Yong Shi1, Liang Zhou, Jie Tian, Yang Wang. 1. Department of Otolaryngology-Head and Neck Surgery, Fudan University Affiliated Eye, Ear, Nose and Throat Hospital, Shanghai, China.
Abstract
CONCLUSION: Combining neurotrophic factor support and neural stem cell (NSC) transplantation may improve regeneration of the peripheral nervous system. Objectives. We constructed a biodegradable nerve conduit (NC) filled with NSCs overexpressing glia-derived neurotrophic factor (GDNF), which is known to protect facial motoneurons, and tested the effect of this NC on facial nerve regeneration. MATERIALS AND METHODS: Primary cultured NSCs were transduced with a lentiviral vector encoding enhanced green fluorescent protein (EGFP) and GDNF. GDNF expression was confirmed by Western blotting and ELISA. Sprague Dawley (SD) rats were subjected to right facial nerve transection, and polyglycolic/polyglycolic acid (PLGA) NCs filled with NSCs-GDNF were used to bridge the nerve gap (n=24). In vivo GDNF expression was confirmed by real-time PCR. NCs containing NSCs, transgenic Schwann cells (SCs-GDNF), or empty NCs served as controls (n=24 per group). Facial nerve regeneration was assessed 2-12 weeks after surgery, by electrophysiological testing, immunohistochemical staining, and morphometric analysis of axons. RESULTS: NSCs exhibited sustained and robust GDNF expression in culture and following implantation. Nerve action potential amplitude, axonal area, and axonal number were significantly greater in the NSCs-GDNF group than in the NSCs or empty NC groups. Axonal area and number were also greater in the NSCs-GDNF group than the SCs-GDNF group, although this was not statistically significant. The enhanced regeneration observed in the NSCs-GDNF group was accompanied by increased labeling for S100, NF, and βIII tubulin.
CONCLUSION: Combining neurotrophic factor support and neural stem cell (NSC) transplantation may improve regeneration of the peripheral nervous system. Objectives. We constructed a biodegradable nerve conduit (NC) filled with NSCs overexpressing glia-derived neurotrophic factor (GDNF), which is known to protect facial motoneurons, and tested the effect of this NC on facial nerve regeneration. MATERIALS AND METHODS: Primary cultured NSCs were transduced with a lentiviral vector encoding enhanced green fluorescent protein (EGFP) and GDNF. GDNF expression was confirmed by Western blotting and ELISA. Sprague Dawley (SD) rats were subjected to right facial nerve transection, and polyglycolic/polyglycolic acid (PLGA) NCs filled with NSCs-GDNF were used to bridge the nerve gap (n=24). In vivo GDNF expression was confirmed by real-time PCR. NCs containing NSCs, transgenic Schwann cells (SCs-GDNF), or empty NCs served as controls (n=24 per group). Facial nerve regeneration was assessed 2-12 weeks after surgery, by electrophysiological testing, immunohistochemical staining, and morphometric analysis of axons. RESULTS: NSCs exhibited sustained and robust GDNF expression in culture and following implantation. Nerve action potential amplitude, axonal area, and axonal number were significantly greater in the NSCs-GDNF group than in the NSCs or empty NC groups. Axonal area and number were also greater in the NSCs-GDNF group than the SCs-GDNF group, although this was not statistically significant. The enhanced regeneration observed in the NSCs-GDNF group was accompanied by increased labeling for S100, NF, and βIII tubulin.
Authors: Ricardo Ferreira Bento; Raquel Salomone; Silvia Bona do Nascimento; Ricardo Jose Rodriguez Ferreira; Ciro Ferreira da Silva; Heloisa Juliana Zabeu Rossi Costa Journal: Int Arch Otorhinolaryngol Date: 2014-05-23
Authors: Robert Gaudin; Christian Knipfer; Anders Henningsen; Ralf Smeets; Max Heiland; Tessa Hadlock Journal: Biomed Res Int Date: 2016-07-31 Impact factor: 3.411