Abstract
Electrospun nanofibrous scaffolds have received a great deal of attention in tissue engineering in recent years. Bridging larger nerve gaps between proximal and distal ends requires exogenous tubular constructs with uniaxially aligned topographical cues to promote the axonal re-growth due to the lack of fibrin cable formation. In this study, we have designed and developed a collector to obtain aligned nanofibers of PLGA–PCL. The average diameter of the fibers obtained is 230 ± 63 nm and the alignment of fibers is quantified by calculating relative angle of each fiber. The tensile strength, porosity, contact angle, and biodegradation of the uniaxial PLGA–PCL nanofibers are measured and compared with the corresponding random fibers. Pore size, Young’s modulus, and degradation of the aligned scaffold are significantly lesser than random fibers (p < 0.05). The in vitro cell adhesion and proliferation of Schwann cells on the aligned nanofibers are evaluated and compared with random nanofibers. Our results demonstrate that the alignment of nanofibers has a significant influence on the adhesion and proliferation of Schwann cells. Thus, the axially aligned nanofibers may mimic the fibrin cable architecture; thereby it may represent an ideal scaffold for extending the growth of axonal processes.
Similar content being viewed by others
References
Agarwal, S., A. Greiner, and J. H. Wendorff. Electrospinning of manmade and biopolymer nanofibers-progress in techniques, materials, and applications. Adv. Funct. Mater. 19:2863–2879, 2009.
Amado, S., M. J. Simoes, P. A. S. Armada da Silva, A. L. Luýs, Y. Shirosaki, M. A. Lopes, J. D. Santos, F. Fregnan, G. Gambarotta, S. Raimondo, M. Fornaro, A. P. Veloso, A. S. P. Varejao, A. C. Maurýcio, and S. Geuna. Use of hybrid chitosan membranes and N1E-115 cells for promoting nerve regeneration in an axonotmesis rat model. Biomaterials 29:4409–4419, 2008.
Balgude, A. P., X. Yu, A. Szymanski, and R. V. Bellamkonda. Agarose gel stiffness determines rate of DRG neurite extension in 3D cultures. Biomaterials 22:1077–1084, 2001.
Bellamkonda, R. V. Peripheral nerve regeneration: an opinion on channels, scaffolds and anisotropy. Biomaterials 27:3515–3518, 2006.
Bhang, S. H., J. S. Lim, C. Y. Choi, Y. K. Kwon, and B.-S. Kim. The behavior of neural stem cells on biodegradable synthetic polymers. J. Biomater. Sci. Polym. Ed. 18:223–239, 2007.
Bhattarai, S. R., N. Bhattarai, H. K. Yi, P. H. Hwang, D. I. Cha, and H. Y. Kim. Novel biodegradable electrospun membrane: scaffold for tissue engineering. Biomaterials 25:2595–2602, 2004.
Bhattarai, N., D. Edmondson, O. Veiseh, F. A. Matsen, and M. Zhang. Electrospun chitosan-based nanofibers and their cellular compatibility. Biomaterials 26:6176–6184, 2005.
Bini, T. B., S. Gao, S. Wang, and S. Ramakrishna. Development of fibrous biodegradable polymer conduits for guided nerve regeneration. J. Mater. Sci. Mater. Med. 2005(16):367–375, 2005.
Cao, H., T. Liu, and S. Y. Chew. The application of nanofibrous scaffolds in neural tissue engineering. Adv. Drug Deliv. Rev. 61:1055–1064, 2009.
Chew, S. Y., R. Mi, A. Hoke, and K. W. Leong. The effect of the alignment of electrospun fibrous scaffolds on Schwann cell maturation. Biomaterials 29:653–661, 2008.
Dhandayuthapani, B., U. M. Krishnan, and S. Sethuraman. Fabrication & characterization of chitosan–gelatin blend nanofibers for skin tissue engineering. J. Biomed. Mater. Res. B Appl. Biomater. 94B:264–272, 2010.
Duan, B., X. Yuan, Y. Zhu, Y. Zhang, X. Li, Y. Zhang, and K. Yao. A nanofibrous composite membrane of PLGA–chitosan/PVA prepared by electrospinning. J. Eur. Polym. 42:2013–2022, 2006.
Ellis, M. J., and J. B. Chaudhuri. Poly (lactic-co-glycolic acid) hollow fiber membranes for use as a Tissue engineering Scaffold. Biotechnol. Bioeng. 96:177–187, 2007.
Gupta, D., J. Venugopal, M. P. Prabhakaran, V. R. Giri Dev, S. Low, A. T. Choon, and S. Ramakrishan. Aligned and random nanofibrous substrate for in vitro culture of Schwann cells for neural tissue engineering. Acta Biomater. 5:2560–2569, 2009.
Hadlock, T., J. Elisseeff, R. Langer, J. Vacanti, and M. Cheney. A tissue engineered conduit for peripheral nerve repair. Arch. Otolaryngol. Head Neck Surg. 124:1081–1086, 1998.
Hiep, N. T., and B. T. Lee. Electrospinning of PLGA/PCL blends for tissue engineering and their biocompatibility. J. Mater. Sci. Mater. Med. 21:1969–1978, 2010.
Hutmacher, D. W. Scaffold in tissue engineering bone and cartilage. Biomaterials 20:2529–2543, 2000.
IJkema-Paassen, J., K. Jansen, A. Gramsbergen, and M. F. Meek. Transection of peripheral nerves, bridging strategies and effect evaluation. Biomaterials 25:1583–1592, 2004.
Jayaraman, K., M. Kotaki, Y. Z. Zhang, X. M. Mo, and S. Ramakrishna. Recent advances in polymer nanofibers. J. Nanosci. Nanotechnol. 4:52–65, 2004.
Jones, L. L., M. Oudega, M. B. Bunge, and M. H. Tuszynski. Neurotrophic factors, cellular bridges and gene therapy for spinal cord injury. J. Physiol. 533:83–89, 2001.
Kang, Y. O., I. Yoon, S. Y. Lee, D. Kim, S. J. Lee, W. H. Park, and S. M. Hudson. Chitosan-coated poly(vinyl alcohol) nanofibers for wound dressings. J. Biomed. Mater. Res. B Appl. Biomater. 92B:568–576, 2010.
Kim, J. Y., and D. W. Cho. Blended PCL/PLGA scaffold fabrication using multi-head deposition system. Microelectron. Eng. 86:1447–1450, 2009.
Kim, Y. T., V. K. Haftel, S. Kumar, and R. V. Bellamkonda. The role of aligned polymer fiber-based constructs in the bridging of long peripheral nerve gaps. Biomaterials 29:3117–3127, 2008.
Kuppan, P., S. Sethuraman, and U. M. Krishnan. Tissue engineering interventions for esophageal disorders—promises and challenges. Biotechnol. Adv., 2012. doi:10.1016/j.biotechadv.2012.03.005.
Kuppan, P., K. S. Vasanthan, D. Sundaramurthi, U. M. Krishnan, and S. Sethuraman. Development of poly(3-hydroxybutyrate-co-3-hydroxyvalerate) fibers for skin tissue engineering: effects of topography, mechanical, and chemical stimuli. Biomacromolecules 12:3156–3165, 2011.
Liang, D., B. S. Hsiao, and B. Chu. Functional electron nanofibrous scaffolds for biomedical application. Adv. Drug Deliv. Rev. 59:1392–1412, 2007.
Liu, Y., H. Jiang, Y. Li, and K. Zhu. Control of dimensional stability and degradation rate in electrospun composite scaffolds composed of poly(d,l-lactide-co-glycolide) and poly (ε-caprolactone). Chin. J. Polym. Sci. 26:63–71, 2008.
Mobarakeh, L. G., M. P. Prabhakaran, M. Morshed, M. H. Nasr-Esfahani, and S. Ramakrishna. Electrospun poly (3-caprolactone)/gelatin nanofibrous scaffolds for nerve tissue engineering. Biomaterials 29:4532–4539, 2008.
Nichols, C. M., M. J. Brenner, I. K. Fox, D. A. Hunter, S. R. Rickman, and S. E. Mackinnon. Effects of motor versus sensory nerve grafts on peripheral nerve regeneration. Exp. Neurol. 190:347–355, 2004.
Pearse, D. D., F. C. Pereira, A. E. Marcillo, M. L. Bates, Y. A. Berrocal, M. T. Filbin, and M. B. Bunge. cAMP and Schwann cells promote axonal growth and functional recovery after spinal cord injury. Nat. Med. 10:610–616, 2004.
Rutkowski, G. E., and C. A. Heath. Development of a bioartificial nerve graft. II. Nerve regeneration in vitro. Biotechnol. Prog. 18:373–379, 2002.
Salifu, A. A., B. D. Nury, and C. Lekakou. Electrospinning of nanocomposite fibrillar tubular and flat scaffolds with controlled fiber orientation. Ann. Biomed. Eng. 39:2510–2520, 2011.
Sangsanoh, P., S. Waleetorncheepsawat, O. Suwantong, P. Wutticharoenmongkol, O. Weeranantanapan, B. Chuenjitbuntaworn, P. Cheepsunthorn, P. Pavasant, and P. Supaphol. In vitro biocompatibility of Schwann cells on surfaces of biocompatible polymeric electrospun fibrous and solution-cast film scaffolds. Biomacromolecules 8:1587–1594, 2007.
Schnell, E., K. Klinkhammer, S. Balzer, G. Brook, D. Klee, P. Dalton, and J. Mey. Guidance of glial cell migration and axonal growth on electrospun nanofibers of poly-ε-caprolactone and a collagen/poly-ε-caprolactone blend. Biomaterials 28:3012–3025, 2007.
Sethuraman, S., L. S. Nair, S. El-Amin, M. Nguyen, A. Singh, Y. E. Greish, H. R. Allcock, P. W. Brown, and C. T. Laurencin. Nanocomposite injectabls for orthopaedic applications based on polyphosphazenes. J. Biomater. Sci. Polym. Ed. 22:733–752, 2011.
Sethuraman, S., L. S. Nair, S. El-Amin, M. Nguyen, A. Singh, N. Krogman, Y. E. Greish, H. R. Allcock, P. W. Brown, and C. T. Laurencin. Mechanical properties and osteocompatibility of novel biodegradable alanine based polyphosphazenes: side group effects. Acta Biomater. 6:1931–1937, 2010.
Sill, T. J., and H. A. Von Recum. Electrospinning: applications in drug delivery and tissue engineering. Biomaterials 29:1989–2006, 2008.
Subramanian, A., U. M. Krishnan, and S. Sethuraman. Axially aligned electrically conducting biodegradable nanofibers for neural regeneration. J. Mater. Sci. Mater. Med., 2012. doi:10.1007/s10856-012-4654-y.
Subramanian, A., U. M. Krishnan, and S. Sethuraman. Development of biomaterial scaffold for nerve tissue engineering: biomaterial mediated neural regeneration. J. Biomed. Sci. 16:108, 2009.
Subramanian, A., U. M. Krishnan, and S. Sethuraman. Fabrication of uniaxially aligned 3D electrospun scaffolds for neural regeneration. Biomed. Mater. 6:025004, 2011.
Supaphol, P., and S. Chuangchote. On the electrospinning of poly(vinyl alcohol) nanofiber mats: a revisit. J. Appl. Polym. Sci. 108:969–978, 2008.
Teo, W. E., M. Kotaki, X. M. Mo, and S. Ramakrishna. Porous tubular structures with controlled fibre orientation using a modified electrospinning method. Nanotechnology 16:918–924, 2005.
Teo, W. E., and S. Ramakrishna. Electrospun fibre bundle made of aligned nanofibres over two fixed points. Nanotechnology 16:1878–1884, 2005.
Teo, W. E., and S. Ramakrishna. A review on electrospinning design and nanofibre assemblies. Nanotechnology 17:R89–R106, 2006.
Vasanthan, K. S., A. Subramanian, U. M. Krishnan, and S. Sethuraman. Role of biomaterials, therapeutic molecules and cells for hepatic tissue engineering. Biotechnol. Adv. 30:742–752, 2012.
Wang, W., S. Itoh, K. Konno, T. Kikkawa, S. Ichinose, K. Sakai, T. Ohkuma, and K. Watabe. Effects of Schwann cell alignment along the oriented electrospun chitosan nanofibers on nerve regeneration. J. Biomed. Mater. Res. A 91:994–1005, 2009.
Wen, X., and P. A. Tresco. Fabrication and characterization of permeable degradable poly(dl–lactide-co-glycolide) (PLGA) hollow fiber phase inversion membranes for use as nerve tract guidance channels. Biomaterials 27:3800–3809, 2006.
Xie, J., M. R. MacEwan, S. M. Willerth, X. Li, D. W. Moran, S. E. Sakiyama-Elbert, and Y. Xia. Conductive core-sheath nanofibers and their potential application in neural tissue engineering. Adv. Funct. Mater. 19:2312–2318, 2009.
Xu, X. M., A. Chen, V. Guenard, N. Kleitman, and M. B. Bunge. Bridging Schwann cell transplants promote axonal regeneration from both the rostral and caudal stumps of transected adult rat spinal cord. J. Neurocytol. l26:1–16, 1997.
Xu, C. Y., R. Inai, M. Kotaki, and S. Ramakrishna. Aligned biodegradable nanofibrous structure: a potential scaffold for blood vessel engineering. Biomaterials 25:877–886, 2004.
Yang, F., R. Murugan, S. Ramakrishna, X. Wang, Y. X. Ma, and S. Wang. Fabrication of nano-structured porous PLLA scaffold intended for nerve tissue engineering. Biomaterials 25:1891–1900, 2004.
Yang, F., R. Murugan, S. Wang, and S. Ramakrishna. Electrospinning of nano/micro scale poly(l-lactic acid) aligned fibers and their potential in neural tissue engineering. Biomaterials 26:2603–2610, 2005.
Yeo, S., C. Choi, J. Yang, and D. Jung. Patterned amine surfaces with reduced background nonspecific protein adsorption fabricated by using inductively coupled plasma chemical vapour deposition. J. Korean Phys. Soc. 51:1000–1006, 2007.
Yu, X., and R. V. Bellamkonda. Tissue engineered scaffolds are effective alternatives to autografts in bridging peripheral nerve gaps in rodents. Tissue Eng. 9:421–430, 2003.
Zussman, E., D. Rittel, and A. L. Yarin. Failure modes of electrospun nanofibers. Appl. Phys. Lett. 82:3958–3960, 2003.
Acknowledgments
This work is funded by the Indian Council for Medical Research (35/12/2009–BMS). The authors wish to acknowledge the infrastructure support provided by the Nano Mission Council (SR/S5/NM-07/2006 & SR/NM/PG-16/2007) and the FIST program (SR/FST/LSI-327/2007) of the Department of Science & Technology, Govt. of India.
Author information
Authors and Affiliations
Corresponding author
Additional information
Associate Editor Smadar Cohen oversaw the review of this article.
Rights and permissions
About this article
Cite this article
Subramanian, A., Krishnan, U.M. & Sethuraman, S. Fabrication, Characterization and In Vitro Evaluation of Aligned PLGA–PCL Nanofibers for Neural Regeneration. Ann Biomed Eng 40, 2098–2110 (2012). https://doi.org/10.1007/s10439-012-0592-6
Received:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s10439-012-0592-6