In vivo integration of poly(e-caprolactone)/gelatin nanofibrous nerve guide seeded with teeth derived stem cells for peripheral nerve regeneration Mohammad-Hossein Beigi, 1,2 Laleh Ghasemi-Mobarakeh, 3 Molamma P. Prabhakaran, 4 Khadijeh Karbalaie, 1 Hamid Azadeh, 5 Seeram Ramakrishna, 4 Hossein Baharvand, 6 Mohammad-Hossein Nasr-Esfahani 1 1 Department of Cellular Biotechnology at Cell Science Research Center, Royan Institute for Biotechnology, ACECR, Isfahan, Iran 2 Materials Engineering Department, Najafabad Branch, Islamic Azad University, Najafabad, Iran 3 Department of Textile Engineering, Isfahan University of Technology, Isfahan, 84156-83111, Iran 4 Center for Nanofibers and Nanotechnology, E3-05-14, Nanoscience and Nanotechnology Initiative, Faculty of Engineering, National University of Singapore, Singapore 117576 5 Department of Physiotherapy, School of Rehabilitation Science, Isfahan University of Medical Sciences, Isfahan, Iran 6 Department of Stem Cells and Developmental Biology at Cell Science Research Center, Royan Institute for Stem Cell Biology and Technology, ACECR, Tehran, Iran Received 5 November 2013; revised 29 January 2014; accepted 4 February 2014 Published online 00 Month 2014 in Wiley Online Library (wileyonlinelibrary.com). DOI: 10.1002/jbm.a.35119 Abstract: Artificial nanofiber nerve guides have gained huge interest in bridging nerve gaps and associated peripheral nerve regeneration due to its high surface area, flexibility and porous structure. In this study, electrospun poly (e-caprolac- tone)/gelatin (PCL/Gel) nanofibrous mats were fabricated, rolled around a copper wire and fixed by medical grade adhesive to obtain a tubular shaped bio-graft, to bridge 10 mm sciatic nerve gap in in vivo rat models. Stem cells from human exfoliated deciduous tooth (SHED) were transplanted to the site of nerve injury through the nanofibrous nerve guides. In vivo experiments were performed in animal mod- els after creating a sciatic nerve gap, such that the nerve gap was grafted using (i) nanofiber nerve guide (ii) nanofiber nerve guide seeded with SHED (iii) suturing, while an untreated nerve gap remained as the negative control. In vitro cell culture study was carried out for primary investiga- tion of SHED-nanofiber interaction and its viability within the nerve guides after 2 and 16 weeks of implantation time. Walking track analysis, plantar test, electrophysiology and immunohistochemistry were performed to evaluate func- tional recovery during nerve regeneration. Vascularization was also investigated by hematoxilin/eosine (H&E) staining. Overall results showed that the SHED seeded on nanofibrous nerve guide could survive and promote axonal regeneration in rat sciatic nerves, whereby the biocompatible PCL/Gel nerve guide with cells can support axonal regeneration and could be a promising tissue engineered graft for peripheral nerve regeneration. V C 2014 Wiley Periodicals, Inc. J Biomed Mater Res Part A: 00A:000–000, 2014. Key Words: peripheral nerve regeneration, nerve guide, electro- spinning, stem cells How to cite this article: Beigi M-H, Ghasemi-Mobarakeh L, Prabhakaran MP, Karbalaie K, Azadeh H, Ramakrishna S, Baharvand H, Nasr-Esfahani M-H. 2014. In vivo integration of poly(e-caprolactone)/gelatin nanofibrous nerve guide seeded with teeth derived stem cells for peripheral nerve regeneration. J Biomed Mater Res Part A 2014:00A:000–000. INTRODUCTION The regeneration of defective or damaged peripheral nerve has been a difficult and challenging problem in reconstruc- tive surgery and is considered as a common cause of per- manent functional loss and post traumatic morbidity, mainly because nerve regeneration is a complex biological phenom- enon. 1–3 Direct end-to-end suturing of the damaged nerve ends is a commonly utilized method for peripheral nerve regenera- tion (PNS). For PNS injuries of relatively short distances (<5 mm), axonal regeneration occurs spontaneously. How- ever, functional recovery of nerve repair remains unsatisfac- tory especially when the nerve defect or gap is too long. 4 As a promising alternative method to suturing, implantation of an autologous nerve graft harvested from functionally less important nerves such as sural nerves, superficial cutaneous nerves or lateral and medial antebrachii cutaneous nerves have been used traditionally to bridge peripheral nerve defects. It provides structural support for sprouting axons and bridges the proximal and distal nerve stumps, Correspondence to: M.-H. Nasr-Esfahani; e-mail:mh.nasr-esfahani@royaninstitute.org V C 2014 WILEY PERIODICALS, INC. 1