  37. Youling Yuan, Betsy M. Chesnutt, Warren O. Haggard and Joel D. Bumgardner, "Deacetylation of Chitosan: Material Characterization and in vitro Evaluation via Albumin Adsorption and Pre-Osteoblastic Cell Cultures" Materials, vol. 4, pp. 1399-1416, 2011.
  38. Renata Czechowska-Biskup, Diana Jarosi?ska, Bo?ena Rokita, Piotr Ula?ski, Janusz M. Rosiak, "Determination of Degree of Deacetylation of Chitosan - Comparision of methods" Progress on chemistry and application of chitin and its derivatives, vol. 17, pp. 5-20, 2012.
  39. Md Rabiul Hussain, Murshid Iman and Tarun K. Maji, "Determination of Degree of Deacetylation of Chitosan and Their effect on the Release Behavior of Essential Oil from Chitosan and Chitosan- Gelatin Complex Microcapsules" Advanced Engineering Applications, vol. 2, nº 4, pp. 4-12, 2013.
  40. Tanveer Ahmad Khan Kulliyyah, Kok Khiang Peh, Hung Seng Chang "Reporting degree of deacetylation values of chitosan: the influence of analytical methods" Pharm Pharmaceut Sci., vol. 5, nº 3, pp. 205-212, 2002.
  41. Maria G. Carrero, James J. Posada, Marcos A. Sabino “Intelligent Copolymers Based on Poly (N-Iopropylacrylamide) pNIPAm with potentialuse in Biomedical Applications. Part. I pNIPAm Functionalization with 3-Butenoic Acid and Piperazine” International Journal of Advances in Medical Biotechnology, vol. 1, nº 1, pp. 1-8, 2018.
  42. Steve Rimmer, "Biomedical Hydrogels, Oxford, Cambridge, Philadephia and New: Woodhead Publishing, pp. 35-61, 2011.
  43. Brian J. Tindall, Johannes Sikorski, Robert A. Smibert, and Noel R. Krieg "Phenotypic Characterization and the Principles of Comparative Systematics" de Methods for General and Molecular Microbiology, Washington, ASM Press, 2007, pp. 335-386.
  44. Encyclopedia, "World of Microbiology and Immunology", Available: https://www.encyclopedia.com/science/encyclopedias-almanacs-transcripts-and-maps/blood-agar-hemolysis-and-hemolytic-reactions. [last access: 7/4/2018].
  45. G. P. Box and D. R. Cox, "An Analysis of Transformations" Research Methods Meeting of the Society, pp. 211-252, 1964.
  46. Kenneth P. Burnham and David R. Anderson, "Model Selection and Multimodel Inference: A Practical Information- Theoretic Approach", Berlin, Heidelberg, Barcelona, Hong Kong, London, Milan and Paris: Springer, pp. 49-65 y 98-121. 2002.
  47. A. Carpi, "Progress in Molecular and Environmental Bioengineering - From Analysis and Modeling to Technology Applications", InTech, pp. 117-141. 2011.
  48. John K. Gillham, "Formation and Properties of Thermosetting and High Tg Polymeric Materials" Polymer Engineering and Science, vol. 26, nº 20, pp. 1429-1433, 1986.
  49. H. S. V. y. R. J. W. Jean Pierre Pscault, Thermosetting Polymers, New York: Marcel Dekker, pp. 1-10. 2002.
  50. M. José Moura, M. Margarida Figueiredo y M. Helena Gil, "Rheological study of genipin cross-linked chitosan hydrogels" Biomacromolecules, vol. 8, pp. 3823-3829, 2007.
  51. George Odian, "Principles of Polymerization" New York, John Wiley & Sons, 2004, pp. 39-50.
  52. Nand A.V., Rohindra D.R. y Khurma, J.R., "Characterization of genipin crosslinked hydrogels composed of chitosan and partially hydrolyzed poly(vinyl alcohol)" E-Polymers, pp. 30–38, 2007.
  53. Robert M. Silverstein, Francis X. Webster, David J. Kiemle "Spectrometric identification of organic compounds", New York: John Wiley & Sons, 81-90. 2005.
  54. Tao Wang, Mahir Turhan and Sundaram Gunasekaran "Selected properties of pH-sensitive, biodegradable chitosan–poly(vinyl alcohol) hydrogel" Polymer International, vol. 53, pp. 911-918, 2004.
  55. A. M. Shehap, "Thermal and Spectroscopic Studies of Polyvinyl" Egypt. J. Solids, vol. 31, nº 1, 2008.
  56. Christie M. Hassan, and Nikolaos A. Peppas, "Structure and Applications of Poly(vinyl alcohol) Hydrogels Produced by Conventional Crosslinking or by Freezing/Thawing Methods" de Biopolymers PVA Hydrogels, Anionic Polymerisation Nanocomposites, Berlin Heidelberg, Springer-Verlag, 2000, pp. 37-65.
  57. Long Bi, Zheng Cao,Yunyu Hu, Yang Song, Long Yu, Bo Yang, Jihong Mu, Zhaosong Huang y Yisheng Han, "Effects of different cross-linking conditions on the properties of genipin-cross-linked chitosan/collagen scaffolds for cartilage tissue engineering" J Mater Sci: Mater Med, vol. 22, pp. 51-62, 2011.
  58. Simona Dimida, Amilcare Barca, Nadia Cancelli, Vincenzo De Benedictis, Maria Grazia Raucci, y Christian Demitri, "Effects of Genipin Concentration on Cross-Linked Chitosan Scaffolds for Bone Tissue Engineering: Structural Characterization and Evidence of Biocompatibility Features" International Journal of Polymer Science, pp. 1-7, 2017.
  59. Jiabing Ran, Lingjun Xie, Guanglin Sun, Jingxiao Hu, Si Chen, Pei Jiang, Xinyu Shen, Hua Tong "A facile method for the preparation of chitosan-based scaffolds with anisotropic pores for tissue engineering applications" Carbohydrate Polymers, vol. 152, p. 615–623, 2016.
  60. Tayser Sumer Gaaz, Abu Bakar Sulong, Majid Niaz Akhtar, Abdul Amir H. Kadhum, "Properties and Applications of Polyvinyl Alcohol, Halloysite Nanotubes and Their Nanocomposites" Molecules, vol. 20, p. 22833–22847, 2015.
  61. Sundaram Gunasekaran, Tao Wang, Chunxiang Chai, "Swelling of pH-Sensitive Chitosan–Poly(vinyl alcohol) Hydrogels" Journal ofApplied Polymer Science, vol. 102, p. 4665–4671, 2006.
  62. Mingzhen Wang, Yu Fang , Daodao Hu"Preparation and properties of chitosan-poly(N-isopropylacrylamide) full-IPN hydrogels" Reactive & Functional Polymers, vol. 48, pp. 215-221, 2001.
  63. Kamiya Jain, Raman Verdarajan, Masaki Watanabe, Masaki Ishikiriyama, and Noriyoshi Matsumi, "Tunable LCST behavior of poly(N-isopropylacrylamide/ionic liquid) copolymers" Polymer Chemistry, nº 38, pp. 6819-6825 , 2015.
  64. I. G. Pedley and B. J. Tighe, "Water Binding Properties of Hydrogel Polymers for Reverse Osmosis and Related Applications" Brit. Polym., vol. 11, pp. 130-136, 1979.
  65. Hossein Omidian and Kinam Park, "Introduction to Hydrogels" de Biomedical Applications of Hidrogels Handbook, New York, Dordrecht, Heidelberg and London, Springer, 2010, pp. 1-15.
  66. Victor T. Wyatt, "Effects of Swelling on the Viscoelastic Properties of Polyester Films Made from Glycerol and Glutaric Acid" Journal of Applied Polymer Science, vol. 1, pp. 1-10, 2012.
  67. H. Bendaikha, S. Chaoui, S. Ould Kada, and A. Krallafa,"Swelling behavior and viscoelastic properties of a Polyacetal Hydrogel" AIP Conference Proceedings, vol. 64, 2008.
  68. Kristi S. Anseth, Christopher N. Bowman and Lisa Brannon-Peppas, "Mechanical properties of hydrogels and their experimental determination" Biomaterials, vol. 17, pp. 1647-5167, 1996.
  69. Hye Yun Kim, Ha Neul Kim, So Jin Lee, Jeong Eun Song, Soon Yong Kwon, Jin Wha Chung, Dongwon Lee and Gilson Khang,"Effect of pore sizes of PLGA scaffolds on mechanical properties and cell behaviour for nucleus pulposus regeneration in vivo" Tissue Engineering and Regenerative Medicina, vol. 11, nº 1, pp. 44-57, 2014.
  70. Jorge M. Sobral, Sofia G. Caridade, Rui A. Sousa, João F. Mano, "Three-dimensional plotted scaffolds with controlled pore size gradients: Effect of scaffold geometry on mechanical performance and cell seeding efficiency" Acta Biomaterialia, vol. 7, pp. 1009-1018, 2011.
  71. Laxman S. Desai and Laurence Lister, "Biocompatibility Safety Assessment of Medical Devices: FDA/ISO and Japanese Guidelines" Toxikon Corp., pp. 1-19, 2016.
  72. Marta A. Cooperstein and Heather E. Canavan, "Assessment of cytotoxicity of (N-isopropyl acrylamide) and Poly(N-isopropyl acrylamide)coated surfaces" Biointerphases , vol. 8, nº 19, pp. 1-12, 2013.
  73. J.L. Bron, G.H. Koenderink, V. Everts, T.H. Smit, "Rheological Characterization of the Nucleus Pulposus and Dense Collagen Scaffolds Intended for Functional Replacement" Journal of Orthopadic Research, pp. 620-626, 2009.
  74. F. Causa, L. Manto, A. Borzacchiello, R. De Santis, P. A. Netti, L. Ambrosio, L Nicolais, "Spatial and Structura Dependence of Mechanical Properties of Porcine Intervertebral Disc" Journal of Material Science: Materials in Medicina, vol. 13, pp. 1277-1280, 2002.
  75. Sung-Wook Choi, Yu Zhang y Younan Xia, "Three-dimensional Scaffolds for Tissue Engineering: The Importance of Uniformity in Pore Size and Structure" Langmuir, vol. 26, nº 24, p. 19001–19006, 2010.

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